Evaluation of the expression of lymphocyte surface activation markers and production of secretory immunoglobulin A in the formation of immunity against cholera

Filippenko A.V., Ivanova I.A., Omelchenko N.D., Trufanova A.A.

Rostov-on-Don Anti-Plague Institute of Rospotrebnadzor, Rostov-on-Don

The aim of the study was to evaluate the expression of markers of activation of peripheral blood lymphocytes and the production of secretory immunoglobulin A (sIgA) in the small intestine of white mice vaccinated with cholera vaccine and receiving immunomodulators. To do this, white mice were vaccinated with azoximer bromide, glucosaminylmuramyldipeptide, and sodium deoxyribonucleate once. The expression of markers of lymphocyte activation in laboratory animals was carried out on 3-21 days of the post-vaccination period. Cells after lysis were stained with MCA to CD45, CD23, CD38 and CD69 ("Invitrogen", USA) mice and analyzed on a flow cytometer "NaviosTM" ("Beckman Coulter", USA). The amounts of sIgA were determined in the washing waters of the small intestine of mice using the "Enzyme-linked immunosorbent assay kit for sIgA" (USA) on a multifunctional reader "SynergyTM 2" for 5-21 days after vaccination. The protective activity of the vaccine and the effect of immunomodulators on this process were evaluated 21 days after immunization, causing a generalized form of cholera in white mice. An increase in the expression of CD69, CD38 and CD23 on the surface of lymphocytes in vaccinated experimental animals was revealed. The introduction of azoximer bromide and glucosaminylmuramyldipeptide led to an increase in the number of CD69, starting from the first stages of the study and up to the end of the experiment. Under the influence of sodium deoxyribonucleate, the synthesis of this receptor was also enhanced, but less intensively. To a greater extent and earlier terms of azoximer bromide and glucosaminylmuramyldipeptide contributed to an increase in the number of CD23 on the membranes of immunocompetent cells of vaccinated white mice. A significant increase in the number of CD38 in relation to immunized animals was observed by us only under the influence of azoximer bromide, starting from the first week and until the end of the observation period. It has been shown that the use of immunomodulators, especially azoximer bromide and glucosaminylmuramyldipeptide, contributes to an increase in the synthesis of sIgA, the main effector of mucosal immunity, in the small intestine of vaccinated animals. Correlative links were found between the intensity of anti-cholera immunity, sIgA production and the level of expression of activation markers, especially CD69 and CD23, and when using azoximer bromide and glucosaminylmuramyldipeptide.