TY  - JOUR
AU  - Kuczkowska, Katarzyna
AU  - Kleiveland, Charlotte R.
AU  - Minić, Rajna
AU  - Moen, Lars F.
AU  - Overland, Lise
AU  - Tjaland, Rannei
AU  - Carlsen, Harald
AU  - Lea, Tor
AU  - Mathiesen, Geir
AU  - Eijsink, Vincent G.H.
PY  - 2017
UR  - http://intor.torlakinstitut.com/handle/123456789/492
AB  - Tuberculosis (TB) remains among the most deadly diseases in the world. The only available vaccine against tuberculosis is the bacille Calmette-Guerin (BCG) vaccine, which does not ensure full protection in adults. There is a global urgency for the development of an effective vaccine for preventing disease transmission, and it requires novel approaches. We are exploring the use of lactic acid bacteria (LAB) as a vector for antigen delivery to mucosal sites. Here, we demonstrate the successful expression and surface display of a Mycobacterium tuberculosis fusion antigen (comprising Ag85B and ESAT-6, referred to as AgE6) on Lactobacillus plantarum. The AgE6 fusion antigen was targeted to the bacterial surface using two different anchors, a lipoprotein anchor directing the protein to the cell membrane and a covalent cell wall anchor. AgE6-producing L. plantarum strains using each of the two anchors induced antigen-specific proliferative responses in lymphocytes purified from TB-positive donors. Similarly, both strains induced immune responses in mice after nasal or oral immunization. The impact of the anchoring strategies was reflected in dissimilarities in the immune responses generated by the two L. plantarum strains in vivo. The present study comprises an initial step toward the development of L. plantarum as a vector for M. tuberculosis antigen delivery. IMPORTANCE This work presents the development of Lactobacillus plantarum as a candidate mucosal vaccine against tuberculosis. Tuberculosis remains one of the top infectious diseases worldwide, and the only available vaccine, bacille Calmette-Guerin (BCG), fails to protect adults and adolescents. Direct antigen delivery to mucosal sites is a promising strategy in tuberculosis vaccine development, and lactic acid bacteria potentially provide easy, safe, and low-cost delivery vehicles for mucosal immunization. We have engineered L. plantarum strains to produce a Mycobacterium tuberculosis fusion antigen and to anchor this antigen to the bacterial cell wall or to the cell membrane. The recombinant strains elicited proliferative antigenspecific T-cell responses in white blood cells from tuberculosis-positive humans and induced specific immune responses after nasal and oral administrations in mice.
PB  - Amer Soc Microbiology, Washington
T2  - Applied and Environmental Microbiology
T1  - Immunogenic Properties of Lactobacillus plantarum Producing Surface-Displayed Mycobacterium tuberculosis Antigens
IS  - 2
VL  - 83
DO  - 10.1128/AEM.02782-16
ER  - 

@article{
author = "Kuczkowska, Katarzyna and Kleiveland, Charlotte R. and Minić, Rajna and Moen, Lars F. and Overland, Lise and Tjaland, Rannei and Carlsen, Harald and Lea, Tor and Mathiesen, Geir and Eijsink, Vincent G.H.",
year = "2017",
abstract = "Tuberculosis (TB) remains among the most deadly diseases in the world. The only available vaccine against tuberculosis is the bacille Calmette-Guerin (BCG) vaccine, which does not ensure full protection in adults. There is a global urgency for the development of an effective vaccine for preventing disease transmission, and it requires novel approaches. We are exploring the use of lactic acid bacteria (LAB) as a vector for antigen delivery to mucosal sites. Here, we demonstrate the successful expression and surface display of a Mycobacterium tuberculosis fusion antigen (comprising Ag85B and ESAT-6, referred to as AgE6) on Lactobacillus plantarum. The AgE6 fusion antigen was targeted to the bacterial surface using two different anchors, a lipoprotein anchor directing the protein to the cell membrane and a covalent cell wall anchor. AgE6-producing L. plantarum strains using each of the two anchors induced antigen-specific proliferative responses in lymphocytes purified from TB-positive donors. Similarly, both strains induced immune responses in mice after nasal or oral immunization. The impact of the anchoring strategies was reflected in dissimilarities in the immune responses generated by the two L. plantarum strains in vivo. The present study comprises an initial step toward the development of L. plantarum as a vector for M. tuberculosis antigen delivery. IMPORTANCE This work presents the development of Lactobacillus plantarum as a candidate mucosal vaccine against tuberculosis. Tuberculosis remains one of the top infectious diseases worldwide, and the only available vaccine, bacille Calmette-Guerin (BCG), fails to protect adults and adolescents. Direct antigen delivery to mucosal sites is a promising strategy in tuberculosis vaccine development, and lactic acid bacteria potentially provide easy, safe, and low-cost delivery vehicles for mucosal immunization. We have engineered L. plantarum strains to produce a Mycobacterium tuberculosis fusion antigen and to anchor this antigen to the bacterial cell wall or to the cell membrane. The recombinant strains elicited proliferative antigenspecific T-cell responses in white blood cells from tuberculosis-positive humans and induced specific immune responses after nasal and oral administrations in mice.",
publisher = "Amer Soc Microbiology, Washington",
journal = "Applied and Environmental Microbiology",
title = "Immunogenic Properties of Lactobacillus plantarum Producing Surface-Displayed Mycobacterium tuberculosis Antigens",
number = "2",
volume = "83",
doi = "10.1128/AEM.02782-16"
}

Kuczkowska, K., Kleiveland, C. R., Minić, R., Moen, L. F., Overland, L., Tjaland, R., Carlsen, H., Lea, T., Mathiesen, G.,& Eijsink, V. G.H.. (2017). Immunogenic Properties of Lactobacillus plantarum Producing Surface-Displayed Mycobacterium tuberculosis Antigens. in Applied and Environmental Microbiology
Amer Soc Microbiology, Washington., 83(2).
https://doi.org/10.1128/AEM.02782-16

Kuczkowska K, Kleiveland CR, Minić R, Moen LF, Overland L, Tjaland R, Carlsen H, Lea T, Mathiesen G, Eijsink VG. Immunogenic Properties of Lactobacillus plantarum Producing Surface-Displayed Mycobacterium tuberculosis Antigens. in Applied and Environmental Microbiology. 2017;83(2).
doi:10.1128/AEM.02782-16 .

Kuczkowska, Katarzyna, Kleiveland, Charlotte R., Minić, Rajna, Moen, Lars F., Overland, Lise, Tjaland, Rannei, Carlsen, Harald, Lea, Tor, Mathiesen, Geir, Eijsink, Vincent G.H., "Immunogenic Properties of Lactobacillus plantarum Producing Surface-Displayed Mycobacterium tuberculosis Antigens" in Applied and Environmental Microbiology, 83, no. 2 (2017),
https://doi.org/10.1128/AEM.02782-16 . .

TY  - JOUR
AU  - Ovchinnikov, Kirill V.
AU  - Kristiansen, Per E.
AU  - Uzelac, Gordana
AU  - Topisirović, Ljubiša
AU  - Kojić, Milan
AU  - Nissen-Meyer, Jon
AU  - Nes, Ingolf F.
AU  - Diep, Dzung B.
PY  - 2014
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/750
UR  - http://intor.torlakinstitut.com/handle/123456789/728
AB  - Background: The bacteriocin LsbB targets a membrane-bound zinc-dependent peptidase. Results: The structure of LsbB was resolved by NMR. The C-terminal unstructured domains of LsbB and several other related bacteriocins were responsible for receptor binding. Conclusion: A subgroup of leaderless bacteriocins has been found to share a similar mechanism in receptor recognition. Significance: The study highlights the structure-function relationship of LsbB. LsbB is a class II leaderless lactococcal bacteriocin of 30 amino acids. In the present work, the structure and function relationship of LsbB was assessed. Structure determination by NMR spectroscopy showed that LsbB has an N-terminal -helix, whereas the C-terminal of the molecule remains unstructured. To define the receptor binding domain of LsbB, a competition assay was performed in which a systematic collection of truncated peptides of various lengths covering different parts of LsbB was used to inhibit the antimicrobial activity of LsbB. The results indicate that the outmost eight-amino acid sequence at the C-terminal end is likely to contain the receptor binding domain because only truncated fragments from this region could antagonize the antimicrobial activity of LsbB. Furthermore, alanine substitution revealed that the tryptophan in position 25 (Trp(25)) is crucial for the blocking activity of the truncated peptides, as well as for the antimicrobial activity of the full-length bacteriocin. LsbB shares significant sequence homology with five other leaderless bacteriocins, especially at their C-terminal halves where all contain a conserved KXXXGXXPWE motif, suggesting that they might recognize the same receptor as LsbB. This notion was supported by the fact that truncated peptides with sequences derived from the C-terminal regions of two LsbB-related bacteriocins inhibited the activity of LsbB, in the same manner as found with the truncated version of LsbB. Taken together, these structure-function studies provide strong evidence that the receptor-binding parts of LsbB and sequence-related bacteriocins are located in their C-terminal halves.
PB  - Amer Soc Biochemistry Molecular Biology Inc, Bethesda
T2  - Journal of Biological Chemistry
T1  - Defining the Structure and Receptor Binding Domain of the Leaderless Bacteriocin LsbB
EP  - 23845
IS  - 34
SP  - 23838
VL  - 289
DO  - 10.1074/jbc.M114.579698
ER  - 

@article{
author = "Ovchinnikov, Kirill V. and Kristiansen, Per E. and Uzelac, Gordana and Topisirović, Ljubiša and Kojić, Milan and Nissen-Meyer, Jon and Nes, Ingolf F. and Diep, Dzung B.",
year = "2014",
abstract = "Background: The bacteriocin LsbB targets a membrane-bound zinc-dependent peptidase. Results: The structure of LsbB was resolved by NMR. The C-terminal unstructured domains of LsbB and several other related bacteriocins were responsible for receptor binding. Conclusion: A subgroup of leaderless bacteriocins has been found to share a similar mechanism in receptor recognition. Significance: The study highlights the structure-function relationship of LsbB. LsbB is a class II leaderless lactococcal bacteriocin of 30 amino acids. In the present work, the structure and function relationship of LsbB was assessed. Structure determination by NMR spectroscopy showed that LsbB has an N-terminal -helix, whereas the C-terminal of the molecule remains unstructured. To define the receptor binding domain of LsbB, a competition assay was performed in which a systematic collection of truncated peptides of various lengths covering different parts of LsbB was used to inhibit the antimicrobial activity of LsbB. The results indicate that the outmost eight-amino acid sequence at the C-terminal end is likely to contain the receptor binding domain because only truncated fragments from this region could antagonize the antimicrobial activity of LsbB. Furthermore, alanine substitution revealed that the tryptophan in position 25 (Trp(25)) is crucial for the blocking activity of the truncated peptides, as well as for the antimicrobial activity of the full-length bacteriocin. LsbB shares significant sequence homology with five other leaderless bacteriocins, especially at their C-terminal halves where all contain a conserved KXXXGXXPWE motif, suggesting that they might recognize the same receptor as LsbB. This notion was supported by the fact that truncated peptides with sequences derived from the C-terminal regions of two LsbB-related bacteriocins inhibited the activity of LsbB, in the same manner as found with the truncated version of LsbB. Taken together, these structure-function studies provide strong evidence that the receptor-binding parts of LsbB and sequence-related bacteriocins are located in their C-terminal halves.",
publisher = "Amer Soc Biochemistry Molecular Biology Inc, Bethesda",
journal = "Journal of Biological Chemistry",
title = "Defining the Structure and Receptor Binding Domain of the Leaderless Bacteriocin LsbB",
pages = "23845-23838",
number = "34",
volume = "289",
doi = "10.1074/jbc.M114.579698"
}

Ovchinnikov, K. V., Kristiansen, P. E., Uzelac, G., Topisirović, L., Kojić, M., Nissen-Meyer, J., Nes, I. F.,& Diep, D. B.. (2014). Defining the Structure and Receptor Binding Domain of the Leaderless Bacteriocin LsbB. in Journal of Biological Chemistry
Amer Soc Biochemistry Molecular Biology Inc, Bethesda., 289(34), 23838-23845.
https://doi.org/10.1074/jbc.M114.579698

Ovchinnikov KV, Kristiansen PE, Uzelac G, Topisirović L, Kojić M, Nissen-Meyer J, Nes IF, Diep DB. Defining the Structure and Receptor Binding Domain of the Leaderless Bacteriocin LsbB. in Journal of Biological Chemistry. 2014;289(34):23838-23845.
doi:10.1074/jbc.M114.579698 .

Ovchinnikov, Kirill V., Kristiansen, Per E., Uzelac, Gordana, Topisirović, Ljubiša, Kojić, Milan, Nissen-Meyer, Jon, Nes, Ingolf F., Diep, Dzung B., "Defining the Structure and Receptor Binding Domain of the Leaderless Bacteriocin LsbB" in Journal of Biological Chemistry, 289, no. 34 (2014):23838-23845,
https://doi.org/10.1074/jbc.M114.579698 . .