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Study of Antigenic properties of Human immunodeficiency Virus NEF protein Recombinant Analog

https://doi.org/10.31631/2073-3046-2016-15-3-83-93

Abstract

Protein p27/p25 is a product of HIV nef gene and is a multifunctional factor of HIV pathogenicity. Antigenic properties of recombinant NEF polypeptide which includes N-terminal fragment of HIV-1 p27 protein fused to E. coli ß-galactosidase were studied by serological and virological methods. The interaction was shown between recombinant NEF antigen and serums of HIV-positive individuals by ELISA, Western blot and line immunoassay. There was no interaction with sera of healthy individuals. The antigen specificity of recombinant antigen was shown in the reactions with commercial HIV-1 p27/p25 protein analog and with polyclonal antibodies to synthetic peptides corresponding to N-, C-terminal regions of HIV-1 p27/p25 protein. The interaction between IgG of rabbits immunized with recombinant NEF antigen and viral antigens was shown by indirect immunofluorescence and neutralizing assays. Thus it was proven the possibility of using recombinant NEF protein as an antigen for diagnostic and experimental purposes.

About the Authors

G. I. Alatortseva
Mechnikov Research Institute of Vaccines and Sera
Russian Federation


M. N. Nosik
Mechnikov Research Institute of Vaccines and Sera
Russian Federation


L. N. Nesterenko
Mechnikov Research Institute of Vaccines and Sera
Russian Federation


I. I. Amiantova
Mechnikov Research Institute of Vaccines and Sera
Russian Federation


V. V. Dotsenko
Mechnikov Research Institute of Vaccines and Sera
Russian Federation


L. N. Luhverchik
Mechnikov Research Institute of Vaccines and Sera
Russian Federation


M. V. Zhuckina
Mechnikov Research Institute of Vaccines and Sera
Russian Federation


V. U. Kabargina
Mechnikov Research Institute of Vaccines and Sera
Russian Federation


E. N. Kudryavtseva
State Budgetary Institution of Public Health of Moscow Region «M.F. Vladimirsky Moscow Regional Research Clinical Institute»
Russian Federation


V. V. Zverev
Mechnikov Research Institute of Vaccines and Sera
Russian Federation


References

1. Methodological letter No. 4174-PX dated 04.08.2006. Laboratory testing for HIV infection (including the study of immunity and viral load in HIV infection): 1 – 19 (in Russia).

2. Kim S.Y., Byrn R., Groopman J., Baltimore D. Temporal aspects of DNA and RNA synthesis during human immunodeficiency virus infection: Evidence for differential gene expression. J. Virol. 1989. 63: 3708 – 3713.

3. Pandori M.W., Fitch N.J., Craig H.M. Richman D.D., Spina C.A., Guatelli J.C.. Producer-cell modification of human immunodeficiency virus type 1: Nef is a virion protein. J. Virol. 1996. 70: 4283 – 4290.

4. Wieland U., Kühn J.E., Jassoy C., Rübsamen-Waigmann H., Wolber V., Braun R. Antibodies to recombinant HIV-1 NEF, tat, and nef proteins in human sera Medical Microbiology and Immunology. 1990. 179. 1: 1 – 11.

5. O'Neil C., Lee D., Clewley G., Johnson M.A., Emery V.C. Prevalence of anti-NEF antibodies in HIV-1 infected individuals assessed using recombinant baculovirus expressed NEF protein. J. Med. Virol. 1997: 139 – 44.

6. Frahma N., Lindea C., Brander C. Identification of HIV-Derived, HLA Class I Restricted CTL Epitopes: Insights into TCR Repertoire, CTL Escape and Viral Fitness.HIV Molecular Immunology. 2006: 3 – 28.

7. Alatortseva G.I., Nesterenko L.N., Volynskaya E.A., Amiantova I.I., Kabargina V.Y., Zukina M.V., Dotsenko V.V., Kudryavtseva E.N., Chartchenko O.S., Goltsov V.A., Zverev V.V. Production of recombinant polypeptide containing antigenic determinants of HIV-1 nef protein and its application for diagnosis of HIV infection by the line blot. Рroceedings of the XI international Congress on modern problems of аllergology, immunology and immunopharmacology. Russian Allergology J. 2011. 1 (4): 12 – 14 (in Russia).

8. Alatortsev V.E., Alatortseva G.I. Strain of bacterium Escherichia coli used for ecombinant proteins preparing. Patent RF, № 2043409; 1995 (in Russian).

9. Alatortsev V.E., Alatortseva G.I. Vector pel5а designated for foreign DNA expression. Patent RF, № 2071501; 1992 (in Russian).

10. News in DNA cloning. A practical approach. Ed. Glover D.M. Moscow, 1989: 122 – 128 (in Russian).

11. Sameh Magdeldin and Annette Moser. Affinity Cromatography: Principles and Applications, Affinity Chromatography. Ed.: S. Magdeldin. In Tech. 2012: 70 – 75.

12. Virca G.D., Travis J., Hall P.K., Roberts R.C. Purification of α-2-Macroglobulin by Chromatography on Cibacron Blue Sepharose. Analytical Biochemistry. 1978. 8: 274 – 276.

13. Rattana Wongchuphan, Beng Ti Tey, Wen Siang Tan, Senthil Kumar Subramanian, Farah Saleena Taip, Tau Chuan Ling. Purification of rabbit polyclonal immunoglobulin G using anion exchangers. Process Biochemistry 2011. 46: 101 – 107.

14. Practical Protein Chemistry. А Handbook. Ed.: А. Darbre. Moscow, 1989: 1 – 621 (in Russian).

15. Towbin H., Staehlin T., Gordon Y. Proc. nat. Acad. Sci. USA. 1979; 76: 4350 – 4359.

16. Nosik M.N., Ryzhov K.A., Kiseleva I.A., Kravtchenko A.V., Pokrovsky V.V. Biological properties of Human Immunodeficiency Virus strains isolated from HIVinfected individuals living in the territory of Russia over period of 2009-2010 year. ZniSO. 2011; 4 (217): 31 – 34 (in Russia).

17. End point neutralization assay. WHO Guidelines for Standart HIV Isolation Procedures 1994, 1998 update.

18. McCoy L.E., Weiss R.A. Neutralizing antibodies to HIV-1 induced by immunization. J. Exp. Med. 2013; 210 (2): 209 – 223.

19. Pleguezuelos O., Stoloff G.A., Caparrós-Wanderley W. Synthetic immunotherapy induces HIV virus specific Th1 cytotoxic response and death of an HIV-1 infected human cell line through classic complement activation. Virology Journal. 2013; 10: 107 – 120.

20. Chen Y.M., Lin R.H., Lee C.M., Fu C.Y., Chen S.C., Syu W.J. Decreasing levels of anti-Nef antibody correlate with increasing HIV type 1 viral loads and AIDS disease progression. AIDS Res. Hum Retroviruses. 1999; 15 (1): 43 – 50.

21. Duri K., Miller F., Gumbo F.Z., Kurewa N.E., Rusakaniko S., Chirenje M.Z. et al. Human immunodeficiency virus (HIV) types Western blot (WB) band profiles as potential surrogate markers of HIV disease progression and predictors of vertical transmission in a cohort of infected but antiretroviral therapy naïve pregnant women in Harare, Zimbabwe. BMC Infect. Dis. 2011; 11: 7.

22. Opendra Narayan, Zhenqian Liu. HIV DNA vaccine сomposition and methods of use. Patent USA. 2011. № 8,003,113 B2.


Review

For citations:


Alatortseva G.I., Nosik M.N., Nesterenko L.N., Amiantova I.I., Dotsenko V.V., Luhverchik L.N., Zhuckina M.V., Kabargina V.U., Kudryavtseva E.N., Zverev V.V. Study of Antigenic properties of Human immunodeficiency Virus NEF protein Recombinant Analog. Epidemiology and Vaccinal Prevention. 2016;15(3):83-93. (In Russ.) https://doi.org/10.31631/2073-3046-2016-15-3-83-93

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ISSN 2073-3046 (Print)
ISSN 2619-0494 (Online)