THE IMMUNOCOMPETENT CELLS RECEPTORS RESEARCH UNDER EXPERIMENTAL INFLUENZA INFECTION IN VITRO

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Abstract

Introduction. It is known that interferon is a cytokine and is a substantial part of the immune system necessary for antigenic challenge immune response full expression. Also it is considered that every antigen is an interferon inducer. Interferon induces antivirus response via binding to specific receptors, this receptors can be revealed straight on cell membranes of immune cells. Research objective. To evaluate the interferon inducer ability of some Influenza A virus strains upon indications of receptors functional activity (capacity) to alpha and gamma interferons on peripheral mononuclear blood cells (PBMC) induced in vitro by different Influenza A virus strains. Material and methods. The method is based on lymphocytes separation from the venous heparinized blood, with followed by in vitro lymphocytes inducing at temperature 36.5°С in the presence of 5% CO2. Blood samples were taken in different time intervals, labelled by mouse anti-idiotipyc FITCconjugated antibodies, structurally simulated human alpha and gamma interferon, samples were fixed with paraformaldehyde. Interferon receptors expression were performed by flow cytometer. Results. The in vitro experiments have determined the interferon-inducing ability of three influenza virus strains: A/PR8/34 (H1N1), A/Krasnodar/101/59 (H2N2) and A/ Ryazan/6103/87 (H3N2). MPBC blood sample (blood group was 0, Rh factor – positive) was induced by irradiated noninfectious allantoic fluid with hemagglutinating activity. Expression of alpha and gamma interferon receptors (alpha and gamma IFNR) on MPBC was determined by IFNR markers labelled with FITC and it (expression) was estimated by flow cytometer. In parallel we compared expression of alpha and gamma IFNR on MPBC in primed and non primed cells by low doses of human alpha interferon. It was found that expression of alpha and gamma IFNR on MPBC, induced influenza A/ PR8/34 (H1N1) antigen, with high hemagglutinating activity was higher in primed MPBC in comparison with non primed and higher then expression was induced by influenza virus A/Krasnodar/101/59 (H2N2) and A/Ryazan/6103/87 (H3N2) with lower hemagglutinating activity. It should be noted that IFN alpha receptor (IFNAR) expression on induced by influenza virus strain A/PR8/34 (H1N1) and primed by low doses of alpha interferon, repose on high level from induction point (1 hour) and protract during high level during 5 hours. Evaluation of gamma IFNR (IFNGR) expression level on MPBC induced by different influenza virus strains testify that firstly up-regulation IFNGR expression on MPBC primed by low doses of alpha interferon is absent and secondly up-regulation IFNGR on MPBC bear no relation with hemagglutinating activity of Influenza virus antigen. Conclusion. Experiment results clearly suggest that all influenza strains used carry the interferon induced ability which is possible to see by expression of IFNAR and IFNGR on MPBC induced by described above virus antigens. Rate of interferon induce ability in different influenza virus A connected on one side with virus hemagglutinating activity level in estimating IFNAR and on the other side with virus strains in estimating IFNGR. 

About the authors

A. N. Lisakov

Research Institute Vaccine and Serum named after I.I. Mechnikov, Moscow, Russian Federation

Email: fake@neicon.ru
Junior Researcher, Laboratory of Hybrid Cell Cultures, Department of Virology, Mechnikov Research Institute of Vaccines and Sera, Moscow, Russian Federation; Russian Federation

F. G. Nagieva

Research Institute Vaccine and Serum named after I.I. Mechnikov, Moscow, Russian Federation

Author for correspondence.
Email: fgn42@yandex.ru

PhD, MD (Medicine), Associate Professor, Head of the Laboratory of Hybrid Cell Cultures, Department of Virology, Mechnikov Research Institute of Vaccines and Sera, Moscow, Russian Federation; 

Russian Federation

E. P. Barkova

Research Institute Vaccine and Serum named after I.I. Mechnikov, Moscow, Russian Federation

Email: fake@neicon.ru

PhD (Biology), Leading Researcher, Laboratory of Hybrid Cell Cultures, Department of Virology, Mechnikov Research Institute of Vaccines and Sera, Moscow, Russian Federation; 

Russian Federation

L. A. Gaiderova

Research Center for Expertise of Means Medical Application, Moscow, Russian Federation

Email: fake@neicon.ru

PhD (Medicine), Head of the Laboratory of Immunology, Scientific Center for Expertise of Means Medical Application, Ministry of Health Care, Russian Federation, Moscow, Russian Federation;

Russian Federation

A. Y. Fedotov

Research Institute Vaccine and Serum named after I.I. Mechnikov, Moscow, Russian Federation

Email: fake@neicon.ru

Junior Researcher, Laboratory of Hybrid Cell Cultures, Department of Virology, Mechnikov Research Institute of Vaccines and Sera, Moscow, Russian Federation;

Russian Federation

V. G. Nikulina

Research Institute Vaccine and Serum named after I.I. Mechnikov, Moscow, Russian Federation

Email: fake@neicon.ru

PhD (Medicine), Associate Professor, Leading Researcher, Laboratory of Hybrid Cell Cultures, Department of Virology, Mechnikov Research Institute of Vaccines and Sera, Moscow, Russian Federation;

Russian Federation

M. V. Gavrilova

Research Institute Vaccine and Serum named after I.I. Mechnikov, Moscow, Russian Federation

Email: fake@neicon.ru

Researcher, Laboratory of Synthesis of Immunoglobulins, Mechnikov Research Institute of Vaccines and Sera, Moscow, Russian Federation. 

Russian Federation

References

  1. Баркова Е.П., Вдовина Е.Т., Нагиева Ф.Г., Ющук Н.В., Знойко О.О., Никулина В.Г., Воробьев А.С. Функциональная активность интерфероновых рецепторов мононуклеаров периферической крови пациентов с вирусными гепатитами // Биопрепараты. 2001. № 4. С. 18–21. [Barkova E.P., Vdovina E.T., Nagieva F.G., Ushuk N.V., Znoiko O.O., Nikulina V.G., Vorobev A.S. Functional activity (capacity) interferon receptors peripheral blood mononuclear cells of patients with viral hepatitis. Biopreparaty = Biopharmaceuticals, 2001, no. 4, pp. 18–21. (In Russ.)]
  2. Баркова Е.П., Нагиева Ф.Г., Кузнецов В.П., Беляев Д.Л., Никулина В.Г., Бабаянц А.А., Крель П.Е., Анджапаридзе О.Г. Экспрессия рецепторов для человеческих интерферонов альфа и гамма на поверхности мононуклеарных клеток периферической крови при некоторых вирусных инфекциях // Вопросы вирусологии. 1998. Т. 43, № 6. С. 16–18. [Barkova E.P., Nagieva F.G., Kuznetsov V.P., Belyaev D.L., Nikulina V.G., Babayans A.A., Krel P.E., Andgaparidze O.G. Expression receptors to human alpha and gamma interferon on surface of peripheral blood mononuclear cells in some virus infections. Voprosy virusologii = Problems of Virology, 1998, vol. 43, no. 6, pp. 16–18. (In Russ.)]
  3. Лимфоциты. Методы / Под. ред. Дж. Клаус. М.: Мир, 1990. 395 с. [Limfotsity. Metody. [Lymphocytes. Methods / Ed. Klaus G.]. Moscow: Mir, 1990, 395 p. (In Russ.)]
  4. Cоловьев В.Д., Бектемиров Т.А. Интерфероны в теории и практике медицины. М.: Медицина, 1981. 283 с. [Solovev V.D., Bektemirov T.A. Interferony v teorii i praktike meditsiny [Interferons in theory and practice in medicine]. Moscow: Meditsina, 1981. 283 p. (In Russ.)]
  5. Mizukoshi E., Kaneko S., Yanagi M., Ohno H., Kaji K., Terasaki S., Shimoda A., Matsushita E., Kobayashi K. Expression of interferon alpha/beta receptor in the liver of chronic hepatitis C patients. J. Med. Virol., 1998, vol. 56, no. 3, pp. 217–223.
  6. Reik L.M., Maines S.L., Ryan D.E., Levin W., Bandiera S., Thomas P.E. A simple, non-chromatographic purification procedure for monoclonal antibodies. Isolation of monoclonal antibodies against cytochrome P450 isozymes. J. Immunol. Methods, 1987, vol. 100, no. 1–2, pp. 123–130.
  7. Ren J., Liu T., Pang L., Li K., Garofalo R.P., Casola A., Bao X. A novel mechanism for the inhibition of interferon regulatory factor-3-dependent gene expression by human respiratory syncytial virus NS1 protein. J. Gen. Virol., 2011, 92 (Pt. 9), pp. 2153–2159. doi: 10.1099/vir.0.032987-0
  8. Skurkovich S.V., Eremkina E.I. The probable role of interferon in allergy. Ann. Allergy, 1975, vol. 35, no. 6, pp. 356–360.
  9. Woo H.M., Kim K.S., Lee J.M., Shim H.S., Cho S.J., Lee W.K., Ko H.W., Keum Y.S., Kim S.Y., Pathinayake P., Kim C.J., Jeong Y.J. Single-stranded DNA aptamer that specifically binds to the influenza virus NS1 protein suppresses interferonantagonism. Antiviral Res., 2013, vol. 100, no. 2, pp. 337–345. doi: 10.1016/j.antiviral.2013.09.004.

Copyright (c) 2015 Lisakov A.N., Nagieva F.G., Barkova E.P., Gaiderova L.A., Fedotov A.Y., Nikulina V.G., Gavrilova M.V.

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This work is licensed under a Creative Commons Attribution 4.0 International License.

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