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  - JOUR
AU  - Ćuruvija, Ivana
AU  - Blagojević, Veljko
AU  - Dragačević, Luka
AU  - Vujić, Vesna
AU  - Lukić, Ivana
AU  - Stanojević, Stanislava
PY  - 2022
UR  - http://intor.torlakinstitut.com/handle/123456789/693
AB  - BCG vaccination induces a memory-like response in innate immune cells known as
trained immunity. In this study, we investigated the modification of innate immune
cells by BCG vaccination in acute peritoneal inflammation. We induced peritonitis with
thioglycollate (TG) in young Albino Oxford male rats which were immunised s.c. with
a BCG vaccine (BCG group) or saline (control group) 7 days prior. Peritoneal cells were
examined for 7 days after TG injection by flow cytometry, and for NO production and
peroxidase activity. Prior in vivo BCG priming altered TG-elicited peritoneal lavage
cells as following: increased in vitro LPS and BCG stimulated NO production from total
cells compared to adherent cells (day 1); increased cell number (days 3 and 5);
increased percentage of inflammatory monocytes (SSCmidCD43lowCD11bmid) and
eosinophils (SSCHihiS48+CD43hi), and a higher level of surface CD11b expression on
CD163+ macrophages (day 5); increased in vitro LPS and BCG stimulated peroxidase
activity (days 5 and 7); and increased percentage of CD163+MHCII+ cells (day 7). On
day 7, cells from both experimental groups showed no production of NO in response to
in vitro stimulation. We conclude that BCG vaccination had a substantial effect on the
acute phase of sterile inflammation, which may lead to the later observed phenotypic
and functional changes that could be seen as accelerated resolution of inflammation
and possibly point to trained immune response.
PB  - Wiley
T1  - BCG vaccination induced alterations of thioglycollate-elicited peritoneal phagocytes: a case of trained immunity?
EP  - 127
IS  - S2
SP  - 127
VL  - 52
UR  - https://hdl.handle.net/21.15107/rcub_intor_693
ER  - 

@article{
author = "Ćuruvija, Ivana and Blagojević, Veljko and Dragačević, Luka and Vujić, Vesna and Lukić, Ivana and Stanojević, Stanislava",
year = "2022",
abstract = "BCG vaccination induces a memory-like response in innate immune cells known as
trained immunity. In this study, we investigated the modification of innate immune
cells by BCG vaccination in acute peritoneal inflammation. We induced peritonitis with
thioglycollate (TG) in young Albino Oxford male rats which were immunised s.c. with
a BCG vaccine (BCG group) or saline (control group) 7 days prior. Peritoneal cells were
examined for 7 days after TG injection by flow cytometry, and for NO production and
peroxidase activity. Prior in vivo BCG priming altered TG-elicited peritoneal lavage
cells as following: increased in vitro LPS and BCG stimulated NO production from total
cells compared to adherent cells (day 1); increased cell number (days 3 and 5);
increased percentage of inflammatory monocytes (SSCmidCD43lowCD11bmid) and
eosinophils (SSCHihiS48+CD43hi), and a higher level of surface CD11b expression on
CD163+ macrophages (day 5); increased in vitro LPS and BCG stimulated peroxidase
activity (days 5 and 7); and increased percentage of CD163+MHCII+ cells (day 7). On
day 7, cells from both experimental groups showed no production of NO in response to
in vitro stimulation. We conclude that BCG vaccination had a substantial effect on the
acute phase of sterile inflammation, which may lead to the later observed phenotypic
and functional changes that could be seen as accelerated resolution of inflammation
and possibly point to trained immune response.",
publisher = "Wiley",
title = "BCG vaccination induced alterations of thioglycollate-elicited peritoneal phagocytes: a case of trained immunity?",
pages = "127-127",
number = "S2",
volume = "52",
url = "https://hdl.handle.net/21.15107/rcub_intor_693"
}

Ćuruvija, I., Blagojević, V., Dragačević, L., Vujić, V., Lukić, I.,& Stanojević, S.. (2022). BCG vaccination induced alterations of thioglycollate-elicited peritoneal phagocytes: a case of trained immunity?. 
Wiley., 52(S2), 127-127.
https://hdl.handle.net/21.15107/rcub_intor_693

Ćuruvija I, Blagojević V, Dragačević L, Vujić V, Lukić I, Stanojević S. BCG vaccination induced alterations of thioglycollate-elicited peritoneal phagocytes: a case of trained immunity?. 2022;52(S2):127-127.
https://hdl.handle.net/21.15107/rcub_intor_693 .

Ćuruvija, Ivana, Blagojević, Veljko, Dragačević, Luka, Vujić, Vesna, Lukić, Ivana, Stanojević, Stanislava, "BCG vaccination induced alterations of thioglycollate-elicited peritoneal phagocytes: a case of trained immunity?", 52, no. S2 (2022):127-127,
https://hdl.handle.net/21.15107/rcub_intor_693 .

TY  - CONF
AU  - Blagojević, Veljko
AU  - Ćuruvija, Ivana
AU  - Dragačević, Luka
AU  - Vujić, Vesna
AU  - Lukić, Ivana
AU  - Stanojević, Stanislava
PY  - 2022
UR  - http://intor.torlakinstitut.com/handle/123456789/694
AB  - Inflammation is a redistribution of immune cells, providing a more efficient elimination of the inflammatory offense. However, it is not limited to local microenvironment. In this study, the interaction of the effect of BCG priming and peritoneal inflammation on the remote inflammatory milieu of infected lung was investigated. Young male AO rats infected with Mycoplasma spp. were s.c. injected with BCG (3x105 CFU) or saline, and 7 days later received an i.p. injection of 7ml of thioglycollate (TG) or saline. Up to 7 days after TG injection, a broncho-alveolar lavage (BAL) was performed, and cells were analysed for their surface marker expression and NO production. Infected rats had a high percentage of HIS48HiCD11bHi neutrophils. BCG priming didn’t alter BAL cells phenotype, while TG injection increased the proportion of MHCII+CD11blow activated alveolar macrophages (aAMFs) on day 7. However, the BCG+TG group showed significant changes – percentage of HIS48HiCD11bHi neutrophils decreased from day 3, the share of aAMFs increased from day 5 and the share of MHCII+CD11b-AMFs increased on days 3-5. However, the percentage of B220+FSClow B lymphocytes were increased from day 1. Production of NO from BAL fluid cells was low in all groups. We conclude that BCG vaccination likely increased the number of circulating B lymphocytes, while TG-induced peritoneal inflammation potentially prevented their entry into the peritoneal cavity, forcing them into permissive tissues, such as lungs.
PB  - Wiley
T1  - Modifying Mycoplasma-infected lung immune cells through an intriguing interplay of BCG priming and peritoneal inflammation
EP  - 55
IS  - S2
SP  - 55
VL  - 52
UR  - https://hdl.handle.net/21.15107/rcub_intor_694
ER  - 

@conference{
author = "Blagojević, Veljko and Ćuruvija, Ivana and Dragačević, Luka and Vujić, Vesna and Lukić, Ivana and Stanojević, Stanislava",
year = "2022",
abstract = "Inflammation is a redistribution of immune cells, providing a more efficient elimination of the inflammatory offense. However, it is not limited to local microenvironment. In this study, the interaction of the effect of BCG priming and peritoneal inflammation on the remote inflammatory milieu of infected lung was investigated. Young male AO rats infected with Mycoplasma spp. were s.c. injected with BCG (3x105 CFU) or saline, and 7 days later received an i.p. injection of 7ml of thioglycollate (TG) or saline. Up to 7 days after TG injection, a broncho-alveolar lavage (BAL) was performed, and cells were analysed for their surface marker expression and NO production. Infected rats had a high percentage of HIS48HiCD11bHi neutrophils. BCG priming didn’t alter BAL cells phenotype, while TG injection increased the proportion of MHCII+CD11blow activated alveolar macrophages (aAMFs) on day 7. However, the BCG+TG group showed significant changes – percentage of HIS48HiCD11bHi neutrophils decreased from day 3, the share of aAMFs increased from day 5 and the share of MHCII+CD11b-AMFs increased on days 3-5. However, the percentage of B220+FSClow B lymphocytes were increased from day 1. Production of NO from BAL fluid cells was low in all groups. We conclude that BCG vaccination likely increased the number of circulating B lymphocytes, while TG-induced peritoneal inflammation potentially prevented their entry into the peritoneal cavity, forcing them into permissive tissues, such as lungs.",
publisher = "Wiley",
title = "Modifying Mycoplasma-infected lung immune cells through an intriguing interplay of BCG priming and peritoneal inflammation",
pages = "55-55",
number = "S2",
volume = "52",
url = "https://hdl.handle.net/21.15107/rcub_intor_694"
}

Blagojević, V., Ćuruvija, I., Dragačević, L., Vujić, V., Lukić, I.,& Stanojević, S.. (2022). Modifying Mycoplasma-infected lung immune cells through an intriguing interplay of BCG priming and peritoneal inflammation. 
Wiley., 52(S2), 55-55.
https://hdl.handle.net/21.15107/rcub_intor_694

Blagojević V, Ćuruvija I, Dragačević L, Vujić V, Lukić I, Stanojević S. Modifying Mycoplasma-infected lung immune cells through an intriguing interplay of BCG priming and peritoneal inflammation. 2022;52(S2):55-55.
https://hdl.handle.net/21.15107/rcub_intor_694 .

Blagojević, Veljko, Ćuruvija, Ivana, Dragačević, Luka, Vujić, Vesna, Lukić, Ivana, Stanojević, Stanislava, "Modifying Mycoplasma-infected lung immune cells through an intriguing interplay of BCG priming and peritoneal inflammation", 52, no. S2 (2022):55-55,
https://hdl.handle.net/21.15107/rcub_intor_694 .

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  - 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  - 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 . .

TY  - CONF
AU  - Inić-Kanada, Aleksandra
AU  - Stojanović, Marijana
AU  - Marinković, Emilija
AU  - Stein, Elisabeth
AU  - Lukić, Ivana
AU  - Belij, Sandra
AU  - Schuerer, Nadine
AU  - Montanaro, Jacqueline
AU  - Ghasemian, Ehsan
AU  - Đokić, Radmila
AU  - Barisani-Asenbauer, Talin
PY  - 2016
UR  - http://intor.torlakinstitut.com/handle/123456789/459
PB  - Assoc Research Vision Ophthalmology Inc, Rockville
C3  - Investigative Ophthalmology & Visual Science
T1  - Immune Responses Induced by chlamydial Polymorphic Membrane Protein C formulated with probiotic Lactobacillus rhamnosus as an Adjuvant in Conjunctival Immunization
IS  - 12
VL  - 57
UR  - https://hdl.handle.net/21.15107/rcub_intor_459
ER  - 

@conference{
author = "Inić-Kanada, Aleksandra and Stojanović, Marijana and Marinković, Emilija and Stein, Elisabeth and Lukić, Ivana and Belij, Sandra and Schuerer, Nadine and Montanaro, Jacqueline and Ghasemian, Ehsan and Đokić, Radmila and Barisani-Asenbauer, Talin",
year = "2016",
publisher = "Assoc Research Vision Ophthalmology Inc, Rockville",
journal = "Investigative Ophthalmology & Visual Science",
title = "Immune Responses Induced by chlamydial Polymorphic Membrane Protein C formulated with probiotic Lactobacillus rhamnosus as an Adjuvant in Conjunctival Immunization",
number = "12",
volume = "57",
url = "https://hdl.handle.net/21.15107/rcub_intor_459"
}

Inić-Kanada, A., Stojanović, M., Marinković, E., Stein, E., Lukić, I., Belij, S., Schuerer, N., Montanaro, J., Ghasemian, E., Đokić, R.,& Barisani-Asenbauer, T.. (2016). Immune Responses Induced by chlamydial Polymorphic Membrane Protein C formulated with probiotic Lactobacillus rhamnosus as an Adjuvant in Conjunctival Immunization. in Investigative Ophthalmology & Visual Science
Assoc Research Vision Ophthalmology Inc, Rockville., 57(12).
https://hdl.handle.net/21.15107/rcub_intor_459

Inić-Kanada A, Stojanović M, Marinković E, Stein E, Lukić I, Belij S, Schuerer N, Montanaro J, Ghasemian E, Đokić R, Barisani-Asenbauer T. Immune Responses Induced by chlamydial Polymorphic Membrane Protein C formulated with probiotic Lactobacillus rhamnosus as an Adjuvant in Conjunctival Immunization. in Investigative Ophthalmology & Visual Science. 2016;57(12).
https://hdl.handle.net/21.15107/rcub_intor_459 .

Inić-Kanada, Aleksandra, Stojanović, Marijana, Marinković, Emilija, Stein, Elisabeth, Lukić, Ivana, Belij, Sandra, Schuerer, Nadine, Montanaro, Jacqueline, Ghasemian, Ehsan, Đokić, Radmila, Barisani-Asenbauer, Talin, "Immune Responses Induced by chlamydial Polymorphic Membrane Protein C formulated with probiotic Lactobacillus rhamnosus as an Adjuvant in Conjunctival Immunization" in Investigative Ophthalmology & Visual Science, 57, no. 12 (2016),
https://hdl.handle.net/21.15107/rcub_intor_459 .

TY  - JOUR
AU  - Inić-Kanada, Aleksandra
AU  - Stojanović, Marijana
AU  - Marinković, Emilija
AU  - Becker, Elisabeth
AU  - Stein, Elisabeth
AU  - Lukić, Ivana
AU  - Đokić, Radmila
AU  - Schuerer, Nadine
AU  - Hegemann, Johannes H.
AU  - Barisani-Asenbauer, Talin
PY  - 2016
UR  - http://intor.torlakinstitut.com/handle/123456789/455
AB  - Recent advances in the development of chlamydia vaccines, using live-attenuated or ultraviolet light-inactivated chlamydia, are paving the way for new possibilities to oppose the societal challenges posed by chlamydia-related diseases, such as blinding trachoma. An effective subunit vaccine would mitigate the risks associated with the use of a whole-cell vaccine. Our rationale for the design of an efficient subunit vaccine against Chlamydia trachomatis (Ct) is based on the membrane proteins involved in the initial Ct-host cell contact and on the route of immunization that mimics the natural infection process (i.e., via the ocular mucosa). The first aim of our study was to characterize the specific conjunctival and vaginal immune responses following eye drop immunization in BALB/c mice, using the N-terminal portion of the Ct serovar E polymorphic membrane protein C (N-PmpC) as the subunit vaccine antigen. Second, we aimed to examine the adjuvant properties of the probiotic Lactobacillus rhamnosus (LB) when formulated with N-PmpC. N-PmpC applied alone stimulated the production of N-PmpC-and Ct serovar B-specific antibodies in serum, tears and vaginal washes, whereas the combination with LB significantly enhanced these responses. The N-PmpC/LB combination initiated a T cell response characterized by an elevated percentage of CD25+ T cells and CD8+ effector T cells, enhanced CD4+ T-helper 1 skewing, and increased regulatory T cell responses. Together, these results show that eye drop vaccination with combined use of N-PmpC and a live probiotic LB stimulates specific cellular and humoral immune responses, not only locally in the conjunctiva but also in the vaginal mucosa, which could be a promising approach in Ct vaccine development.
PB  - Public Library Science, San Francisco
T2  - PLoS One
T1  - A Probiotic Adjuvant Lactobacillus rhamnosus Enhances Specific Immune Responses after Ocular Mucosal Immunization with Chlamydial Polymorphic Membrane Protein C
IS  - 9
VL  - 11
DO  - 10.1371/journal.pone.0157875
ER  - 

@article{
author = "Inić-Kanada, Aleksandra and Stojanović, Marijana and Marinković, Emilija and Becker, Elisabeth and Stein, Elisabeth and Lukić, Ivana and Đokić, Radmila and Schuerer, Nadine and Hegemann, Johannes H. and Barisani-Asenbauer, Talin",
year = "2016",
abstract = "Recent advances in the development of chlamydia vaccines, using live-attenuated or ultraviolet light-inactivated chlamydia, are paving the way for new possibilities to oppose the societal challenges posed by chlamydia-related diseases, such as blinding trachoma. An effective subunit vaccine would mitigate the risks associated with the use of a whole-cell vaccine. Our rationale for the design of an efficient subunit vaccine against Chlamydia trachomatis (Ct) is based on the membrane proteins involved in the initial Ct-host cell contact and on the route of immunization that mimics the natural infection process (i.e., via the ocular mucosa). The first aim of our study was to characterize the specific conjunctival and vaginal immune responses following eye drop immunization in BALB/c mice, using the N-terminal portion of the Ct serovar E polymorphic membrane protein C (N-PmpC) as the subunit vaccine antigen. Second, we aimed to examine the adjuvant properties of the probiotic Lactobacillus rhamnosus (LB) when formulated with N-PmpC. N-PmpC applied alone stimulated the production of N-PmpC-and Ct serovar B-specific antibodies in serum, tears and vaginal washes, whereas the combination with LB significantly enhanced these responses. The N-PmpC/LB combination initiated a T cell response characterized by an elevated percentage of CD25+ T cells and CD8+ effector T cells, enhanced CD4+ T-helper 1 skewing, and increased regulatory T cell responses. Together, these results show that eye drop vaccination with combined use of N-PmpC and a live probiotic LB stimulates specific cellular and humoral immune responses, not only locally in the conjunctiva but also in the vaginal mucosa, which could be a promising approach in Ct vaccine development.",
publisher = "Public Library Science, San Francisco",
journal = "PLoS One",
title = "A Probiotic Adjuvant Lactobacillus rhamnosus Enhances Specific Immune Responses after Ocular Mucosal Immunization with Chlamydial Polymorphic Membrane Protein C",
number = "9",
volume = "11",
doi = "10.1371/journal.pone.0157875"
}

Inić-Kanada, A., Stojanović, M., Marinković, E., Becker, E., Stein, E., Lukić, I., Đokić, R., Schuerer, N., Hegemann, J. H.,& Barisani-Asenbauer, T.. (2016). A Probiotic Adjuvant Lactobacillus rhamnosus Enhances Specific Immune Responses after Ocular Mucosal Immunization with Chlamydial Polymorphic Membrane Protein C. in PLoS One
Public Library Science, San Francisco., 11(9).
https://doi.org/10.1371/journal.pone.0157875

Inić-Kanada A, Stojanović M, Marinković E, Becker E, Stein E, Lukić I, Đokić R, Schuerer N, Hegemann JH, Barisani-Asenbauer T. A Probiotic Adjuvant Lactobacillus rhamnosus Enhances Specific Immune Responses after Ocular Mucosal Immunization with Chlamydial Polymorphic Membrane Protein C. in PLoS One. 2016;11(9).
doi:10.1371/journal.pone.0157875 .

Inić-Kanada, Aleksandra, Stojanović, Marijana, Marinković, Emilija, Becker, Elisabeth, Stein, Elisabeth, Lukić, Ivana, Đokić, Radmila, Schuerer, Nadine, Hegemann, Johannes H., Barisani-Asenbauer, Talin, "A Probiotic Adjuvant Lactobacillus rhamnosus Enhances Specific Immune Responses after Ocular Mucosal Immunization with Chlamydial Polymorphic Membrane Protein C" in PLoS One, 11, no. 9 (2016),
https://doi.org/10.1371/journal.pone.0157875 . .

TY  - JOUR
AU  - Inić-Kanada, Aleksandra
AU  - Stojanović, Marijana
AU  - Schlacher, Simone
AU  - Stein, Elisabeth
AU  - Belij-Rammerstorfer, Sandra
AU  - Marinković, Emilija
AU  - Lukić, Ivana
AU  - Montanaro, Jacqueline
AU  - Schuerer, Nadine
AU  - Bintner, Nora
AU  - Kovačević-Jovanović, Vesna
AU  - Krnjaja, Ognjen
AU  - Mayr, Ulrike Beate
AU  - Lubitz, Werner
AU  - Barisani-Asenbauer, Talin
PY  - 2015
UR  - http://intor.torlakinstitut.com/handle/123456789/434
AB  - Trachoma, caused by the intracellular bacterium Chlamydia trachomatis (Ct), remains the world's leading preventable infectious cause of blindness. Recent attempts to develop effective vaccines rely on modified chlamydial antigen delivery platforms. As the mechanisms engaged in the pathology of the disease are not fully understood, designing a subunit vaccine specific to chlamydial antigens could improve safety for human use. We propose the delivery of chlamydia-specific antigens to the ocular mucosa using particulate carriers, bacterial ghosts (BGs). We therefore characterized humoral and cellular immune responses after conjunctival and subcutaneous immunization with a N-terminal portion (amino acid 1-893) of the chlamydial polymorphic membrane protein C (PmpC) of Ct serovar B, expressed in probiotic Escherichia coli Nissle 1917 bacterial ghosts (EcN BGs) in BALB/cmice. Three immunizations were performed at two-week intervals, and the immune responses were evaluated two weeks after the final immunization in mice. In a guinea pig model of ocular infection animals were immunized in the same manner as the mice, and protection against challenge was assessed two weeks after the last immunization. N-PmpC was successfully expressed within BGs and delivery to the ocularmucosa was well tolerated without signs of inflammation. N-PmpC- specific mucosal IgA levels in tears yielded significantly increased levels in the group immunized via the conjunctiva compared with the subcutaneously immunized mice. Immunization with N-PmpC EcN BGs via both immunization routes prompted the establishment of an N-PmpC-specific IFN gamma immune response. Immunization via the conjunctiva resulted in a decrease in intensity of the transitional inflammatory reaction in conjunctiva of challenged guinea pigs compared with subcutaneously and non-immunized animals. The delivery of the chlamydial subunit vaccine to the ocular mucosa using a particulate carrier, such as BGs, induced both humoral and cellular immune responses. Further investigations are needed to improve the immunization scheme and dosage.
PB  - Public Library Science, San Francisco
T2  - PLoS One
T1  - Delivery of a Chlamydial Adhesin N-PmpC Subunit Vaccine to the Ocular Mucosa Using Particulate Carriers
IS  - 12
VL  - 10
DO  - 10.1371/journal.pone.0144380
ER  - 

@article{
author = "Inić-Kanada, Aleksandra and Stojanović, Marijana and Schlacher, Simone and Stein, Elisabeth and Belij-Rammerstorfer, Sandra and Marinković, Emilija and Lukić, Ivana and Montanaro, Jacqueline and Schuerer, Nadine and Bintner, Nora and Kovačević-Jovanović, Vesna and Krnjaja, Ognjen and Mayr, Ulrike Beate and Lubitz, Werner and Barisani-Asenbauer, Talin",
year = "2015",
abstract = "Trachoma, caused by the intracellular bacterium Chlamydia trachomatis (Ct), remains the world's leading preventable infectious cause of blindness. Recent attempts to develop effective vaccines rely on modified chlamydial antigen delivery platforms. As the mechanisms engaged in the pathology of the disease are not fully understood, designing a subunit vaccine specific to chlamydial antigens could improve safety for human use. We propose the delivery of chlamydia-specific antigens to the ocular mucosa using particulate carriers, bacterial ghosts (BGs). We therefore characterized humoral and cellular immune responses after conjunctival and subcutaneous immunization with a N-terminal portion (amino acid 1-893) of the chlamydial polymorphic membrane protein C (PmpC) of Ct serovar B, expressed in probiotic Escherichia coli Nissle 1917 bacterial ghosts (EcN BGs) in BALB/cmice. Three immunizations were performed at two-week intervals, and the immune responses were evaluated two weeks after the final immunization in mice. In a guinea pig model of ocular infection animals were immunized in the same manner as the mice, and protection against challenge was assessed two weeks after the last immunization. N-PmpC was successfully expressed within BGs and delivery to the ocularmucosa was well tolerated without signs of inflammation. N-PmpC- specific mucosal IgA levels in tears yielded significantly increased levels in the group immunized via the conjunctiva compared with the subcutaneously immunized mice. Immunization with N-PmpC EcN BGs via both immunization routes prompted the establishment of an N-PmpC-specific IFN gamma immune response. Immunization via the conjunctiva resulted in a decrease in intensity of the transitional inflammatory reaction in conjunctiva of challenged guinea pigs compared with subcutaneously and non-immunized animals. The delivery of the chlamydial subunit vaccine to the ocular mucosa using a particulate carrier, such as BGs, induced both humoral and cellular immune responses. Further investigations are needed to improve the immunization scheme and dosage.",
publisher = "Public Library Science, San Francisco",
journal = "PLoS One",
title = "Delivery of a Chlamydial Adhesin N-PmpC Subunit Vaccine to the Ocular Mucosa Using Particulate Carriers",
number = "12",
volume = "10",
doi = "10.1371/journal.pone.0144380"
}

Inić-Kanada, A., Stojanović, M., Schlacher, S., Stein, E., Belij-Rammerstorfer, S., Marinković, E., Lukić, I., Montanaro, J., Schuerer, N., Bintner, N., Kovačević-Jovanović, V., Krnjaja, O., Mayr, U. B., Lubitz, W.,& Barisani-Asenbauer, T.. (2015). Delivery of a Chlamydial Adhesin N-PmpC Subunit Vaccine to the Ocular Mucosa Using Particulate Carriers. in PLoS One
Public Library Science, San Francisco., 10(12).
https://doi.org/10.1371/journal.pone.0144380

Inić-Kanada A, Stojanović M, Schlacher S, Stein E, Belij-Rammerstorfer S, Marinković E, Lukić I, Montanaro J, Schuerer N, Bintner N, Kovačević-Jovanović V, Krnjaja O, Mayr UB, Lubitz W, Barisani-Asenbauer T. Delivery of a Chlamydial Adhesin N-PmpC Subunit Vaccine to the Ocular Mucosa Using Particulate Carriers. in PLoS One. 2015;10(12).
doi:10.1371/journal.pone.0144380 .

Inić-Kanada, Aleksandra, Stojanović, Marijana, Schlacher, Simone, Stein, Elisabeth, Belij-Rammerstorfer, Sandra, Marinković, Emilija, Lukić, Ivana, Montanaro, Jacqueline, Schuerer, Nadine, Bintner, Nora, Kovačević-Jovanović, Vesna, Krnjaja, Ognjen, Mayr, Ulrike Beate, Lubitz, Werner, Barisani-Asenbauer, Talin, "Delivery of a Chlamydial Adhesin N-PmpC Subunit Vaccine to the Ocular Mucosa Using Particulate Carriers" in PLoS One, 10, no. 12 (2015),
https://doi.org/10.1371/journal.pone.0144380 . .

TY  - JOUR
AU  - Lukić, Ivana
AU  - Marinković, Emilija
AU  - Filipović, Ana
AU  - Krnjaja, Ognjen
AU  - Kosanović, Dejana
AU  - Inić-Kanada, Aleksandra
AU  - Stojanović, Marijana
PY  - 2015
UR  - http://intor.torlakinstitut.com/handle/123456789/425
AB  - Antibodies capable to neutralize tetanus toxin (TeNT) are key factors in protection against tetanus disease. Although antibody-based therapeutics for treatment of tetanus exist on the market its production is tedious. Hence, the tetanus-specific antibodies preparation that could be easily produced in large scale in vitro would be beneficial. Monoclonal antibodies (MAbs) are considered for a long time as a reagent of choice, but the core drawback is how to select a MAb that would be safe in providing efficacious protection. In this study we have investigated the parameters crucial for a single MAb to be assigned as protective. Eight murine MAbs were characterized in vitro for their reactivity toward TeNT and assessed in vivo for protectiveness against TeNT intoxication. Correlation of in vitro and in vivo data has revealed that in vitro selection of MAb that is protective in vivo could be performed by a combination of two assays: the measurement of MAb affinity toward TeNT taking Ka 1 x 10(8) M-1 as a threshold level, and the evaluation of its capability to prevent TeNT-ganglioside interaction. Single MAb could be taken into consideration as a potential therapeutic only if it has a capacity to completely inhibits TeNT-ganglioside complex formation. (C) 2015 Elsevier Ltd. All rights reserved.
PB  - Pergamon-Elsevier Science Ltd, Oxford
T2  - Toxicon
T1  - Key protection factors against tetanus: Anti-tetanus toxin antibody affinity and its ability to prevent tetanus toxin - ganglioside interaction
EP  - 144
SP  - 135
VL  - 103
DO  - 10.1016/j.toxicon.2015.06.025
ER  - 

@article{
author = "Lukić, Ivana and Marinković, Emilija and Filipović, Ana and Krnjaja, Ognjen and Kosanović, Dejana and Inić-Kanada, Aleksandra and Stojanović, Marijana",
year = "2015",
abstract = "Antibodies capable to neutralize tetanus toxin (TeNT) are key factors in protection against tetanus disease. Although antibody-based therapeutics for treatment of tetanus exist on the market its production is tedious. Hence, the tetanus-specific antibodies preparation that could be easily produced in large scale in vitro would be beneficial. Monoclonal antibodies (MAbs) are considered for a long time as a reagent of choice, but the core drawback is how to select a MAb that would be safe in providing efficacious protection. In this study we have investigated the parameters crucial for a single MAb to be assigned as protective. Eight murine MAbs were characterized in vitro for their reactivity toward TeNT and assessed in vivo for protectiveness against TeNT intoxication. Correlation of in vitro and in vivo data has revealed that in vitro selection of MAb that is protective in vivo could be performed by a combination of two assays: the measurement of MAb affinity toward TeNT taking Ka 1 x 10(8) M-1 as a threshold level, and the evaluation of its capability to prevent TeNT-ganglioside interaction. Single MAb could be taken into consideration as a potential therapeutic only if it has a capacity to completely inhibits TeNT-ganglioside complex formation. (C) 2015 Elsevier Ltd. All rights reserved.",
publisher = "Pergamon-Elsevier Science Ltd, Oxford",
journal = "Toxicon",
title = "Key protection factors against tetanus: Anti-tetanus toxin antibody affinity and its ability to prevent tetanus toxin - ganglioside interaction",
pages = "144-135",
volume = "103",
doi = "10.1016/j.toxicon.2015.06.025"
}

Lukić, I., Marinković, E., Filipović, A., Krnjaja, O., Kosanović, D., Inić-Kanada, A.,& Stojanović, M.. (2015). Key protection factors against tetanus: Anti-tetanus toxin antibody affinity and its ability to prevent tetanus toxin - ganglioside interaction. in Toxicon
Pergamon-Elsevier Science Ltd, Oxford., 103, 135-144.
https://doi.org/10.1016/j.toxicon.2015.06.025

Lukić I, Marinković E, Filipović A, Krnjaja O, Kosanović D, Inić-Kanada A, Stojanović M. Key protection factors against tetanus: Anti-tetanus toxin antibody affinity and its ability to prevent tetanus toxin - ganglioside interaction. in Toxicon. 2015;103:135-144.
doi:10.1016/j.toxicon.2015.06.025 .

Lukić, Ivana, Marinković, Emilija, Filipović, Ana, Krnjaja, Ognjen, Kosanović, Dejana, Inić-Kanada, Aleksandra, Stojanović, Marijana, "Key protection factors against tetanus: Anti-tetanus toxin antibody affinity and its ability to prevent tetanus toxin - ganglioside interaction" in Toxicon, 103 (2015):135-144,
https://doi.org/10.1016/j.toxicon.2015.06.025 . .

TY  - JOUR
AU  - Barisani-Asenbauer, Talin
AU  - Inić-Kanada, Aleksandra
AU  - Belij, Sandra
AU  - Marinković, Emilija
AU  - Lukić, Ivana
AU  - Montanaro, Jacqueline
AU  - Stein, Elisabeth
AU  - Bintner, Nora
AU  - Stojanović, Marijana
PY  - 2013
UR  - http://intor.torlakinstitut.com/handle/123456789/393
AB  - Background: In a quest for a needle-free vaccine administration strategy, we evaluated the ocular conjunctiva as an alternative mucosal immunization route by profiling and comparing the local and systemic immune responses to the subcutaneous or conjunctival administration of tetanus toxoid (TTd), a model antigen. Materials and methods: BALB/c and C57BL/6 mice were immunized either subcutaneously with TTd alone or via the conjunctiva with TTd alone, TTd mixed with 2% glycerol or TTd with merthiolate-inactivated whole-cell B. pertussis (wBP) as adjuvants. Mice were immunized on days 0, 7 and 14 via both routes, and an evaluation of the local and systemic immune responses was performed two weeks after the last immunization. Four weeks after the last immunization, the mice were challenged with a lethal dose (2 x LD50) of tetanus toxin. Results: The conjunctival application of TTd in BALB/c mice induced TTd-specific secretory IgA production and skewed the TTd-specific immune response toward a Th1/Th17 profile, as determined by the stimulation of IFN gamma and IL-17A secretion and/or the concurrent pronounced reduction of IL-4 secretion, irrespective of the adjuvant. In conjunctivaly immunized C57BL/6 mice, only TTd administered with wBP promoted the establishment of a mixed Th1/Th17 TTd-specific immune response, whereas TTd alone or TTd in conjunction with glycerol initiated a dominant Th1 response against TTd. Immunization via the conjunctiva with TTd plus wBP adjuvant resulted in a 33% survival rate of challenged mice compared to a 0% survival rate in non-immunized animals (p lt 0.05). Conclusion: Conjunctival immunization with TTd alone or with various adjuvants induced TTd-specific local and systemic immune responses, predominantly of the Th1 type. The strongest immune responses developed in mice that received TTd together with wBP, which implies that this alternative route might tailor the immune response to fight intracellular bacteria or viruses more effectively.
PB  - Public Library Science, San Francisco
T2  - PLoS One
T1  - The Ocular Conjunctiva as a Mucosal Immunization Route: A Profile of the Immune Response to the Model Antigen Tetanus Toxoid
IS  - 4
VL  - 8
DO  - 10.1371/journal.pone.0060682
ER  - 

@article{
author = "Barisani-Asenbauer, Talin and Inić-Kanada, Aleksandra and Belij, Sandra and Marinković, Emilija and Lukić, Ivana and Montanaro, Jacqueline and Stein, Elisabeth and Bintner, Nora and Stojanović, Marijana",
year = "2013",
abstract = "Background: In a quest for a needle-free vaccine administration strategy, we evaluated the ocular conjunctiva as an alternative mucosal immunization route by profiling and comparing the local and systemic immune responses to the subcutaneous or conjunctival administration of tetanus toxoid (TTd), a model antigen. Materials and methods: BALB/c and C57BL/6 mice were immunized either subcutaneously with TTd alone or via the conjunctiva with TTd alone, TTd mixed with 2% glycerol or TTd with merthiolate-inactivated whole-cell B. pertussis (wBP) as adjuvants. Mice were immunized on days 0, 7 and 14 via both routes, and an evaluation of the local and systemic immune responses was performed two weeks after the last immunization. Four weeks after the last immunization, the mice were challenged with a lethal dose (2 x LD50) of tetanus toxin. Results: The conjunctival application of TTd in BALB/c mice induced TTd-specific secretory IgA production and skewed the TTd-specific immune response toward a Th1/Th17 profile, as determined by the stimulation of IFN gamma and IL-17A secretion and/or the concurrent pronounced reduction of IL-4 secretion, irrespective of the adjuvant. In conjunctivaly immunized C57BL/6 mice, only TTd administered with wBP promoted the establishment of a mixed Th1/Th17 TTd-specific immune response, whereas TTd alone or TTd in conjunction with glycerol initiated a dominant Th1 response against TTd. Immunization via the conjunctiva with TTd plus wBP adjuvant resulted in a 33% survival rate of challenged mice compared to a 0% survival rate in non-immunized animals (p lt 0.05). Conclusion: Conjunctival immunization with TTd alone or with various adjuvants induced TTd-specific local and systemic immune responses, predominantly of the Th1 type. The strongest immune responses developed in mice that received TTd together with wBP, which implies that this alternative route might tailor the immune response to fight intracellular bacteria or viruses more effectively.",
publisher = "Public Library Science, San Francisco",
journal = "PLoS One",
title = "The Ocular Conjunctiva as a Mucosal Immunization Route: A Profile of the Immune Response to the Model Antigen Tetanus Toxoid",
number = "4",
volume = "8",
doi = "10.1371/journal.pone.0060682"
}

Barisani-Asenbauer, T., Inić-Kanada, A., Belij, S., Marinković, E., Lukić, I., Montanaro, J., Stein, E., Bintner, N.,& Stojanović, M.. (2013). The Ocular Conjunctiva as a Mucosal Immunization Route: A Profile of the Immune Response to the Model Antigen Tetanus Toxoid. in PLoS One
Public Library Science, San Francisco., 8(4).
https://doi.org/10.1371/journal.pone.0060682

Barisani-Asenbauer T, Inić-Kanada A, Belij S, Marinković E, Lukić I, Montanaro J, Stein E, Bintner N, Stojanović M. The Ocular Conjunctiva as a Mucosal Immunization Route: A Profile of the Immune Response to the Model Antigen Tetanus Toxoid. in PLoS One. 2013;8(4).
doi:10.1371/journal.pone.0060682 .

Barisani-Asenbauer, Talin, Inić-Kanada, Aleksandra, Belij, Sandra, Marinković, Emilija, Lukić, Ivana, Montanaro, Jacqueline, Stein, Elisabeth, Bintner, Nora, Stojanović, Marijana, "The Ocular Conjunctiva as a Mucosal Immunization Route: A Profile of the Immune Response to the Model Antigen Tetanus Toxoid" in PLoS One, 8, no. 4 (2013),
https://doi.org/10.1371/journal.pone.0060682 . .

TY  - JOUR
AU  - Živković, Irena
AU  - Petrušić, Vladimir
AU  - Stojanović, Marijana
AU  - Inić-Kanada, Aleksandra
AU  - Lukić, Ivana
AU  - Dimitrijević, Ljiljana
PY  - 2012
UR  - http://intor.torlakinstitut.com/handle/123456789/363
AB  - It has already been shown that tetanus toxoid (TTd) hyper-immunization is a suitable experimental method for creating the animal model of antiphospholipid syndrome (APS) in BALB/c mice. The severity of APS pathology in BALB/c mice mainly correlates to the affinity of anti-beta(2) glycoprotein I (beta(2)GPI) antibodies. In this study we have investigated reproductive pathology induced in C57BL/6 mice by TTd hyper-immunization using a combination of different pretreatments (complete Freund's adjuvant or glycerol) and adjuvants (alhydrogel or glycerol). A decrease in fecundity was recorded in only C57BL/6 mice immunized with alhydrogel adjuvant, irrespective of the kind of applied pretreatment; it was associated with an increase in abundance of low affinity anti-beta(2)GPI IgG antibodies and Th1 prevalence.
PB  - Sage Publications Ltd, London
T2  - Innate Immunity
T1  - Induction of decreased fecundity by tetanus toxoid hyper-immunization in C57BL/6 mice depends on the applied adjuvant
EP  - 342
IS  - 2
SP  - 333
VL  - 18
DO  - 10.1177/1753425911407361
ER  - 

@article{
author = "Živković, Irena and Petrušić, Vladimir and Stojanović, Marijana and Inić-Kanada, Aleksandra and Lukić, Ivana and Dimitrijević, Ljiljana",
year = "2012",
abstract = "It has already been shown that tetanus toxoid (TTd) hyper-immunization is a suitable experimental method for creating the animal model of antiphospholipid syndrome (APS) in BALB/c mice. The severity of APS pathology in BALB/c mice mainly correlates to the affinity of anti-beta(2) glycoprotein I (beta(2)GPI) antibodies. In this study we have investigated reproductive pathology induced in C57BL/6 mice by TTd hyper-immunization using a combination of different pretreatments (complete Freund's adjuvant or glycerol) and adjuvants (alhydrogel or glycerol). A decrease in fecundity was recorded in only C57BL/6 mice immunized with alhydrogel adjuvant, irrespective of the kind of applied pretreatment; it was associated with an increase in abundance of low affinity anti-beta(2)GPI IgG antibodies and Th1 prevalence.",
publisher = "Sage Publications Ltd, London",
journal = "Innate Immunity",
title = "Induction of decreased fecundity by tetanus toxoid hyper-immunization in C57BL/6 mice depends on the applied adjuvant",
pages = "342-333",
number = "2",
volume = "18",
doi = "10.1177/1753425911407361"
}

Živković, I., Petrušić, V., Stojanović, M., Inić-Kanada, A., Lukić, I.,& Dimitrijević, L.. (2012). Induction of decreased fecundity by tetanus toxoid hyper-immunization in C57BL/6 mice depends on the applied adjuvant. in Innate Immunity
Sage Publications Ltd, London., 18(2), 333-342.
https://doi.org/10.1177/1753425911407361

Živković I, Petrušić V, Stojanović M, Inić-Kanada A, Lukić I, Dimitrijević L. Induction of decreased fecundity by tetanus toxoid hyper-immunization in C57BL/6 mice depends on the applied adjuvant. in Innate Immunity. 2012;18(2):333-342.
doi:10.1177/1753425911407361 .

Živković, Irena, Petrušić, Vladimir, Stojanović, Marijana, Inić-Kanada, Aleksandra, Lukić, Ivana, Dimitrijević, Ljiljana, "Induction of decreased fecundity by tetanus toxoid hyper-immunization in C57BL/6 mice depends on the applied adjuvant" in Innate Immunity, 18, no. 2 (2012):333-342,
https://doi.org/10.1177/1753425911407361 . .

TY  - JOUR
AU  - Petrušić, Vladimir
AU  - Živković, Irena
AU  - Stojanović, Marijana
AU  - Lukić, Ivana
AU  - Marinković, Emilija
AU  - Dimitrijević, Ljiljana
PY  - 2012
UR  - http://intor.torlakinstitut.com/handle/123456789/349
AB  - Tetanus neurotoxin (TeNT) represents a potent toxin that binds to its receptors on neurons and inhibits the release of neurotransmitters. Additionally, its fragments are used to transport pharmacological substances to neuronal cell bodies. The main objective of this study was the development of a suitable model system to study internalization of the TeNT. We have produced a monoclonal antibody (MoAb) specific for TeNT by hybridoma technology, after immunization of BALB/c mice with tetanus toxoid, and have named it T-62. The immunochemical characteristics of MoAb T-62 were tested using ELISA, PAGE and immunoblotting. Finally, we have used an immunohistochemical method to detect specific binding of MoAb T-62 to TeNT bound to PC 12 cells. Our results show that MoAb T-62 is highly specific for TeNT, even when it is bound to its receptor, and that it could be of considerable importance in studies regarding fundamental research on TeNT receptors, intracellular transport of TeNT, as well as retrograde transport of pharmaceutical substances and non-invasive delivery of polypeptides through the blood brain barrier. In addition, MoAb T-62 is an invaluable tool in TeNT vaccine production as it can be used for the detection of reverse toxicity, which could drastically reduce the need to use animals in these experiments. (C) 2011 Elsevier GmbH. All rights reserved.
PB  - Elsevier Gmbh, Urban & Fischer Verlag, Jena
T2  - Acta Histochemica
T1  - Production, characterization and applications of a tetanus toxin specific monoclonal antibody T-62
EP  - 486
IS  - 5
SP  - 480
VL  - 114
DO  - 10.1016/j.acthis.2011.09.001
ER  - 

@article{
author = "Petrušić, Vladimir and Živković, Irena and Stojanović, Marijana and Lukić, Ivana and Marinković, Emilija and Dimitrijević, Ljiljana",
year = "2012",
abstract = "Tetanus neurotoxin (TeNT) represents a potent toxin that binds to its receptors on neurons and inhibits the release of neurotransmitters. Additionally, its fragments are used to transport pharmacological substances to neuronal cell bodies. The main objective of this study was the development of a suitable model system to study internalization of the TeNT. We have produced a monoclonal antibody (MoAb) specific for TeNT by hybridoma technology, after immunization of BALB/c mice with tetanus toxoid, and have named it T-62. The immunochemical characteristics of MoAb T-62 were tested using ELISA, PAGE and immunoblotting. Finally, we have used an immunohistochemical method to detect specific binding of MoAb T-62 to TeNT bound to PC 12 cells. Our results show that MoAb T-62 is highly specific for TeNT, even when it is bound to its receptor, and that it could be of considerable importance in studies regarding fundamental research on TeNT receptors, intracellular transport of TeNT, as well as retrograde transport of pharmaceutical substances and non-invasive delivery of polypeptides through the blood brain barrier. In addition, MoAb T-62 is an invaluable tool in TeNT vaccine production as it can be used for the detection of reverse toxicity, which could drastically reduce the need to use animals in these experiments. (C) 2011 Elsevier GmbH. All rights reserved.",
publisher = "Elsevier Gmbh, Urban & Fischer Verlag, Jena",
journal = "Acta Histochemica",
title = "Production, characterization and applications of a tetanus toxin specific monoclonal antibody T-62",
pages = "486-480",
number = "5",
volume = "114",
doi = "10.1016/j.acthis.2011.09.001"
}

Petrušić, V., Živković, I., Stojanović, M., Lukić, I., Marinković, E.,& Dimitrijević, L.. (2012). Production, characterization and applications of a tetanus toxin specific monoclonal antibody T-62. in Acta Histochemica
Elsevier Gmbh, Urban & Fischer Verlag, Jena., 114(5), 480-486.
https://doi.org/10.1016/j.acthis.2011.09.001

Petrušić V, Živković I, Stojanović M, Lukić I, Marinković E, Dimitrijević L. Production, characterization and applications of a tetanus toxin specific monoclonal antibody T-62. in Acta Histochemica. 2012;114(5):480-486.
doi:10.1016/j.acthis.2011.09.001 .

Petrušić, Vladimir, Živković, Irena, Stojanović, Marijana, Lukić, Ivana, Marinković, Emilija, Dimitrijević, Ljiljana, "Production, characterization and applications of a tetanus toxin specific monoclonal antibody T-62" in Acta Histochemica, 114, no. 5 (2012):480-486,
https://doi.org/10.1016/j.acthis.2011.09.001 . .

TY  - JOUR
AU  - Petrušić, Vladimir
AU  - Živković, Irena
AU  - Stojanović, Marijana
AU  - Lukić, Ivana
AU  - Marinković, Emilija
AU  - Inić-Kanada, Aleksandra
AU  - Dimitrijevic, Ljiliana
PY  - 2011
UR  - http://intor.torlakinstitut.com/handle/123456789/339
AB  - Natural antibodies (NAbs) are present in circulation even before the exposure to antigen and they exert various biological functions. They are polyreactive and mainly represented by immunoglobulin M (IgM), which is the first antibody produced in an ongoing immune response to infection and/or immunization. IgM is always secreted as a polymer with predominant pentameric structure, although other polymeric forms such as hexamer can be also formed. The biological functions of hexameric IgM are still not known and it is proposed that its existence as a NAb could be deleterious. However, the nature of IgM hexamers has not been investigated yet. In this paper, we have tested the expression of natural idiotope and antigenic specificities of pentameric and hexameric IgM polymers originating from sera of patients with Waldenstrom's macroglobulinemia, as well as patients suffering from recurrent urinary bacterial infections. We demonstrate that although pentameric IgM polymers can exist as natural and immune antibodies, IgM hexamers are exclusively immune and do not exist as NAbs.
PB  - Humana Press Inc, Totowa
T2  - Immunologic Research
T1  - Antigenic specificity and expression of a natural idiotope on human pentameric and hexameric IgM polymers
EP  - 107
IS  - 1
SP  - 97
VL  - 51
DO  - 10.1007/s12026-011-8236-8
ER  - 

@article{
author = "Petrušić, Vladimir and Živković, Irena and Stojanović, Marijana and Lukić, Ivana and Marinković, Emilija and Inić-Kanada, Aleksandra and Dimitrijevic, Ljiliana",
year = "2011",
abstract = "Natural antibodies (NAbs) are present in circulation even before the exposure to antigen and they exert various biological functions. They are polyreactive and mainly represented by immunoglobulin M (IgM), which is the first antibody produced in an ongoing immune response to infection and/or immunization. IgM is always secreted as a polymer with predominant pentameric structure, although other polymeric forms such as hexamer can be also formed. The biological functions of hexameric IgM are still not known and it is proposed that its existence as a NAb could be deleterious. However, the nature of IgM hexamers has not been investigated yet. In this paper, we have tested the expression of natural idiotope and antigenic specificities of pentameric and hexameric IgM polymers originating from sera of patients with Waldenstrom's macroglobulinemia, as well as patients suffering from recurrent urinary bacterial infections. We demonstrate that although pentameric IgM polymers can exist as natural and immune antibodies, IgM hexamers are exclusively immune and do not exist as NAbs.",
publisher = "Humana Press Inc, Totowa",
journal = "Immunologic Research",
title = "Antigenic specificity and expression of a natural idiotope on human pentameric and hexameric IgM polymers",
pages = "107-97",
number = "1",
volume = "51",
doi = "10.1007/s12026-011-8236-8"
}

Petrušić, V., Živković, I., Stojanović, M., Lukić, I., Marinković, E., Inić-Kanada, A.,& Dimitrijevic, L.. (2011). Antigenic specificity and expression of a natural idiotope on human pentameric and hexameric IgM polymers. in Immunologic Research
Humana Press Inc, Totowa., 51(1), 97-107.
https://doi.org/10.1007/s12026-011-8236-8

Petrušić V, Živković I, Stojanović M, Lukić I, Marinković E, Inić-Kanada A, Dimitrijevic L. Antigenic specificity and expression of a natural idiotope on human pentameric and hexameric IgM polymers. in Immunologic Research. 2011;51(1):97-107.
doi:10.1007/s12026-011-8236-8 .

Petrušić, Vladimir, Živković, Irena, Stojanović, Marijana, Lukić, Ivana, Marinković, Emilija, Inić-Kanada, Aleksandra, Dimitrijevic, Ljiliana, "Antigenic specificity and expression of a natural idiotope on human pentameric and hexameric IgM polymers" in Immunologic Research, 51, no. 1 (2011):97-107,
https://doi.org/10.1007/s12026-011-8236-8 . .

TY  - JOUR
AU  - Petrušić, Vladimir
AU  - Živković, Irena
AU  - Stojanović, Marijana
AU  - Lukić, Ivana
AU  - Marinković, Emilija
AU  - Dimitrijević, Ljiljana
PY  - 2011
UR  - http://intor.torlakinstitut.com/handle/123456789/334
AB  - Immunoglobulin M (IgM) is the first antibody produced upon infection, and is often suggested as the first line of defense of human immune system. In addition to being present on the surface of naive B cells as a monomeric molecule, IgM is always secreted as a polymer. The most abundant IgM polymer in humans is pentamer, composed of five monomeric units, joined together by so-called joining on chain. On the other hand, it is well known that hexameric IgM can be also found in human sera. Its presence is often related to different dissorders (Waldenstrom's macroglobulinemia, cold agglutinin, and recurrent urinary bacterial infections), although it is believed that small amounts of hexamer are present in normal human sera as well. Unlike pentamer, IgM hexamer contains six monomeric blocks and completely lacks J chain. Although it has been decades since its discovery, the precise function of IgM hexamer is still unknown. Since it was documented that hexamer is very potent in activating complement, it is suggested that its production in humans must be under strict control, and that it is produced in special conditions, when strong activation of complement is absolutely needed. However, the question is whether hexameric IgM is really a secret weapon or just an undesirable molecule in humans. According to structural and known functional characteristics of both pentamers and hexamers of IgM, it can be concluded that hexamers are, in addition to being maybe too reactive to be around, probably not that efficient in protecting us from bacterial and viral infections. (C) 2011 Elsevier Ltd. All rights reserved.
PB  - Churchill Livingstone, Edinburgh
T2  - Medical Hypotheses
T1  - Hexameric immunoglobulin M in humans: Desired or unwanted?
EP  - 961
IS  - 6
SP  - 959
VL  - 77
DO  - 10.1016/j.mehy.2011.08.018
ER  - 

@article{
author = "Petrušić, Vladimir and Živković, Irena and Stojanović, Marijana and Lukić, Ivana and Marinković, Emilija and Dimitrijević, Ljiljana",
year = "2011",
abstract = "Immunoglobulin M (IgM) is the first antibody produced upon infection, and is often suggested as the first line of defense of human immune system. In addition to being present on the surface of naive B cells as a monomeric molecule, IgM is always secreted as a polymer. The most abundant IgM polymer in humans is pentamer, composed of five monomeric units, joined together by so-called joining on chain. On the other hand, it is well known that hexameric IgM can be also found in human sera. Its presence is often related to different dissorders (Waldenstrom's macroglobulinemia, cold agglutinin, and recurrent urinary bacterial infections), although it is believed that small amounts of hexamer are present in normal human sera as well. Unlike pentamer, IgM hexamer contains six monomeric blocks and completely lacks J chain. Although it has been decades since its discovery, the precise function of IgM hexamer is still unknown. Since it was documented that hexamer is very potent in activating complement, it is suggested that its production in humans must be under strict control, and that it is produced in special conditions, when strong activation of complement is absolutely needed. However, the question is whether hexameric IgM is really a secret weapon or just an undesirable molecule in humans. According to structural and known functional characteristics of both pentamers and hexamers of IgM, it can be concluded that hexamers are, in addition to being maybe too reactive to be around, probably not that efficient in protecting us from bacterial and viral infections. (C) 2011 Elsevier Ltd. All rights reserved.",
publisher = "Churchill Livingstone, Edinburgh",
journal = "Medical Hypotheses",
title = "Hexameric immunoglobulin M in humans: Desired or unwanted?",
pages = "961-959",
number = "6",
volume = "77",
doi = "10.1016/j.mehy.2011.08.018"
}

Petrušić, V., Živković, I., Stojanović, M., Lukić, I., Marinković, E.,& Dimitrijević, L.. (2011). Hexameric immunoglobulin M in humans: Desired or unwanted?. in Medical Hypotheses
Churchill Livingstone, Edinburgh., 77(6), 959-961.
https://doi.org/10.1016/j.mehy.2011.08.018

Petrušić V, Živković I, Stojanović M, Lukić I, Marinković E, Dimitrijević L. Hexameric immunoglobulin M in humans: Desired or unwanted?. in Medical Hypotheses. 2011;77(6):959-961.
doi:10.1016/j.mehy.2011.08.018 .

Petrušić, Vladimir, Živković, Irena, Stojanović, Marijana, Lukić, Ivana, Marinković, Emilija, Dimitrijević, Ljiljana, "Hexameric immunoglobulin M in humans: Desired or unwanted?" in Medical Hypotheses, 77, no. 6 (2011):959-961,
https://doi.org/10.1016/j.mehy.2011.08.018 . .

TY  - JOUR
AU  - Lukić, Ivana
AU  - Dimitrijević, Ljiljana
AU  - Dovezenski, Nebojsa
AU  - Živković, Irena
AU  - Petrušić, Vladimir
AU  - Marinković, Emilija
AU  - Inić-Kanada, Aleksandra
AU  - Stojanović, Marijana
PY  - 2011
UR  - http://intor.torlakinstitut.com/handle/123456789/319
AB  - Given an existing demand to establish a process of tetanus vaccine production in a way that allows its complete validation and standardization, this paper focuses on tetanus toxoid purification step. More precisely, we were looking at a possibility to replace the widely used ammonium-sulphate precipitation by a chromatographic method. Based on the tetanus toxin's biochemical characteristics, we have decided to examine the possibility of tetanus toxoid purification by hydrophobic chromatography, and by chromatographic techniques based on interaction with immobilized metal ions, i.e. chelating chromatography and immobilized metal affinity chromatography. We used samples obtained from differently fragmented crude tetanus toxins by formaldehyde treatment (assigned as TTd-A and TTd-B) as starting material for tetanus toxoid purification. Obtained results imply that purification of tetanus toxoid by hydrophobic chromatography represents a good alternative to ammonium-sulphate precipitation. Tetanus toxoid preparations obtained by hydrophobic chromatography were similar to those obtained by ammonium-sulphate precipitation in respect to yield, purity and immunogenicity. In addition, their immunogenicity was similar to standard tetanus toxoid preparation (NIBSC, Potters Bar, UK). Furthermore, the characteristics of crude tetanus toxin preparations had the lowest impact on the final purification product when hydrophobic chromatography was the applied method of tetanus toxoid purification. On the other hand, purifications of tetanus toxoid by chelating chromatography or immobilized metal affinity chromatography generally resulted in a very low yield due to not satisfactory tetanus toxoid binding to the column, and immunogenicity of the obtained tetanus toxoid-containing preparations was poor. (C) 2011 Elsevier BM. All rights reserved.
PB  - Elsevier Science Bv, Amsterdam
T2  - Journal of Chromatography B-Analytical Technologies in the Biomedical and Life Sciences
T1  - Tetanus toxoid purification: Chromatographic procedures as an alternative to ammonium-sulphate precipitation
EP  - 2219
IS  - 23
SP  - 2213
VL  - 879
DO  - 10.1016/j.jchromb.2011.06.003
ER  - 

@article{
author = "Lukić, Ivana and Dimitrijević, Ljiljana and Dovezenski, Nebojsa and Živković, Irena and Petrušić, Vladimir and Marinković, Emilija and Inić-Kanada, Aleksandra and Stojanović, Marijana",
year = "2011",
abstract = "Given an existing demand to establish a process of tetanus vaccine production in a way that allows its complete validation and standardization, this paper focuses on tetanus toxoid purification step. More precisely, we were looking at a possibility to replace the widely used ammonium-sulphate precipitation by a chromatographic method. Based on the tetanus toxin's biochemical characteristics, we have decided to examine the possibility of tetanus toxoid purification by hydrophobic chromatography, and by chromatographic techniques based on interaction with immobilized metal ions, i.e. chelating chromatography and immobilized metal affinity chromatography. We used samples obtained from differently fragmented crude tetanus toxins by formaldehyde treatment (assigned as TTd-A and TTd-B) as starting material for tetanus toxoid purification. Obtained results imply that purification of tetanus toxoid by hydrophobic chromatography represents a good alternative to ammonium-sulphate precipitation. Tetanus toxoid preparations obtained by hydrophobic chromatography were similar to those obtained by ammonium-sulphate precipitation in respect to yield, purity and immunogenicity. In addition, their immunogenicity was similar to standard tetanus toxoid preparation (NIBSC, Potters Bar, UK). Furthermore, the characteristics of crude tetanus toxin preparations had the lowest impact on the final purification product when hydrophobic chromatography was the applied method of tetanus toxoid purification. On the other hand, purifications of tetanus toxoid by chelating chromatography or immobilized metal affinity chromatography generally resulted in a very low yield due to not satisfactory tetanus toxoid binding to the column, and immunogenicity of the obtained tetanus toxoid-containing preparations was poor. (C) 2011 Elsevier BM. All rights reserved.",
publisher = "Elsevier Science Bv, Amsterdam",
journal = "Journal of Chromatography B-Analytical Technologies in the Biomedical and Life Sciences",
title = "Tetanus toxoid purification: Chromatographic procedures as an alternative to ammonium-sulphate precipitation",
pages = "2219-2213",
number = "23",
volume = "879",
doi = "10.1016/j.jchromb.2011.06.003"
}

Lukić, I., Dimitrijević, L., Dovezenski, N., Živković, I., Petrušić, V., Marinković, E., Inić-Kanada, A.,& Stojanović, M.. (2011). Tetanus toxoid purification: Chromatographic procedures as an alternative to ammonium-sulphate precipitation. in Journal of Chromatography B-Analytical Technologies in the Biomedical and Life Sciences
Elsevier Science Bv, Amsterdam., 879(23), 2213-2219.
https://doi.org/10.1016/j.jchromb.2011.06.003

Lukić I, Dimitrijević L, Dovezenski N, Živković I, Petrušić V, Marinković E, Inić-Kanada A, Stojanović M. Tetanus toxoid purification: Chromatographic procedures as an alternative to ammonium-sulphate precipitation. in Journal of Chromatography B-Analytical Technologies in the Biomedical and Life Sciences. 2011;879(23):2213-2219.
doi:10.1016/j.jchromb.2011.06.003 .

Lukić, Ivana, Dimitrijević, Ljiljana, Dovezenski, Nebojsa, Živković, Irena, Petrušić, Vladimir, Marinković, Emilija, Inić-Kanada, Aleksandra, Stojanović, Marijana, "Tetanus toxoid purification: Chromatographic procedures as an alternative to ammonium-sulphate precipitation" in Journal of Chromatography B-Analytical Technologies in the Biomedical and Life Sciences, 879, no. 23 (2011):2213-2219,
https://doi.org/10.1016/j.jchromb.2011.06.003 . .