Monitoring of cytogenetic instability by micronuclei assay of both immunocompetent and non-immunocompetent cells in tick-borne encephalitis patients depending on variants of glutathione-S-transferase genes in the genotype

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Abstract

Aim of this study was to study the dynamics of the frequency of cytokinesis-blocked T-lymphocytes with micronuclei in peripheral blood and the frequency of buccal micronucleated epithelium cells for a period of half a year in patients with acute tick-borne encephalitis, depending on burden of active and inactive variants of glutathione-S-transferase genes (GSTM1 and GSTT1) in the patient's genotype. We carried out micronucleus assay in immunocompetent and non-immunocompetent cells in 54 patients with acute tick-borne encephalitis and 35 healthy persons (control) residing in the Tomsk and Tyumen regions. To analyze the frequency of cytokinesis-blocked micronucleated T-lymphocytes was used venous peripheral blood as material for phytohemagglutinin-stimulated cultures, and to study the frequency of buccal micronucleated cells, samples of the buccal mucous membrane epithelial cells were obtained. To carried out both techniques of micronucleus assay, cytological preparations were prepared, which were stained using the Giemsa or Felgen methods. The material for the study was obtained repeatedly during admission of patients to treatment, and also after 1 week, 1, 3 and 6 months.  Polymerase chain reaction was used to analyze the alleles of the GSTM1 and GSTT1 genes. As a result of this analysis was found a significant increase in the frequency of micronucleated cells in tick-borne encephalitis patients compared with the control group. In addition, the frequency of cytokinesis-blocked micronucleated T-lymphocytes was increased significantly higher than the one of micronucleated buccal cells. The most significant and prolonged increase in the frequency of micronucleated cells was associated with the mutant inactive variants of the genes GSTM1 (0/0) and GSTT1 (0/0). In the patients with burden the inactive forms of these genes, the cytogenetic instability of the cytokinesis-blocked blood T-lymphocytes could persist for up to six months. In case of buccal cells, the frequency of micronucleated cells was close to the one in the control group as early as 1-3 months after a course of treatment. Conclusion. It was found that the most increased and prolonged frequency of cytogenetically instable cells persisted in cytokinesis-blocked T-lymphocytes of peripheral blood of patients with tick-borne encephalitis who were carriers of the genotype with inactive variants of  both GSTM1 (0/0) and GSTT1 (0/0 ) glutathione-S-transferase genes.

About the authors

Nikolay Nikolaevich Ilyinskikh

Siberian State Medical University, National Research Tomsk State University

Author for correspondence.
Email: nauka-tomsk@yandex.ru
ORCID iD: 0000-0003-1014-1096

PhD, Doctor of Biological Sciences, Professor, Professor of Department of Biology and Genetics, Siberian State Medical University, Professor of Department of Ecology, Nature Management and Environmental Engineering, National Research Tomsk State University

Россия

Ekaterina Nikolaevna Ilyinskikh

Siberian State Medical University, National Research Tomsk State University

Email: infconf2009@mail.ru
ORCID iD: 0000-0001-7646-6905
PhD, MD, Associate Professor, Professor of Department of Infectious Diseases and Epidemiology, Siberian State Medical University, Professor of Department of Ecology, Nature Management and Environmental Engineering, National Research Tomsk State University Россия

Evgenia Vladimirovna Zamyatina

Siberian State Medical University

Email: e_zamyt@mail.ru
ORCID iD: 0000-0001-7579-8975

Assistant of Department of Infectious Diseases and Epidemiology, Siberian State Medical University

Россия

Svetoslava Vyacheslavovna Lee

Siberian State Medical University

Email: infconf2009@mail.ru

5th year student of the Pediatric faculty, Siberian State Medical University

Россия

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Copyright (c) 2019 Ilyinskikh N.N., Ilyinskikh E.N., Zamyatina E.V., Lee S.V.

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