Резюме. Возрастная динамика формирования Th17 и Treg у здоровых людей недостаточно изучена. Целью настоящего исследования было изучить количественные характеристики субпопуляций Th17 и Treg у здоровых людей в зависимости от возраста.Было обследовано 352 человека (168 лиц женского пола и 184 -мужского), в возрасте от 1 месяца до 85 лет, в том числе младше 1 года -79, в возрасте 1-2 года 11 мес. -34, 3-4 года 11 мес. -24, 5-6 лет 11 мес. -28, 7-8 лет 11 мес. -25, 9-11 лет 11мес. -36, 12-14 лет 11 мес. -39, 15-18 лет -26, 20-35 лет -25, 36-49 лет -11, 50-70 лет -16 и в возрасте более 70 лет -9 человек. У детей в возрасте до 2 лет иссле-дование проводилось из капиллярной крови, в более старшем возрасте -из венозной. Определение субпопуляций лимфоцитов крови проводили методом проточной цитофлюорометрии с четырехцвет-ным окрашиванием в реакции прямой иммунофлюоресценции. Окрашивание проводили в цельной крови с последующим лизированием эритроцитов. Для определения субпопуляций лимфоцитов ис-пользовали следующие поверхностные маркеры: CD3, CD4, CD8, CD25, CD127, CD161, CD45R0.Показано, что процент Treg в гейте CD3 + CD4 + практически не зависит от возраста обследуемых лиц и аппроксимируется линейной функцией. В абсолютном выражении уровень Treg в детском возрасте прогрессивно снижается и после 10 лет выходит на плато и аппроксимируется логарифми-ческой кривой. При исследовании возрастной динамики уровня Th17 обнаружен устойчивый рост этой субпопуляции как в процентном, так и в абсолютном выражении. Процент этих клеток от гейта CD3 + CD4 + и абсолютное количество аппроксимируется квадратными уравнениями. Возраст 10-12 лет является, по-видимому, критическим в процессе формирования иммунной системы, к этому воз-расту завершаются основные процессы развития различных звеньев иммунитета и дальше происхо- Адрес для переписки:Топтыгина Анна Павловна ФБУН «Московский научно-исследовательский институт эпидемиологии и микробиологии им. Г.Н. Габричевского» Роспотребнадзора 125212, Россия, Москва, ул. Адмирала Макарова, 10. Тел.: 8 (495) 452-18-01. Факс: 8 (495) 452-18-30.

The question of the duration and effectiveness of post-infection immunity to SARS-CoV-2 and its comparison with post-vaccination remains at the center of study by many researchers. The aim of the work was to study the duration of maintaining post-infection and post-vaccination immunity to the SARS-CoV-2 virus, as well as the formation of hybrid (vaccination after infection) and breakthrough (repeated disease or disease after vaccination) immunity in the context of an ongoing pandemic. 107 adults who had mild or moderate COVID-19 3-18 months after the disease and 30 people vaccinated twice with the Sputnik V vaccine were examined 1-6 times. Antibodies to the SARS-CoV-2 virus were determined by ELISA on the SARS-CoV-2-IgG quantitative-ELISA-BEST test systems. The avidity of antibodies was determined by additional incubation with and without denaturing solution. Mononuclear cells were isolated from blood by gradient centrifugation, incubated with and without coronavirus S-protein for 20 hours, stained with fluorescently labeled antibodies, and the percentage of CD8highCD107a+ was counted on a FACSCanto II cytometer. It was shown that in the group of reconvalescent and vaccinated, the level of antibodies specific to the virus decreased more pronounced in individuals with an initially high humoral response, but after 9 months the decrease slowed down and reached a plateau. The avidity of antibodies rose to 50% and persisted for 18 months. Cellular immunity in recovered patients did not change for 1.5 years, while in vaccinated patients it gradually decreased after 6 months, but remained at a detectable level. After revaccination of the vaccinated, a significant increase in the level of antibodies, avidity to 67.6% and cellular immunity to the initial level was noted. Hybrid immunity turned out to be significantly higher than post-infection and post-vaccination immunity. The level of antibodies increased to 1218.2 BAU/ml, avidity to 69.85%, and cellular immunity to 9.94%. Breakthrough immunity was significantly higher than after the first disease. The level of antibodies rose to 1601 BAU/ml, avidity - up to 81.6%, cellular immunity - up to 13.71%. Using the example of dynamic observation of four COVID-19 reconvalescents, it has been shown that in the context of the ongoing pandemic and active mutation of the coronavirus, natural boosting occurs both asymptomatically and as a result of a mild re-infection, which prevents the disappearance of humoral and cellular immunity specific to SARS-CoV -2.

Psoriasis is considered an autoimmune disease with a predominantly cellular mechanism for the development of disorder. Studies in immune pathogenesis of psoriasis were performed either in animal model, which is not just similar to humans, or the data were obtained in patients by means of skin window method, which is traumatic, or by examining venous blood. However, it is difficult to discern parameters of the local immune response in venous blood samples. We have attempted to find an adequate method which would be convenient both for the patient and for the researcher, in order to assess local immune processes occurring in the skin affected by psoriasis. We examined 20 patients with a verified diagnosis of psoriasis, the average age was 44.3 years. The control group included 15 healthy adults, with average age of 46.6 years. Capillary blood was taken by fingerprick, whereas, in psoriatic patients, the samples were taken near the psoriatic lesion at a final volume of 400 μL in two microvettes. Venous blood (3 mL) was taken from the cubital vein into a vacuum tube with EDTA. Clinical analysis of venous and capillary blood was performed in automated hematological analyzer. Immunophenotyping was performed by four-color staining of whole capillary and venous blood followed by lysis of erythrocytes. Cytofluorometry was performed using techniques and reagents from BD Biosciences (USA). Plasma cytokines were determined by multiplex approach (MagPix, BioRad, USA). Upon clinical analysis of blood, the difference between capillary and venous blood was not found, either in healthy group, or among patients with psoriasis. In healthy people, the subsets of mononuclear cells, did not differ between venous and capillary blood. The samples of capillary and venous blood in the patients with psoriasis showed significantly increased levels of double-positive lymphocytes (CD45RA+/CD45R0+), B lymphocytes and NKT lymphocytes (both for relative and and absolute values). A significant increase in the percentage of naive T lymphocytes, activated helpers (Thact) and Treg, as well as B1 cells and Breg, and a significant decrease in B2 lymphocytes was registered in capillary blood of the patients with psoriasis. In venous blood samples from psoriatic patients, only a significant increase in Thact, Treg, and Breg was revealed. In the capillary blood of patients with psoriasis, we found a significant increase in the levels of non-classical M2 monocytes and inflammatory Minfl monocytes, and a decrease in classical M1 monocyte levels; in venous blood of psoriatic patients, only an increase in inflammatory Minfl monocytes was revealed. In capillary blood, all the studied cytokines in psoriasis patients significantly exceeded the levels of corresponding cytokines in healthy controls, except of IL-10. The levels of this cytokine did not differ from healthy group. In venous blood, the levels of most studied cytokines in the group of patients with psoriasis did not differ from the group of healthy ones. Approximately two-fold increase was revealed for IL-4, IL-21, IL-23 and TNF. First, the subsets of mononuclear cells and the cytokine profile of capillary and venous blood of healthy people did not differ significantly. Secondly, our proposed method for determining the subsets of mononuclear cells and capillary blood cytokines profile from the area of psoriatic lesions may be used to monitor local immunity in the patients with psoriasis. This approach is significantly less traumatic than the skin window method and more informative than the studies of venous blood.

The SARS-CoV-2 virus that caused the COVID-19 pandemic is related to the SARS-CoV-1 and MERS coronaviruses, which were the cause of epidemics in 2003 and 2012. Antibodies in patients with COVID-19 appear 7-14 days after the onset of symptoms and gradually increase. As the COVID-19 pandemic continues, it is difficult to say how long the immunological memory to the SARS-CoV-2 virus will last. The aim of this study was to study the ratio of humoral and cellular immunity to the S-protein of the SARS-CoV-2 virus in COVID-19 convalescents. The study involved 60 adults with mild to moderate COVID-19 2 to 12 months prior to the examination. The control group consisted of 15 adults who did not have COVID-19 and were not vaccinated against this infection. Specific antibodies to the SARS-CoV-2 virus were determined by ELISA using the SARS-CoV-2-IgG-ELISA-BEST kit. To determine the specific IgG and IgA subclasses, the anti-IgG conjugate from the kit was replaced with a conjugate against the IgG subclasses and IgA. Additional incubation with or without denaturing urea solution was used to determine the avidity of antibodies. Mononuclear cells were isolated from blood by gradient centrifugation, incubated with or without coronavirus S antigen for 20 hours, stained with fluorescently labeled antibodies, and the percentage of CD8highCD107a were counted on a flow cytometer Canto II. In the control group, neither humoral nor cellular immunity to the S-protein of the SARS-CoV-2 was found. In the group of those who had recovered, the level of IgG antibodies to the S-protein of the SARS-CoV-2 virus varies greatly and was not strictly associated with the duration of the disease, 57% of COVID-19 patients had a high level of humoral response, and 43% - low level. The correlation between the levels of specific IgG and IgA was r = 0.43. The avidity of antibodies increased over time after the disease, amounting to 49.9% at a period of 6-12 months. There were no specific IgG subclasses IgG2 and IgG4, and the percentage of IgG1 increased over time and amounted to 100% after 6-12 months after illness. 50% of the examined had high cellular immunity, and the same number - low, no correlations with the level of humoral immunity were found. We identified 4 combinations of humoral and cellular immunity to the S-protein of the SARS-CoV-2: high humoral and cellular, low humoral and cellular, high humoral and low cellular, and vice versa, low humoral and high cellular immunity.