Abstract
Vibrio parahaemolyticus is one of the major pathogens responsible for vibriosis and zoonotic infections in teleosts, marine invertebrates, and also humans through consumption of contaminated or unprocessed seafood. Emergence of resistance against current accessible antibiotics and spread to the food chain and environment necessitate the development of safe and effective subunit vaccine against this bacterium. Many bacteria including V. parahaemolyticus produce extracellular curli fibrils, heteropolymeric filaments of major and minor subunit, which have been implicated in adhesion, biofilm formation, and virulence. Adhesins are the primary contact points with the host which help in establishing infection and colonization. CsgA, an adhesin, is the major structural component of the curli fiber that forms homopolymers of several hundred units. Due to their exposure on the cell surface, the curli fibers are recognized by the host’s immune system, would generate high immune response, and therefore can serve as effective vaccine candidate. In the present study, we describe characterization of the csgA gene, and preparation of recombinant soluble CsgA of V. parahaemolyticus (rVpCsgA), and evaluation of its vaccine potential. Immunization of BALB/c mice with the rVpCsgA mounted a strong immune response. Cellular immune assays such as antibody isotyping, in vitro splenocyte proliferation analysis, and cytokine profiling revealed mixed T-helper cell immune response. The anti-rVpCsgA antiserum was agglutination positive and specifically cross-reacted with the curli CsgA present on the outer membrane of V. parahaemolyticus cells, thus demonstrating its neutralization potential. One hundred percent survival of the immunized mice upon challenge with the lethal dosage of the bacterium established that the rVpCsgA could serve as an effective vaccine against the bacterium.
Key points
• Recombinant histidine-tagged CsgA of V. parahaemolyticus, rVpCsgA, was purified.
• The rVpCsgA immunization produced mixed immune response and agglutinating antibodies.
• Immunization with the rVpCsgA protected mice against V. parahaemolyticus challenge.
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Data availability
The authors declare that the data supporting the findings of this study are available within the article and its supplementary information files.
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Acknowledgments
The Indian Council of Medical Research, New Delhi is acknowledged for research fellowship to SK. Prof. M. R. Chapman, Molecular, Cellular and Developmental Biology, University of Michigan, Ann Arbor, MI, USA is gratefully acknowledged for kindly providing with the E. coli C600 csg knockout strain, E. coli LSR12.
Funding
The present study has been carried out with the financial support from the Department of Science and Technology PURSE grant (SR/PURSE/Phase2/11(C) 2015) to the Jawaharlal Nehru University, New Delhi and the intramural funding from the JNU, New Delhi, India.
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AD, DC, and SK conceived and designed the study. SK performed the experiments and acquired the data. SK, DC, and AD analyzed and interpreted the data. SK, DC, and AD drafted and revised the manuscript. All authors approved the final version of the manuscript.
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The work presented in this manuscript does not contain any study comprising human participants. The use of animals (BALB/c mice) for immunization was approved by the Institutional Animal Ethics Committee (IAEC) of the University (IAEC project code 08/2017), New Delhi. The guidelines prescribed by the IAEC were followed for the use of animals.
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Karan, S., Choudhury, D. & Dixit, A. Immunogenic characterization and protective efficacy of recombinant CsgA, major subunit of curli fibers, against Vibrio parahaemolyticus. Appl Microbiol Biotechnol 105, 599–616 (2021). https://doi.org/10.1007/s00253-020-11038-4
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DOI: https://doi.org/10.1007/s00253-020-11038-4