Evaluation of antibody responses to outer membrane vesicles (OMVs) and killed whole cell of Vibrio cholerae O1 El Tor in immunized mice
Background and Objectives: Cholera disease remains an important global health problem affecting 3-5 million subjects worldwide. Outer membrane vesicles (OMVs) have been found in a variety of Gram-negative bacteria and act as protective transport vesicles. The aim of this study was to evaluate Immune responses against Vibrio cholerae O1 El Tor clinical strain OMV and compare it with killed whole cell (KWC), complex of (KWC-OMV) as well as the internationally licensed oral cholera vaccine, Dukoral, in serum and intestinal secretions of mice.
Materials and Methods: OMVs were prepared by using modified detergent-centrifugation procedure from V. cholerae O1 El Tor clinical strain from 2005 outbreak. The ultrastructure and content of OMVs were investigated via the Scanning Electron Microscopy (SEM) and SDS-PAGE analysis. Three doses of oral immunization were adjusted and total IgG and IgA in serum and intestinal secretion were measured by enzyme-linked immunosorbent assay (ELISA).
Results: Extracted OMVs from the V. cholerae were spherical vesicles with a size ranging from 10 to 300 nm. OMV-immunized mice showed an increased level of total IgG and IgA both in serum and intestinal secretion when compared to the negative controls. Also, there existed a higher level of secretory IgA than the total IgG, suggesting the most of protection against V. cholerae colonization provided by sIgA.
Conclusion: Our findings revealed that oral immunization with V. cholerae OMVs might induce a long-term immunity, especially when administered in combination with KWC. This study tested the adjuvant activity of OMVs and may be useful in future nano vaccine research.
2. Sedaghat M, Rahimi F, Talebi M, Pourshafie MR. Serotyping, antibiotics susceptibility pattern and detection of hlyA gene among cholera patients in Iran. Jundishapur J Microbiol 2013; 6: 20-23.
3. Ranjbar R, Rahbar M, Naghoni A, Farshad S, Davari A, Shahcheraghi F. A cholera outbreak associated with drinking contaminated well water. Arch Iran Med 2011; 14:339-340.
4. Schild S, Nelson EJ, Andrew Camilli. Immunization with Vibrio cholerae outer membrane vesicles induces protective immunity in mice. Infect Immun 2008; 76: 4554-4563.
5. Levine MM, Black RE, Clements ML, Cisneros L, Nalin DR, Young CR. Duration of infection-derived immunity to cholera. J Infect Dis 1981; 143:818-820.
6. Kang SS, Yang JS, Kim KW, Yun CH, Holmgren J, Czerkinsky C. Anti-bacterial and anti-toxic immunity induced by a killed whole-cell-cholera toxin B subunit cholera vaccine is essential for protection against lethal bacterial infection in mouse pulmonary cholera model. Mucosal Immunol 2013; 6: 826-837.
7. Xu G, Wang S, Zhuang L, Hackett A, Gu L, Zhang L, et al. Intramuscular delivery of a cholera DNA vaccine primes both systemic and mucosal protective antibody responses against cholera. Vaccine 2009; 27:3821-3830.
8. Chatterjee SN, Chaudhuri K. Outer Membrane Vesicles of Bacteria. Springer Briefs in Microbiology. e book,2012.
9. Roy N, Barman S, Ghosh A, Pal A, Chakraborty K. Immunogenicity and protective efficacy of Vibrio cholerae outer membrane vesicles in rabbit model. FEMS Immunol Med Microbiol 2010; 60: 18-27.
10. Pourshafie MR, Bakhshi B, Ranjbar R, Sedaghat M, Sadeghifard N. Dissemination of a single Vibrio cholerae clone in cholera outbreaks during 2005 in Iran. J Med Microbiol 2007; 56: 1615-1619.
11. Siadat SD, Behzadiyannejad Q, Tabaraie B, Ahmadi H, Norouzian D, Pirayeeh SN. Evaluation of serum bactericidal activity specific for Neisseria meningitidis serogroup A and B: effect of immunization with Neisseria meningitidis serogroup A polysaccharide and serogroup B outer membrane vesicle conjugate as a bivalent meningococcus vac. Res J Microbiol 2007;2:436-444.
12. Claassen I, Meylis J, leyf PVD, Peetersf C, Brons H, Robert J, et al. Production, characterization and control of a Neisseria meningitides hexavalent class 1 outer membrane protein containing vesicle vaccine. Vaccine 1996; 14: 1001-1008.
13. Lebens M, Karlsson SL, Källgård S, Blomquist M, Ekman A, Erik Nygren. Construction of novel vaccine strains of Vibrio cholerae co-expressing the Inaba and Ogawa serotype antigens. Vaccine 2011;29:7505-7513.
14. Chatterjee D, Chaudhuri K. Association of cholera toxin with outer membrane vesicles which are internalized by human intestinal epithelial cells. FEBS Lett 2011; 585:1357-1362.
15. Beniac DR, Siemens CG, Wright CJ, Booth TF. A filtration based technique for simultaneous SEM and TEM sample preparation for the rapid detection of pathogens. Viruses 2014; 6: 3458-3471.
16. Elson CO, Ealding W, Lefkowitz J. A lavage technique allowing repeated measurement of IgA antibody in mouse intestinal secretions. J Immunol Methods 1984; 67:101-108.
17. Sinha R, Koley H, Nag D, Mitra S, Asish K, Chattopadhyay MB. Pentavalent outer membrane vesicles of Vibrio cholerae induce adaptive, immune response and protective efficacy in both adult and passive suckling mice models. Microbes Infect 2015; 17: 215-227.
18. Ali M, Nelson AR, Lopez AL, Sack DA. Updated global burden of cholera in endemic countries. PLoS Negl Trop Dis 2015; 9(6): e0003832.
19. Ellis TN, Kuehn MJ. Virulence and immunomodulatory roles of bacterial outer membrane vesicles. Microbiol Mol Biol Rev 2010; 74: 81-94.
20. Bjune GEA, Hoiby JK, Gronnesby O, Arnesen JH, Fredriksen A, Halstensen E, et al. Effect of outer membrane vesicle vaccine against group B meningococcal disease in Norway. Lancet 1991; 338: 1093-1096.
21. WHO. Oral cholera vaccine use in complex emergencies: World Health Organization, Geneva 2005.
22. Boustanshenas M, Bakhshi B, Ghorbani M. Investigation into immunological responses against a native recombinant CTB whole-cell Vibrio cholerae vaccine in a rabbit model. J Appl Microbiol 2013; 114:509-515.
23. Nagaputra JC, Rollier CS, Sadarangani M. Neisseria meningitidis native outer membrane vesicles containing different lipopolysaccharide glycoforms as adjuvants for meningococcal and nonmeningococcal antigens. Clin Vaccine Immunol 2014; 21: 234-242.
24. Schild S, Nelson E J, Bishop AL, Camilli A. Characterization of Vibrio cholerae outer membrane vesicles as a candidate vaccine for cholera. Infect Immun 2009;77:472-484.
25. Sliva MJB, Carneiro MBH, Pultz BA, Silva DP, Moura Lopes ME, Liliane Martins dos Santos. The Multifaceted role of commensal microbiota in homeostasis and gastrointestinal diseases. J Immunol Res 2015; 2015:321241.