Abstract
The possibility of post-vaccination anti-plague immunity evaluation was researched using antigen-stimulated cells tests in vitro and cytometry analysis. The object of study — the blood samples of 17 people immunised by the live plague vaccine (Yersinia pestis EV) epicutaneously. Blood taking was carried out before vaccination and after immunisation on 7 and on 21 days, in 3 and in 6 months. Intensity antigen reactivity of lymphocytes was detected by cell tests in vitro, analysing markers of early (CD45+CD3+CD25+) and late (CD45+CD3+HLA-DR+) lymphocyte activation using flow cytometry. The complex of water-soluble Y. pestis antigens and allergen — pestin PP was tested as antigen. The high stimulating potential was defined of the water-soluble antigens Y. pestis complex. It is shown that coefficient of stimulation of relative level T- lymphocytes which express receptors for IL-2 was positive for all observation times after immunisation. The coefficient of stimulation had maximum values at 21 days (56.37%) and at 3 (47.41%) months. In identifying HLADR-positive lymphocytes before vaccination, the negative coefficient of stimulation was indicated on 7 and 21 days and the positive coefficient of stimulation was indicated at 3 and at 6 months. Analysis of intensity expression of early and late lymphocyte activation markers dynamics showed the possibility and prospect of application of cellular in vitro tests for the laboratory evaluation of specific reactivity of cellular immunity in both the early (7 days) and late (6 months) periods after vaccination. The results can be the basis for developing a new algorithm for assessment of immunological effectiveness of vaccination people against plague. It is the algorithm based on the identification of lymphocyte activation markers by antigen stimulation in conditions in vitro.
Highlights
The possibility of post-vaccination anti-plague immunity evaluation was researched using antigen-stimulated cells tests in vitro and cytometry analysis
Intensity antigen reactivity of lymphocytes was detected by cell tests in vitro, analysing markers of early (CD45+CD3+CD25+) and late (CD45+CD3+HLA-DR+) lymphocyte activation using flow cytometry
The complex of water-soluble Y. pestis antigens and allergen — pestin PP was tested as antigen
Summary
Что коэффициент стимуляции относительного содержания Т-лимфоцитов, экспрессирующих рецепторы к IL-2, после вакцинации во все сроки наблюдения был положительный, при этом наибольших значений он достигал на 21 сутки (56,37%) и через 3 месяца (47,41%). При выявлении HLA-DR-позитивных лимфоцитов до вакцинации, на 7 и 21 сутки отмечается отрицательный коэффициент стимуляции, через 3 и 6 месяцев коэффициент стимуляции был положительный. Анализ динамики интенсивности экспрессии маркеров ранней и поздней активации лимфоцитов показал возможность и перспективу применения клеточных тестов in vitro для лабораторной оценки специфической реактивности клеточного иммунитета как на ранних (7 суток), так и поздних (6 месяцев) сроках после вакцинации. Полученные результаты могут быть основанием для разработки нового алгоритма оценки иммунологической эффективности вакцинации контингентов против чумы, основанного на выявлении маркеров активации лимфоцитов при стимуляции антигеном в условиях in vitro. Ключевые слова: специфическая профилактика чумы, клеточный иммунитет, оценка эффективности вакцинации, маркеры активации лимфоцитов, антигены Yersinia pestis, коэффициент стимуляции.
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