Immunogenicity of recombinant Omp16 protein of Pasteurella multocida B:2 in mouse model

S B SHIVACHANDRA; A KUMAR; N N MOHANTY; R YOGISHARADHYA

doi:10.56093/ijans.v87i1.66834

Authors

S B SHIVACHANDRA ICAR-Indian Veterinary Research Institute, Mukteswar, Uttarakhand 263 138 India
A KUMAR ICAR-Indian Veterinary Research Institute, Mukteswar, Uttarakhand 263 138 India
N N MOHANTY ICAR-Indian Veterinary Research Institute, Mukteswar, Uttarakhand 263 138 India
R YOGISHARADHYA ICAR-Indian Veterinary Research Institute, Mukteswar, Uttarakhand 263 138 India

https://doi.org/10.56093/ijans.v87i1.66834

Keywords:

Immunogenicity, Mouse model, Pasteurella multocida, Peptidoglycan-associated lipoprotein (PAL), Recombinant Omp16

Abstract

Bacterial peptidoglycan-associated lipoproteins (PAL) are potential targets for the development of diagnostics/subunit vaccines for infectious diseases. Most commonly prevalent Omp16 lipoprotein is absolutely conserved among Pasteurella multocida strains, which are involved in multiple infectious diseases of livestock worldwide. In the present study, we cloned omp16 gene encoding for mature Omp16 of P. multocida B:2 strain P52 and overexpressed as a fusion protein in Escherichia coli. Mice immunized with purified recombinant non-lipidated Omp16 fusion protein (~32 kDa) resulted in elicitation of significant antigen specific serum antibody titres (total IgG and subtypes). A more pronounced increase in Th2 response (IgG1) was noticed. The study indicated the potential possibilities to use lipidated recombinant Omp16 protein in developing a composite subunit vaccine along with suitable adjuvant for haemorrhagic septicaemia/ pasteurellosis in livestock.

Downloads

Download data is not yet available.

References

Ahmad T A, Rammah S S, Sheweita S A, Haroun M and El-Sayed L H. 2014. Development of immunization trials against Pasteurella multocida. Vaccine 32: 909–17. DOI: https://doi.org/10.1016/j.vaccine.2013.11.068

Biswas A, Shivachandra S B, Saxena M K, Kumar A A, Singh V P and Srivastava S K. 2004. Molecular variability among strains of Pasteurella multocida isolated from an outbreak of haemorrhagic septicaemia in India. Veterinary Research Communications 28: 287–98. DOI: https://doi.org/10.1023/B:VERC.0000026656.77847.5b

Cox J C and Coulter A R. 1997. Adjuvants-a classification and review of their modes of action. Vaccine 15: 248–56. DOI: https://doi.org/10.1016/S0264-410X(96)00183-1

Godlewska R, Wisniewska K, Pietras Z and Jagusztyn-Krynicka E K. 2009. Peptidoglycan-associated lipoprotein (Pal) of Gram-negative bacteria: function, structure, role in pathogenesis and potential application in immunoprophylaxis. FEMS Microbiology Letters 298: 1–11. DOI: https://doi.org/10.1111/j.1574-6968.2009.01659.x

Goswami P P, Chaudhuri P, Tiwari V, Parihar N S and Harbola P C. 2004. Cloning and sequencing of a 16 kDa outer membrane protein gene of Pasteurella multocida P52. Veterinary Research Communications 28: 17–25. DOI: https://doi.org/10.1023/B:VERC.0000009529.26278.21

Hatfaludi T, Al-Hasani K, Boyce J D and Adler B. 2010. Outer membrane proteins of Pasteurella multocida. Veterinary Microbiology 144: 1–17. DOI: https://doi.org/10.1016/j.vetmic.2010.01.027

Kasten R W, Hansen L M, Hinojoza J, Bieber D, Ruehl W W and Hirshi D C. 1995. Pasteurella multocida produces a protein with homology to the P6 outer membrane protein of Haemophilus influenzae. Infection and Immunity 63: 989–93. DOI: https://doi.org/10.1128/iai.63.3.989-993.1995

Kasten R W, Wakenell P S, Ahmad S, Yilma T D and Hirsh D C. 1997. Lack of protection against avian cholera by vaccination with recombinant P6-like protein from Pasteurella multocida. Avian Diseases 41: 972–76. DOI: https://doi.org/10.2307/1592355

Kovacs-Simon A, Titball R W and Michell S L. 2011. Lipoproteins of bacterial pathogens. Infection and Immunity 79: 548–61. DOI: https://doi.org/10.1128/IAI.00682-10

Kumar A A, Shivachandra S B, Biswas A, Singh V P, Singh V P and Srivastava S K. 2004. Prevalent serotypes of Pasteurella multocida isolated from different animal and avian species in India. Veterinary Research Communications 28: 657–67. DOI: https://doi.org/10.1023/B:VERC.0000045959.36513.e9

Kumar A, Mohanty N N, Yogisharadhya R, Chacko N and Shivachandra S B. 2014. Structural features of a highly conserved Omp16 protein of Pasteurella multocida and comparison with related peptidoglycans-associated lipoproteins (PAL). Indian Journal of Microbiology 55(1): 50– 56. DOI: https://doi.org/10.1007/s12088-014-0489-1

Kumar A, Yogisharadhya R, Ramakrishnan M A, Viswas K N and Shivachandra S B. 2013. Structural analysis and cross- protective efficacy of recombinant 87 kDa outer membrane protein (Omp87) of Pasteurella multocida serogroup B:2. Microbial Pathogenesis 65: 48–56. DOI: https://doi.org/10.1016/j.micpath.2013.09.007

Lo M, Boyce J D, Wilkie I W and Adler B. 2004. Characterization of two lipoproteins in Pasteurella multocida. Microbiology and Infection 6: 58–67. DOI: https://doi.org/10.1016/j.micinf.2003.09.023

Lugade AA, Bianchi-Smiraglia A, Pradhan V, Elkin G, Murphy T F and Thanavala Y. 2011. Lipid motif of a bacterial antigen mediates immune responses via TLR2 signaling. PLoS One 6: e19781. DOI: https://doi.org/10.1371/journal.pone.0019781

Shivachandra S B, Kumar A A, Gautam R, Singh V P, Saxena M K and Srivastava S K. 2006. Identification of avian strains of Pasteurella multocida in India by conventional and PCR assays. Veterinary Journal 172: 561–64. DOI: https://doi.org/10.1016/j.tvjl.2005.05.007

Shivachandra S B, Viswas K N and Kumar A A. 2011. A review of haemorrhagic septicaemia in cattle and buffalo. Animal Health Research Reviews 12: 67–82. DOI: https://doi.org/10.1017/S146625231100003X

Shivachandra S B, Yogisharadhya R, Ahuja A and Bhanuprakash V. 2012. Expression and purification of recombinant type IV fimbrial subunit protein of Pasteurella multocida serogroup B:2 in Escherichia coli. Research in Veterinary Science 93: 1128–31. DOI: https://doi.org/10.1016/j.rvsc.2012.02.010

Shivachandra S B, Kumar A, Yogisharadhya R, Ramakrishnan M A and Viswas K N. 2013. Carboxyl terminus heterogeneity of type IV fimbrial subunit protein of Pasteurella multocida isolates. Veterinary Research Communications 37: 269–75. DOI: https://doi.org/10.1007/s11259-013-9569-7

Shivachandra S B, Kumar A, Mohanty N N, Yogisharadhya R, Chacko N, Viswas K N and Ramakrishnan M A. 2014a. Homogeneity of VacJ outer membrane lipoproteins among Pasteurella multocida strains and heterogeneity among members of Pasteurellaceae. Research in Veterinary Science 96: 415–21. DOI: https://doi.org/10.1016/j.rvsc.2014.03.016

Shivachandra S B, Kumar A, Yogisharadhya R and Viswas K N. 2014b. Immunogenicity of highly conserved recombinant VacJ outer membrane lipoprotein of Pasteurella multocida. Vaccine 32: 290–96. DOI: https://doi.org/10.1016/j.vaccine.2013.10.075

Shivachandra S B, Yogisharadhya R, Kumar A, Mohanty N N and Nagaleekar V K. 2015. Recombinant transferrin binding protein A (TbpA) fragments of Pasteurella multocida serogroup B:2 provide variable protection following homologous challenge in mouse model. Research in Veterinary Science 98 (1): 1–6. DOI: https://doi.org/10.1016/j.rvsc.2014.11.013

Wheeler R. 2009. Outer membrane proteomics of Pasteurella multocida isolates to identify putative host-specificity determinants. Bioscience Horizons 2: 1–12. DOI: https://doi.org/10.1093/biohorizons/hzp002

Wilkie I W, Harper M, Boyce J D and Adler B. 2012. Pasteurella multocida: diseases and pathogenesis. Current Topics in Microbiology and Immunology 361: 1–22. DOI: https://doi.org/10.1007/82_2012_216