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Russian Journal of Parasitology, 2017, V.40, Iss.2
DOI:
Received: 11.06.2016
Accepted: 27.02.2017
STUDY OF MARKERS OF BLOOD LYMPHOCYTES APOPTOSIS IN ANIMALS AT HELMINTHIASES
Grishina E.A.
Medical Academy of Postgraduate Education, Ministry of Health of the Russian Federation, 125993 Moscow, Barrikadnaya St. 2/1, Build. 1, e-mail: gelana2010@yandex.ru
Abstract
Studying the role of apoptosis of blood cells allows to determine more precisely the mechanisms of immunopathology in general, and immunosuppression - in particular. The aim of that research was to study morphological and molecular indicators of apoptosis of peripheral blood lymphocytes in mice infected with Trichocephalus muris and rabbits infected with Passalurus ambiguus to determine possible mechanisms of secondary immunosuppression development under chronisation of helminthiasis.
Materials and methods. During the experiment, the animals were divided into the following groups: 1) intact mice (control group) - 15 ind.; 2) intact rabbits (control group) - 10 ind.; 3) mice infected with Trichocephalus muris (Schrank, 1788) – 60 ind.; 4) rabbits infected with Passalurus ambiguous (Rudolphi, 1819) - 30 ind. Blood from the tail vein of mice, and the ear vein of rabbits was used to prepare smears 1, 2, 3, 6, 7, 8 weeks after infestation. Light microscopy of blood smears stained by Romanovsky’s method [2] was applied for leucogram determination and investigation of morphological indicators of apoptosis. For molecular studies, mononuclear leukocytes were isolated from venous whole blood by density gradient centrifugation using Ficoll-Paque media (= 1,077 g / cm3, «Pharmacia», Sweden) [12]. The concentration of the pro-apoptotic protein caspase-3 and anti-apoptotic protein Bcl-2 was determined in lysates of lymphocytes by ELISA using kits Human Caspase-3 instant ELISA and the Human Bcl-2 ELISA of the company «Bender MedSystems GmbH» (Vienna, Austria).
Results and discussion. The analysis of obtained data shows that at helminth infection of animals and under chronisation of helminthiasis, statistically significant changes occur within 8 weeks in the total number of leukocytes, the total number of lymphocytes and lymphocytes with morphological signs of apoptosis, as well as in pro-apoptotic protein caspase-3 and anti-apoptotic Bcl-2 protein.
This comprehensive study convincingly shown that a gradual and significant increase in apoptotic activity of lymphocytes on the receptor and cellular levels is observed in animals in the period from the beginning of infestation up to the chronic stage of helminthiasis. This is manifested in the increased number of lymphocytes with morphological features of apoptosis against the background of the increased level of pro-apoptotic protein caspase-3, and the reduced level of anti-apoptotic protein Bcl-2.
Key words: helminthiasis, immunity, immunosuppression, apoptosis of lymphocytes, inductors and inhibitors of apoptosis.
References
1. Bekish V.Ya., Durnev A.D. In vitro genotoxic and cytotoxic effects of protein somatic Products from helminths on donor blood. Byulleten' Eksperimental'noy Biologii i Meditsiny [Bull. exp. biol. and med.], 2004, vol. 138, no. 8, pp. 198-201. (In Russian)
2. Ryabykina N. V. The study of apoptosis of white blood cells and changes in leukocyte formula in older male mice under the influence of hypohydration stress and α– tocopherol acetate. Molodoy uchenyj [Young Scientist], 2010, no.12, vol.1, pp. 57-59. (In Russian)
3. Fil'chenkov A.A., Stoyka R.S. Apoptoz i rak [Apoptosis in cancer]. Kiev, Morion, 1999. 184 p. (In Russian)
4. Yarilin A.A. Immunologiya: uchebnik [Immunology. Textbook], 2010. 752 p. (In Russian)
5. Alam M. S., Chowdhury M. A. Z., Khaliq Q. A., Yasmin S. Stability analysis for seed yield and its components in soybean (Glycýne max (L.) Merrill). Annals of Bangladesh Agriculture, 1999, no. 9, pp. 43-48.
6. Bekish V.J. The alterations in genetic apparatus of somatic and generative cells of the host caused by helminthes metabolites. Wiadomosci Parazytologiczne (Poland), 2001, vol. 47, no. 4, pp. 891-896.
7. Chow S.C., Brown A., Pritchard D. The human hookworm pathogen Necator americanus induces apoptosis in T lymphocytes. Parasite Immunology, 2000, vol. 22, pp. 29-37.
8. Eymin В., Claverie P. Salon C. PI4arf Triggers G2 Arrest through. Oncogene, 1999, vol. 18, pp. 1411-1418.
9. Estaquier J., Marguerite M., Sahuc F. Interleukin 10-mediated T cell apoptosis during the T helper type 2 cytokine response in murine Schistosoma mansoni parasite infection. Eur. Cytokine Netw., 1997, vol. 8, pp. 153-160.
10. Fadeel, B., Orrenius S., Zhivotovsky B. Apoptosis in human disease: a new skin for the old ceremony?” Biochem Biophys Res Commun., 1999, vol. 266, no.3, pp. 699-717.
11. Fallon P.G., Smith P., Dunne D.W. Type 1 type 2 cytokine producing mouse CD4+ and CD8+ cells in acute Schistosoma mansoni infection. Eur. J. Immunol., 1998, vol. 28, pp. 1408-1416.
12. Fernando H.C., Luketich J.D., Christie N.A., Ikramuddin S., Schauer P.R. Outcomes of laparoscopic Toupet compared to laparoscopic Nissen fundoplication. Surg. Endose., 2002, vol.1, pp. 905-908.
13. Gondal, M.A., Laser photoacoustic spectrometer for remote monitoring of atmospheric pollutants, Applied Optics, 1997, vol. 36, no. 15ISSN: 1559-128X (print) ISSN: 2155-3165 (online)
14. Kennedy P. M., Lowry, J. B., Conlan, L. L., Isolation of grass cell walls as neutral detergent fibre increases their fermentability for rumen micro-organisms. J. Sci. Food Agric., 1999, vol. 79, no. 4, pp. 544-548.
15. Los M., Stroh C., Jänicke R.U. Caspases: more than just killers? Trends Immunol., 2001, vol. 22, no. 1, pp. 31-34.
16. Lundy S.K., Lerman S.P., Boros D.L. Soluble egg antigen-stimulated T helper lymphocyte apoptosis and evidence for cell death mediated by FasL+ T and B cells during murine Schistosoma mansoni infection. Infection and Immunity, 2001, vol. 69, no. 1, pp. 271-280.
17. Robertson J.D., Orrenius S., Zhivotovsky B.J. Struct. Biol., 2000, no.129, pp. 346–358.
18. Rumbley C.A., Zekavat S.A., Sugaya H. The schistosome granulema: characterization of lymphocyte migration, activation and cytokine production. J. Immunol. 1998, vol. 161, pp. 4129- 4137.
19. Tato P., Fernandez A.M., Solano S. A cysteine protease from Taenia solium metacestodes induce apoptosis in human CD4+ T-cells. Parasitol. Res., 2004, vol. 92, no. 3, pp. 197-204.
20. Zhou D., Lambert, S., Malen P.L., Carpenter S., Boland L.M., Bennett V. J. Cell Biol., 1998, no. 143, pp.1295-1304.
21. Zhang J, Rosenberg HF, Nei M. Positive Darwinian selection after gene duplication in primate ribonuclease genes. Proceedings of the National Academy of Sciences (USA), 1998, no. 95, pp. 3708-3713.
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