Email this article ПРИМЕНЕНИЕ БИОФЛУОРЕСЦЕНТНОГО ШТАММА YERSINIA PESTIS И ПРОТОЧНОЙ ЦИТОМЕТРИИ ДЛЯ ОЦЕНКИ ФАГОЦИТАРНОЙ АКТИВНОСТИ ЛЕЙКОЦИТОВ КРОВИ В ТЕСТЕ IN VITRO
- Authors: Kravtsov A.L.1, Budanova A.A.1, Klyueva S.N.1, Kozhevnikov V.A.1, Bugorkova S.A.1
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Affiliations:
- FKUN Russian Anti-Plague Institute "Microbe" of Rospotrebnadzor, Saratov, Russia
- Section: SHORT COMMUNICATIONS
- Submitted: 08.07.2024
- Accepted: 24.07.2025
- URL: https://rusimmun.ru/jour/article/view/17010
- DOI: https://doi.org/10.46235/1028-7221-17010-AOA
- ID: 17010
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Abstract
Abstract
Phagocytic activity (PA) of blood leukocytes in relation to Yersinia pestis reflects the state of cellular anti-plague immunity in humans and laboratory animals. The purpose of this work was to evaluate the stimulating effect of anti-plague vaccination on the PA of peripheral blood leukocytes in an in vitro test using flow cytometry and the biofluorescent strain of Y. pestis EV NIIEG pTurboGFP-B (KM2115), expressing the green fluorescent protein GFP. Materials and methods. We studied leukocytes in 100 μl of heparinized whole blood of mice, guinea pigs and people vaccinated and not vaccinated with live plague vaccine. Suspensions of living cells of strain KM2115 were mixed with blood diluted with physiological solution. The result of the phagocytic reaction was taken into account after 15 minutes by flow cytometry, determining the proportion of active phagocytes in the granulocyte gate that have intense fluorescence in the green region of the spectrum. The analysis was carried out according to the protocol we had previously developed in experiments with killed plague microbes labeled with FITC dye. Results. Experimental data have been obtained indicating the possibility of using a biofluorescent Y. pestis strain for the rapid determination of granulocyte PA by flow cytometry in microvolumes of whole blood of humans and animals without lengthy preliminary staining of microbes with a fluorescent dye. Increased values of phagocytic indices were registered for blood cells of people and animals vaccinated against plague, which is consistent with data on the stimulating effect of anti-plague vaccination on the phagocytic function of leukocytes. Conclusion. The use of a biofluorescent strain simplified the procedure for modeling the interaction of macroorganism cells with plague microbes and subsequent assessment of neutrophil PA in samples of whole peripheral blood, which can be used to characterize the strength of cellular post-vaccination anti-plague immunity in an in vitro test.
About the authors
Alexander Leonidovich Kravtsov
FKUN Russian Anti-Plague Institute "Microbe" of Rospotrebnadzor, Saratov, Russia
Email: kravzov195723@gmail.com
ORCID iD: 0000-0002-9016-6578
PhD, MD (Biology), Leading Research, Department Immunology
Russian Federation, 410005, Russian Federation, Saratov, Universitetskaya str., 46Angelina Andreevna Budanova
FKUN Russian Anti-Plague Institute "Microbe" of Rospotrebnadzor, Saratov, Russia
Email: rusrapi@microbe.ru
ORCID iD: 0000-0002-5092-432X
PhD (Biology), Senior Researcher, Department Immunology
Russian Federation, 410005, Russian Federation, Saratov, Universitetskaya str., 46Svetlana Nikolaevna Klyueva
FKUN Russian Anti-Plague Institute "Microbe" of Rospotrebnadzor, Saratov, Russia
Email: klyueva.cvetlana@mail.ru
ORCID iD: 0000-0002-5550-6063
PhD (Biology), Research Associate, Department Immunology
Russian Federation, 410005, Russian Federation, Saratov, Universitetskaya str., 46Vitaly Aleksandrovich Kozhevnikov
FKUN Russian Anti-Plague Institute "Microbe" of Rospotrebnadzor, Saratov, Russia
Email: rusrapi@microbe.ru
ORCID iD: 0000-0001-7267-7027
Junior Researcher, Department of Immunology
Russian Federation, 410005, Russian Federation, Saratov, Universitetskaya str., 46Svetlana Aleksandrovna Bugorkova
FKUN Russian Anti-Plague Institute "Microbe" of Rospotrebnadzor, Saratov, Russia
Author for correspondence.
Email: rusrapi@microbe.ru
ORCID iD: 0000-0001-7548-4845
PhD, MD (Medicine), Chief Researcher, Department of Immunology
Russian Federation, 410005, Russian Federation, Saratov, Universitetskaya str., 46References
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СМИ зарегистрировано Федеральной службой по надзору в сфере связи, информационных технологий и массовых коммуникаций (Роскомнадзор).