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Recombinant Lactococcus Lactis Displaying Omp31 Antigen of Brucella melitensis Can Induce an Immunogenic Response in BALB/c Mice

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Abstract

Since Brucella infection mostly occurs through the mucosal surfaces, immune response induced by vaccine that is delivered by a way of mucosal route can be drastically enhanced to control the brucellosis. Omp31is the major outer membrane protein of Brucella, and is considered as a protective antigen against Brucella infection. Accordingly, Lactococcus lactis has been used as an antigen-delivering vector to develop a vaccine-induced mucosal response for having a safer vaccination against brucellosis. A designed omp31 gene fused to the usp45 signal peptide and M6 cell wall anchor was sub cloned in the pNZ7021 expression vector, and a recombinant L. lactis displaying Omp31 was constructed. Omp31 protein expression was confirmed using Western blotting and immunofluorescence analysis. Animals were orally and intraperitoneally immunized with live or killed L. lactis expressing Omp31, respectively. The humoral and cellular immune responses were evaluated by measuring the specific cytokines and antibodies. sIgA, serum IgA, IgM, and total IgG antibodies significantly increased in the mice immunized with live recombinant L. lactis expressing Omp31 and also serum IgM, and total IgG antibodies significantly increased in mice immunized with killed recombinant L. lactis expressing Omp31. Among IgG subtypes, IgG2a response was significantly higher in both groups compared to IgG1. In mice groups immunized with recombinant L. lactis, the IFN-γ and IL-10 level elevated; however, there was no change in the level of IL-4. These results indicated that recombinants L. lactis induce both humoral and cellular immune responses in mice, and also vaccines based on L. lactis-derived live carriers are promising interventions against Brucella melitensis infections.

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Acknowledgments

The present study was supported by the grant from Zanjan University of Medical Sciences (grant NO. A- 12-873-7).

The authors thank Dr. Negar Seyed, Dr.Yeganeh Talebkhan at Pasteur Institute of Iran and Dr.Narges Nazifi, Dr.Soheil Yousefi at Ferdowsi University of Mashhad for their technical guidance and constant support.

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Authors and Affiliations

  1. Department of Medical Biotechnology, Faculty of Medicine, Zanjan University of Medical Sciences, Zanjan, Iran

    Hoda Shirdast, Fatemeh Ebrahimzadeh, Amir Hossein Taromchi, Yousef Mortazavi & Keivan Nedaei

  2. Cancer Gene Therapy Research Center, Zanjan University of Medical Sciences, Zanjan, Iran

    Amir Hossein Taromchi, Yousef Mortazavi & Abdolreza Esmaeilzadeh

  3. Department of Immunology, Faculty of Medicine, Zanjan University of Medical Sciences, Zanjan, Iran

    Abdolreza Esmaeilzadeh

  4. Department of Animal Science, Ferdowsi University of Mashhad, Mashhad, Iran

    Mohammad Hadi Sekhavati

  5. Department of Molecular Medicine, Biotechnology Research Center, Pasteur Institute of Iran, Tehran, Iran

    Esmat Mirabzadeh

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  1. Hoda Shirdast
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Contributions

All authors contributed to the study conception and design. Amirhossein Taromchi and Hoda Shirdast: Material preparation; Methodology: Hoda Shirdast and Fatemeh Ebrahimzadeh; data collection and analysis: Hoda Shirdast and Amirhossein Taromchi; Rabbit immunization: Esmat Mirabzadeh; Mice immunization: Hoda Shirdast and Keivan Nedaei; Writing - original draft preparation: Hoda Shirdast; Writing - review and editing: Amirhossein Taromchi, Yousef Mortazavi, Mohammad Hadi Sekhavati and Abdolreza Esmaeilzadeh; Supervision: Amirhossein Taromchi. All authors commented on previous versions of the manuscript. All authors read and approved the final manuscript.

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Correspondence to Amir Hossein Taromchi.

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The study was approved by the Animal Experimentation Ethics Committee of Zanjan University of Medical Sciences (ZUMS.REC.1396.146).

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Shirdast, H., Ebrahimzadeh, F., Taromchi, A.H. et al. Recombinant Lactococcus Lactis Displaying Omp31 Antigen of Brucella melitensis Can Induce an Immunogenic Response in BALB/c Mice. Probiotics & Antimicro. Prot. 13, 80–89 (2021). https://doi.org/10.1007/s12602-020-09684-1

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