CHIMERIC PROTEIN HBcAg CONTAINING THE EPITOPE MIMETIC RECOGNIZED BY MONOCLONAL ANTIBODY VRC01

DOI: https://doi.org/10.29296/25877313-2018-04-08
Issue: 
4
Year: 
2018

A.P. Rudometov Post-graduate Student, State Research Center of Virology and Biotechnology «Vector» Rospotrebnadzor (Koltsovo, Novosibirsk) E-mail: rudometov_ap@vector.nsc.ru A.N. Chikaev Ph.D. (Biol.), Institute of Molecular and Cellular Biology of the Siberian Branch of the Russian Academy of Sciences (Novosibirsk) N.B. Andreeva Post-graduate Student, State Research Center of Virology and Biotechnology «Vector» Rospotrebnadzor (Koltsovo, Novosibirsk) N.S. Shcherbakova Research Scientist, State Research Center of Virology and Biotechnology «Vector» Rospotrebnadzor (Koltsovo, Novosibirsk) L.R. Lebedev Dr.Sc. (Med.), State Research Center of Virology and Biotechnology «Vector» Rospotrebnadzor (Koltsovo, Novosibirsk) O.N. Kaplina Senior Research Scientist, State Research Center of Virology and Biotechnology «Vector» Rospotrebnadzor (Koltsovo, Novosibirsk) A.A. Ilyichev Dr.Sc. (Biol.), State Research Center of Virology and Biotechnology «Vector» Rospotrebnadzor (Koltsovo, Novosibirsk) L.I. Karpenko Dr.Sc. (Biol.), State Research Center of Virology and Biotechnology «Vector» Rospotrebnadzor (Koltsovo, Novosibirsk)

The HIV-1 epidemic is one of the most acute problems of the modern healthcare. The discovery of broadly neutralizing antibodies (bnAbs) which possesses neutralizing activity against most primary isolates of HIV-1, has added an optimism to the vaccine researchers. According to several authors, the design of an immunogen capable of inducing such antibodies can provide protection against the wide genetic diversity of HIV-1 and thus contribute to the development of an effective vaccine against this virus. The aim of this work was the construction of an HIV-immunogen capable of inducing virus-neutralizing antibodies having the properties of bnAbs VRC01. This unique monoclonal antibody can neutralize about 91% of the different HIV-1 isolates. HBcAg is one of the promising systems for the presentation of foreign epitopes (peptides) for the development of highly immunogenic vaccines. In this work, HBcAg was used as a carrier of the SWTLLGY peptide (an epitope mimic recognized by the monoclonal antibody VRC01, previously selected using a phage display library). The recombinant plasmid pUC/HBcAg-mimicVRC01 was obtained; it contained the sequence HBcAg, with the inserted sequence encoding the SWTLLGY peptide in the region of the main antigenic determinant of the core. BALB/c mice were immunized with purified protein preparations. Using the ELISA it was shown that sera from immunized animals contained antibodies that specifically recognized the recombinant protein HBcAg-mimicVRC01. IgG fractions were isolated to assess the viral neutralizing activity from the sera of immunized animals. Neutralizing activity of IgGs was analyzed using the recombinant HIV-1 virus 92BR025. The results of neutralizing analysis showed that IgGs from sera of animals, immunized with HBcAg-mimicVRC01 protein, can neutralize the recombinant 92BR025 virus.

Keywords: 
HIV-1
HIV-immunogens
broadly neutralizing antibodies
epitope mimetic
chimeric HBcAg

References: 
  1. Haynes B.F. New approaches to HIV vaccine development // Current Opinion in Immunology. 2015. № 35. R. 39−47.
  2. Rerks-Ngarm S., Pitisuttithum P., Nitayaphan S., et al. Vaccination with ALVAC and AIDSVAX to Prevent HIV-1 Infection in Thailand // New England Journal of Medicine. 2009. № 361. R. 2209−2220.
  3. Shcherbakov D.N., Bakulina A.Y., Karpenko L.I., Ilyichev A.A. Broadly Neutralizing Antibodies against HIV-1 As a Novel Aspect of the Immune Response // Acta Naturae. 2015. V. 7. № 4. P. 11-21.
  4. Karpenko L.I., Scherbakova N.S., Chikaev A.N., et al. Polyepitope protein incorporated the HIV-1 mimotope recognized by monoclonal antibody 2G12 // Molecular Immunology. 2012. № 50. R. 193–199.
  5. Shcherbakova N.S., Shcherbakov D.N., Bakulina A.Y., et al. Artificial polyepitope HIV-1 immunogen containing mimotope of 2F5 epitope // Protein & Peptides Letters. 2016. № 23. R. 159–168.
  6. Veremejko T.A., Lebedev L.R., Chikaev N.A. i dr. Gumoral'nyj immunnyj otvet u myshej linii BALB/c, immunizirovannyh himernymi belkami HBcAg, nesuschimiepitopy poverhnostnogo belka virusa gepatita V //Voprosy virusologii. 2007. № 1. S. 40–45.
  7. Medina-Ramírez M., Sa´nchez-Merino V., Sa´nchez-Palomino S.,et al. Broadly Cross-Neutralizing Antibodies in HIV-1 Patients with Undetectable Viremia // Journal of virology. 2011. № 85. R. 5804–5813.
  8. Zhou T., Georgiev I., Wu X., et al. Structural basis for broad and potent neutralization of HIV-1 by antibody VRC01 // Science. 2010. № 329. R. 811–817.
  9. Chikaev A.N., Bakulina A.Y., Burdicket R.C., et al. Selection of peptide mimics of HIV-1 epitope recognized by neutralizing antibody VRC01 // PloS One. 2015. № 10. R. 1–13.
  10. Pumpens P., Grens E. The true story and advantages of the famous Hepatitis B virus core particles: Outlook 2016 // Molecular Biology. 2016. № 50. R. 489–509.