Analysis of main and small populations of lymphocytes of rhesus macaque peripheral blood using the method of flow cytometry

Abstract

Introduction. When assessing the immune status of humans and animals, along with the main populations of lymphocytes, in some cases, it is necessary to take into account data on small subpopulations of lymphocytes and pools of activated cells.

The aim - determination presence and intervals of main and small lymphocytes populations distribution in the peripheral blood of healthy rhesus macaque using multicolored cytometric analysis and comparison of the date obtained with the corresponding characteristics of humans.

Material and methods. Venous blood of 17 healthy Macaca mulatta males 2.0-2.5 years old was used in the experiments. Whole blood of animals was stained with fluorochrome-conjugated mouse anti-human monoclonal antibodies that cross-react with rhesus monkey lymphocyte antigens. Determination of the populations of monkey lymphocytes was carried out using the flow cytometry method. The results obtained were compared with similar data for children and adolescents published in Russian scientific journals.

Results. We identified 20 major and small populations of rhesus monkey lymphocytes, studied the spontaneous expression level of markers of early (CD25) and late (HLA-DR) lymphocyte activation on T-cell subpopulations. In rhesus monkeys, a significantly lower relative content of T-lymphocytes is observed; NKT cells, T-helpers, cytotoxic T-lymphocytes, double negative T-lymphocytes, T-lymphocytes expressing an early activation marker, T-helpers expressing an early activation marker, T-regulatory cells, but higher - B-lymphocytes, natural killer cells, CD8 positive natural killer cells, double-positive T-lymphocytes, T-lymphocytes expressing a late activation marker, T-helpers expressing a marker of late activation, and cytotoxic T-lymphocytes expressing a marker of late activation (p < 0.05).

Conclusion. Determination by flow cytometry of indicators of the main, small and activated lymphocytes of rhesus monkeys will allow, when simulating human diseases in animals, to assess not only the period of the onset of the immune response and its nature, but also make it possible to predict the course of the disease, carry out laboratory monitoring of therapy, and in the future will allow the use of lymphocytes as targets of therapeutic action.

Keywords:lymphocytes; rhesus macaques; flow cytometry; markers of early and late activation

For citation: Borisevich G.V., Kirillova S.L., Petrov A.A., Lebedev V.N., Pashchenko Yu.I., Kirillov V.B., Sizikova T.E., Paveliev D.I., Pantyukhov V.B., Kutaev D.A., Borisevich S.V. Analysis of main and small populations of lymphocytes of rhesus macaque peripheral blood using the method of flow cytometry. Immunologiya. 2020; 41 (5): 411-20. DOI: https://doi.org/10.33029/0206-4952-2020-41-5-411-420 (in Russian)

Funding. The study had no sponsor support.

Conflict of interests. The authors declare no conflict of interests.

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