Article history: Received: 16-03-2017, Accepted: 13-07-2017, Published online: 19-08-2017
Corresponding author: Mumtesh Kumar Saxena
E-mail: firstname.lastname@example.orgCitation: Saxena A, Kumar R, Saxena MK (2017) Vaccination with Salmonella Typhi recombinant outer membrane protein 28 induces humoral but non-protective immune response in rabbit, Veterinary World, 10(8): 946-949.
Aim: Typhoid is one of the most important food and water borne disease causing millions of deaths over the world. Presently, there is no cost effective vaccine available in India. The outer-membrane proteins (Omps) of Salmonella have been exhibited as a potential candidate for development of subunit vaccine against typhoid. The objective of the present study was to evaluate the use of recombinant Omp 28 protein for immunization of rabbit to elucidate its protection against virulent Salmonella Typhi.
Materials and Methods: Immune potential of recombinant Omp28 was tested in New Zealand Rabbits. Rabbits were divided into two groups, i.e., control and test group. Control group was injected with phosphate buffer saline with adjuvant while test group were injected with recombinant Omp28 along with adjuvant. Rabbits were bleed and serum was collected from each rabbit. Serum was tested by Enzyme-linked immunosorbent assay (ELISA) for humoral response. Rabbits were challenged with virulent culture to test the protective immunity.
Results: Humoral response was provoked at 15th day and maintained till 30th day. The mean ELISA titer at 15th day was 1 : 28000 (mean titer log 10 : 4.4472) and on the 30th day was 1 : 25866 (mean titer log 10 : 4.4127). Protective immune potential of Omp 28 was assessed by challenge studies in rabbits for which vaccinated and control rabbits were challenged with 109 cells of virulent culture of S. Typhi. In control group, out of six, no rabbit could survive after 48 days while in vaccinated group, three out of six rabbit were survived.
Conclusion: Immunization of rabbit with recombinant Omp 28 induced a strong humoral response which was exhibited by high antibody titer in ELISA. Subsequently, intraperitoneal homologous challenge of the immunized New Zealand rabbit resulted in lack of significant protection. These findings indicate that Omp 28 though provoked the humoral immunity but could not provide the protective immunity in rabbit model.
Keywords: adjuvant, outer-membrane proteins, recombinant protein, Salmonella, vaccine.
1. CDC. (2014) Typhoid Fever Surveillance and Vaccine use South East Asia and Western Pacific Regions 2009-2012. Morbidity and Mortality Weekly Report. NO. 39. p855-860. [PubMed]
2. Shivshankara, N., Saxena, M.K. and Singh V.P. (2001) Rapid diagnosis of hemorrhagicsepticemia by PCR assay. Indian Vet. J., 78: 101-103.
3. Saxena, M.K., Singh, V.P., Kumar, A.A., Choudhuri, P., Singh, V.P., Shivachandra, S.B., Biswas A. and Sharma, B. (2006) REP-PCR analysis of P. multocida isolates from wild and domestic animals in India. Vet. Res. Commun., 30: 851-861. [Crossref] [PubMed]
4. Shivachandra, S.B., Kumar, A.A., Gautam, R., Joseph, S., Chaudhuri, P., Saxena, M.K. and Srivastava, S.K. (2006) Detection of Pasteurella multocida in experimental infectedembryonated chicken eggs by PCR assay. Indian J. Exp. Biol., 44: 321-324. [PubMed]
5. Saxena, M.K., Singh, V.P., Lakhchura, B.D., Saxena, A. and Shrama, B. (2004) Detection of virulence in Indian isolates of Salmonella by polymerase chain reaction. Indian J. Biotechnol., 3: 37-40.
6. Naggapa, K., Singh, S.P., Tamuly, S. and Saxena, M.K. (2007) Isolation of Salmonella Typhimurium from poultry eggs and meat of Tarai region of Uttaranchal. Indian J. Biotechnol., 6: 407-408.
7. Rahman, B.A., Wasfy, M.O., Maksoud, M.A., Hanna, N., Dueger, E. and House, B. (2014) Multidrug resistance and reduced susceptibility to ciprofloxacin among Salmonella enterica serovar Typhi isolated from the middle east and central Asia. New Microbes New Infect., 2: 88-92. [Crossref] [PubMed] [PMC]
10. Hamid, N. and Jain, S.K. (2010) Immunogenic evaluation of a recombinant 49-kilodalton outer membrane protein of Salmonella Typhi as a candidate for a subunit vaccine against typhoid. J. Infect. Dis. Immun., 2: 30-40.
11. Yang, Y., Wan, C., Xu, H., Aquilar, Z.P., Tan, Q., Xu, F., Lai, W., Xiong, Y. and Wei, H. (2013) Identification of an outer membrane protein of Salmonella enterica serovar Typhimurium as a potential vaccine candidate for salmonellosis in mice. Microbes Infect., 15(5): 388-398. [Crossref] [PubMed]
12. Kaur, J. and Jain, S.K. (2013) High level expression of 49 kDa outer membrane protein of Salmonella enterica serovar Typhi. Ann. Biol. Res., 4(1): 107-117.
13. Andrade, D., Ferreira, A.G., Silva, D.J.D., Nascimento, H.J. and Silva, J.G. (1998) Chemical and immunological characterization of a low molecular weight outer membrane protein of Salmonella Typhi. Microbiol. Immunol., 42: 521-526. [Crossref] [PubMed]
14. Neves, F.A.G., Andrade, D., Vannier, S.M.A., Jonas, P., Hilton, J.N. and Silva, J.G. (2004) Complete amino acid sequence and location of Omp 28 an important immunogenic protein from Salmonella enterica serovar Typhi. Protein J., 23: 71-77. [Crossref]
15. Saxena, A., Tamuly, S. and Saxena, M.K. (2012) Cloning, sequencing and in silico characterization of Omp 28 gene of Salmonella Typhi (strain MTCC 733). J. Nat. Sci. Biol. Med., 3: 133-138. [Crossref] [PubMed] [PMC]
16. Pandey, M. and Saxena, M.K. (2015) Cloning and immune-potential analysis of Omp 28 of Salmonella Typhimurium for the development of subunit vaccine for poultry salmonellosis. Indian J. Poult. Sci., 50: 138-142.
17. Engvall, E. and Perlman, P. (1971) Enzyme-linked immunosorbent assay (ELISA). Quantitative assay of immunoglobulin G. Immunochemistry., 8: 871-874. [Crossref]
18. Hanes, D.E., Robr, M.G., Schneider, C.M. and Burr, D.H. (2001) New Zealand white rabbit as a nonsurgical experimental model for Salmonella Enteric gastroenteritis. Infect. Immun., 69: 6523-6526. [Crossref] [PubMed] [PMC]
19. Toobak, H., Rasooli, I., Talei, D., Jahangiri, A., Owlia, P. and Darvish, A.A.S. (2013) Immune response variation to Salmonella enteric serovar Typhi recombinant porinprotein in mice. Biologicals., 41: 224-230. [Crossref] [PubMed]
20. Moreno, E.M.A., Tenorio, C.A., Pastelin, P.R., Perez, S.C., Gill, C.C., Lopez, S.R., Baeza, I., Fernandez, M., Bonifaz, L., Isibasi, A., Calva, E. and Lopez, M.C. (2013) Salmonella Typhi Omp S1 and Omp S2 porins are potent protective immunogens with adjuvant properties. Immunology., 139: 459-471. [Crossref] [PubMed] [PMC]
21. Yang, Y., Wan, C., Xu, H. and Wei, H. (2013) Identification and characterization of Omp L as a potential vaccine candidate for immune-protection against salmonellosis in mice. Vaccine., 31(28): 2930-2936. [Crossref] [PubMed]
22. Ghosh, S., Chakraborty, K., Nagaraja, T., Basak, S., Koley, H., Dutta, S., Mitra, U. and Das, S. (2011) An adhesion protein of Salmonella enterica serovar Typhi is requitred for pathogenesis and potential target for vaccine development. Proc. Natl. Acad. Sci., 108(8): 3348-3353. [Crossref] [PubMed] [PMC]
23. Prejit ., Agarawal, R.K., Porteen, K., Dubal, Z.B., Singh, A.K. and Biswas, R. (2013) Evaluation of recombinant outer membrane protein based vaccine against Salmonella Typhimurium in birds. Biologicals., 41(3): 162-168. [Crossref] [PubMed]24. Tamuly, S. and Saxena, M.K. (2012) Preparation of calcium phosphate nano-particle sand evaluation its effect on muscle of rat. Curr. Sci., 102(4): 610-612.