Abstract
Objective. The aim was revealing gender differences in morphological and functional changes of lymphoid organs (thymus and spleen), changes of cytokine production and subpopulation composition of peripheral blood lymphocytes in Wistar rats of three age groups with endotoxemia. Materials and methods. We used male and female Wistar rats of three age groups: newborns, prepubertal and sexually mature adult rats. A day after the injection of 15 mg/kg of O26:B6 E. coli lipopolysaccharide (LPS), the volume fraction of the functional zones of the thymus and spleen, the number of AnnexinV + apoptotically dying cells in the thymus, the relative and absolute number of lymphocyte subpopulations (CD3+CD4+, CD3+CD8a+, CD4+CD25+Foxp3+, CD3-CD45R+) in peripheral blood and ex vivo production of IL-2, IL-4, IFN-γ and TNF-α were estimated. Results. Sex differences in the response of the immune system after the LPS injection in different age periods are expressed differently: in the neonatal period, there is immunosuppression in females (decrease in the ex vivo production of IL-2, TNF-α and IFN-γ), and there is activation of pro-inflammatory reactions in males (increase in ex vivo production of IL-2 and TNF-α). As compared with other age periods at prepubertal age, LPS-induced immunological disorders are more pronounced, and gender differences are minimal and related only to the number of T-regulatory and B-lymphocytes. In the adults, the LPS-induced immunosuppression is most pronounced in males - they have a decrease in the production of all the cytokines studied and a decrease in the number of cytotoxic and regulatory T-lymphocytes and B-cells. Conclusion. Thus, in each of the studied age periods - newborn, prepubertal and adult, the sexual differences in the immune system reactions are expressed differently and, apparently, these differences are determined by the content of sex steroid hormones, the concentration of which varies with age.
Highlights
The LPS-induced immunosuppression is most pronounced in males – they have a decrease in the production of all the cytokines studied and a decrease in the number of cytotoxic and regulatory T-lymphocytes and B-cells
In each of the studied age periods – newborn, prepubertal and adult, the sexual differences in the immune system reactions are expressed differently and, apparently, these differences are determined by the content of sex steroid hormones, the concentration of which varies with age
SOURCE OF FINANCING The study was performed as a part of a state assignment for Research Institute of Human Morphology (AAAA-A19119021490067-4)
Summary
Исследования проводили на самцах (n=60) и самках (n=60) крыс Вистар трех возрастных групп. Для разведения животных использовали 25 половозрелых животных (15 самок и 10 самцов). Через 24 ч после введения ЛПС крыс выводили из эксперимента передозировкой диэтилового эфира. Количество апоптотически гибнущих клеток (AnnexinV+PI-) в тимусе оценивали с помощью проточной цитофлуориметрии на приборе Cytomics FC 500 («Beckman Coulter») с использованием Annexin V FITC Kit (Beckman Coulter). Для индукции синтеза и секреции цитокинов суспензию клеток селезенки в концентрации 106/мл культивировали 20 ч в 1 мл полной ростовой среды с добавлением конканавалина А (5 мкг/мл) в 24-луночных культуральных планшетах при 37°С и 5% СО2. Относительное и абсолютное количество основных субпопуляций лимфоцитов в периферической крови определяли на приборе Cytomics FC 500 («Beckman Coulter») с использованием следующих антител («eBioscience»): antiRat CD3 (маркер Т-лимфоцитов); anti-Rat CD4 (маркер Т-хелперов); аnti-Rat CD8a (маркер цитотоксических Т-клеток); аnti-Rat CD45R (маркер В-лимфоцитов); аnti-Rat CD25
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