EVALUATION OF IMMUNOTOXICITY OF THE THERAPEUTIC DRUG PROLONGED ACTION FOR MULTIPLE SCLEROSIS ON RHESUS MONKEYS

This article presents the results of immunotoxicity study of a novel slow-release drug for multiple sclerosis treatment based on recombinant human interferon beta-1а. The test article is polyethylene glycol (PEG)-conjugated interferon beta-1a. Performed modification allows to improve pharmacokinetic parameters, decrease immunogenicity and elevate tolerance that significantly increases safety of the test article. The study is performed in nonhuman primates – rhesus monkeys (Macaca mulatta). The species, used in this study, is susceptible to human interferon beta-1a that has previously been shown in specific activity studies. Dynamics of peripheral blood lymphocyte subsets composition, activated lymphocyte count (based on the presence of early activation marker), serum antibodies (IgM, IgG, IgA and IgE) level and ratio were assessed within in vivo experiments. The effect of interferon beta-1a on CD69 expression was examined in mononuclear cells culture. It was shown that the test article causes changes in lymphocyte subsets ratio (decrease of NK-cells relative count with T-lymphocytes relative count elevation) in primates’ peripheral blood. Revealed changes were reversible and dose-independent. It was not shown that the test article have reliable effect on CD69 expression rate. There was no evidence of test article effect on level and ratio of serum antibodies and polymorphonucleocytes phagocytic rate in the absence of additional antigenic exposure. The results obtained during the experiment indicate the absence of pathological effect of the test article on the nonhuman primates’ immune system.

1. Беленький М.Л. Элементы количественной оценки фармакологического эффекта. Ленинград: Мед-гиз, 1963. С. 81-117. [Belenkiy M.L. Elements of quantitative assessment of pharmacological effect]. Leningrad: Medgis, 1963, pp. 81-117.

2. Бородулин В.И., Ланцман М.Н. Справочник: Болезни. Синдромы. Симптомы. М.: Оникс, 2009. C. 896. Borodulin V.I., Lanzman M.N. Reference: Diseases. Syndromes. Symptoms]. Moscow: Onyx, 2009, p. 896.

3. Куксова М.И. Кроветворная система обезьян в норме и патологии. М.: Медицина, 1972. C. 128. [Kuksova M.I. Hematopoietic system of monkeys in health and disease]. Moscow: Medicine, 1972, pp. 128.

4. Никитин И.Г., Сторожаков Т.Н. Пегилированные лекарственные препараты: современное состояние проблемы и перспективы // Информационный бюллетень «Вирусные гепатиты: достижения и перспективы», 2001. № 3 (13). [Nikitin I.G., Storozhakov T.N. PEGylated drugs: state of the art and perspectives. Information Bulletin «Viral hepatitis: Achievements and Prospects», 2001, no. 3 (13). (In Russ.)]

5. Bon A.L., Schiavoni G., Agostino G., Gresser I., Belardeli F., Tough D.F. Type I Interferons Potently Enhance Humoral Immunity and Can Promote Isotype Switching by Stimulating Dendritic Cells In Vivo. Immunity, 2001, Vol. 14, pp. 461-470.

6. Brodeur, B.R., Merigan, T.C. Suppressive effect of interferon on the humoral immune response to sheep red blood cells in mice. J. Immunol., 1974, no. 113, pp. 1319-1325.

7. Вrucе А. Clinical considerations in pegylated protein therapy. From Research to Practice, 2001, no. 3 (1), pp. 3-9.

8. Cebriаn M., Redondo J.M., Lоpez-Rivas A., Rodriguez-Tarduchy G., De Landazuri M.O., Sánchez-Madrid F. Expression and function of AIM, an activation inducer molecule of human lymphocytes, is dependent on the activation of protein kinase C. Eur. J. Immunol., 1989, May, no. 19 (5), pp. 809-815.

9. De Maeyer E., De Maeyer-Guignard J. Host genotype influences immunomodulation by interferon. Nature, 1980, no. 284, pp. 173-175.

10. Delgado С., Francis G.E., Fisher D. The uses and properties of PEG – linked proteins. Crit. Rev. Ther. Drug Carrier Syst., 1992, no. 9 (3, 4), pp. 249-304.

11. Dhib-Jalbut S., Marks S. Interferon-β mechanisms of action in multiple sclerosis. Neurology, 2010, pp. 74-17.

12. Hartrich L., Weinstock-Guttmanb, Hallc D., Badgettc D., Baierd M., Patrickb K., Feichterb J., Hongc J., Ramanathan M. Dynamics of immune cell trafficking in interferon-h treated multiple sclerosis patients. Journal of Neuroimmunology, 2003, no. 139, pp. 84-92.

13. Horwitz, D.A., Gray, J.D., Ohtsuda, K., Hirokawa, M., Takahashi, T. The immunoregulatory effects of NK cells: the role of TGFβ and implications for autoimmunity. Immunol. Today, 1997, no. 18 (11), pp. 538-542.

14. Huang Y.M., Hussien Y., Jin Y-P, Söderstrom M., Link H. Multiple sclerosis: deficient in vitro responses of blood mononuclear cells to IFN-β. Acta Neurol Scand, 2001, no. 104, pp. 249-256.

15. Kos F.J., Engleman E.G. Immune regulation: a critical link between NK cells and CTLs. Immunol. Today, 1996, no. 17(4), pp. 174-176.

16. Muggia F. The Benefits of pegylation in cancer and antiviral therapy From Research to Practise, 2001, no. 3 (1), pp. 1-3.

17. Munschauer, F.E., Hartrich, L.A., Stewart, C.C., Jacobs, L. Circulating natural killer but not cytotoxic T lymphocytes are reduced in patients with active relapsing multiple sclerosis and little clinical disability as compared to controls. J. Neuroimmunol., 1995, no. 62 (2), pp. 81-177.

18. Natarajan K., Sawicki M. V., Margulies D., Mariuzza R. Crystal Structure of Human CD69: A C-Type Lectin-Like Activation Marker of Hematopoietic Cells. Biochemistry, 2000, no. 39, pp. 14779-14786.

19. Northfeld D.W., Dezube В.J., Thommes J.A. Pegylated – liposomal doxorubicin versus doxorubicin, bleomicin and vincristine in the treatment of AIDS – related Kaposhi’s sarcoma: results of a randomized, phase 111 clinical trial. J. Clin. Oncol., 1998, no. 16, pp. 2445-2451.

20. Perini P., Wadhwa M., Buttarello M., Meager A., Facchinetti A., Thorpe R., Biasi G., Galloa P. Effect of IFNβ and anti-IFNb antibodies on NK cells in multiple sclerosis patients. Journal of Neuroimmunology, 2000, no. 105 (1), pp. 5-91.

21. Rani S., Shrock J., Appachi S., Rudick A., Williams B., Ransohoff R. Novel interferon-beta-induced gene expression in peripheral blood cells. Journal of Leukocyte Biology, 2007, Vol. 82, pp. 1353-1360.

22. Takahashi K., Aranami T., Endoh M., Miyake T., Yamamur T. The regulatory role of natural killer cells in multiple sclerosis. Brain, 2004, Vol. 127, no. 9, pp. 1917-1927.

23. Testi R., D’Ambrosio D., De Maria R., Santoni A. The CD69 receptor: a multipurpose cell-surface trigger for hematopoietic cells. Immunol. Today, 1994, no. 15, pp. 479-483.

24. Vega A., Pugslay M.K. An overview of colometric assay methods used to assess survival or proliferation of mammalian cells. Proc. West. Pharmacol. Soc., 2011, no. 54, pp. 10-14.