Characterisation of M2e Antigenicity using anti-M2 Monoclonal  Antibody and anti-M2e Polyclonal Antibodies

Sumarningsih Sumarningsih; Simson Tarigan; Hemmatzadeh Farhid; Jagoda Ignjatovic

doi:10.14334/jitv.v24i3.1987

Characterisation of M2e Antigenicity using anti-M2 Monoclonal Antibody and anti-M2e Polyclonal Antibodies

Sumarningsih Sumarningsih, Simson Tarigan, Hemmatzadeh Farhid, Jagoda Ignjatovic

DOI

https://doi.org/10.14334/jitv.v24i3.1987

Abstract

Matrix 2 ectodomain (M2e) protein is a potential antigen for detection of influenza A virus infection in vaccinated poultry (DIVA test). However the M2e antigenicity and immune response it induces in either humans or animals are poorly understood. Seventeen M2e peptides and sixteen recombinant M2e (rM2e) proteins with amino acid (aa) changes introduced at position 10, 11, 12, 13 14, 16, 18 and 20 were compared by western blot (WB) and enzyme-linked immunosorbent assay (ELISA) using mouse anti-M2 monoclonal antibody (mAb) 14C2, and anti-M2e peptide chicken and rabbit polyclonal antibody (pAb). The mAb 14C had the best discriminating power and indicated that all six positions contributed to the M2e antigenicity. Position 11 was the important immunodominant and affected Mab14C binding to a greatest degree. Changes in the adjacent position 14, 16 and 18 also influenced the binding, and it detected regardless of the method (WB or ELISA), or the antigen used (M2e peptide or rM2e). For chicken pAb and rabbit pAb, the immunodominant aa was position 10 and the antibody reaction was not affected by aa change at 11. The binding of rabbit pAb was also affected by changes at 14 and 16, which confirm the contribution of these positions to the M2e antigenicity. Position 10 was the only important position for the binding of chicken pAb to M2e. Overall, the study showed that the M2e antigenic sites are located between residues 10 â€“ 18 and that aa changes at position 10, 11, 12, 14, 16 and 18 may all affect the antibody binding within the M2e protein.

Keywords

Influenza A Virus, M2e epitope, antigenicity

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References

De Filette, M., Ramne, A., Birkett, A., Lycke, N., LÃ¶wenadler, B., Min Jou, W., Saelens, X., Fiers, W. 2006. The universal influenza vaccine M2e-HBc administered intranasally in combination with the adjuvant CTA1-DD provides complete protection. Vaccine 24: 544-551.

Feng, J., Zhang, M., Mozdzanowska, K., Zharikova, D., Hoff, H., Wunner, W., Couch, R., Gerhard, W. 2006. Influenza A virus infection engenders a poor antibody response against the ectodomain of matrix protein 2. Virology Journal 3: 102.

Fiers, W., De Filette, M., Birkett, A., Neirynck, S., Min Jou, W. 2004. A "universal" human influenza A vaccine. Virus Research 103: 173-176.

Frace, A.M., Klimov, A.I., Rowe, T., Black, R.A., Katz, J.M. 1999. Modified M2 proteins produce heteropypic immunity against influenza A virus. Vaccine 17: 2237-2244.

Furuse, Y. 2009. Evolution of the M gene of the influenza A virus in different host species large scale analysis. Virology Journal 6.

Fu, T.-M., Freed, D.C., Horton, M.S., Fan, J., Citron, M.P., Joyce, J.G., Garsky, V.M., Casimiro, D.R., Zhao, Q., Shiver, J.W., Liang, X. 2009. Characterizations of four monoclonal antibodies against M2 protein ectodomain of influenza A virus. Virology 385: 218-226.

Heinen, P.P., de Boer-Luijtze, E.A., Bianchi, A.T.J. 2001. Respiratory and systemic humoral and cellular immune responses of pigs to a heterosubtypic influenza A virus infection. Journal of General Virology 82: 2697-2707.

Hemmatzadeh. F., S. Sumarningsih, S. Tarigan, R. Indriani, N.L.P.I. Dharmayanti, E. Ebrahimie, J. Ignjatovic. 2013. Recombinant M2e protein-based ELISA: a novel and inexpensive approach for differentiating avian influenza infected chickens from vaccinated ones. PloS one 8: e56801.

Ho, S.N., Hunt, H.D., Horton, R.M., Pullen, J.K., Pease, L.R.1989. Site-directed mutagenesis by overlap extension using the polymerase chain reaction. Gene 77: 51-59.

Horton, R.M., Ho, S.N., Pullen, J.K., Hunt, H.D., Cai, Z., Pease, L.R., Ray, W. 1993. Gene splicing by overlap extension, In: Methods in Enzymology. Academic Press.: 270-279.

Ito, T., Gorman, O.T., Kawaoka, Y., Bean, W.J., Webster, R.G. 1991. Evolutionary analysis of the influenza A virus M gene with comparison of the M1 and M2 proteins. Journal of Virology 65: 5491-5498.

Jackson, D.C., Tang, X.-L., Murti, K.G., Webster, R.G., Tregear, G.W., Bean, W.J. 1991. Electron microscopic evidence for the association of M2 protein with the influenza virion. Archives of Virology 118: 199-207.

KIM, M. C., J.G. Choi, J.S. Kwon, H.M. Kang, M.R. Paek, O.M. Jeong, J.H. Kwon, Y.J. Lee. 2010. Field application of the H9M2e enzyme-linked immunosorbent assay for differentiation of H9N2 avian influenza virus-infected chickens from vaccinated chickens. Clinical and vaccine immunology : CVI 17: 1977-1984.

Kitikoon, P., Strait, E.L., Thacker, E.L. 2008. The antibody responses to swine influenza virus (SIV) recombinant matrix 1 (rM1), matrix 2 (M2), and hemagglutinin (HA) proteins in pigs with different SIV exposure. Veterinary Microbiology 126: 51-62.

Lam, T.T.-Y., Hon, C.-C., Pybus, O.G., Kosakovsky Pond, S.L., Wong, R.T.-Y., Yip, C.-W., Zeng, F., Leung, F.C.-C. 2008. Evolutionary and transmission dynamics of reassortant h5n1 influenza virus in Indonesia. PLoS Pathogens 4: e1000130.

Lambrecht, B., Steensels, M., Borm, S.V., Meulemans, G., Berg, T.v.d. 2007. Development of an M2e-specific enzyme-linked immunosorbent assay for differentiating infected from vaccinated animals. Avian Diseases 51: 221-226.

Li, R., Lim, A., Ow, S.T.L., Phoon, M.C., Locht, C., Chow, V.T., Alonso, S. 2011. Development of live attenuated Bordetella pertussis strains expressing the universal influenza vaccine candidate M2e. Vaccine 29: 5502-5511.

Liu, W., Chen, Y.H. 2005. High epitope density in a single protein molecule significantly enhances antigenicity as well as immunogenecity: A novel strategy for modern vaccine development and a preliminary investigation about B cell discrimination of monomeric proteins. European Journal of Immunology 35: 505-514.

MarchÃ©, S., Lambrecht, B., Van Den Berg, T. 2010. Evaluation of different serologic markers for the early detection of avian influenza infection in chickens. Avian Diseases 54: 690-698.

Mozdzanowska, K., Feng, J., Eid, M., Kragol, G., Cudic, M., Otvos Jr, L., Gerhard, W. 2003. Induction of influenza type A virus-specific resistance by immunization of mice with a synthetic multiple antigenic peptide vaccine that contains ectodomains of matrix protein 2. Vaccine 21: 2616-2626.

Reikofski, J., Tao, B.Y. 1992. Polymerase chain reaction (PCR) techniques for site-directed mutagenesis. Biotechnology Advances 10: 535-547.

Schotsaert, M., De Filette, M., Fiers, W., Saelens, X. 2009. Universal M2 ectodomain-based influenza A vaccines: preclinical and clinical developments. Expert Review of Vaccines 8: 499-508.

Wang, R., Song, A., Levin, J., Dennis, D., Zhang, N.J., Yoshida, H., Koriazova, L., Madura, L., Shapiro, L., Matsumoto, A., Mikayama, T., Kubo, R.T., Sarawar, S., Cheroutre, H., Kato, S. 2008. Therapeutic potential of a fully human monoclonal antibody against influenza A virus M2 protein. Antiviral Research 80: 168-177.

Wang, Y., Zhou, L., Shi, H., Xu, H., Yao, H., Xi, X.G., Toyoda, T., Wang, X., Wang, T. 2009. Monoclonal antibody recognizing SLLTEVET epitope of M2 protein potently inhibited the replication of influenza A viruses in MDCK cells. Biochemical and Biophysical Research Communications 385: 118-122.

Wu, F., Huang, J.H., Yuan, X.Y., Huang, W.S., Chen, Y.H. 2007. Characterization of immunity induced by M2e of influenza virus. Vaccine 25: 8868-8873.

Xia, M., Tan, M., Wei, C., Zhong, W., Wang, L., McNeal, M., Jiang, X. 2011. A candidate dual vaccine against influenza and noroviruses. Vaccine 29: 7670-7677.

Zhang, M., Zharikova, D., Mozdzanowska, K., Otvos, L., Gerhard, W. 2006. Fine specificity and sequence of antibodies directed against the ectodomain of matrix protein 2 of influenza A virus. Molecular Immunology 43: 2195-2206.

Zebedee, S.L., Lamb, R.A. 1988. Influenza A virus M2 protein: Monoclonal antibody restriction of virus growth and detection of M2 in virions. Journal of Virology 62: 2762-2772.

Zharikova, D., Mozdzanowska, K., Feng, J.Q., Zhang, M.X., Gerhard, W. 2005. Influenza type a virus escape mutants emerge in vivo in the presence of antibodies to the ectodomain of matrix protein 2. Journal of Virology 79: 6644-6654.

Zou, P., Liu, W., Chen, Y.H. 2005. The epitope recognized by a monoclonal antibody in influenza A virus M2 protein is immunogenic and confers immune protection. International Immunopharmacology 5: 631-635.

Zou, P., Liu, W., Wu, F., Chen, Y.-H. 2008. Fine-epitope mapping of an antibody that binds the ectodomain of influenza matrix protein 2. FEMS Immunology and Medical Microbiology 53: 79-84.