TY  - DATA
AU  - Mladenović Stokanić, Maja
AU  - Simović, Ana
AU  - Jovanović, Vesna
AU  - Radomirović, Mirjana
AU  - Udovički, Božidar
AU  - Krstić Ristivojević, Maja
AU  - Djukić, Teodora
AU  - Vasović, Tamara
AU  - Aćimović, Jelena
AU  - Sabljić, Ljiljana
AU  - Lukić, Ivana
AU  - Kovačević, Ana
AU  - Cujic, Danica
AU  - Gnjatović, Marija
AU  - Smiljanić, Katarina
AU  - Stojadinović, Marija
AU  - Radosavljević, Jelena
AU  - Stanić-Vučinić, Dragana
AU  - Stojanović, Marijana
AU  - Rajković, Andreja
AU  - Ćirkovic Veličković, Tanja
PY  - 2024
UR  - http://intor.torlakinstitut.com/handle/123456789/859
AB  - S1.1. Checking of N protein purity Recombinant N protein purity was checked after imidazole removal and buffer exchange by SDS PAGE (Figure 6.). For comparison, commercial high-purity HSA was also analyzed. S1.2. Identification of N protein Tandem mass spectrometry identification of proteins in an in-gel digested band of N protein (Figure S1, lane 3), confirmed the identity of N protein with high scores and peptide coverage (Fig. S2.). S2. Purification of polyclonal antibodies from mice and rabbit sera For the development of an ELISA test specific for the detection of SARS-CoV-2 N protein, recombinantly produced N protein was used for the immunization of mice and rabbits. Sera obtained from rabbits and mice were then tested for titer and specificity (Figure S3 and Figure 1). To determine the titer of polyclonal sera required to detect N protein in samples, we use wells coated with N protein and serial dilution of sera pools from different animals. After multiple washing steps, we detected the binding of rabbit and mice antibodies using secondary biotinylated antibodies and streptavidin-alkaline phosphatase chimaera or secondary antibodies with previously coupled alkaline phosphatase, where the amount of enzymes’ substrate converted to the product was measured as an increase in absorbance at 405 nm. As shown in Figure S3A, unpurified sera pools from both animals showed very high titers and expected logarithmic decrease of signal with dilution. Based on the obtained data titer for unpurified sera was determined to be X. The same trend was observed for pools purified using AS precipitation and rabbit sera purified using protein A affinity chromatography (Figure S3B and S3C). As shown in Figure S3D, clear bands from antibodies could be observed in both full and purified samples. Western blot analysis showed only one protein band on mass around 40 kDa, a Accession number / Protein Name Score Coverage (%) Unique peptides P0DTC9|NCAP_SARS2 Nucleoprotein OS=Severe acute respiratory syndrome coronavirus 2, 46 kDa 504.9 74.22 183 mass of purified N protein suggesting that the obtained sera is highly specific for N protein (Figure 2). Section S3 Diagnostic validationS3.1. Stabilization of capture antibodies Pre-coated ELISA plates were prepared for usage in clinical practice. To ensure the preservation of the biofunctionality of the surface-bound capture antibodies, the commonly used stabilizing excipient, 3% sucrose with 10% glycerol in MilliQ water was used. The plates were incubated with 300 μL per well of a stabilizing agent for 1 hour at room temperature. After an hour of incubation, the solution was carefully aspirated from each well. The plate was then blotted against clear paper towels to remove any remaining liquid, and the plates were allowed to air dry for 3 hours at RT. Dried plates were wrapped in parafilm and stored at 4 °C for later use. To remove the stabilizing agent coating, wells were washed with slightly acidic distilled water (pH of 6) three times, leaving the plate prepared for subsequent assay steps. Section S4. Characterization of N protein by HRMS S4.1. SDS PAGE and in-gel digestion Characterization of the produced recombinant N protein was done by HRMS after its in-gel digestion. A total of 10 μg of purified protein(s) were loaded in a 0.5 cm wide well and after SDSPAGE gel was stained with Coomassie Brilliant Blue R-250 (CBB). Protein gel bands were washed, reduced with dithiothreitol, and alkylated with iodoacetamide, followed by in-gel trypsin digestion1 (Shevchenko et al. 2006) with some minor modifications. The amount of trypsin was leveled to a trypsin/sample ratio of 1:30 (w/w). The final concentration of MS-grade trypsin (diluted in 25 mM ammonium bicarbonate buffer) was 1 ng/μL. Sample clean-up was performed using zip tips HyperSep C18 (Thermo Fisher Scientific Inc., Bremen, Germany). S5.1 Immunization of rabbits and mice Mice immunization Swiss Webster mice (n=10) were immunized subcutaneously with N protein formulated with Complete Freund`s adjuvant (CFA; 1st dose, 100 μg N protein / dose) or Incomplete Freund`s adjuvant (IFA; 2nd and 3rd doses, 50 μg N protein / dose) in three-week intervals. Mice were housed in small groups of up to six animals and had access to commercial mice food and water ad libitum. N protein solution (500ug/ml in PBS) was sterilized by filtering through 0.22 um filters. Sterile N protein solution was mixed with CFA (Sigma, Cat. No. F5881) at ratio 1:1 (v/v) under aseptic conditions. In total 400 ul of N protein-CFA emulsion (N protein final concentration 250ug/ml) was applied per immunization per mouse. Initial immunization was done by injection of N protein in CFA given subcutaneously (SC) in four sites (thigh pocket, base of tail, and mediastinum) with a 100 ul using 23-25 gauge needle. In total 100 ug of N protein was applied per mouse (25 ug per site). Subsequent immunizations with booster doses were done in the same way, but using IFA (Sigma, Cat. No. F5506) instead of CFA and N protein final concentration was 125 ug/ml. . In total 50 ug of N protein was applied per mouse (12.5 ug per site). Immunizations were done every three weeks. Mice immunization scheme: 1. day 0 – N protein in PBS: CFA = 1:1 (v/v); N protein final concentration was 250 μg/mL; 400 μL per mice (4x100 μL), e.g. 100 μg per mice 2. day 21 - N protein in PBS: IFA = 1:1 (v/v); N protein final concentration was 125 μg/mL; 400 μL per mice (4x100 μL), e.g. 50 μg per mice 3. day 42 - N protein in PBS: IFA = 1:1 (v/v); N protein final concentration was 125 μg/mL; 400 μl per mice (4x100 μL), e.g. 50 μg per mice First bleeding was performed two weeks after the 3rd dose, and then in intervals not shorter than two weeks. The sera obtained after the first bleeding was tested for the production of specific anti-N protein antibodies.
PB  - MDPI
T2  - International Journal of Molecular Sciences
T1  - Supplementary information for the article:       Mladenovic Stokanic, M.; Simovic, A.; Jovanovic, V.; Radomirovic, M.; Udovicki, B.; Krstic Ristivojevic, M.; Djukic, T.; Vasovic, T.; Acimovic, J.; Sabljic, L.; Lukic, I.; Kovacevic, A.; Cujic, D.; Gnjatovic, M.; Smiljanic, K.; Stojadinovic, M.; Radosavljevic, J.; Stanic-Vucinic, D.; Stojanovic, M.; Rajkovic, A.; Cirkovic Velickovic, T. Sandwich ELISA for the Quantification of Nucleocapsid Protein of SARS-CoV-2 Based on Polyclonal Antibodies from Two Different Species. International Journal of Molecular Sciences 2024, 25 (1), 333. https://doi.org/10.3390/ijms25010333.
IS  - 1
VL  - 25
UR  - https://hdl.handle.net/21.15107/rcub_intor_859
ER  - 

@misc{
author = "Mladenović Stokanić, Maja and Simović, Ana and Jovanović, Vesna and Radomirović, Mirjana and Udovički, Božidar and Krstić Ristivojević, Maja and Djukić, Teodora and Vasović, Tamara and Aćimović, Jelena and Sabljić, Ljiljana and Lukić, Ivana and Kovačević, Ana and Cujic, Danica and Gnjatović, Marija and Smiljanić, Katarina and Stojadinović, Marija and Radosavljević, Jelena and Stanić-Vučinić, Dragana and Stojanović, Marijana and Rajković, Andreja and Ćirkovic Veličković, Tanja",
year = "2024",
abstract = "S1.1. Checking of N protein purity Recombinant N protein purity was checked after imidazole removal and buffer exchange by SDS PAGE (Figure 6.). For comparison, commercial high-purity HSA was also analyzed. S1.2. Identification of N protein Tandem mass spectrometry identification of proteins in an in-gel digested band of N protein (Figure S1, lane 3), confirmed the identity of N protein with high scores and peptide coverage (Fig. S2.). S2. Purification of polyclonal antibodies from mice and rabbit sera For the development of an ELISA test specific for the detection of SARS-CoV-2 N protein, recombinantly produced N protein was used for the immunization of mice and rabbits. Sera obtained from rabbits and mice were then tested for titer and specificity (Figure S3 and Figure 1). To determine the titer of polyclonal sera required to detect N protein in samples, we use wells coated with N protein and serial dilution of sera pools from different animals. After multiple washing steps, we detected the binding of rabbit and mice antibodies using secondary biotinylated antibodies and streptavidin-alkaline phosphatase chimaera or secondary antibodies with previously coupled alkaline phosphatase, where the amount of enzymes’ substrate converted to the product was measured as an increase in absorbance at 405 nm. As shown in Figure S3A, unpurified sera pools from both animals showed very high titers and expected logarithmic decrease of signal with dilution. Based on the obtained data titer for unpurified sera was determined to be X. The same trend was observed for pools purified using AS precipitation and rabbit sera purified using protein A affinity chromatography (Figure S3B and S3C). As shown in Figure S3D, clear bands from antibodies could be observed in both full and purified samples. Western blot analysis showed only one protein band on mass around 40 kDa, a Accession number / Protein Name Score Coverage (%) Unique peptides P0DTC9|NCAP_SARS2 Nucleoprotein OS=Severe acute respiratory syndrome coronavirus 2, 46 kDa 504.9 74.22 183 mass of purified N protein suggesting that the obtained sera is highly specific for N protein (Figure 2). Section S3 Diagnostic validationS3.1. Stabilization of capture antibodies Pre-coated ELISA plates were prepared for usage in clinical practice. To ensure the preservation of the biofunctionality of the surface-bound capture antibodies, the commonly used stabilizing excipient, 3% sucrose with 10% glycerol in MilliQ water was used. The plates were incubated with 300 μL per well of a stabilizing agent for 1 hour at room temperature. After an hour of incubation, the solution was carefully aspirated from each well. The plate was then blotted against clear paper towels to remove any remaining liquid, and the plates were allowed to air dry for 3 hours at RT. Dried plates were wrapped in parafilm and stored at 4 °C for later use. To remove the stabilizing agent coating, wells were washed with slightly acidic distilled water (pH of 6) three times, leaving the plate prepared for subsequent assay steps. Section S4. Characterization of N protein by HRMS S4.1. SDS PAGE and in-gel digestion Characterization of the produced recombinant N protein was done by HRMS after its in-gel digestion. A total of 10 μg of purified protein(s) were loaded in a 0.5 cm wide well and after SDSPAGE gel was stained with Coomassie Brilliant Blue R-250 (CBB). Protein gel bands were washed, reduced with dithiothreitol, and alkylated with iodoacetamide, followed by in-gel trypsin digestion1 (Shevchenko et al. 2006) with some minor modifications. The amount of trypsin was leveled to a trypsin/sample ratio of 1:30 (w/w). The final concentration of MS-grade trypsin (diluted in 25 mM ammonium bicarbonate buffer) was 1 ng/μL. Sample clean-up was performed using zip tips HyperSep C18 (Thermo Fisher Scientific Inc., Bremen, Germany). S5.1 Immunization of rabbits and mice Mice immunization Swiss Webster mice (n=10) were immunized subcutaneously with N protein formulated with Complete Freund`s adjuvant (CFA; 1st dose, 100 μg N protein / dose) or Incomplete Freund`s adjuvant (IFA; 2nd and 3rd doses, 50 μg N protein / dose) in three-week intervals. Mice were housed in small groups of up to six animals and had access to commercial mice food and water ad libitum. N protein solution (500ug/ml in PBS) was sterilized by filtering through 0.22 um filters. Sterile N protein solution was mixed with CFA (Sigma, Cat. No. F5881) at ratio 1:1 (v/v) under aseptic conditions. In total 400 ul of N protein-CFA emulsion (N protein final concentration 250ug/ml) was applied per immunization per mouse. Initial immunization was done by injection of N protein in CFA given subcutaneously (SC) in four sites (thigh pocket, base of tail, and mediastinum) with a 100 ul using 23-25 gauge needle. In total 100 ug of N protein was applied per mouse (25 ug per site). Subsequent immunizations with booster doses were done in the same way, but using IFA (Sigma, Cat. No. F5506) instead of CFA and N protein final concentration was 125 ug/ml. . In total 50 ug of N protein was applied per mouse (12.5 ug per site). Immunizations were done every three weeks. Mice immunization scheme: 1. day 0 – N protein in PBS: CFA = 1:1 (v/v); N protein final concentration was 250 μg/mL; 400 μL per mice (4x100 μL), e.g. 100 μg per mice 2. day 21 - N protein in PBS: IFA = 1:1 (v/v); N protein final concentration was 125 μg/mL; 400 μL per mice (4x100 μL), e.g. 50 μg per mice 3. day 42 - N protein in PBS: IFA = 1:1 (v/v); N protein final concentration was 125 μg/mL; 400 μl per mice (4x100 μL), e.g. 50 μg per mice First bleeding was performed two weeks after the 3rd dose, and then in intervals not shorter than two weeks. The sera obtained after the first bleeding was tested for the production of specific anti-N protein antibodies.",
publisher = "MDPI",
journal = "International Journal of Molecular Sciences",
title = "Supplementary information for the article:       Mladenovic Stokanic, M.; Simovic, A.; Jovanovic, V.; Radomirovic, M.; Udovicki, B.; Krstic Ristivojevic, M.; Djukic, T.; Vasovic, T.; Acimovic, J.; Sabljic, L.; Lukic, I.; Kovacevic, A.; Cujic, D.; Gnjatovic, M.; Smiljanic, K.; Stojadinovic, M.; Radosavljevic, J.; Stanic-Vucinic, D.; Stojanovic, M.; Rajkovic, A.; Cirkovic Velickovic, T. Sandwich ELISA for the Quantification of Nucleocapsid Protein of SARS-CoV-2 Based on Polyclonal Antibodies from Two Different Species. International Journal of Molecular Sciences 2024, 25 (1), 333. https://doi.org/10.3390/ijms25010333.",
number = "1",
volume = "25",
url = "https://hdl.handle.net/21.15107/rcub_intor_859"
}

Mladenović Stokanić, M., Simović, A., Jovanović, V., Radomirović, M., Udovički, B., Krstić Ristivojević, M., Djukić, T., Vasović, T., Aćimović, J., Sabljić, L., Lukić, I., Kovačević, A., Cujic, D., Gnjatović, M., Smiljanić, K., Stojadinović, M., Radosavljević, J., Stanić-Vučinić, D., Stojanović, M., Rajković, A.,& Ćirkovic Veličković, T.. (2024). Supplementary information for the article:       Mladenovic Stokanic, M.; Simovic, A.; Jovanovic, V.; Radomirovic, M.; Udovicki, B.; Krstic Ristivojevic, M.; Djukic, T.; Vasovic, T.; Acimovic, J.; Sabljic, L.; Lukic, I.; Kovacevic, A.; Cujic, D.; Gnjatovic, M.; Smiljanic, K.; Stojadinovic, M.; Radosavljevic, J.; Stanic-Vucinic, D.; Stojanovic, M.; Rajkovic, A.; Cirkovic Velickovic, T. Sandwich ELISA for the Quantification of Nucleocapsid Protein of SARS-CoV-2 Based on Polyclonal Antibodies from Two Different Species. International Journal of Molecular Sciences 2024, 25 (1), 333. https://doi.org/10.3390/ijms25010333.. in International Journal of Molecular Sciences
MDPI., 25(1).
https://hdl.handle.net/21.15107/rcub_intor_859

Mladenović Stokanić M, Simović A, Jovanović V, Radomirović M, Udovički B, Krstić Ristivojević M, Djukić T, Vasović T, Aćimović J, Sabljić L, Lukić I, Kovačević A, Cujic D, Gnjatović M, Smiljanić K, Stojadinović M, Radosavljević J, Stanić-Vučinić D, Stojanović M, Rajković A, Ćirkovic Veličković T. Supplementary information for the article:       Mladenovic Stokanic, M.; Simovic, A.; Jovanovic, V.; Radomirovic, M.; Udovicki, B.; Krstic Ristivojevic, M.; Djukic, T.; Vasovic, T.; Acimovic, J.; Sabljic, L.; Lukic, I.; Kovacevic, A.; Cujic, D.; Gnjatovic, M.; Smiljanic, K.; Stojadinovic, M.; Radosavljevic, J.; Stanic-Vucinic, D.; Stojanovic, M.; Rajkovic, A.; Cirkovic Velickovic, T. Sandwich ELISA for the Quantification of Nucleocapsid Protein of SARS-CoV-2 Based on Polyclonal Antibodies from Two Different Species. International Journal of Molecular Sciences 2024, 25 (1), 333. https://doi.org/10.3390/ijms25010333.. in International Journal of Molecular Sciences. 2024;25(1).
https://hdl.handle.net/21.15107/rcub_intor_859 .

Mladenović Stokanić, Maja, Simović, Ana, Jovanović, Vesna, Radomirović, Mirjana, Udovički, Božidar, Krstić Ristivojević, Maja, Djukić, Teodora, Vasović, Tamara, Aćimović, Jelena, Sabljić, Ljiljana, Lukić, Ivana, Kovačević, Ana, Cujic, Danica, Gnjatović, Marija, Smiljanić, Katarina, Stojadinović, Marija, Radosavljević, Jelena, Stanić-Vučinić, Dragana, Stojanović, Marijana, Rajković, Andreja, Ćirkovic Veličković, Tanja, "Supplementary information for the article:       Mladenovic Stokanic, M.; Simovic, A.; Jovanovic, V.; Radomirovic, M.; Udovicki, B.; Krstic Ristivojevic, M.; Djukic, T.; Vasovic, T.; Acimovic, J.; Sabljic, L.; Lukic, I.; Kovacevic, A.; Cujic, D.; Gnjatovic, M.; Smiljanic, K.; Stojadinovic, M.; Radosavljevic, J.; Stanic-Vucinic, D.; Stojanovic, M.; Rajkovic, A.; Cirkovic Velickovic, T. Sandwich ELISA for the Quantification of Nucleocapsid Protein of SARS-CoV-2 Based on Polyclonal Antibodies from Two Different Species. International Journal of Molecular Sciences 2024, 25 (1), 333. https://doi.org/10.3390/ijms25010333." in International Journal of Molecular Sciences, 25, no. 1 (2024),
https://hdl.handle.net/21.15107/rcub_intor_859 .

TY  - JOUR
AU  - Mladenović Stokanić, Maja
AU  - Simović, Ana
AU  - Jovanović, Vesna
AU  - Radomirović, Mirjana
AU  - Udovički, Božidar
AU  - Krstić Ristivojević, Maja
AU  - Djukić, Teodora
AU  - Vasović, Tamara
AU  - Aćimović, Jelena
AU  - Sabljić, Ljiljana
AU  - Lukić, Ivana
AU  - Kovačević, Ana
AU  - Cujic, Danica
AU  - Gnjatović, Marija
AU  - Smiljanić, Katarina
AU  - Stojadinović, Marija
AU  - Radosavljević, Jelena
AU  - Stanić-Vučinić, Dragana
AU  - Stojanović, Marijana
AU  - Rajković, Andreja
AU  - Ćirkovic Veličković, Tanja
PY  - 2024
UR  - http://intor.torlakinstitut.com/handle/123456789/858
AB  - In this study, a cost-effective sandwich ELISA test, based on polyclonal antibodies, for routine quantification SARS-CoV-2 nucleocapsid (N) protein was developed. The recombinant N protein was produced and used for the production of mice and rabbit antisera. Polyclonal N protein-specific antibodies served as capture and detection antibodies. The prototype ELISA has LOD 0.93 ng/mL and LOQ 5.3 ng/mL, with a linear range of 1.52–48.83 ng/mL. N protein heat pretreatment (56 °C, 1 h) decreased, while pretreatment with 1% Triton X-100 increased analytical ELISA sensitivity. The diagnostic specificity of ELISA was 100% (95% CI, 91.19–100.00%) and sensitivity was 52.94% (95% CI, 35.13–70.22%) compared to rtRT-PCR (Ct < 40). Profoundly higher sensitivity was obtained using patient samples mostly containing Wuhan-similar variants (Wuhan, alpha, and delta), 62.50% (95% CI, 40.59 to 81.20%), in comparison to samples mostly containing Wuhan-distant variants (Omicron) 30.00% (6.67–65.25%). The developed product has relatively high diagnostic sensitivity in relation to its analytical sensitivity due to the usage of polyclonal antibodies from two species, providing a wide repertoire of antibodies against multiple N protein epitopes. Moreover, the fast, simple, and inexpensive production of polyclonal antibodies, as the most expensive assay components, would result in affordable antigen tests.
PB  - MDPI
T2  - International Journal of Molecular Sciences
T1  - Sandwich ELISA for the Quantification of Nucleocapsid Protein of SARS-CoV-2 Based on Polyclonal Antibodies from Two Different Species
IS  - 1
SP  - 333
VL  - 25
DO  - 10.3390/ijms25010333
ER  - 

@article{
author = "Mladenović Stokanić, Maja and Simović, Ana and Jovanović, Vesna and Radomirović, Mirjana and Udovički, Božidar and Krstić Ristivojević, Maja and Djukić, Teodora and Vasović, Tamara and Aćimović, Jelena and Sabljić, Ljiljana and Lukić, Ivana and Kovačević, Ana and Cujic, Danica and Gnjatović, Marija and Smiljanić, Katarina and Stojadinović, Marija and Radosavljević, Jelena and Stanić-Vučinić, Dragana and Stojanović, Marijana and Rajković, Andreja and Ćirkovic Veličković, Tanja",
year = "2024",
abstract = "In this study, a cost-effective sandwich ELISA test, based on polyclonal antibodies, for routine quantification SARS-CoV-2 nucleocapsid (N) protein was developed. The recombinant N protein was produced and used for the production of mice and rabbit antisera. Polyclonal N protein-specific antibodies served as capture and detection antibodies. The prototype ELISA has LOD 0.93 ng/mL and LOQ 5.3 ng/mL, with a linear range of 1.52–48.83 ng/mL. N protein heat pretreatment (56 °C, 1 h) decreased, while pretreatment with 1% Triton X-100 increased analytical ELISA sensitivity. The diagnostic specificity of ELISA was 100% (95% CI, 91.19–100.00%) and sensitivity was 52.94% (95% CI, 35.13–70.22%) compared to rtRT-PCR (Ct < 40). Profoundly higher sensitivity was obtained using patient samples mostly containing Wuhan-similar variants (Wuhan, alpha, and delta), 62.50% (95% CI, 40.59 to 81.20%), in comparison to samples mostly containing Wuhan-distant variants (Omicron) 30.00% (6.67–65.25%). The developed product has relatively high diagnostic sensitivity in relation to its analytical sensitivity due to the usage of polyclonal antibodies from two species, providing a wide repertoire of antibodies against multiple N protein epitopes. Moreover, the fast, simple, and inexpensive production of polyclonal antibodies, as the most expensive assay components, would result in affordable antigen tests.",
publisher = "MDPI",
journal = "International Journal of Molecular Sciences",
title = "Sandwich ELISA for the Quantification of Nucleocapsid Protein of SARS-CoV-2 Based on Polyclonal Antibodies from Two Different Species",
number = "1",
pages = "333",
volume = "25",
doi = "10.3390/ijms25010333"
}

Mladenović Stokanić, M., Simović, A., Jovanović, V., Radomirović, M., Udovički, B., Krstić Ristivojević, M., Djukić, T., Vasović, T., Aćimović, J., Sabljić, L., Lukić, I., Kovačević, A., Cujic, D., Gnjatović, M., Smiljanić, K., Stojadinović, M., Radosavljević, J., Stanić-Vučinić, D., Stojanović, M., Rajković, A.,& Ćirkovic Veličković, T.. (2024). Sandwich ELISA for the Quantification of Nucleocapsid Protein of SARS-CoV-2 Based on Polyclonal Antibodies from Two Different Species. in International Journal of Molecular Sciences
MDPI., 25(1), 333.
https://doi.org/10.3390/ijms25010333

Mladenović Stokanić M, Simović A, Jovanović V, Radomirović M, Udovički B, Krstić Ristivojević M, Djukić T, Vasović T, Aćimović J, Sabljić L, Lukić I, Kovačević A, Cujic D, Gnjatović M, Smiljanić K, Stojadinović M, Radosavljević J, Stanić-Vučinić D, Stojanović M, Rajković A, Ćirkovic Veličković T. Sandwich ELISA for the Quantification of Nucleocapsid Protein of SARS-CoV-2 Based on Polyclonal Antibodies from Two Different Species. in International Journal of Molecular Sciences. 2024;25(1):333.
doi:10.3390/ijms25010333 .

Mladenović Stokanić, Maja, Simović, Ana, Jovanović, Vesna, Radomirović, Mirjana, Udovički, Božidar, Krstić Ristivojević, Maja, Djukić, Teodora, Vasović, Tamara, Aćimović, Jelena, Sabljić, Ljiljana, Lukić, Ivana, Kovačević, Ana, Cujic, Danica, Gnjatović, Marija, Smiljanić, Katarina, Stojadinović, Marija, Radosavljević, Jelena, Stanić-Vučinić, Dragana, Stojanović, Marijana, Rajković, Andreja, Ćirkovic Veličković, Tanja, "Sandwich ELISA for the Quantification of Nucleocapsid Protein of SARS-CoV-2 Based on Polyclonal Antibodies from Two Different Species" in International Journal of Molecular Sciences, 25, no. 1 (2024):333,
https://doi.org/10.3390/ijms25010333 . .

TY  - GEN
AU  - Ćirković-Veličković, Tanja
AU  - Gnjatović, Marija
AU  - Ćujić, Danica
AU  - Todorović, Aleksandra
AU  - Stanić-Vučinić, Dragana
AU  - Đukić, Teodora
AU  - Mladenović, Maja
AU  - Vasović, Tamara
AU  - Stojadinović, Marija
AU  - Krstić-Ristivojević, Maja
AU  - Jovanović, Vesna
AU  - Simović, Ana
AU  - Radosavljević, Jelena
AU  - Aćimović, Jelena M.
AU  - Radomirović, Mirjana Ž.
AU  - Stojanović, Marijana
PY  - 2023
UR  - http://intor.torlakinstitut.com/handle/123456789/860
AB  - Novi korona virus (SARS CoV-2) koji se pojavio u Vuhanu 2019. godine pripada grupi jednolančanih RNK virusa [1]. Predstavlja novi infektivni agens za humanu populaciju i veoma je brzo detektovan u velikom broju zemalja. Uzročnik je respiratornih infekcija koje mogu da budu praćene i veoma teškom kliničkom slikom. Brzo širenje, odsustvo imuniteta na ovaj virus i odsustvo pouzdanih testova za detekciju virusa u trenutku izbijanja pandemije su bolest izazvanu ovim virusom brzo pretvorili u zdravstveni i društveni problem najvišeg prioriteta na globalnom nivou. Iako su najveće biotehnološke kompanije ubrzano počele sa razvojem i masovnom proizvodnjom dijagnostičkih testova i vakcina, njihova dostupnost u trenucima najveće potražnje je i dalje nedovoljna, a cene istih su limitirajući faktor za bolju kontrolu bolesti i širenja pandemije [2]. Razvoj sopstvenih i održiva proizvodnja testova i vakcina za COVID-19 su od velikog društvenog značaja. Važan preduslov za održivu proizvodnju testova je dostupnost rekombinantnih antigena virusa i mogućnost proizvodnje istih na velikoj skali za potrebe proizvodnje domaćih testova. Ovim tehničkim rešenjem se opisuje dobijanje dva ključna antigena novog korona virusa rekombinantnom tehnologijom i njihova primena u serološkom ELISA testu koji proizvodi Institut za primenu nuklearne energije, INEP, kao i za dobijanje reagenasa za detekciju antigena novog korona virusa (specifičnih antitela). U prvoj fazi, optimizovane su sekvence proteina koje su podigle osetljivost postojećih seroloških testova. Inovativnost našeg pristupa se ogleda i u razrađenim eksperimentalnim protokolima za dobijanje rekombinantnih proteina nukleokapsida na velikoj skali, kao i u solubilnoj formi, što olakšava postupak prečišćavanja. Izbor fragmenta nukleokapsida koji se heterologo eksprimira u solubilnoj formi, a specifično detektuje antitela i generiše jak imuni odgovor tokom imunizacije životinja (imunogenost) na osnovu pregleda poznatih epitopskih sekvenci je ključna inovacija ovog tehničkog rešenja. Ovo je prvi primer uspešno primenjenog rekombinatnog proteina proizvedenog u Srbiji u dijagnostičkom testu koji je registrovankod Agencije za lekove i medicinska sredstva Republike Srbije (broj rešenja 515-02-02370-21-002), a koji je primenu našao i na međunarodnom nivou.
T1  - Dobijanje rekombinantnog imunogenog fragmenta proteina nukleokapsida SARS-CoV-2 virusa za proizvodnju reagenasa i dijagnostičkih testova na novi korona virus
UR  - https://hdl.handle.net/21.15107/rcub_intor_860
ER  - 

@misc{
author = "Ćirković-Veličković, Tanja and Gnjatović, Marija and Ćujić, Danica and Todorović, Aleksandra and Stanić-Vučinić, Dragana and Đukić, Teodora and Mladenović, Maja and Vasović, Tamara and Stojadinović, Marija and Krstić-Ristivojević, Maja and Jovanović, Vesna and Simović, Ana and Radosavljević, Jelena and Aćimović, Jelena M. and Radomirović, Mirjana Ž. and Stojanović, Marijana",
year = "2023",
abstract = "Novi korona virus (SARS CoV-2) koji se pojavio u Vuhanu 2019. godine pripada grupi jednolančanih RNK virusa [1]. Predstavlja novi infektivni agens za humanu populaciju i veoma je brzo detektovan u velikom broju zemalja. Uzročnik je respiratornih infekcija koje mogu da budu praćene i veoma teškom kliničkom slikom. Brzo širenje, odsustvo imuniteta na ovaj virus i odsustvo pouzdanih testova za detekciju virusa u trenutku izbijanja pandemije su bolest izazvanu ovim virusom brzo pretvorili u zdravstveni i društveni problem najvišeg prioriteta na globalnom nivou. Iako su najveće biotehnološke kompanije ubrzano počele sa razvojem i masovnom proizvodnjom dijagnostičkih testova i vakcina, njihova dostupnost u trenucima najveće potražnje je i dalje nedovoljna, a cene istih su limitirajući faktor za bolju kontrolu bolesti i širenja pandemije [2]. Razvoj sopstvenih i održiva proizvodnja testova i vakcina za COVID-19 su od velikog društvenog značaja. Važan preduslov za održivu proizvodnju testova je dostupnost rekombinantnih antigena virusa i mogućnost proizvodnje istih na velikoj skali za potrebe proizvodnje domaćih testova. Ovim tehničkim rešenjem se opisuje dobijanje dva ključna antigena novog korona virusa rekombinantnom tehnologijom i njihova primena u serološkom ELISA testu koji proizvodi Institut za primenu nuklearne energije, INEP, kao i za dobijanje reagenasa za detekciju antigena novog korona virusa (specifičnih antitela). U prvoj fazi, optimizovane su sekvence proteina koje su podigle osetljivost postojećih seroloških testova. Inovativnost našeg pristupa se ogleda i u razrađenim eksperimentalnim protokolima za dobijanje rekombinantnih proteina nukleokapsida na velikoj skali, kao i u solubilnoj formi, što olakšava postupak prečišćavanja. Izbor fragmenta nukleokapsida koji se heterologo eksprimira u solubilnoj formi, a specifično detektuje antitela i generiše jak imuni odgovor tokom imunizacije životinja (imunogenost) na osnovu pregleda poznatih epitopskih sekvenci je ključna inovacija ovog tehničkog rešenja. Ovo je prvi primer uspešno primenjenog rekombinatnog proteina proizvedenog u Srbiji u dijagnostičkom testu koji je registrovankod Agencije za lekove i medicinska sredstva Republike Srbije (broj rešenja 515-02-02370-21-002), a koji je primenu našao i na međunarodnom nivou.",
title = "Dobijanje rekombinantnog imunogenog fragmenta proteina nukleokapsida SARS-CoV-2 virusa za proizvodnju reagenasa i dijagnostičkih testova na novi korona virus",
url = "https://hdl.handle.net/21.15107/rcub_intor_860"
}

Ćirković-Veličković, T., Gnjatović, M., Ćujić, D., Todorović, A., Stanić-Vučinić, D., Đukić, T., Mladenović, M., Vasović, T., Stojadinović, M., Krstić-Ristivojević, M., Jovanović, V., Simović, A., Radosavljević, J., Aćimović, J. M., Radomirović, M. Ž.,& Stojanović, M.. (2023). Dobijanje rekombinantnog imunogenog fragmenta proteina nukleokapsida SARS-CoV-2 virusa za proizvodnju reagenasa i dijagnostičkih testova na novi korona virus. .
https://hdl.handle.net/21.15107/rcub_intor_860

Ćirković-Veličković T, Gnjatović M, Ćujić D, Todorović A, Stanić-Vučinić D, Đukić T, Mladenović M, Vasović T, Stojadinović M, Krstić-Ristivojević M, Jovanović V, Simović A, Radosavljević J, Aćimović JM, Radomirović MŽ, Stojanović M. Dobijanje rekombinantnog imunogenog fragmenta proteina nukleokapsida SARS-CoV-2 virusa za proizvodnju reagenasa i dijagnostičkih testova na novi korona virus. 2023;.
https://hdl.handle.net/21.15107/rcub_intor_860 .

Ćirković-Veličković, Tanja, Gnjatović, Marija, Ćujić, Danica, Todorović, Aleksandra, Stanić-Vučinić, Dragana, Đukić, Teodora, Mladenović, Maja, Vasović, Tamara, Stojadinović, Marija, Krstić-Ristivojević, Maja, Jovanović, Vesna, Simović, Ana, Radosavljević, Jelena, Aćimović, Jelena M., Radomirović, Mirjana Ž., Stojanović, Marijana, "Dobijanje rekombinantnog imunogenog fragmenta proteina nukleokapsida SARS-CoV-2 virusa za proizvodnju reagenasa i dijagnostičkih testova na novi korona virus" (2023),
https://hdl.handle.net/21.15107/rcub_intor_860 .

TY  - JOUR
AU  - Mögling, Ramona
AU  - Reimerink, Johan
AU  - Stanoeva, Kamelia R.
AU  - Keramarou, Maria
AU  - Guiomar, Raquel
AU  - Costa, Inês
AU  - Haveri, Anu
AU  - Holzer, Barbara
AU  - Korukluoğlu, Gülay
AU  - Nguyen, Trung
AU  - Pakarna, Gatis
AU  - Pancer, Katarzyna
AU  - Trilar, Katarina Prosenc
AU  - Protić, Jelena
AU  - Stojanović, Marijana
AU  - De Santis, Riccardo
AU  - Lista, Florigio
AU  - Vremera, Teodora
AU  - Leustean, Mihaela
AU  - Pistol, Adriana
AU  - Zelena, Hana
AU  - Reusken, Chantal
AU  - Broberg, Eeva K.
PY  - 2023
UR  - http://intor.torlakinstitut.com/handle/123456789/805
AB  - One consequence of the ongoing coronavirus disease pandemic was the rapid development of both in-house and commercial serological assays detecting anti-SARS-CoV-2 antibodies, in an effort to reliably detect acute and past SARS-CoV-2 infections. It is crucial to evaluate the quality of these serological tests and consequently the sero-epidemiological studies that are performed with the respective tests. Here, we describe the set-up and results of a comparative study, in which a laboratory contracted by the European Centre for Disease Prevention and Control offered a centralised service to EU/EEA Member and pre-accession Member States to test representative serum specimens with known serological results, with the gold standard technique (virus neutralisation tests) to determine the presence of neutralising antibodies. Laboratories from 12 European countries shared 719 serum specimens with the contractor laboratory. We found that in-house serological tests detecting neutralising antibodies showed the highest percent agreement, both positive and negative, with the virus neutralisation test results. Despite extensive differences in virus neutralisation protocols neutralisation titres showed a strong correlation. From the commercial assays, the best positive percent agreement was found for SARS-CoV-2 IgG (sCOVG) (Siemens - Atellica IM Analyzer). Despite lower positive percent agreement of LIAISON SARS-CoV-2 TrimericS IgG kit (Diasorin Inc.), the obtained results showed relatively good correlation with neutralisation titres. The set-up of this study allowed for high comparability between laboratories and enabled laboratories that do not have the capacity or capability to perform VNTs themselves. Given the variety of in-house protocols detecting SARS-CoV-2 specific neutralising antibodies, including the virus strain, it could be of interest to select reference isolates for SARS-CoV-2 diagnostic to be made available for interested EU Member States and pre-accession countries.
PB  - Elsevier
T2  - Journal of Virological Methods
T1  - Comparative study between virus neutralisation testing and other serological methods detecting anti-SARS-CoV-2 antibodies in Europe, 2021
SP  - 114825
VL  - 322
DO  - 10.1016/j.jviromet.2023.114825
ER  - 

@article{
author = "Mögling, Ramona and Reimerink, Johan and Stanoeva, Kamelia R. and Keramarou, Maria and Guiomar, Raquel and Costa, Inês and Haveri, Anu and Holzer, Barbara and Korukluoğlu, Gülay and Nguyen, Trung and Pakarna, Gatis and Pancer, Katarzyna and Trilar, Katarina Prosenc and Protić, Jelena and Stojanović, Marijana and De Santis, Riccardo and Lista, Florigio and Vremera, Teodora and Leustean, Mihaela and Pistol, Adriana and Zelena, Hana and Reusken, Chantal and Broberg, Eeva K.",
year = "2023",
abstract = "One consequence of the ongoing coronavirus disease pandemic was the rapid development of both in-house and commercial serological assays detecting anti-SARS-CoV-2 antibodies, in an effort to reliably detect acute and past SARS-CoV-2 infections. It is crucial to evaluate the quality of these serological tests and consequently the sero-epidemiological studies that are performed with the respective tests. Here, we describe the set-up and results of a comparative study, in which a laboratory contracted by the European Centre for Disease Prevention and Control offered a centralised service to EU/EEA Member and pre-accession Member States to test representative serum specimens with known serological results, with the gold standard technique (virus neutralisation tests) to determine the presence of neutralising antibodies. Laboratories from 12 European countries shared 719 serum specimens with the contractor laboratory. We found that in-house serological tests detecting neutralising antibodies showed the highest percent agreement, both positive and negative, with the virus neutralisation test results. Despite extensive differences in virus neutralisation protocols neutralisation titres showed a strong correlation. From the commercial assays, the best positive percent agreement was found for SARS-CoV-2 IgG (sCOVG) (Siemens - Atellica IM Analyzer). Despite lower positive percent agreement of LIAISON SARS-CoV-2 TrimericS IgG kit (Diasorin Inc.), the obtained results showed relatively good correlation with neutralisation titres. The set-up of this study allowed for high comparability between laboratories and enabled laboratories that do not have the capacity or capability to perform VNTs themselves. Given the variety of in-house protocols detecting SARS-CoV-2 specific neutralising antibodies, including the virus strain, it could be of interest to select reference isolates for SARS-CoV-2 diagnostic to be made available for interested EU Member States and pre-accession countries.",
publisher = "Elsevier",
journal = "Journal of Virological Methods",
title = "Comparative study between virus neutralisation testing and other serological methods detecting anti-SARS-CoV-2 antibodies in Europe, 2021",
pages = "114825",
volume = "322",
doi = "10.1016/j.jviromet.2023.114825"
}

Mögling, R., Reimerink, J., Stanoeva, K. R., Keramarou, M., Guiomar, R., Costa, I., Haveri, A., Holzer, B., Korukluoğlu, G., Nguyen, T., Pakarna, G., Pancer, K., Trilar, K. P., Protić, J., Stojanović, M., De Santis, R., Lista, F., Vremera, T., Leustean, M., Pistol, A., Zelena, H., Reusken, C.,& Broberg, E. K.. (2023). Comparative study between virus neutralisation testing and other serological methods detecting anti-SARS-CoV-2 antibodies in Europe, 2021. in Journal of Virological Methods
Elsevier., 322, 114825.
https://doi.org/10.1016/j.jviromet.2023.114825

Mögling R, Reimerink J, Stanoeva KR, Keramarou M, Guiomar R, Costa I, Haveri A, Holzer B, Korukluoğlu G, Nguyen T, Pakarna G, Pancer K, Trilar KP, Protić J, Stojanović M, De Santis R, Lista F, Vremera T, Leustean M, Pistol A, Zelena H, Reusken C, Broberg EK. Comparative study between virus neutralisation testing and other serological methods detecting anti-SARS-CoV-2 antibodies in Europe, 2021. in Journal of Virological Methods. 2023;322:114825.
doi:10.1016/j.jviromet.2023.114825 .

Mögling, Ramona, Reimerink, Johan, Stanoeva, Kamelia R., Keramarou, Maria, Guiomar, Raquel, Costa, Inês, Haveri, Anu, Holzer, Barbara, Korukluoğlu, Gülay, Nguyen, Trung, Pakarna, Gatis, Pancer, Katarzyna, Trilar, Katarina Prosenc, Protić, Jelena, Stojanović, Marijana, De Santis, Riccardo, Lista, Florigio, Vremera, Teodora, Leustean, Mihaela, Pistol, Adriana, Zelena, Hana, Reusken, Chantal, Broberg, Eeva K., "Comparative study between virus neutralisation testing and other serological methods detecting anti-SARS-CoV-2 antibodies in Europe, 2021" in Journal of Virological Methods, 322 (2023):114825,
https://doi.org/10.1016/j.jviromet.2023.114825 . .

TY  - JOUR
AU  - Ljubic, Verica
AU  - Milosevic, Milena
AU  - Cvetkovic, Slobodan
AU  - Stojanović, Marijana
AU  - Novovic, Katarina
AU  - Dinic, Miroslav
AU  - Popovic, Mina
PY  - 2022
UR  - http://intor.torlakinstitut.com/handle/123456789/631
AB  - As one of the most promising groups of microbes, lactic acid bacteria (LAB) can synthesize metabolites that can be used in different industries over the world, mainly in the pharmaceutical, food, and dairy industries. In this study, a novel exopolysaccharide was extracted and isolated from the probiotic strain Lactobacillus reuteri B2, and assessed on biological activity and its possible application as a biosorbent for the removal of Ni2+ ions from contaminated water. New exopolysaccharide was characterized using FTIR, SEM, XRD, NMR, MALDI-TOF MS, and TGA/DTG analysis. Biological assays included antioxidative activity, cytotoxic assay, and adhesion assay of L. reuteri B2 to HT29 cells. Our hypothesis was that if this exopolysaccharide is nontoxic, it can be used as a novel biomaterial for the possible application of the removal of Ni2+ ions from contaminated water. The scavenging effect of nontoxic exopolysaccharide was 76% at 2 mg/mL using ABTS assay, in biological assays, while the removal efficiency of nickel from the aqueous solution was 92.96% in biosorption study. According to these results, this exopolysaccharide can be considered a very promising biomaterial for potential application in different industries, from pharmacy to wastewater treatments.
PB  - Springer
T2  - Biomass Conversion and Biorefinery
T1  - The new exopolysaccharide produced by the probiotic strain L. reuteri B2: extraction, biological properties, and possible application for Ni2+ ion removal from the contaminated water
DO  - 10.1007/s13399-022-03292-5
ER  - 

@article{
author = "Ljubic, Verica and Milosevic, Milena and Cvetkovic, Slobodan and Stojanović, Marijana and Novovic, Katarina and Dinic, Miroslav and Popovic, Mina",
year = "2022",
abstract = "As one of the most promising groups of microbes, lactic acid bacteria (LAB) can synthesize metabolites that can be used in different industries over the world, mainly in the pharmaceutical, food, and dairy industries. In this study, a novel exopolysaccharide was extracted and isolated from the probiotic strain Lactobacillus reuteri B2, and assessed on biological activity and its possible application as a biosorbent for the removal of Ni2+ ions from contaminated water. New exopolysaccharide was characterized using FTIR, SEM, XRD, NMR, MALDI-TOF MS, and TGA/DTG analysis. Biological assays included antioxidative activity, cytotoxic assay, and adhesion assay of L. reuteri B2 to HT29 cells. Our hypothesis was that if this exopolysaccharide is nontoxic, it can be used as a novel biomaterial for the possible application of the removal of Ni2+ ions from contaminated water. The scavenging effect of nontoxic exopolysaccharide was 76% at 2 mg/mL using ABTS assay, in biological assays, while the removal efficiency of nickel from the aqueous solution was 92.96% in biosorption study. According to these results, this exopolysaccharide can be considered a very promising biomaterial for potential application in different industries, from pharmacy to wastewater treatments.",
publisher = "Springer",
journal = "Biomass Conversion and Biorefinery",
title = "The new exopolysaccharide produced by the probiotic strain L. reuteri B2: extraction, biological properties, and possible application for Ni2+ ion removal from the contaminated water",
doi = "10.1007/s13399-022-03292-5"
}

Ljubic, V., Milosevic, M., Cvetkovic, S., Stojanović, M., Novovic, K., Dinic, M.,& Popovic, M.. (2022). The new exopolysaccharide produced by the probiotic strain L. reuteri B2: extraction, biological properties, and possible application for Ni2+ ion removal from the contaminated water. in Biomass Conversion and Biorefinery
Springer..
https://doi.org/10.1007/s13399-022-03292-5

Ljubic V, Milosevic M, Cvetkovic S, Stojanović M, Novovic K, Dinic M, Popovic M. The new exopolysaccharide produced by the probiotic strain L. reuteri B2: extraction, biological properties, and possible application for Ni2+ ion removal from the contaminated water. in Biomass Conversion and Biorefinery. 2022;.
doi:10.1007/s13399-022-03292-5 .

Ljubic, Verica, Milosevic, Milena, Cvetkovic, Slobodan, Stojanović, Marijana, Novovic, Katarina, Dinic, Miroslav, Popovic, Mina, "The new exopolysaccharide produced by the probiotic strain L. reuteri B2: extraction, biological properties, and possible application for Ni2+ ion removal from the contaminated water" in Biomass Conversion and Biorefinery (2022),
https://doi.org/10.1007/s13399-022-03292-5 . .

TY  - DATA
AU  - Ljubić, Verica
AU  - Milošević, Milena
AU  - Cvetković, Slobodan
AU  - Stojanović, Marijana
AU  - Novović, Katarina
AU  - Dinić, Miroslav
AU  - Popović, Mina
PY  - 2022
UR  - http://intor.torlakinstitut.com/handle/123456789/644
AB  - Using the universal primers (UNI16Sfw and UNI16Srev) [1], the representative isolates were identified by 16S rDNA sequencing. Amplification was carried out in a thermal cycler (Applied Biosystems, ThermoFisher Scientific) and DNA fragments were amplified as follows: initial denaturation at 94 °C for 5 min, followed by 30 cycles consisting of denaturation at 94 °C for 1 min, annealing at 55°C for 1 min, and polymerization at 72°C for 1 min, and a final extension at 72°C for 7 min. The expected length was 1549 bp. Aliquots (5 µl) of the amplified products were subjected to electrophoresis in 1% agarose gel (ThermoFisher Scientific) in TAE buffer (40 mM Tris acetate, 1 mM EDTA, pH 8.2). Gels were stained with ethidium bromide (500 ng/mL) and visualized under UV light (BioDoc Analyze). All amplicons were eluted and purified using GeneJet PCR Purification Kit (ThermoScientific) by following the manufacturer`s protocol. The PCR products that we obtained were sequenced by the Macrogen Sequencing Service (Macrogen, Amsterdam, The Netherlands) and analyzed by using BLAST algorithm (http://www.ncbi.nlm.nih.gov/index.html). Selected isolates were identified as follows: isolate B2 - Lacotbacillus reuteri, isolate H10 - Lactobacillus murinus, and isolate J7 - Klebsiella oxytoca [2]. The most numerous colonies belong to isolate B2, hence it was chosen for further characterization as a potential source for exopolysaccharide (EPS) production.
PB  - Springer
T2  - Biomass Conversion and Biorefinery
T1  - Supplementary information for the article: Ljubic, V.; Milosevic, M.; Cvetkovic, S.; Stojanovic, M.; Novovic, K.; Dinic, M.; Popovic, M. The New Exopolysaccharide Produced by the Probiotic Strain L. Reuteri B2: Extraction, Biological Properties, and Possible Application for Ni2+ Ion Removal from the Contaminated Water. Biomass Conversion and Biorefinery 2022. https://doi.org/10.1007/s13399-022-03292-5.
UR  - https://hdl.handle.net/21.15107/rcub_intor_644
ER  - 

@misc{
author = "Ljubić, Verica and Milošević, Milena and Cvetković, Slobodan and Stojanović, Marijana and Novović, Katarina and Dinić, Miroslav and Popović, Mina",
year = "2022",
abstract = "Using the universal primers (UNI16Sfw and UNI16Srev) [1], the representative isolates were identified by 16S rDNA sequencing. Amplification was carried out in a thermal cycler (Applied Biosystems, ThermoFisher Scientific) and DNA fragments were amplified as follows: initial denaturation at 94 °C for 5 min, followed by 30 cycles consisting of denaturation at 94 °C for 1 min, annealing at 55°C for 1 min, and polymerization at 72°C for 1 min, and a final extension at 72°C for 7 min. The expected length was 1549 bp. Aliquots (5 µl) of the amplified products were subjected to electrophoresis in 1% agarose gel (ThermoFisher Scientific) in TAE buffer (40 mM Tris acetate, 1 mM EDTA, pH 8.2). Gels were stained with ethidium bromide (500 ng/mL) and visualized under UV light (BioDoc Analyze). All amplicons were eluted and purified using GeneJet PCR Purification Kit (ThermoScientific) by following the manufacturer`s protocol. The PCR products that we obtained were sequenced by the Macrogen Sequencing Service (Macrogen, Amsterdam, The Netherlands) and analyzed by using BLAST algorithm (http://www.ncbi.nlm.nih.gov/index.html). Selected isolates were identified as follows: isolate B2 - Lacotbacillus reuteri, isolate H10 - Lactobacillus murinus, and isolate J7 - Klebsiella oxytoca [2]. The most numerous colonies belong to isolate B2, hence it was chosen for further characterization as a potential source for exopolysaccharide (EPS) production.",
publisher = "Springer",
journal = "Biomass Conversion and Biorefinery",
title = "Supplementary information for the article: Ljubic, V.; Milosevic, M.; Cvetkovic, S.; Stojanovic, M.; Novovic, K.; Dinic, M.; Popovic, M. The New Exopolysaccharide Produced by the Probiotic Strain L. Reuteri B2: Extraction, Biological Properties, and Possible Application for Ni2+ Ion Removal from the Contaminated Water. Biomass Conversion and Biorefinery 2022. https://doi.org/10.1007/s13399-022-03292-5.",
url = "https://hdl.handle.net/21.15107/rcub_intor_644"
}

Ljubić, V., Milošević, M., Cvetković, S., Stojanović, M., Novović, K., Dinić, M.,& Popović, M.. (2022). Supplementary information for the article: Ljubic, V.; Milosevic, M.; Cvetkovic, S.; Stojanovic, M.; Novovic, K.; Dinic, M.; Popovic, M. The New Exopolysaccharide Produced by the Probiotic Strain L. Reuteri B2: Extraction, Biological Properties, and Possible Application for Ni2+ Ion Removal from the Contaminated Water. Biomass Conversion and Biorefinery 2022. https://doi.org/10.1007/s13399-022-03292-5.. in Biomass Conversion and Biorefinery
Springer..
https://hdl.handle.net/21.15107/rcub_intor_644

Ljubić V, Milošević M, Cvetković S, Stojanović M, Novović K, Dinić M, Popović M. Supplementary information for the article: Ljubic, V.; Milosevic, M.; Cvetkovic, S.; Stojanovic, M.; Novovic, K.; Dinic, M.; Popovic, M. The New Exopolysaccharide Produced by the Probiotic Strain L. Reuteri B2: Extraction, Biological Properties, and Possible Application for Ni2+ Ion Removal from the Contaminated Water. Biomass Conversion and Biorefinery 2022. https://doi.org/10.1007/s13399-022-03292-5.. in Biomass Conversion and Biorefinery. 2022;.
https://hdl.handle.net/21.15107/rcub_intor_644 .

Ljubić, Verica, Milošević, Milena, Cvetković, Slobodan, Stojanović, Marijana, Novović, Katarina, Dinić, Miroslav, Popović, Mina, "Supplementary information for the article: Ljubic, V.; Milosevic, M.; Cvetkovic, S.; Stojanovic, M.; Novovic, K.; Dinic, M.; Popovic, M. The New Exopolysaccharide Produced by the Probiotic Strain L. Reuteri B2: Extraction, Biological Properties, and Possible Application for Ni2+ Ion Removal from the Contaminated Water. Biomass Conversion and Biorefinery 2022. https://doi.org/10.1007/s13399-022-03292-5." in Biomass Conversion and Biorefinery (2022),
https://hdl.handle.net/21.15107/rcub_intor_644 .

TY  - DATA
AU  - Frohns, Antonia
AU  - Stojanović, Marijana
AU  - Barisani-Asenbauer, Talin
AU  - Kuratli, Jasmin
AU  - Borel, Nicole
AU  - Inić-Kanada, Aleksandra
PY  - 2021
UR  - http://intor.torlakinstitut.com/handle/123456789/639
AB  - Figure S1. Structure of guinea pig retina and cornea. Nuclear staining with DAPI of guinea pig retina and cornea. In the retina, three nuclear layers (ONL-outer nuclear layer, INL-inner nuclear layer, and GCL-ganglion cell layer) are present. In the ONL, the photoreceptor cells are located, with their outer segments (OS) projecting into the adjacent retinal pigment epithelium (RPE). The INL exhibit nuclei of horizontal-, bipolar-, Müller glia, and amacrine cells. The cornea consists of an epithelium (EP), the stroma (S) containing keratinocytes, and the endothelium (EN); Figure S2. Western blots of HSPs in the guinea pig retina and cornea 19 days after wIRA/VIS treatment. Exemplary western blots of HSP40, HSP60, HSP70, and HSP90 in the retina and cornea. Comparison of non-irradiated (no wIRA/VIS) tissues with ones subjected to single (wIRA/VIS 1×) or sequential (wIRA/VIS 2×) irradiation. The constitutively expressed protein vinculin served as a loading control; Figure S3. Localization of Vinculin in the guinea pig retina and cornea 19 days after wIRA/VIS treatment. Immunolabeling of Vinculin in retinal layers (OS-outer segments, ONL-outer nuclear layer, INL-inner nuclear layer, and GCL-ganglion cell layer) and corneal layers (EP-epithelium, S-Stroma, and EN-endothelium). Comparison of non-irradiated (no wIRA/VIS) tissues with ones subjected to single (wIRA/VIS 1×) or sequential (wIRA/VIS 2×) irradiation. Inserts in cornea images show magnified sections of EP and EN. Scale bars = 10 μm retina, 20 μm cornea; Figure S4. Western blots of stress-responsive proteins in the guinea pig retina and cornea 19 days after wIRA/VIS treatment. Exemplary western blots of Akt, pAkt, pPTEN, pcRaf, Erk1/2, and pErk1/2 in the retina and cornea. Comparison of non-irradiated (no wIRA/VIS) tissues with ones subjected to single (wIRA/VIS 1×) or sequential (wIRA/VIS 2×) irradiation. The constitutively expressed protein vinculin served as a loading control.
PB  - Elsevier
T2  - Journal of Photochemistry and Photobiology B: Biology
T1  - Supplementary material for: Frohns, A.; Stojanović, M.; Barisani-Asenbauer, T.; Kuratli, J.; Borel, N.; Inić-Kanada, A. Effects of Water-Filtered Infrared A and Visible Light (WIRA/VIS) Radiation on Heat- and Stress-Responsive Proteins in the Retina and Cornea of Guinea Pigs. Journal of Photochemistry and Photobiology B: Biology 2021, 224, 112306. https://doi.org/10.1016/j.jphotobiol.2021.112306.
SP  - 112306
VL  - 224
UR  - https://hdl.handle.net/21.15107/rcub_intor_639
ER  - 

@misc{
author = "Frohns, Antonia and Stojanović, Marijana and Barisani-Asenbauer, Talin and Kuratli, Jasmin and Borel, Nicole and Inić-Kanada, Aleksandra",
year = "2021",
abstract = "Figure S1. Structure of guinea pig retina and cornea. Nuclear staining with DAPI of guinea pig retina and cornea. In the retina, three nuclear layers (ONL-outer nuclear layer, INL-inner nuclear layer, and GCL-ganglion cell layer) are present. In the ONL, the photoreceptor cells are located, with their outer segments (OS) projecting into the adjacent retinal pigment epithelium (RPE). The INL exhibit nuclei of horizontal-, bipolar-, Müller glia, and amacrine cells. The cornea consists of an epithelium (EP), the stroma (S) containing keratinocytes, and the endothelium (EN); Figure S2. Western blots of HSPs in the guinea pig retina and cornea 19 days after wIRA/VIS treatment. Exemplary western blots of HSP40, HSP60, HSP70, and HSP90 in the retina and cornea. Comparison of non-irradiated (no wIRA/VIS) tissues with ones subjected to single (wIRA/VIS 1×) or sequential (wIRA/VIS 2×) irradiation. The constitutively expressed protein vinculin served as a loading control; Figure S3. Localization of Vinculin in the guinea pig retina and cornea 19 days after wIRA/VIS treatment. Immunolabeling of Vinculin in retinal layers (OS-outer segments, ONL-outer nuclear layer, INL-inner nuclear layer, and GCL-ganglion cell layer) and corneal layers (EP-epithelium, S-Stroma, and EN-endothelium). Comparison of non-irradiated (no wIRA/VIS) tissues with ones subjected to single (wIRA/VIS 1×) or sequential (wIRA/VIS 2×) irradiation. Inserts in cornea images show magnified sections of EP and EN. Scale bars = 10 μm retina, 20 μm cornea; Figure S4. Western blots of stress-responsive proteins in the guinea pig retina and cornea 19 days after wIRA/VIS treatment. Exemplary western blots of Akt, pAkt, pPTEN, pcRaf, Erk1/2, and pErk1/2 in the retina and cornea. Comparison of non-irradiated (no wIRA/VIS) tissues with ones subjected to single (wIRA/VIS 1×) or sequential (wIRA/VIS 2×) irradiation. The constitutively expressed protein vinculin served as a loading control.",
publisher = "Elsevier",
journal = "Journal of Photochemistry and Photobiology B: Biology",
title = "Supplementary material for: Frohns, A.; Stojanović, M.; Barisani-Asenbauer, T.; Kuratli, J.; Borel, N.; Inić-Kanada, A. Effects of Water-Filtered Infrared A and Visible Light (WIRA/VIS) Radiation on Heat- and Stress-Responsive Proteins in the Retina and Cornea of Guinea Pigs. Journal of Photochemistry and Photobiology B: Biology 2021, 224, 112306. https://doi.org/10.1016/j.jphotobiol.2021.112306.",
pages = "112306",
volume = "224",
url = "https://hdl.handle.net/21.15107/rcub_intor_639"
}

Frohns, A., Stojanović, M., Barisani-Asenbauer, T., Kuratli, J., Borel, N.,& Inić-Kanada, A.. (2021). Supplementary material for: Frohns, A.; Stojanović, M.; Barisani-Asenbauer, T.; Kuratli, J.; Borel, N.; Inić-Kanada, A. Effects of Water-Filtered Infrared A and Visible Light (WIRA/VIS) Radiation on Heat- and Stress-Responsive Proteins in the Retina and Cornea of Guinea Pigs. Journal of Photochemistry and Photobiology B: Biology 2021, 224, 112306. https://doi.org/10.1016/j.jphotobiol.2021.112306.. in Journal of Photochemistry and Photobiology B: Biology
Elsevier., 224, 112306.
https://hdl.handle.net/21.15107/rcub_intor_639

Frohns A, Stojanović M, Barisani-Asenbauer T, Kuratli J, Borel N, Inić-Kanada A. Supplementary material for: Frohns, A.; Stojanović, M.; Barisani-Asenbauer, T.; Kuratli, J.; Borel, N.; Inić-Kanada, A. Effects of Water-Filtered Infrared A and Visible Light (WIRA/VIS) Radiation on Heat- and Stress-Responsive Proteins in the Retina and Cornea of Guinea Pigs. Journal of Photochemistry and Photobiology B: Biology 2021, 224, 112306. https://doi.org/10.1016/j.jphotobiol.2021.112306.. in Journal of Photochemistry and Photobiology B: Biology. 2021;224:112306.
https://hdl.handle.net/21.15107/rcub_intor_639 .

Frohns, Antonia, Stojanović, Marijana, Barisani-Asenbauer, Talin, Kuratli, Jasmin, Borel, Nicole, Inić-Kanada, Aleksandra, "Supplementary material for: Frohns, A.; Stojanović, M.; Barisani-Asenbauer, T.; Kuratli, J.; Borel, N.; Inić-Kanada, A. Effects of Water-Filtered Infrared A and Visible Light (WIRA/VIS) Radiation on Heat- and Stress-Responsive Proteins in the Retina and Cornea of Guinea Pigs. Journal of Photochemistry and Photobiology B: Biology 2021, 224, 112306. https://doi.org/10.1016/j.jphotobiol.2021.112306." in Journal of Photochemistry and Photobiology B: Biology, 224 (2021):112306,
https://hdl.handle.net/21.15107/rcub_intor_639 .

TY  - DATA
AU  - Popović, Mina
AU  - Stojanović, Marijana
AU  - Veličković, Zlate
AU  - Kovačević, Ana
AU  - Miljković, Radmila
AU  - Mirković, Nemanja
AU  - Marinković, Aleksandar D.
PY  - 2021
UR  - https://cer.ihtm.bg.ac.rs/handle/123456789/4596
UR  - http://intor.torlakinstitut.com/handle/123456789/627
AB  - Preparation of materials for encapsulation (Materials, Laboratory isolation of ricinoleic acid, Laboratory preparation of starch maleate monoester, Characterization, Statistical analysis). Additional results (Antimicrobial activity, Optimization of encapsulation yield, Size distribution of alginate beads in acidic conditions). additional references
PB  - Elsevier
T2  - International Journal of Biological Macromolecules
T1  - Supplementary material for: Popović, M.; Stojanović, M.; Veličković, Z.; Kovačević, A.; Miljković, R.; Mirković, N.; Marinković, A. D. Characterization of Potential Probiotic Strain, L. Reuteri B2, and Its Microencapsulation Using Alginate-Based Biopolymers. International Journal of Biological Macromolecules 2021, 183, 423–434. https://doi.org/10.1016/j.ijbiomac.2021.04.177.
UR  - https://hdl.handle.net/21.15107/rcub_intor_627
ER  - 

@misc{
author = "Popović, Mina and Stojanović, Marijana and Veličković, Zlate and Kovačević, Ana and Miljković, Radmila and Mirković, Nemanja and Marinković, Aleksandar D.",
year = "2021",
abstract = "Preparation of materials for encapsulation (Materials, Laboratory isolation of ricinoleic acid, Laboratory preparation of starch maleate monoester, Characterization, Statistical analysis). Additional results (Antimicrobial activity, Optimization of encapsulation yield, Size distribution of alginate beads in acidic conditions). additional references",
publisher = "Elsevier",
journal = "International Journal of Biological Macromolecules",
title = "Supplementary material for: Popović, M.; Stojanović, M.; Veličković, Z.; Kovačević, A.; Miljković, R.; Mirković, N.; Marinković, A. D. Characterization of Potential Probiotic Strain, L. Reuteri B2, and Its Microencapsulation Using Alginate-Based Biopolymers. International Journal of Biological Macromolecules 2021, 183, 423–434. https://doi.org/10.1016/j.ijbiomac.2021.04.177.",
url = "https://hdl.handle.net/21.15107/rcub_intor_627"
}

Popović, M., Stojanović, M., Veličković, Z., Kovačević, A., Miljković, R., Mirković, N.,& Marinković, A. D.. (2021). Supplementary material for: Popović, M.; Stojanović, M.; Veličković, Z.; Kovačević, A.; Miljković, R.; Mirković, N.; Marinković, A. D. Characterization of Potential Probiotic Strain, L. Reuteri B2, and Its Microencapsulation Using Alginate-Based Biopolymers. International Journal of Biological Macromolecules 2021, 183, 423–434. https://doi.org/10.1016/j.ijbiomac.2021.04.177.. in International Journal of Biological Macromolecules
Elsevier..
https://hdl.handle.net/21.15107/rcub_intor_627

Popović M, Stojanović M, Veličković Z, Kovačević A, Miljković R, Mirković N, Marinković AD. Supplementary material for: Popović, M.; Stojanović, M.; Veličković, Z.; Kovačević, A.; Miljković, R.; Mirković, N.; Marinković, A. D. Characterization of Potential Probiotic Strain, L. Reuteri B2, and Its Microencapsulation Using Alginate-Based Biopolymers. International Journal of Biological Macromolecules 2021, 183, 423–434. https://doi.org/10.1016/j.ijbiomac.2021.04.177.. in International Journal of Biological Macromolecules. 2021;.
https://hdl.handle.net/21.15107/rcub_intor_627 .

Popović, Mina, Stojanović, Marijana, Veličković, Zlate, Kovačević, Ana, Miljković, Radmila, Mirković, Nemanja, Marinković, Aleksandar D., "Supplementary material for: Popović, M.; Stojanović, M.; Veličković, Z.; Kovačević, A.; Miljković, R.; Mirković, N.; Marinković, A. D. Characterization of Potential Probiotic Strain, L. Reuteri B2, and Its Microencapsulation Using Alginate-Based Biopolymers. International Journal of Biological Macromolecules 2021, 183, 423–434. https://doi.org/10.1016/j.ijbiomac.2021.04.177." in International Journal of Biological Macromolecules (2021),
https://hdl.handle.net/21.15107/rcub_intor_627 .

TY  - JOUR
AU  - Popović, Mina
AU  - Stojanović, Marijana
AU  - Veličković, Zlate
AU  - Kovačević, Ana
AU  - Miljković, Radmila
AU  - Mirković, Nemanja
AU  - Marinković, Aleksandar D.
PY  - 2021
UR  - https://cer.ihtm.bg.ac.rs/handle/123456789/4595
UR  - http://intor.torlakinstitut.com/handle/123456789/628
AB  - In this study, Lactobacillus reuteri B2was isolated fromthe feces of C57BL/6 mice and assessed on probiotic activity.L. reuteri B2was identified by 16S rDNA sequencing, which the cell viability in acidic conditions at pH 2.0was64% after 2 h, and in the presents of 0.30% of the bile salts, after 6 h, was 37%. Antimicrobial assay with L. reuteri B2showed maximumdiameters against Klebsiela oxytoca J7 (12.5±0.71mm).Wefurther hypothesized if L. reuteriB2 strain in the free form can survive all conditions in the gastrointestinal tract (GIT) then the utilization of theappropriate biomaterials would ameliorate its stability and viability in GIT. L. reuteri B2 was microencapsulatedinto sodium alginate-(Na-alg) and different content of Na-alg and sodium maleate (SM) beads. Characterizationmaterials enveloped their thermal characteristics (TGA/DTA analysis) and structure using: scanning electron microscopy(SEM), FTIR, and particle size distribution. The high survival rate of L. reuteri B2 at lowpH from2.0 to 4.0and in the presence of the bile salts, at concentrations up to 0.30%, was obtained. L. reuteri B2 showed strong antimicrobialactivity and the best protection microencapsulated with Na-alg + SM in simulated gastric juices(SGJ).
PB  - Elsevier
T2  - International Journal of Biological Macromolecules
T1  - Characterization of potential probiotic strain, L. reuteri B2, and its microencapsulation using alginate-based biopolymers
EP  - 434
SP  - 423
VL  - 183
DO  - 10.1016/j.ijbiomac.2021.04.177
ER  - 

@article{
author = "Popović, Mina and Stojanović, Marijana and Veličković, Zlate and Kovačević, Ana and Miljković, Radmila and Mirković, Nemanja and Marinković, Aleksandar D.",
year = "2021",
abstract = "In this study, Lactobacillus reuteri B2was isolated fromthe feces of C57BL/6 mice and assessed on probiotic activity.L. reuteri B2was identified by 16S rDNA sequencing, which the cell viability in acidic conditions at pH 2.0was64% after 2 h, and in the presents of 0.30% of the bile salts, after 6 h, was 37%. Antimicrobial assay with L. reuteri B2showed maximumdiameters against Klebsiela oxytoca J7 (12.5±0.71mm).Wefurther hypothesized if L. reuteriB2 strain in the free form can survive all conditions in the gastrointestinal tract (GIT) then the utilization of theappropriate biomaterials would ameliorate its stability and viability in GIT. L. reuteri B2 was microencapsulatedinto sodium alginate-(Na-alg) and different content of Na-alg and sodium maleate (SM) beads. Characterizationmaterials enveloped their thermal characteristics (TGA/DTA analysis) and structure using: scanning electron microscopy(SEM), FTIR, and particle size distribution. The high survival rate of L. reuteri B2 at lowpH from2.0 to 4.0and in the presence of the bile salts, at concentrations up to 0.30%, was obtained. L. reuteri B2 showed strong antimicrobialactivity and the best protection microencapsulated with Na-alg + SM in simulated gastric juices(SGJ).",
publisher = "Elsevier",
journal = "International Journal of Biological Macromolecules",
title = "Characterization of potential probiotic strain, L. reuteri B2, and its microencapsulation using alginate-based biopolymers",
pages = "434-423",
volume = "183",
doi = "10.1016/j.ijbiomac.2021.04.177"
}

Popović, M., Stojanović, M., Veličković, Z., Kovačević, A., Miljković, R., Mirković, N.,& Marinković, A. D.. (2021). Characterization of potential probiotic strain, L. reuteri B2, and its microencapsulation using alginate-based biopolymers. in International Journal of Biological Macromolecules
Elsevier., 183, 423-434.
https://doi.org/10.1016/j.ijbiomac.2021.04.177

Popović M, Stojanović M, Veličković Z, Kovačević A, Miljković R, Mirković N, Marinković AD. Characterization of potential probiotic strain, L. reuteri B2, and its microencapsulation using alginate-based biopolymers. in International Journal of Biological Macromolecules. 2021;183:423-434.
doi:10.1016/j.ijbiomac.2021.04.177 .

Popović, Mina, Stojanović, Marijana, Veličković, Zlate, Kovačević, Ana, Miljković, Radmila, Mirković, Nemanja, Marinković, Aleksandar D., "Characterization of potential probiotic strain, L. reuteri B2, and its microencapsulation using alginate-based biopolymers" in International Journal of Biological Macromolecules, 183 (2021):423-434,
https://doi.org/10.1016/j.ijbiomac.2021.04.177 . .

TY  - JOUR
AU  - Popović, Mina
AU  - Stojanović, Marijana
AU  - Veličković, Zlate
AU  - Kovačević, Ana
AU  - Miljković, Radmila
AU  - Mirković, Nemanja
AU  - Marinković, Aleksandar D.
PY  - 2021
UR  - https://cer.ihtm.bg.ac.rs/handle/123456789/4594
UR  - http://intor.torlakinstitut.com/handle/123456789/626
AB  - In this study, Lactobacillus reuteri B2was isolated fromthe feces of C57BL/6 mice and assessed on probiotic activity.L. reuteri B2was identified by 16S rDNA sequencing, which the cell viability in acidic conditions at pH 2.0was64% after 2 h, and in the presents of 0.30% of the bile salts, after 6 h, was 37%. Antimicrobial assay with L. reuteri B2showed maximumdiameters against Klebsiela oxytoca J7 (12.5±0.71mm).Wefurther hypothesized if L. reuteriB2 strain in the free form can survive all conditions in the gastrointestinal tract (GIT) then the utilization of theappropriate biomaterials would ameliorate its stability and viability in GIT. L. reuteri B2 was microencapsulatedinto sodium alginate-(Na-alg) and different content of Na-alg and sodium maleate (SM) beads. Characterizationmaterials enveloped their thermal characteristics (TGA/DTA analysis) and structure using: scanning electron microscopy(SEM), FTIR, and particle size distribution. The high survival rate of L. reuteri B2 at lowpH from2.0 to 4.0and in the presence of the bile salts, at concentrations up to 0.30%, was obtained. L. reuteri B2 showed strong antimicrobialactivity and the best protection microencapsulated with Na-alg + SM in simulated gastric juices(SGJ).
PB  - Elsevier
T2  - International Journal of Biological Macromolecules
T1  - Characterization of potential probiotic strain, L. reuteri B2, and its microencapsulation using alginate-based biopolymers
EP  - 434
SP  - 423
VL  - 183
DO  - 10.1016/j.ijbiomac.2021.04.177
ER  - 

@article{
author = "Popović, Mina and Stojanović, Marijana and Veličković, Zlate and Kovačević, Ana and Miljković, Radmila and Mirković, Nemanja and Marinković, Aleksandar D.",
year = "2021",
abstract = "In this study, Lactobacillus reuteri B2was isolated fromthe feces of C57BL/6 mice and assessed on probiotic activity.L. reuteri B2was identified by 16S rDNA sequencing, which the cell viability in acidic conditions at pH 2.0was64% after 2 h, and in the presents of 0.30% of the bile salts, after 6 h, was 37%. Antimicrobial assay with L. reuteri B2showed maximumdiameters against Klebsiela oxytoca J7 (12.5±0.71mm).Wefurther hypothesized if L. reuteriB2 strain in the free form can survive all conditions in the gastrointestinal tract (GIT) then the utilization of theappropriate biomaterials would ameliorate its stability and viability in GIT. L. reuteri B2 was microencapsulatedinto sodium alginate-(Na-alg) and different content of Na-alg and sodium maleate (SM) beads. Characterizationmaterials enveloped their thermal characteristics (TGA/DTA analysis) and structure using: scanning electron microscopy(SEM), FTIR, and particle size distribution. The high survival rate of L. reuteri B2 at lowpH from2.0 to 4.0and in the presence of the bile salts, at concentrations up to 0.30%, was obtained. L. reuteri B2 showed strong antimicrobialactivity and the best protection microencapsulated with Na-alg + SM in simulated gastric juices(SGJ).",
publisher = "Elsevier",
journal = "International Journal of Biological Macromolecules",
title = "Characterization of potential probiotic strain, L. reuteri B2, and its microencapsulation using alginate-based biopolymers",
pages = "434-423",
volume = "183",
doi = "10.1016/j.ijbiomac.2021.04.177"
}

Popović, M., Stojanović, M., Veličković, Z., Kovačević, A., Miljković, R., Mirković, N.,& Marinković, A. D.. (2021). Characterization of potential probiotic strain, L. reuteri B2, and its microencapsulation using alginate-based biopolymers. in International Journal of Biological Macromolecules
Elsevier., 183, 423-434.
https://doi.org/10.1016/j.ijbiomac.2021.04.177

Popović M, Stojanović M, Veličković Z, Kovačević A, Miljković R, Mirković N, Marinković AD. Characterization of potential probiotic strain, L. reuteri B2, and its microencapsulation using alginate-based biopolymers. in International Journal of Biological Macromolecules. 2021;183:423-434.
doi:10.1016/j.ijbiomac.2021.04.177 .

Popović, Mina, Stojanović, Marijana, Veličković, Zlate, Kovačević, Ana, Miljković, Radmila, Mirković, Nemanja, Marinković, Aleksandar D., "Characterization of potential probiotic strain, L. reuteri B2, and its microencapsulation using alginate-based biopolymers" in International Journal of Biological Macromolecules, 183 (2021):423-434,
https://doi.org/10.1016/j.ijbiomac.2021.04.177 . .

TY  - JOUR
AU  - Frohns, Antonia
AU  - Stojanović, Marijana
AU  - Barisani-Asenbauer, Talin
AU  - Kuratli, Jasmin
AU  - Borel, Nicole
AU  - Inić-Kanada, Aleksandra
PY  - 2021
UR  - http://intor.torlakinstitut.com/handle/123456789/617
AB  - Water-filtered infrared A and visible light (wIRA/VIS), shown to reduce chlamydial infections in vitro and in vivo, might represent an innovative therapeutic approach against trachoma, a neglected tropical disease caused by ocular infection with the bacterium C. trachomatis. In this in vivo study, we assessed the impact of wIRA radiation in combination with VIS (wavelength range 595–1400 nm, intensity 2100 W/m2) on the retina and cornea in a guinea pig animal model of inclusion conjunctivitis. We investigated the effects 19 days after wIRA/VIS irradiation by comparing a single and double wIRA/VIS treatment with a sham control. By immunolabeling and western blot analyses of critical heat- and stress-responsive proteins, we could not detect wIRA/VIS-induced changes in their expression pattern. Also, immunolabeling of specific retinal marker proteins revealed no changes in their expression pattern caused by the treatment. Our preclinical study suggests wIRA/VIS as a promising and safe therapeutic tool to treat ocular chlamydial infections.
PB  - Elsevier
T2  - Journal of Photochemistry and Photobiology B: Biology
T1  - Effects of water-filtered infrared A and visible light (wIRA/VIS) radiation on heat- and stress-responsive proteins in the retina and cornea of guinea pigs
SP  - 112306
VL  - 224
DO  - 10.1016/j.jphotobiol.2021.112306
ER  - 

@article{
author = "Frohns, Antonia and Stojanović, Marijana and Barisani-Asenbauer, Talin and Kuratli, Jasmin and Borel, Nicole and Inić-Kanada, Aleksandra",
year = "2021",
abstract = "Water-filtered infrared A and visible light (wIRA/VIS), shown to reduce chlamydial infections in vitro and in vivo, might represent an innovative therapeutic approach against trachoma, a neglected tropical disease caused by ocular infection with the bacterium C. trachomatis. In this in vivo study, we assessed the impact of wIRA radiation in combination with VIS (wavelength range 595–1400 nm, intensity 2100 W/m2) on the retina and cornea in a guinea pig animal model of inclusion conjunctivitis. We investigated the effects 19 days after wIRA/VIS irradiation by comparing a single and double wIRA/VIS treatment with a sham control. By immunolabeling and western blot analyses of critical heat- and stress-responsive proteins, we could not detect wIRA/VIS-induced changes in their expression pattern. Also, immunolabeling of specific retinal marker proteins revealed no changes in their expression pattern caused by the treatment. Our preclinical study suggests wIRA/VIS as a promising and safe therapeutic tool to treat ocular chlamydial infections.",
publisher = "Elsevier",
journal = "Journal of Photochemistry and Photobiology B: Biology",
title = "Effects of water-filtered infrared A and visible light (wIRA/VIS) radiation on heat- and stress-responsive proteins in the retina and cornea of guinea pigs",
pages = "112306",
volume = "224",
doi = "10.1016/j.jphotobiol.2021.112306"
}

Frohns, A., Stojanović, M., Barisani-Asenbauer, T., Kuratli, J., Borel, N.,& Inić-Kanada, A.. (2021). Effects of water-filtered infrared A and visible light (wIRA/VIS) radiation on heat- and stress-responsive proteins in the retina and cornea of guinea pigs. in Journal of Photochemistry and Photobiology B: Biology
Elsevier., 224, 112306.
https://doi.org/10.1016/j.jphotobiol.2021.112306

Frohns A, Stojanović M, Barisani-Asenbauer T, Kuratli J, Borel N, Inić-Kanada A. Effects of water-filtered infrared A and visible light (wIRA/VIS) radiation on heat- and stress-responsive proteins in the retina and cornea of guinea pigs. in Journal of Photochemistry and Photobiology B: Biology. 2021;224:112306.
doi:10.1016/j.jphotobiol.2021.112306 .

Frohns, Antonia, Stojanović, Marijana, Barisani-Asenbauer, Talin, Kuratli, Jasmin, Borel, Nicole, Inić-Kanada, Aleksandra, "Effects of water-filtered infrared A and visible light (wIRA/VIS) radiation on heat- and stress-responsive proteins in the retina and cornea of guinea pigs" in Journal of Photochemistry and Photobiology B: Biology, 224 (2021):112306,
https://doi.org/10.1016/j.jphotobiol.2021.112306 . .

TY  - JOUR
AU  - Protić-Rosić, Isidora
AU  - Nešić, Andrijana
AU  - Lukić, Ivana
AU  - Miljković, Radmila
AU  - Popović, Dragan
AU  - Atanasković-Marković, Marina
AU  - Stojanović, Marijana
AU  - Gavrović-Jankulović, Marija
PY  - 2021
UR  - http://intor.torlakinstitut.com/handle/123456789/630
AB  - Allergen-specific immunotherapy (AIT) is a desensitizing treatment for allergic diseases that corrects the underlined pathological immune response to innocuous protein antigens, called allergens. Recombinant allergens employed in the AIT allowed the production of well-defined formulations that possessed consistent quality but were often less efficient than natural allergen extracts. Combining recombinant allergens with an adjuvant or immunomodulatory agent could improve AIT efficacy. This study aimed to perform structural and functional characterization of newly designed recombinant chimera composed of the Bet v 1, the major birch pollen allergen, and Banana Lectin (BanLec), TLR2, and CD14 binding protein, for the application in AIT. rBet v 1-BanLec chimera was designed in silico and expressed as a soluble fraction in Escherichia coli. Purified rBet v 1-BanLec (33.4 kDa) retained BanLec-associated biological activity of carbohydrate-binding and preserved IgE reactive epitopes of Bet v 1. The chimera revealed secondary structures with predominant β sheets. The immunomodulatory capacity of rBet v 1-BanLec tested on macrophages showed changes in myeloperoxidase activity, reduced NO production, and significant alterations in the production of cytokines when compared to both rBanLec and rBet v 1. Comparing to rBet v 1, rBet v 1-BanLec was demonstrated to be more efficient promoter of IL-10 production as well as weaker inducer of NO production and secretion of pro-inflammatory cytokines TNFα, and IL-6. The ability of rBet v 1-BanLec to promote IL-10 in together with the preserved 3D structure of Bet v 1 part implies that the construct might exert a beneficial effect in the allergen-specific immunotherapy.
PB  - Elsevier
T2  - Molecular Immunology
T1  - Recombinant Bet v 1-BanLec chimera modulates functional characteristics of peritoneal murine macrophages by promoting IL-10 secretion
EP  - 67
SP  - 58
VL  - 138
DO  - 10.1016/j.molimm.2021.06.015
ER  - 

@article{
author = "Protić-Rosić, Isidora and Nešić, Andrijana and Lukić, Ivana and Miljković, Radmila and Popović, Dragan and Atanasković-Marković, Marina and Stojanović, Marijana and Gavrović-Jankulović, Marija",
year = "2021",
abstract = "Allergen-specific immunotherapy (AIT) is a desensitizing treatment for allergic diseases that corrects the underlined pathological immune response to innocuous protein antigens, called allergens. Recombinant allergens employed in the AIT allowed the production of well-defined formulations that possessed consistent quality but were often less efficient than natural allergen extracts. Combining recombinant allergens with an adjuvant or immunomodulatory agent could improve AIT efficacy. This study aimed to perform structural and functional characterization of newly designed recombinant chimera composed of the Bet v 1, the major birch pollen allergen, and Banana Lectin (BanLec), TLR2, and CD14 binding protein, for the application in AIT. rBet v 1-BanLec chimera was designed in silico and expressed as a soluble fraction in Escherichia coli. Purified rBet v 1-BanLec (33.4 kDa) retained BanLec-associated biological activity of carbohydrate-binding and preserved IgE reactive epitopes of Bet v 1. The chimera revealed secondary structures with predominant β sheets. The immunomodulatory capacity of rBet v 1-BanLec tested on macrophages showed changes in myeloperoxidase activity, reduced NO production, and significant alterations in the production of cytokines when compared to both rBanLec and rBet v 1. Comparing to rBet v 1, rBet v 1-BanLec was demonstrated to be more efficient promoter of IL-10 production as well as weaker inducer of NO production and secretion of pro-inflammatory cytokines TNFα, and IL-6. The ability of rBet v 1-BanLec to promote IL-10 in together with the preserved 3D structure of Bet v 1 part implies that the construct might exert a beneficial effect in the allergen-specific immunotherapy.",
publisher = "Elsevier",
journal = "Molecular Immunology",
title = "Recombinant Bet v 1-BanLec chimera modulates functional characteristics of peritoneal murine macrophages by promoting IL-10 secretion",
pages = "67-58",
volume = "138",
doi = "10.1016/j.molimm.2021.06.015"
}

Protić-Rosić, I., Nešić, A., Lukić, I., Miljković, R., Popović, D., Atanasković-Marković, M., Stojanović, M.,& Gavrović-Jankulović, M.. (2021). Recombinant Bet v 1-BanLec chimera modulates functional characteristics of peritoneal murine macrophages by promoting IL-10 secretion. in Molecular Immunology
Elsevier., 138, 58-67.
https://doi.org/10.1016/j.molimm.2021.06.015

Protić-Rosić I, Nešić A, Lukić I, Miljković R, Popović D, Atanasković-Marković M, Stojanović M, Gavrović-Jankulović M. Recombinant Bet v 1-BanLec chimera modulates functional characteristics of peritoneal murine macrophages by promoting IL-10 secretion. in Molecular Immunology. 2021;138:58-67.
doi:10.1016/j.molimm.2021.06.015 .

Protić-Rosić, Isidora, Nešić, Andrijana, Lukić, Ivana, Miljković, Radmila, Popović, Dragan, Atanasković-Marković, Marina, Stojanović, Marijana, Gavrović-Jankulović, Marija, "Recombinant Bet v 1-BanLec chimera modulates functional characteristics of peritoneal murine macrophages by promoting IL-10 secretion" in Molecular Immunology, 138 (2021):58-67,
https://doi.org/10.1016/j.molimm.2021.06.015 . .

TY  - DATA
AU  - Stojanović, Marijana
AU  - Lukić, Ivana
AU  - Marinković, Emilija
AU  - Kovačević, Ana
AU  - Miljković, Radmila
AU  - Tobias, Joshua
AU  - Schabussova, Irma
AU  - Zlatović, Mario
AU  - Barisani-Asenbauer, Talin
AU  - Wiedermann, Ursula
AU  - Inić-Kanada, Aleksandra
PY  - 2020
UR  - http://intor.torlakinstitut.com/handle/123456789/643
AB  - Table S1: Characteristics of selected anti-tetanus mAbs [35,36].
PB  - MDPI
T2  - Vaccines
T1  - Supplementary information for the article: Stojanović, M.; Lukić, I.; Marinković, E.; Kovačević, A.; Miljković, R.; Tobias, J.; Schabussova, I.; Zlatović, M.; Barisani-Asenbauer, T.; Wiedermann, U.; Inić-Kanada, A. Cross-Reactive Effects of Vaccines: Heterologous Immunity between Tetanus and Chlamydia. Vaccines 2020, 8 (4), 719. https://doi.org/10.3390/vaccines8040719.
IS  - 4
SP  - 719
VL  - 8
UR  - https://hdl.handle.net/21.15107/rcub_intor_643
ER  - 

@misc{
author = "Stojanović, Marijana and Lukić, Ivana and Marinković, Emilija and Kovačević, Ana and Miljković, Radmila and Tobias, Joshua and Schabussova, Irma and Zlatović, Mario and Barisani-Asenbauer, Talin and Wiedermann, Ursula and Inić-Kanada, Aleksandra",
year = "2020",
abstract = "Table S1: Characteristics of selected anti-tetanus mAbs [35,36].",
publisher = "MDPI",
journal = "Vaccines",
title = "Supplementary information for the article: Stojanović, M.; Lukić, I.; Marinković, E.; Kovačević, A.; Miljković, R.; Tobias, J.; Schabussova, I.; Zlatović, M.; Barisani-Asenbauer, T.; Wiedermann, U.; Inić-Kanada, A. Cross-Reactive Effects of Vaccines: Heterologous Immunity between Tetanus and Chlamydia. Vaccines 2020, 8 (4), 719. https://doi.org/10.3390/vaccines8040719.",
number = "4",
pages = "719",
volume = "8",
url = "https://hdl.handle.net/21.15107/rcub_intor_643"
}

Stojanović, M., Lukić, I., Marinković, E., Kovačević, A., Miljković, R., Tobias, J., Schabussova, I., Zlatović, M., Barisani-Asenbauer, T., Wiedermann, U.,& Inić-Kanada, A.. (2020). Supplementary information for the article: Stojanović, M.; Lukić, I.; Marinković, E.; Kovačević, A.; Miljković, R.; Tobias, J.; Schabussova, I.; Zlatović, M.; Barisani-Asenbauer, T.; Wiedermann, U.; Inić-Kanada, A. Cross-Reactive Effects of Vaccines: Heterologous Immunity between Tetanus and Chlamydia. Vaccines 2020, 8 (4), 719. https://doi.org/10.3390/vaccines8040719.. in Vaccines
MDPI., 8(4), 719.
https://hdl.handle.net/21.15107/rcub_intor_643

Stojanović M, Lukić I, Marinković E, Kovačević A, Miljković R, Tobias J, Schabussova I, Zlatović M, Barisani-Asenbauer T, Wiedermann U, Inić-Kanada A. Supplementary information for the article: Stojanović, M.; Lukić, I.; Marinković, E.; Kovačević, A.; Miljković, R.; Tobias, J.; Schabussova, I.; Zlatović, M.; Barisani-Asenbauer, T.; Wiedermann, U.; Inić-Kanada, A. Cross-Reactive Effects of Vaccines: Heterologous Immunity between Tetanus and Chlamydia. Vaccines 2020, 8 (4), 719. https://doi.org/10.3390/vaccines8040719.. in Vaccines. 2020;8(4):719.
https://hdl.handle.net/21.15107/rcub_intor_643 .

Stojanović, Marijana, Lukić, Ivana, Marinković, Emilija, Kovačević, Ana, Miljković, Radmila, Tobias, Joshua, Schabussova, Irma, Zlatović, Mario, Barisani-Asenbauer, Talin, Wiedermann, Ursula, Inić-Kanada, Aleksandra, "Supplementary information for the article: Stojanović, M.; Lukić, I.; Marinković, E.; Kovačević, A.; Miljković, R.; Tobias, J.; Schabussova, I.; Zlatović, M.; Barisani-Asenbauer, T.; Wiedermann, U.; Inić-Kanada, A. Cross-Reactive Effects of Vaccines: Heterologous Immunity between Tetanus and Chlamydia. Vaccines 2020, 8 (4), 719. https://doi.org/10.3390/vaccines8040719." in Vaccines, 8, no. 4 (2020):719,
https://hdl.handle.net/21.15107/rcub_intor_643 .

TY  - JOUR
AU  - Inić-Kanada, Aleksandra
AU  - Stojanović, Marijana
AU  - Miljković, Radmila
AU  - Stein, Elisabeth
AU  - Filipović, Ana
AU  - Frohns, Antonia
AU  - Zoeller, Nadja
AU  - Kuratli, Jasmin
AU  - Barisani-Asenbauer, Talin
AU  - Borel, Nicole
PY  - 2020
UR  - http://intor.torlakinstitut.com/handle/123456789/564
AB  - Trachoma is a devastating neglected tropical disease caused by Chlamydia trachomatis and the leading global cause of infectious blindness. Although antibiotic treatment against trachoma is efficient (SAFE strategy), additional affordable therapeutic strategies are of high interest. Water-filtered infrared A and visible light (wIRA/VIS) irradiation has proven to reduce chlamydial infectivity in vitro and ex vivo. The aim of this study was to evaluate whether wIRA/VIS can reduce chlamydial infection load and/or ocular pathology in vivo, in a guinea pig model of inclusion conjunctivitis. Guinea pigs were infected with 1 x 10(6) inclusion-forming units/eye of Chlamydia caviae via the ocular conjunctiva on day 0. In infected animals, wIRA/VIS irradiation (2100 W/m(2)) was applied on day 2 (single treatment) and on days 2 and 4 (double treatment) post-infection (pi). wIRA/VIS reduced the clinical pathology score on days 7 and 14 pi and the conjunctival chlamydial load on days 2, 4, 7, and 14 pi in comparison with C. caviae-infected, not irradiated, controls. Furthermore, numbers of chlamydial inclusions were decreased in wIRA/VIS treated C. caviae-infected guinea pigs on day 21 pi compared to C. caviae-infected, non-irradiated, controls. Double treatment with wIRA/VIS (days 2 and 4 pi) was more efficient than a single treatment on day 2 pi. wIRA/VIS treatment did neither induce macroscopic nor histologic changes in ocular tissues. Our results indicate that wIRA/VIS shows promising efficacy to reduce chlamydial infectivity in vivo without causing irradiation related pathologies in the follow-up period. wIRA/VIS irradiation is a promising approach to reduce trachoma transmission and pathology of ocular chlamydial infection.
PB  - Elsevier Science Sa, Lausanne
T2  - Journal of Photochemistry and Photobiology B-Biology
T1  - Water-filtered Infrared A and visible light (wIRA/VIS) treatment reduces Chlamydia caviae-induced ocular inflammation and infectious load in a Guinea pig model of inclusion conjunctivitis
VL  - 209
DO  - 10.1016/j.jphotobiol.2020.111953
ER  - 

@article{
author = "Inić-Kanada, Aleksandra and Stojanović, Marijana and Miljković, Radmila and Stein, Elisabeth and Filipović, Ana and Frohns, Antonia and Zoeller, Nadja and Kuratli, Jasmin and Barisani-Asenbauer, Talin and Borel, Nicole",
year = "2020",
abstract = "Trachoma is a devastating neglected tropical disease caused by Chlamydia trachomatis and the leading global cause of infectious blindness. Although antibiotic treatment against trachoma is efficient (SAFE strategy), additional affordable therapeutic strategies are of high interest. Water-filtered infrared A and visible light (wIRA/VIS) irradiation has proven to reduce chlamydial infectivity in vitro and ex vivo. The aim of this study was to evaluate whether wIRA/VIS can reduce chlamydial infection load and/or ocular pathology in vivo, in a guinea pig model of inclusion conjunctivitis. Guinea pigs were infected with 1 x 10(6) inclusion-forming units/eye of Chlamydia caviae via the ocular conjunctiva on day 0. In infected animals, wIRA/VIS irradiation (2100 W/m(2)) was applied on day 2 (single treatment) and on days 2 and 4 (double treatment) post-infection (pi). wIRA/VIS reduced the clinical pathology score on days 7 and 14 pi and the conjunctival chlamydial load on days 2, 4, 7, and 14 pi in comparison with C. caviae-infected, not irradiated, controls. Furthermore, numbers of chlamydial inclusions were decreased in wIRA/VIS treated C. caviae-infected guinea pigs on day 21 pi compared to C. caviae-infected, non-irradiated, controls. Double treatment with wIRA/VIS (days 2 and 4 pi) was more efficient than a single treatment on day 2 pi. wIRA/VIS treatment did neither induce macroscopic nor histologic changes in ocular tissues. Our results indicate that wIRA/VIS shows promising efficacy to reduce chlamydial infectivity in vivo without causing irradiation related pathologies in the follow-up period. wIRA/VIS irradiation is a promising approach to reduce trachoma transmission and pathology of ocular chlamydial infection.",
publisher = "Elsevier Science Sa, Lausanne",
journal = "Journal of Photochemistry and Photobiology B-Biology",
title = "Water-filtered Infrared A and visible light (wIRA/VIS) treatment reduces Chlamydia caviae-induced ocular inflammation and infectious load in a Guinea pig model of inclusion conjunctivitis",
volume = "209",
doi = "10.1016/j.jphotobiol.2020.111953"
}

Inić-Kanada, A., Stojanović, M., Miljković, R., Stein, E., Filipović, A., Frohns, A., Zoeller, N., Kuratli, J., Barisani-Asenbauer, T.,& Borel, N.. (2020). Water-filtered Infrared A and visible light (wIRA/VIS) treatment reduces Chlamydia caviae-induced ocular inflammation and infectious load in a Guinea pig model of inclusion conjunctivitis. in Journal of Photochemistry and Photobiology B-Biology
Elsevier Science Sa, Lausanne., 209.
https://doi.org/10.1016/j.jphotobiol.2020.111953

Inić-Kanada A, Stojanović M, Miljković R, Stein E, Filipović A, Frohns A, Zoeller N, Kuratli J, Barisani-Asenbauer T, Borel N. Water-filtered Infrared A and visible light (wIRA/VIS) treatment reduces Chlamydia caviae-induced ocular inflammation and infectious load in a Guinea pig model of inclusion conjunctivitis. in Journal of Photochemistry and Photobiology B-Biology. 2020;209.
doi:10.1016/j.jphotobiol.2020.111953 .

Inić-Kanada, Aleksandra, Stojanović, Marijana, Miljković, Radmila, Stein, Elisabeth, Filipović, Ana, Frohns, Antonia, Zoeller, Nadja, Kuratli, Jasmin, Barisani-Asenbauer, Talin, Borel, Nicole, "Water-filtered Infrared A and visible light (wIRA/VIS) treatment reduces Chlamydia caviae-induced ocular inflammation and infectious load in a Guinea pig model of inclusion conjunctivitis" in Journal of Photochemistry and Photobiology B-Biology, 209 (2020),
https://doi.org/10.1016/j.jphotobiol.2020.111953 . .

TY  - JOUR
AU  - Stojanović, Marijana
AU  - Lukić, Ivana
AU  - Marinković, Emilija
AU  - Kovačević, Ana
AU  - Miljković, Radmila
AU  - Tobias, Joshua
AU  - Schabussova, Irma
AU  - Zlatović, Mario
AU  - Barisani-Asenbauer, Talin
AU  - Wiedermann, Ursula
AU  - Inić-Kanada, Aleksandra
PY  - 2020
UR  - http://intor.torlakinstitut.com/handle/123456789/552
AB  - Vaccines can have heterologous effects on the immune system, i.e., effects other than triggering an immune response against the disease targeted by the vaccine. We investigated whether monoclonal antibodies (mAbs) specific for tetanus could cross-react with Chlamydia and confer heterologous protection against chlamydial infection. The capability of two tetanus-specific mAbs, namely mAb26 and mAb51, to prevent chlamydial infection has been assessed: (i) in vitro, by performing a neutralization assay using human conjunctival epithelial (HCjE) cells infected with Chlamydia trachomatis serovar B, and (ii) in vivo, by using a guinea pig model of Chlamydia caviae-induced inclusion conjunctivitis. The mAb26 has been superior in comparison with mAb51 in the prevention of chlamydial infection in HCjE cells. The mAb26 has conferred approximate to 40% inhibition of the infection, compared to less than 5% inhibition in the presence of the mAb51. In vivo, mAb26 significantly diminished ocular pathology intensity in guinea pigs infected with C. caviae compared to either the mAb51-treated or sham-treated guinea pigs. Our data provide insights that tetanus immunization generates antibodies which induce heterologous chlamydial immunity and promote protection beyond the intended target pathogen.
PB  - MDPI, Basel
T2  - Vaccines
T1  - Cross-Reactive Effects of Vaccines: Heterologous Immunity between Tetanus and Chlamydia
IS  - 4
SP  - 719
VL  - 8
DO  - 10.3390/vaccines8040719
ER  - 

@article{
author = "Stojanović, Marijana and Lukić, Ivana and Marinković, Emilija and Kovačević, Ana and Miljković, Radmila and Tobias, Joshua and Schabussova, Irma and Zlatović, Mario and Barisani-Asenbauer, Talin and Wiedermann, Ursula and Inić-Kanada, Aleksandra",
year = "2020",
abstract = "Vaccines can have heterologous effects on the immune system, i.e., effects other than triggering an immune response against the disease targeted by the vaccine. We investigated whether monoclonal antibodies (mAbs) specific for tetanus could cross-react with Chlamydia and confer heterologous protection against chlamydial infection. The capability of two tetanus-specific mAbs, namely mAb26 and mAb51, to prevent chlamydial infection has been assessed: (i) in vitro, by performing a neutralization assay using human conjunctival epithelial (HCjE) cells infected with Chlamydia trachomatis serovar B, and (ii) in vivo, by using a guinea pig model of Chlamydia caviae-induced inclusion conjunctivitis. The mAb26 has been superior in comparison with mAb51 in the prevention of chlamydial infection in HCjE cells. The mAb26 has conferred approximate to 40% inhibition of the infection, compared to less than 5% inhibition in the presence of the mAb51. In vivo, mAb26 significantly diminished ocular pathology intensity in guinea pigs infected with C. caviae compared to either the mAb51-treated or sham-treated guinea pigs. Our data provide insights that tetanus immunization generates antibodies which induce heterologous chlamydial immunity and promote protection beyond the intended target pathogen.",
publisher = "MDPI, Basel",
journal = "Vaccines",
title = "Cross-Reactive Effects of Vaccines: Heterologous Immunity between Tetanus and Chlamydia",
number = "4",
pages = "719",
volume = "8",
doi = "10.3390/vaccines8040719"
}

Stojanović, M., Lukić, I., Marinković, E., Kovačević, A., Miljković, R., Tobias, J., Schabussova, I., Zlatović, M., Barisani-Asenbauer, T., Wiedermann, U.,& Inić-Kanada, A.. (2020). Cross-Reactive Effects of Vaccines: Heterologous Immunity between Tetanus and Chlamydia. in Vaccines
MDPI, Basel., 8(4), 719.
https://doi.org/10.3390/vaccines8040719

Stojanović M, Lukić I, Marinković E, Kovačević A, Miljković R, Tobias J, Schabussova I, Zlatović M, Barisani-Asenbauer T, Wiedermann U, Inić-Kanada A. Cross-Reactive Effects of Vaccines: Heterologous Immunity between Tetanus and Chlamydia. in Vaccines. 2020;8(4):719.
doi:10.3390/vaccines8040719 .

Stojanović, Marijana, Lukić, Ivana, Marinković, Emilija, Kovačević, Ana, Miljković, Radmila, Tobias, Joshua, Schabussova, Irma, Zlatović, Mario, Barisani-Asenbauer, Talin, Wiedermann, Ursula, Inić-Kanada, Aleksandra, "Cross-Reactive Effects of Vaccines: Heterologous Immunity between Tetanus and Chlamydia" in Vaccines, 8, no. 4 (2020):719,
https://doi.org/10.3390/vaccines8040719 . .

TY  - CONF
AU  - Inić-Kanada, Aleksandra
AU  - Lukić, Ivana
AU  - Marinković, Emilija
AU  - Filipović, Ana
AU  - Miljković, Radmila
AU  - Barisani-Asenbauer, Talin
AU  - Wiedermann, Ursula
AU  - Stojanović, Marijana
PY  - 2019
UR  - http://intor.torlakinstitut.com/handle/123456789/528
PB  - Wiley, Hoboken
C3  - European Journal of Immunology
T1  - Beneficial heterologous effects of a tetanus vaccination: the role of molecular mimicry and/or trained immunity
EP  - 1694
SP  - 1693
VL  - 49
UR  - https://hdl.handle.net/21.15107/rcub_intor_528
ER  - 

@conference{
author = "Inić-Kanada, Aleksandra and Lukić, Ivana and Marinković, Emilija and Filipović, Ana and Miljković, Radmila and Barisani-Asenbauer, Talin and Wiedermann, Ursula and Stojanović, Marijana",
year = "2019",
publisher = "Wiley, Hoboken",
journal = "European Journal of Immunology",
title = "Beneficial heterologous effects of a tetanus vaccination: the role of molecular mimicry and/or trained immunity",
pages = "1694-1693",
volume = "49",
url = "https://hdl.handle.net/21.15107/rcub_intor_528"
}

Inić-Kanada, A., Lukić, I., Marinković, E., Filipović, A., Miljković, R., Barisani-Asenbauer, T., Wiedermann, U.,& Stojanović, M.. (2019). Beneficial heterologous effects of a tetanus vaccination: the role of molecular mimicry and/or trained immunity. in European Journal of Immunology
Wiley, Hoboken., 49, 1693-1694.
https://hdl.handle.net/21.15107/rcub_intor_528

Inić-Kanada A, Lukić I, Marinković E, Filipović A, Miljković R, Barisani-Asenbauer T, Wiedermann U, Stojanović M. Beneficial heterologous effects of a tetanus vaccination: the role of molecular mimicry and/or trained immunity. in European Journal of Immunology. 2019;49:1693-1694.
https://hdl.handle.net/21.15107/rcub_intor_528 .

Inić-Kanada, Aleksandra, Lukić, Ivana, Marinković, Emilija, Filipović, Ana, Miljković, Radmila, Barisani-Asenbauer, Talin, Wiedermann, Ursula, Stojanović, Marijana, "Beneficial heterologous effects of a tetanus vaccination: the role of molecular mimicry and/or trained immunity" in European Journal of Immunology, 49 (2019):1693-1694,
https://hdl.handle.net/21.15107/rcub_intor_528 .

TY  - JOUR
AU  - Lukić, Ivana
AU  - Filipović, Ana
AU  - Inić-Kanada, Aleksandra
AU  - Marinković, Emilija
AU  - Miljković, Radmila
AU  - Stojanović, Marijana
PY  - 2018
UR  - http://intor.torlakinstitut.com/handle/123456789/510
AB  - Oligoclonal combinations of several monoclonal antibodies (MAbs) are being considered for the treatment of various infectious pathologies. These combinations are less sensitive to antigen structural changes than individual MAbs; at the same time, their characteristics can be more efficiently controlled than those of polyclonal antibodies. The main goal of this study was to evaluate the binding characteristics of six biclonal equimolar preparations (BEP) of tetanus toxin (TeNT)-specific MAbs and to investigate how the MAb combination influences the BEPs' protective capacity. We show that a combination of TeNT-specific MAbs, which not only bind TeNT but also exert positive cooperative effects, results in a BEP with superior binding characteristics and protective capacity, when compared with the individual component MAbs. Furthermore, we show that a MAb with only partial protective capacity but positive effects on the binding of the other BEP component can be used as a valuable constituent of the BEP. (C) 2018 Elsevier Ltd. All rights reserved.
PB  - Elsevier Sci Ltd, Oxford
T2  - Vaccine
T1  - Cooperative binding of anti-tetanus toxin monoclonal antibodies: Implications for designing an efficient biclonal preparation to prevent tetanus toxin intoxication
EP  - 3771
IS  - 26
SP  - 3764
VL  - 36
DO  - 10.1016/j.vaccine.2018.05.058
ER  - 

@article{
author = "Lukić, Ivana and Filipović, Ana and Inić-Kanada, Aleksandra and Marinković, Emilija and Miljković, Radmila and Stojanović, Marijana",
year = "2018",
abstract = "Oligoclonal combinations of several monoclonal antibodies (MAbs) are being considered for the treatment of various infectious pathologies. These combinations are less sensitive to antigen structural changes than individual MAbs; at the same time, their characteristics can be more efficiently controlled than those of polyclonal antibodies. The main goal of this study was to evaluate the binding characteristics of six biclonal equimolar preparations (BEP) of tetanus toxin (TeNT)-specific MAbs and to investigate how the MAb combination influences the BEPs' protective capacity. We show that a combination of TeNT-specific MAbs, which not only bind TeNT but also exert positive cooperative effects, results in a BEP with superior binding characteristics and protective capacity, when compared with the individual component MAbs. Furthermore, we show that a MAb with only partial protective capacity but positive effects on the binding of the other BEP component can be used as a valuable constituent of the BEP. (C) 2018 Elsevier Ltd. All rights reserved.",
publisher = "Elsevier Sci Ltd, Oxford",
journal = "Vaccine",
title = "Cooperative binding of anti-tetanus toxin monoclonal antibodies: Implications for designing an efficient biclonal preparation to prevent tetanus toxin intoxication",
pages = "3771-3764",
number = "26",
volume = "36",
doi = "10.1016/j.vaccine.2018.05.058"
}

Lukić, I., Filipović, A., Inić-Kanada, A., Marinković, E., Miljković, R.,& Stojanović, M.. (2018). Cooperative binding of anti-tetanus toxin monoclonal antibodies: Implications for designing an efficient biclonal preparation to prevent tetanus toxin intoxication. in Vaccine
Elsevier Sci Ltd, Oxford., 36(26), 3764-3771.
https://doi.org/10.1016/j.vaccine.2018.05.058

Lukić I, Filipović A, Inić-Kanada A, Marinković E, Miljković R, Stojanović M. Cooperative binding of anti-tetanus toxin monoclonal antibodies: Implications for designing an efficient biclonal preparation to prevent tetanus toxin intoxication. in Vaccine. 2018;36(26):3764-3771.
doi:10.1016/j.vaccine.2018.05.058 .

Lukić, Ivana, Filipović, Ana, Inić-Kanada, Aleksandra, Marinković, Emilija, Miljković, Radmila, Stojanović, Marijana, "Cooperative binding of anti-tetanus toxin monoclonal antibodies: Implications for designing an efficient biclonal preparation to prevent tetanus toxin intoxication" in Vaccine, 36, no. 26 (2018):3764-3771,
https://doi.org/10.1016/j.vaccine.2018.05.058 . .

TY  - JOUR
AU  - Schuerer, Nadine
AU  - Stein, Elisabeth
AU  - Inić-Kanada, Aleksandra
AU  - Ghasemian, Ehsan
AU  - Stojanović, Marijana
AU  - Montanaro, Jacqueline
AU  - Bintner, Nora
AU  - Hohenadl, Christine
AU  - Sachsenhofer, Robert
AU  - Barisani-Asenbauer, Talin
PY  - 2018
UR  - http://intor.torlakinstitut.com/handle/123456789/506
AB  - Aim of the study: Chitosan, a partially deacetylated polysaccharide derived from chitin, and chitosan-N-acetylcysteine (C-NAC), a thiolated chitosan, both show enhanced retention times on the ocular surface when compared to other polymers commonly used in eye drops. To evaluate these compounds as adjuvants for ocular drug delivery or uptake of topically administered conjunctival vaccines, biochemical characteristics of both polymers were investigated in vitro and in vivo.

Methods:
Human conjunctival epithelial (HCjE) cells were used to investigate biocompatibility of buffered chitosan and C-NAC containing formulations. Cellular uptake was studied using fluorescein-isothiocyanate (FITC)-labelled polymers. Transepithelial electrical resistance (TEER) measurements were performed to determine effects of chitosan on tight junctions of stratified HCjE cells in vitro. In vivo uptake of topically applied chitosan into conjunctival epithelial cells was investigated in guinea pigs.

Results:
Minimal effects on cell viability were seen with both compounds after application for 30 min in a concentration of 0.1%. In vitro uptake into HCjE cells was only observed with the chitosan containing solution. An effect on tight junctions was demonstrated by significantly (P < .05) decreasing TEER levels 60 min after incubation with chitosan. In vivo, FITC-labelled chitosan was detected within guinea pig conjunctival epithelial cells 120 min after topical administration. No adverse effects were observed.
T2  - Journal of EuCornea
T1  - Effects of chitosan and chitosan Nacetylcysteine solutions on conjunctival epithelial cells
EP  - 18
IS  - 1
SP  - 12
VL  - 1
DO  - 10.1016/j.xjec.2018.04.002
UR  - https://hdl.handle.net/21.15107/rcub_intor_506
ER  - 

@article{
author = "Schuerer, Nadine and Stein, Elisabeth and Inić-Kanada, Aleksandra and Ghasemian, Ehsan and Stojanović, Marijana and Montanaro, Jacqueline and Bintner, Nora and Hohenadl, Christine and Sachsenhofer, Robert and Barisani-Asenbauer, Talin",
year = "2018",
abstract = "Aim of the study: Chitosan, a partially deacetylated polysaccharide derived from chitin, and chitosan-N-acetylcysteine (C-NAC), a thiolated chitosan, both show enhanced retention times on the ocular surface when compared to other polymers commonly used in eye drops. To evaluate these compounds as adjuvants for ocular drug delivery or uptake of topically administered conjunctival vaccines, biochemical characteristics of both polymers were investigated in vitro and in vivo.

Methods:
Human conjunctival epithelial (HCjE) cells were used to investigate biocompatibility of buffered chitosan and C-NAC containing formulations. Cellular uptake was studied using fluorescein-isothiocyanate (FITC)-labelled polymers. Transepithelial electrical resistance (TEER) measurements were performed to determine effects of chitosan on tight junctions of stratified HCjE cells in vitro. In vivo uptake of topically applied chitosan into conjunctival epithelial cells was investigated in guinea pigs.

Results:
Minimal effects on cell viability were seen with both compounds after application for 30 min in a concentration of 0.1%. In vitro uptake into HCjE cells was only observed with the chitosan containing solution. An effect on tight junctions was demonstrated by significantly (P < .05) decreasing TEER levels 60 min after incubation with chitosan. In vivo, FITC-labelled chitosan was detected within guinea pig conjunctival epithelial cells 120 min after topical administration. No adverse effects were observed.",
journal = "Journal of EuCornea",
title = "Effects of chitosan and chitosan Nacetylcysteine solutions on conjunctival epithelial cells",
pages = "18-12",
number = "1",
volume = "1",
doi = "10.1016/j.xjec.2018.04.002",
url = "https://hdl.handle.net/21.15107/rcub_intor_506"
}

Schuerer, N., Stein, E., Inić-Kanada, A., Ghasemian, E., Stojanović, M., Montanaro, J., Bintner, N., Hohenadl, C., Sachsenhofer, R.,& Barisani-Asenbauer, T.. (2018). Effects of chitosan and chitosan Nacetylcysteine solutions on conjunctival epithelial cells. in Journal of EuCornea, 1(1), 12-18.
https://doi.org/10.1016/j.xjec.2018.04.002
https://hdl.handle.net/21.15107/rcub_intor_506

Schuerer N, Stein E, Inić-Kanada A, Ghasemian E, Stojanović M, Montanaro J, Bintner N, Hohenadl C, Sachsenhofer R, Barisani-Asenbauer T. Effects of chitosan and chitosan Nacetylcysteine solutions on conjunctival epithelial cells. in Journal of EuCornea. 2018;1(1):12-18.
doi:10.1016/j.xjec.2018.04.002
https://hdl.handle.net/21.15107/rcub_intor_506 .

Schuerer, Nadine, Stein, Elisabeth, Inić-Kanada, Aleksandra, Ghasemian, Ehsan, Stojanović, Marijana, Montanaro, Jacqueline, Bintner, Nora, Hohenadl, Christine, Sachsenhofer, Robert, Barisani-Asenbauer, Talin, "Effects of chitosan and chitosan Nacetylcysteine solutions on conjunctival epithelial cells" in Journal of EuCornea, 1, no. 1 (2018):12-18,
https://doi.org/10.1016/j.xjec.2018.04.002 .,
https://hdl.handle.net/21.15107/rcub_intor_506 .

TY  - JOUR
AU  - Inić-Kanada, Aleksandra
AU  - Stein, Elisabeth
AU  - Stojanović, Marijana
AU  - Schuerer, Nadine
AU  - Ghasemian, Ehsan
AU  - Filipović, Ana
AU  - Marinković, Emilija
AU  - Kosanović, Dejana
AU  - Barisani-Asenbauer, Talin
PY  - 2018
UR  - http://intor.torlakinstitut.com/handle/123456789/502
AB  - Ocular chlamydial infections with the ocular serovars A, B, Ba, and C of Chlamydia trachomatis represent the world's leading cause of infectious blindness. Carrageenans are naturally occurring, sulfated polysaccharides generally considered safe for food and topical applications. Carrageenans can inhibit infection caused by a variety of viruses and bacteria. To investigate whether iota-carrageenan (I-C) isolated from the red alga Chondrus crispus could prevent ocular chlamydial infection, we assessed if targeted treatment of the conjunctival mucosa with I-C affects chlamydial attachment, entry, and replication in the host cell. Immortalized human conjunctival epithelial cells were treated with I-C prior to C. trachomatis infection and analyzed by flow cytometry and immunofluorescence microscopy. In vivo effects were evaluated in an ocular guinea pig inclusion conjunctivitis model. Ocular pathology was graded daily, and chlamydial clearance was investigated. Our study showed that I-C reduces the infectivity of C. trachomatis in vitro. In vivo results showed a slight reduced ocular pathology and significantly less shedding of infectious elementary bodies by infected animals. Our results indicate that I-C could be a promising agent to reduce the transmission of ocular chlamydial infection and opens perspectives to develop prophylactic approaches to block C. trachomatis entry into the host cell.
PB  - Springer, Dordrecht
T2  - Journal of Applied Phycology
T1  - Effects of iota-carrageenan on ocular Chlamydia trachomatis infection in vitro and in vivo
EP  - 2610
IS  - 4
SP  - 2601
VL  - 30
DO  - 10.1007/s10811-018-1435-0
ER  - 

@article{
author = "Inić-Kanada, Aleksandra and Stein, Elisabeth and Stojanović, Marijana and Schuerer, Nadine and Ghasemian, Ehsan and Filipović, Ana and Marinković, Emilija and Kosanović, Dejana and Barisani-Asenbauer, Talin",
year = "2018",
abstract = "Ocular chlamydial infections with the ocular serovars A, B, Ba, and C of Chlamydia trachomatis represent the world's leading cause of infectious blindness. Carrageenans are naturally occurring, sulfated polysaccharides generally considered safe for food and topical applications. Carrageenans can inhibit infection caused by a variety of viruses and bacteria. To investigate whether iota-carrageenan (I-C) isolated from the red alga Chondrus crispus could prevent ocular chlamydial infection, we assessed if targeted treatment of the conjunctival mucosa with I-C affects chlamydial attachment, entry, and replication in the host cell. Immortalized human conjunctival epithelial cells were treated with I-C prior to C. trachomatis infection and analyzed by flow cytometry and immunofluorescence microscopy. In vivo effects were evaluated in an ocular guinea pig inclusion conjunctivitis model. Ocular pathology was graded daily, and chlamydial clearance was investigated. Our study showed that I-C reduces the infectivity of C. trachomatis in vitro. In vivo results showed a slight reduced ocular pathology and significantly less shedding of infectious elementary bodies by infected animals. Our results indicate that I-C could be a promising agent to reduce the transmission of ocular chlamydial infection and opens perspectives to develop prophylactic approaches to block C. trachomatis entry into the host cell.",
publisher = "Springer, Dordrecht",
journal = "Journal of Applied Phycology",
title = "Effects of iota-carrageenan on ocular Chlamydia trachomatis infection in vitro and in vivo",
pages = "2610-2601",
number = "4",
volume = "30",
doi = "10.1007/s10811-018-1435-0"
}

Inić-Kanada, A., Stein, E., Stojanović, M., Schuerer, N., Ghasemian, E., Filipović, A., Marinković, E., Kosanović, D.,& Barisani-Asenbauer, T.. (2018). Effects of iota-carrageenan on ocular Chlamydia trachomatis infection in vitro and in vivo. in Journal of Applied Phycology
Springer, Dordrecht., 30(4), 2601-2610.
https://doi.org/10.1007/s10811-018-1435-0

Inić-Kanada A, Stein E, Stojanović M, Schuerer N, Ghasemian E, Filipović A, Marinković E, Kosanović D, Barisani-Asenbauer T. Effects of iota-carrageenan on ocular Chlamydia trachomatis infection in vitro and in vivo. in Journal of Applied Phycology. 2018;30(4):2601-2610.
doi:10.1007/s10811-018-1435-0 .

Inić-Kanada, Aleksandra, Stein, Elisabeth, Stojanović, Marijana, Schuerer, Nadine, Ghasemian, Ehsan, Filipović, Ana, Marinković, Emilija, Kosanović, Dejana, Barisani-Asenbauer, Talin, "Effects of iota-carrageenan on ocular Chlamydia trachomatis infection in vitro and in vivo" in Journal of Applied Phycology, 30, no. 4 (2018):2601-2610,
https://doi.org/10.1007/s10811-018-1435-0 . .

TY  - JOUR
AU  - Marinković, Emilija
AU  - Đokić, Radmila
AU  - Lukić, Ivana
AU  - Filipović, Ana
AU  - Inić-Kanada, Aleksandra
AU  - Kosanović, Dejana
AU  - Gavrović-Jankulović, Marija
AU  - Stojanović, Marijana
PY  - 2017
UR  - http://intor.torlakinstitut.com/handle/123456789/493
AB  - We demonstrated that a recombinant banana lectin (rBanLec), which structural characteristics and physiological impacts highly resemble those reported for its natural counterparts, binds murine peritoneal macrophages and specifically modulates their functional characteristics. By using rBanLec in concentrations ranging from 1 mu g to 10 mu g to stimulate resident (RMs) and thioglycollate-elicited (TGMs) peritoneal macrophages from BALB/c and C57BL/6 mice, we have shown that effects of rBanLec stimulation depend on its concentration but also on the functional status of macrophages and their genetic background. rBanLec, in a positive dose-dependent manner, promotes the proliferation of TGMs from both BALB/c and C57BL/6 mice, while its mitogenic influence on RMs is significantly lower (BALB/c mice) or not detectable (C57BL/6 mice). In all peritoneal macrophages, irrespective of their type and genetic background, rBanLec, in a positive dose dependent manner, enhances the secretion of IL-10. rBanLec stimulation of RMs from both BALB/c and C57BL/6 resulted in a positive dose-dependent promotion of proinflammatory phenotype (enhancement of NO production and IL-12 and TNF alpha secretion, reduction of arginase activity). Positive dose-dependent skewing toward proinflammatory phenotype was also observed in TGMs from C57BL/6 mice. However, the enhancement of rBanLec stimulation promotes skewing of TGMs from BALB/c mice towards anti-inflammatory profile (reduction of NO production and IL-12 secretion, enhancement of arginase activity and TGF alpha and IL-4 secretion). Moreover, we established that rBanLec binds oligosaccharide structures of TLR2 and CD14 and that blocking of signaling via these receptors significantly impairs the production of TNFa and NO in BALB/c macrophages. Since the outcome of rBanLec stimulation depends on rBanLec concentration as well as on the functional characteristics of its target cells and their genetic background, further studies are needed to investigate its effects under physiological and specific pathological conditions.
PB  - Public Library Science, San Francisco
T2  - PLoS One
T1  - Modulation of functional characteristics of resident and thioglycollate-elicited peritoneal murine macrophages by a recombinant banana lectin
IS  - 2
VL  - 12
DO  - 10.1371/journal.pone.0172469
ER  - 

@article{
author = "Marinković, Emilija and Đokić, Radmila and Lukić, Ivana and Filipović, Ana and Inić-Kanada, Aleksandra and Kosanović, Dejana and Gavrović-Jankulović, Marija and Stojanović, Marijana",
year = "2017",
abstract = "We demonstrated that a recombinant banana lectin (rBanLec), which structural characteristics and physiological impacts highly resemble those reported for its natural counterparts, binds murine peritoneal macrophages and specifically modulates their functional characteristics. By using rBanLec in concentrations ranging from 1 mu g to 10 mu g to stimulate resident (RMs) and thioglycollate-elicited (TGMs) peritoneal macrophages from BALB/c and C57BL/6 mice, we have shown that effects of rBanLec stimulation depend on its concentration but also on the functional status of macrophages and their genetic background. rBanLec, in a positive dose-dependent manner, promotes the proliferation of TGMs from both BALB/c and C57BL/6 mice, while its mitogenic influence on RMs is significantly lower (BALB/c mice) or not detectable (C57BL/6 mice). In all peritoneal macrophages, irrespective of their type and genetic background, rBanLec, in a positive dose dependent manner, enhances the secretion of IL-10. rBanLec stimulation of RMs from both BALB/c and C57BL/6 resulted in a positive dose-dependent promotion of proinflammatory phenotype (enhancement of NO production and IL-12 and TNF alpha secretion, reduction of arginase activity). Positive dose-dependent skewing toward proinflammatory phenotype was also observed in TGMs from C57BL/6 mice. However, the enhancement of rBanLec stimulation promotes skewing of TGMs from BALB/c mice towards anti-inflammatory profile (reduction of NO production and IL-12 secretion, enhancement of arginase activity and TGF alpha and IL-4 secretion). Moreover, we established that rBanLec binds oligosaccharide structures of TLR2 and CD14 and that blocking of signaling via these receptors significantly impairs the production of TNFa and NO in BALB/c macrophages. Since the outcome of rBanLec stimulation depends on rBanLec concentration as well as on the functional characteristics of its target cells and their genetic background, further studies are needed to investigate its effects under physiological and specific pathological conditions.",
publisher = "Public Library Science, San Francisco",
journal = "PLoS One",
title = "Modulation of functional characteristics of resident and thioglycollate-elicited peritoneal murine macrophages by a recombinant banana lectin",
number = "2",
volume = "12",
doi = "10.1371/journal.pone.0172469"
}

Marinković, E., Đokić, R., Lukić, I., Filipović, A., Inić-Kanada, A., Kosanović, D., Gavrović-Jankulović, M.,& Stojanović, M.. (2017). Modulation of functional characteristics of resident and thioglycollate-elicited peritoneal murine macrophages by a recombinant banana lectin. in PLoS One
Public Library Science, San Francisco., 12(2).
https://doi.org/10.1371/journal.pone.0172469

Marinković E, Đokić R, Lukić I, Filipović A, Inić-Kanada A, Kosanović D, Gavrović-Jankulović M, Stojanović M. Modulation of functional characteristics of resident and thioglycollate-elicited peritoneal murine macrophages by a recombinant banana lectin. in PLoS One. 2017;12(2).
doi:10.1371/journal.pone.0172469 .

Marinković, Emilija, Đokić, Radmila, Lukić, Ivana, Filipović, Ana, Inić-Kanada, Aleksandra, Kosanović, Dejana, Gavrović-Jankulović, Marija, Stojanović, Marijana, "Modulation of functional characteristics of resident and thioglycollate-elicited peritoneal murine macrophages by a recombinant banana lectin" in PLoS One, 12, no. 2 (2017),
https://doi.org/10.1371/journal.pone.0172469 . .

TY  - CONF
AU  - Inić-Kanada, Aleksandra
AU  - Lukić, Ivana
AU  - Stojanović, Marijana
AU  - Stein, Elisabeth
AU  - Marinković, Emilija
AU  - Filipović, Ana
AU  - Đokić, Radmila
AU  - Kosanović, Dejana
AU  - Schuerer, Nadine
AU  - Ghasemian, Ehsan
AU  - Barisani-Asenbauer, Talin
PY  - 2017
UR  - http://intor.torlakinstitut.com/handle/123456789/486
PB  - Assoc Research Vision Ophthalmology Inc, Rockville
C3  - Investigative Ophthalmology & Visual Science
T1  - Tetanus vaccination related to the decline of trachoma in the Western World? Anti-tetanus antibodies confer partial protection against ocular chlamydial infection.
IS  - 8
VL  - 58
UR  - https://hdl.handle.net/21.15107/rcub_intor_486
ER  - 

@conference{
author = "Inić-Kanada, Aleksandra and Lukić, Ivana and Stojanović, Marijana and Stein, Elisabeth and Marinković, Emilija and Filipović, Ana and Đokić, Radmila and Kosanović, Dejana and Schuerer, Nadine and Ghasemian, Ehsan and Barisani-Asenbauer, Talin",
year = "2017",
publisher = "Assoc Research Vision Ophthalmology Inc, Rockville",
journal = "Investigative Ophthalmology & Visual Science",
title = "Tetanus vaccination related to the decline of trachoma in the Western World? Anti-tetanus antibodies confer partial protection against ocular chlamydial infection.",
number = "8",
volume = "58",
url = "https://hdl.handle.net/21.15107/rcub_intor_486"
}

Inić-Kanada, A., Lukić, I., Stojanović, M., Stein, E., Marinković, E., Filipović, A., Đokić, R., Kosanović, D., Schuerer, N., Ghasemian, E.,& Barisani-Asenbauer, T.. (2017). Tetanus vaccination related to the decline of trachoma in the Western World? Anti-tetanus antibodies confer partial protection against ocular chlamydial infection.. in Investigative Ophthalmology & Visual Science
Assoc Research Vision Ophthalmology Inc, Rockville., 58(8).
https://hdl.handle.net/21.15107/rcub_intor_486

Inić-Kanada A, Lukić I, Stojanović M, Stein E, Marinković E, Filipović A, Đokić R, Kosanović D, Schuerer N, Ghasemian E, Barisani-Asenbauer T. Tetanus vaccination related to the decline of trachoma in the Western World? Anti-tetanus antibodies confer partial protection against ocular chlamydial infection.. in Investigative Ophthalmology & Visual Science. 2017;58(8).
https://hdl.handle.net/21.15107/rcub_intor_486 .

Inić-Kanada, Aleksandra, Lukić, Ivana, Stojanović, Marijana, Stein, Elisabeth, Marinković, Emilija, Filipović, Ana, Đokić, Radmila, Kosanović, Dejana, Schuerer, Nadine, Ghasemian, Ehsan, Barisani-Asenbauer, Talin, "Tetanus vaccination related to the decline of trachoma in the Western World? Anti-tetanus antibodies confer partial protection against ocular chlamydial infection." in Investigative Ophthalmology & Visual Science, 58, no. 8 (2017),
https://hdl.handle.net/21.15107/rcub_intor_486 .

TY  - CONF
AU  - Marinković, Emilija
AU  - Đokić, Radmila
AU  - Filipović, Ana
AU  - Lukić, Ivana
AU  - Kosanović, Dejana
AU  - Inić-Kanada, Aleksandra
AU  - Gavrović-Jankulović, Marija
AU  - Stojanović, Marijana
PY  - 2017
UR  - http://intor.torlakinstitut.com/handle/123456789/478
PB  - Wiley, Hoboken
C3  - FEBS Journal
T1  - Prophylactic effect of recombinant banana lectin on TNBS-induced colitis in BALB/c mice
EP  - 125
SP  - 124
VL  - 284
UR  - https://hdl.handle.net/21.15107/rcub_intor_478
ER  - 

@conference{
author = "Marinković, Emilija and Đokić, Radmila and Filipović, Ana and Lukić, Ivana and Kosanović, Dejana and Inić-Kanada, Aleksandra and Gavrović-Jankulović, Marija and Stojanović, Marijana",
year = "2017",
publisher = "Wiley, Hoboken",
journal = "FEBS Journal",
title = "Prophylactic effect of recombinant banana lectin on TNBS-induced colitis in BALB/c mice",
pages = "125-124",
volume = "284",
url = "https://hdl.handle.net/21.15107/rcub_intor_478"
}

Marinković, E., Đokić, R., Filipović, A., Lukić, I., Kosanović, D., Inić-Kanada, A., Gavrović-Jankulović, M.,& Stojanović, M.. (2017). Prophylactic effect of recombinant banana lectin on TNBS-induced colitis in BALB/c mice. in FEBS Journal
Wiley, Hoboken., 284, 124-125.
https://hdl.handle.net/21.15107/rcub_intor_478

Marinković E, Đokić R, Filipović A, Lukić I, Kosanović D, Inić-Kanada A, Gavrović-Jankulović M, Stojanović M. Prophylactic effect of recombinant banana lectin on TNBS-induced colitis in BALB/c mice. in FEBS Journal. 2017;284:124-125.
https://hdl.handle.net/21.15107/rcub_intor_478 .

Marinković, Emilija, Đokić, Radmila, Filipović, Ana, Lukić, Ivana, Kosanović, Dejana, Inić-Kanada, Aleksandra, Gavrović-Jankulović, Marija, Stojanović, Marijana, "Prophylactic effect of recombinant banana lectin on TNBS-induced colitis in BALB/c mice" in FEBS Journal, 284 (2017):124-125,
https://hdl.handle.net/21.15107/rcub_intor_478 .

TY  - JOUR
AU  - Filipović, Ana
AU  - Ghasemian, Ehsan
AU  - Inić-Kanada, Aleksandra
AU  - Lukić, Ivana
AU  - Stein, Elisabeth
AU  - Marinković, Emilija
AU  - Đokić, Radmila
AU  - Kosanović, Dejana
AU  - Schuerer, Nadine
AU  - Chalabi, Hadeel
AU  - Belij-Rammerstorfer, Sandra
AU  - Stojanović, Marijana
AU  - Barisani-Asenbauer, Talin
PY  - 2017
UR  - http://intor.torlakinstitut.com/handle/123456789/477
AB  - Following infection, the balance between protective immunity and immunopathology often depends on the initial infectious load. Several studies have investigated the effect of infectious dose; however, the mechanism by which infectious dose affects disease outcomes and the development of a protective immune response is not known. The aim of this study was to investigate how the infectious dose modulates the local and systemic humoral and the cellular immune responses during primary ocular chlamydial infection in the guinea pig animal model. Guinea pigs were infected by ocular instillation of a Chlamydophila caviae-containing eye solution in the conjunctival sac in three different doses: 1x10(2), 1x10(4), and 1x10(6) inclusion forming units (IFUs). Ocular pathology, chlamydial clearance, local and systemic C. caviae-specific humoral and cellular immune responses were assessed. All inocula of C. caviae significantly enhanced the local production of C. caviae-specific IgA in tears, but only guinea pigs infected with the higher doses showed significant changes in C. caviae-specific IgA levels in vaginal washes and serum. On complete resolution of infection, the low dose of C. caviae did not alter the ratio of CD4(+) and CD8(+) cells within guinea pigs' submandibular lymph node (SMLN) lymphocytes while the higher doses increased the percentages of CD4(+) and CD8(+) cells within the SMLN lymphocytes. A significant negative correlation between pathology intensity and the percentage of CD4(+) and CD8(+) cells within SMLN lymphocyte pool at selected time points post-infection was recorded for both 1x10(4), and 1x10(6) IFU infected guinea pigs. The relevance of the observed dose-dependent differences on the immune response should be further investigated in repeated ocular chlamydial infections.
PB  - Public Library Science, San Francisco
T2  - PLoS One
T1  - The effect of infectious dose on humoral and cellular immune responses in Chlamydophila caviae primary ocular infection
IS  - 7
VL  - 12
DO  - 10.1371/journal.pone.0180551
ER  - 

@article{
author = "Filipović, Ana and Ghasemian, Ehsan and Inić-Kanada, Aleksandra and Lukić, Ivana and Stein, Elisabeth and Marinković, Emilija and Đokić, Radmila and Kosanović, Dejana and Schuerer, Nadine and Chalabi, Hadeel and Belij-Rammerstorfer, Sandra and Stojanović, Marijana and Barisani-Asenbauer, Talin",
year = "2017",
abstract = "Following infection, the balance between protective immunity and immunopathology often depends on the initial infectious load. Several studies have investigated the effect of infectious dose; however, the mechanism by which infectious dose affects disease outcomes and the development of a protective immune response is not known. The aim of this study was to investigate how the infectious dose modulates the local and systemic humoral and the cellular immune responses during primary ocular chlamydial infection in the guinea pig animal model. Guinea pigs were infected by ocular instillation of a Chlamydophila caviae-containing eye solution in the conjunctival sac in three different doses: 1x10(2), 1x10(4), and 1x10(6) inclusion forming units (IFUs). Ocular pathology, chlamydial clearance, local and systemic C. caviae-specific humoral and cellular immune responses were assessed. All inocula of C. caviae significantly enhanced the local production of C. caviae-specific IgA in tears, but only guinea pigs infected with the higher doses showed significant changes in C. caviae-specific IgA levels in vaginal washes and serum. On complete resolution of infection, the low dose of C. caviae did not alter the ratio of CD4(+) and CD8(+) cells within guinea pigs' submandibular lymph node (SMLN) lymphocytes while the higher doses increased the percentages of CD4(+) and CD8(+) cells within the SMLN lymphocytes. A significant negative correlation between pathology intensity and the percentage of CD4(+) and CD8(+) cells within SMLN lymphocyte pool at selected time points post-infection was recorded for both 1x10(4), and 1x10(6) IFU infected guinea pigs. The relevance of the observed dose-dependent differences on the immune response should be further investigated in repeated ocular chlamydial infections.",
publisher = "Public Library Science, San Francisco",
journal = "PLoS One",
title = "The effect of infectious dose on humoral and cellular immune responses in Chlamydophila caviae primary ocular infection",
number = "7",
volume = "12",
doi = "10.1371/journal.pone.0180551"
}

Filipović, A., Ghasemian, E., Inić-Kanada, A., Lukić, I., Stein, E., Marinković, E., Đokić, R., Kosanović, D., Schuerer, N., Chalabi, H., Belij-Rammerstorfer, S., Stojanović, M.,& Barisani-Asenbauer, T.. (2017). The effect of infectious dose on humoral and cellular immune responses in Chlamydophila caviae primary ocular infection. in PLoS One
Public Library Science, San Francisco., 12(7).
https://doi.org/10.1371/journal.pone.0180551

Filipović A, Ghasemian E, Inić-Kanada A, Lukić I, Stein E, Marinković E, Đokić R, Kosanović D, Schuerer N, Chalabi H, Belij-Rammerstorfer S, Stojanović M, Barisani-Asenbauer T. The effect of infectious dose on humoral and cellular immune responses in Chlamydophila caviae primary ocular infection. in PLoS One. 2017;12(7).
doi:10.1371/journal.pone.0180551 .

Filipović, Ana, Ghasemian, Ehsan, Inić-Kanada, Aleksandra, Lukić, Ivana, Stein, Elisabeth, Marinković, Emilija, Đokić, Radmila, Kosanović, Dejana, Schuerer, Nadine, Chalabi, Hadeel, Belij-Rammerstorfer, Sandra, Stojanović, Marijana, Barisani-Asenbauer, Talin, "The effect of infectious dose on humoral and cellular immune responses in Chlamydophila caviae primary ocular infection" in PLoS One, 12, no. 7 (2017),
https://doi.org/10.1371/journal.pone.0180551 . .

TY  - JOUR
AU  - Marinković, Emilija
AU  - Lukić, Ivana
AU  - Kosanović, Dejana
AU  - Inić-Kanada, Aleksandra
AU  - Gavrović-Jankulović, Marija
AU  - Stojanović, Marijana
PY  - 2016
UR  - http://intor.torlakinstitut.com/handle/123456789/471
AB  - Recombinant banana lectin isoform (rBanLec) attaches specifically to the mucosal surface, crosses the epithelial barrier and then directly affects the immune response in mouse colon. Structural characteristics, specificity and physiological impacts of rBanLec reported until now highly resemble those of its natural counterpart. Here, we demonstrated that a dose dependent stimulation of the colon with rBanLec skewed the immune response towards Th1/Th17 direction and this effect was counterbalanced by the rise in IL-10 production. Qualitative and quantitative characteristics of the established cytokine network were dependent on the applied rBanLec concentration. In addition, rBanLec enhanced local NO production and myeloperoxidase activity and promoted an increase in local IgA and IgG production. Stimulation with rBanLec can be beneficial in prevention of pathologies raised due to inappropriate cell-mediated immune response as well as in prevention of the pathogen invasion via the colon. (C) 2015 Elsevier Ltd. All rights reserved.
PB  - Elsevier, Amsterdam
T2  - Journal of Functional Foods
T1  - Recombinantly produced banana lectin isoform promotes balanced pro-inflammatory response in the colon
EP  - 78
SP  - 68
VL  - 20
DO  - 10.1016/j.jff.2015.10.019
ER  - 

@article{
author = "Marinković, Emilija and Lukić, Ivana and Kosanović, Dejana and Inić-Kanada, Aleksandra and Gavrović-Jankulović, Marija and Stojanović, Marijana",
year = "2016",
abstract = "Recombinant banana lectin isoform (rBanLec) attaches specifically to the mucosal surface, crosses the epithelial barrier and then directly affects the immune response in mouse colon. Structural characteristics, specificity and physiological impacts of rBanLec reported until now highly resemble those of its natural counterpart. Here, we demonstrated that a dose dependent stimulation of the colon with rBanLec skewed the immune response towards Th1/Th17 direction and this effect was counterbalanced by the rise in IL-10 production. Qualitative and quantitative characteristics of the established cytokine network were dependent on the applied rBanLec concentration. In addition, rBanLec enhanced local NO production and myeloperoxidase activity and promoted an increase in local IgA and IgG production. Stimulation with rBanLec can be beneficial in prevention of pathologies raised due to inappropriate cell-mediated immune response as well as in prevention of the pathogen invasion via the colon. (C) 2015 Elsevier Ltd. All rights reserved.",
publisher = "Elsevier, Amsterdam",
journal = "Journal of Functional Foods",
title = "Recombinantly produced banana lectin isoform promotes balanced pro-inflammatory response in the colon",
pages = "78-68",
volume = "20",
doi = "10.1016/j.jff.2015.10.019"
}

Marinković, E., Lukić, I., Kosanović, D., Inić-Kanada, A., Gavrović-Jankulović, M.,& Stojanović, M.. (2016). Recombinantly produced banana lectin isoform promotes balanced pro-inflammatory response in the colon. in Journal of Functional Foods
Elsevier, Amsterdam., 20, 68-78.
https://doi.org/10.1016/j.jff.2015.10.019

Marinković E, Lukić I, Kosanović D, Inić-Kanada A, Gavrović-Jankulović M, Stojanović M. Recombinantly produced banana lectin isoform promotes balanced pro-inflammatory response in the colon. in Journal of Functional Foods. 2016;20:68-78.
doi:10.1016/j.jff.2015.10.019 .

Marinković, Emilija, Lukić, Ivana, Kosanović, Dejana, Inić-Kanada, Aleksandra, Gavrović-Jankulović, Marija, Stojanović, Marijana, "Recombinantly produced banana lectin isoform promotes balanced pro-inflammatory response in the colon" in Journal of Functional Foods, 20 (2016):68-78,
https://doi.org/10.1016/j.jff.2015.10.019 . .

TY  - JOUR
AU  - Belij-Rammerstorfer, Sandra
AU  - Inić-Kanada, Aleksandra
AU  - Stojanović, Marijana
AU  - Marinković, Emilija
AU  - Lukić, Ivana
AU  - Stein, Elisabeth
AU  - Montanaro, Jacqueline
AU  - Bintner, Nora
AU  - Schuerer, Nadine
AU  - Ghasemian, Ehsan
AU  - Kundi, Michael
AU  - Barisani-Asenbauer, Talin
PY  - 2016
UR  - http://intor.torlakinstitut.com/handle/123456789/470
AB  - The aim of this study was to determine whether infectious dose of Chlamydia caviae after repeated infections influences the immunological responses and subsequent clearance of pathogen at the ocular surface of guinea pigs. Animals were infected three times via the conjunctiva at six- and twelve-week intervals by applying either 1 x 10(4) or 1 x 10(6) inclusion-forming units (IFUs) of C. caviae. Ocular pathology, infection course, C. caviae-specific serum IgG levels and their capacity to bind and neutralize infection ex vivo were assessed. Animals infected with 1 x 10(4) IFUs had completely diminished ocular infection and pathology after the 2nd infection with increased levels of C. caviae-specific serum IgG and their effective capacity to bind and neutralize C. caviae. Only partial protection was observed in animals infected with 1 x 10(6) IFUs after the 2nd and 3rd infections. Our findings show that full protection was observed in animals repeatedly infected with the lower dose. The lower dose appeared not to compromise the host immune system, thereby enabling fast clearance of the pathogen and the establishment of competent neutralizing antibodies. (C) 2015 Institut Pasteur. Published by Elsevier Masson SAS. All rights reserved.
PB  - Elsevier Science Bv, Amsterdam
T2  - Microbes and Infection
T1  - Infectious dose and repeated infections are key factors influencing immune response characteristics in guinea pig ocular chlamydial infection
EP  - 262
IS  - 4
SP  - 254
VL  - 18
DO  - 10.1016/j.micinf.2015.12.001
ER  - 

@article{
author = "Belij-Rammerstorfer, Sandra and Inić-Kanada, Aleksandra and Stojanović, Marijana and Marinković, Emilija and Lukić, Ivana and Stein, Elisabeth and Montanaro, Jacqueline and Bintner, Nora and Schuerer, Nadine and Ghasemian, Ehsan and Kundi, Michael and Barisani-Asenbauer, Talin",
year = "2016",
abstract = "The aim of this study was to determine whether infectious dose of Chlamydia caviae after repeated infections influences the immunological responses and subsequent clearance of pathogen at the ocular surface of guinea pigs. Animals were infected three times via the conjunctiva at six- and twelve-week intervals by applying either 1 x 10(4) or 1 x 10(6) inclusion-forming units (IFUs) of C. caviae. Ocular pathology, infection course, C. caviae-specific serum IgG levels and their capacity to bind and neutralize infection ex vivo were assessed. Animals infected with 1 x 10(4) IFUs had completely diminished ocular infection and pathology after the 2nd infection with increased levels of C. caviae-specific serum IgG and their effective capacity to bind and neutralize C. caviae. Only partial protection was observed in animals infected with 1 x 10(6) IFUs after the 2nd and 3rd infections. Our findings show that full protection was observed in animals repeatedly infected with the lower dose. The lower dose appeared not to compromise the host immune system, thereby enabling fast clearance of the pathogen and the establishment of competent neutralizing antibodies. (C) 2015 Institut Pasteur. Published by Elsevier Masson SAS. All rights reserved.",
publisher = "Elsevier Science Bv, Amsterdam",
journal = "Microbes and Infection",
title = "Infectious dose and repeated infections are key factors influencing immune response characteristics in guinea pig ocular chlamydial infection",
pages = "262-254",
number = "4",
volume = "18",
doi = "10.1016/j.micinf.2015.12.001"
}

Belij-Rammerstorfer, S., Inić-Kanada, A., Stojanović, M., Marinković, E., Lukić, I., Stein, E., Montanaro, J., Bintner, N., Schuerer, N., Ghasemian, E., Kundi, M.,& Barisani-Asenbauer, T.. (2016). Infectious dose and repeated infections are key factors influencing immune response characteristics in guinea pig ocular chlamydial infection. in Microbes and Infection
Elsevier Science Bv, Amsterdam., 18(4), 254-262.
https://doi.org/10.1016/j.micinf.2015.12.001

Belij-Rammerstorfer S, Inić-Kanada A, Stojanović M, Marinković E, Lukić I, Stein E, Montanaro J, Bintner N, Schuerer N, Ghasemian E, Kundi M, Barisani-Asenbauer T. Infectious dose and repeated infections are key factors influencing immune response characteristics in guinea pig ocular chlamydial infection. in Microbes and Infection. 2016;18(4):254-262.
doi:10.1016/j.micinf.2015.12.001 .

Belij-Rammerstorfer, Sandra, Inić-Kanada, Aleksandra, Stojanović, Marijana, Marinković, Emilija, Lukić, Ivana, Stein, Elisabeth, Montanaro, Jacqueline, Bintner, Nora, Schuerer, Nadine, Ghasemian, Ehsan, Kundi, Michael, Barisani-Asenbauer, Talin, "Infectious dose and repeated infections are key factors influencing immune response characteristics in guinea pig ocular chlamydial infection" in Microbes and Infection, 18, no. 4 (2016):254-262,
https://doi.org/10.1016/j.micinf.2015.12.001 . .