Features of Lymphocyte Subset Composition in Neonates Born to Mothers Suffered from Covid-19 at Different Stages of Pregnancy

The relationship between the incidence of COVID-19 in pregnant women who have had a coronavirus infection at different gestational ages and the health status of paired neonates is of great interest. However, no sufficient convincing data fully reflecting features of subsequent neonatal period, the state of the immune system in this category of children, affecting characteristics of postnatal period have been accumulated. Based on this, it underlies the relevance of the current study aimed at investigating parameters of clinical and immunological state of neonatal health after paired mothers recovered from COVID-19 at different gestational ages. The prospective study included 131 women and 132 children. The main group consisted of women (n = 61) who had COVID-19 during pregnancy and paired newborns (n = 62) at gestational age (GA) of 37-41 weeks, the comparison group - women without laboratory-confirmed COVID-19 during pregnancy (n = 70) and paired newborns (n = 70) of similar gestational age. While analyzing the anamnesis of the patients, no significant differences in somatic and obstetric-gynecological diseases were found. Analyzing course of pregnancy revealed that low molecular weight heparins were significantly more often applied in the main group. The term and frequency of delivery by caesarean section in pregnant women in the main group did not significant differ from that of the control group. No significant difference in the frequency of causes accounting for the severity of the condition of neonates in paired mothers with COVID-19 at different trimester of gestation was found. Investigating lymphocyte subset composition, neutrophil phagocytic activity, and IgG class antibodies specific to SARS-CoV-2 was carried out. It was found that lymphocyte subset profile in newborns from paired mothers with COVID-19 at different trimesters of gestation differed only in the level of NK cells (CD56+) in children born to mothers recovered from COVID-19 in the first trimester. In this study, in general, no severe perinatal outcomes in newborns from paired mothers with COVID-19 during pregnancy were documented. No cases of moderate or severe maternal COVID-19 were observed. Therefore, further prospective studies are needed to assess an impact of COVID-19 severity on maternal and fetal birth outcomes and clarify optimal management of pregnant women in such cases.

FEATURES OF LYMPHOCYTE SUBSET COMPOSITION IN NEONATES BORN TO MOTHERS SUFFERED FROM COVID-19 AT DIFFERENT STAGES OF PREGNANCY.

The relationship between the incidence of COVID-19 in pregnant women who have had a coronavirus infection at different gestational ages and the health status of paired neonates is of great interest. However, no sufficient convincing data fully reflecting features of subsequent neonatal period, the state of the immune system in this category of children, affecting characteristics of postnatal period have been accumulated. Based on this, it underlies the relevance of the current study aimed at investigating parameters of clinical and immunological state of neonatal health after paired mothers recovered from COVID-19 at different gestational ages. The prospective study included 131 women and 132 children. The main group consisted of women (n = 61) who had COVID-19 during pregnancy and paired newborns (n = 62) at gestational age (GA) of 37-41 weeks, the comparison group - women without laboratory-confirmed COVID-19 during pregnancy (n = 70) and paired newborns (n = 70) of similar gestational age. While analyzing the anamnesis of the patients, no significant differences in somatic and obstetric-gynecological diseases were found. Analyzing course of pregnancy revealed that low molecular weight heparins were significantly more often applied in the main group. The term and frequency of delivery by caesarean section in pregnant women in the main group did not significant differ from that of the control group. No significant difference in the frequency of causes accounting for the severity of the condition of neonates in paired mothers with COVID-19 at different trimester of gestation was found. Investigating lymphocyte subset composition, neutrophil phagocytic activity, and IgG class antibodies specific to SARS-CoV-2 was carried out. It was found that lymphocyte subset profile in newborns from paired mothers with COVID-19 at different trimesters of gestation differed only in the level of NK cells (CD56+) in children born to mothers recovered from COVID-19 in the first trimester. In this study, in general, no severe perinatal outcomes in newborns from paired mothers with COVID-19 during pregnancy were documented. No cases of moderate or severe maternal COVID-19 were observed. Therefore, further prospective studies are needed to assess an impact of COVID-19 severity on maternal and fetal birth outcomes and clarify optimal management of pregnant women in such cases.Copyright © 2023 Saint Petersburg Pasteur Institute. All rights reserved.

Post-Gam-Covid-Vac Combined Vector Vaccine Cellular and Humoral Immune Response

Currently, as the SARS-CoV-2 pandemic evolves, there has been increasingly more attention paid to building natural and vaccine-induced immunity against SARS-CoV-2 and related disease known as COVID-19. Widespread pre-ventive vaccination plays an important role in effectively protecting people from viral infections and can reduce national economic costs. Purpose - to study peripheral blood cell subset composition and magnitude of humoral response in vac-cinated Gam-COVID-Vac subjects. The prospective study included 352 patients, of which 194 (119 women and 75 men) underwent an immunogram study and assessed level of anti-SARS-CoV-2 antibodies. In patients, the study of the lym-phocyte subset composition and estimation of anti-SARS-CoV-2 antibodies was carried out at two time points - prior to vaccination and 90 days after inoculated component 1 of the Gam-COVID-Vac vaccine. In general, vaccination was well tolerated by patients, with no serious adverse events after immunization. The reaction to the vaccine (fever, ma-laise, headache, local reactions) was short-term (1-2 days) and more often noted after inoculated vaccine component 2. Comparatively analyzed immunogram parameters in females before and after vaccination revealed increased relative level of T-lymphocytes (CD3+), T-helper cell subset (CD3+CD4+), increased absolute and relative level of activated CD3+CD25+ T-lymphocytes, but decreased absolute and relative level of natural killer (CD3-CD56+CD16+) and natural killer T-cell (CD3+CD56+CD16+) cell subsets as well as decreased CD147 receptor expression on T-lymphocytes. Similar patterns were also found while examining the immunogram in males exepting increased level of lymphocytes and lowered CD147 expression on both T-and B-lymphocytes. No changes in the parameters of the immune T-cell arm was found. The high efficacy of the vaccine was confirmed by development of SARS-CoV-2-specific class G antiviral antibodies in 97.5% and 92.3% of vaccinated females and males, respectively. The data obtained evidence that: 1) vaccination induces a specific humoral immune response determined three months post-vaccination, and 2) it caused no serious disturbances in the im-mune system functioning, which could be reflected in the peripheral blood lymphocyte subset composition. Thus, the data presented allow to conclude that Gam-COVID-Vac is effective vaccine against SARS-CoV-2 infection.

POST-GAM-COVID-VAC COMBINED VECTOR VACCINE CELLULAR AND HUMORAL IMMUNE RESPONSE.

Currently, as the SARS-CoV-2 pandemic evolves, there has been increasingly more attention paid to building natural and vaccine-induced immunity against SARS-CoV-2 and related disease known as COVID-19. Widespread preventive vaccination plays an important role in effectively protecting people from viral infections and can reduce national economic costs. Purpose - to study peripheral blood cell subset composition and magnitude of humoral response in vaccinated Gam-COVID-Vac subjects. The prospective study included 352 patients, of which 194 (119 women and 75 men) underwent an immunogram study and assessed level of anti-SARS-CoV-2 antibodies. In patients, the study of the lymphocyte subset composition and estimation of anti-SARS-CoV-2 antibodies was carried out at two time points - prior to vaccination and 90 days after inoculated component 1 of the Gam-COVID-Vac vaccine. In general, vaccination was well tolerated by patients, with no serious adverse events after immunization. The reaction to the vaccine (fever, malaise, headache, local reactions) was short-term (1-2 days) and more often noted after inoculated vaccine component 2. Comparatively analyzed immunogram parameters in females before and after vaccination revealed increased relative level of T-lymphocytes (CD3+), T-helper cell subset (CD3+CD4+), increased absolute and relative level of activated CD3+CD25+ T-lymphocytes, but decreased absolute and relative level of natural killer (CD3-CD56+CD16+) and natural killer T-cell (CD3+CD56+CD16+) cell subsets as well as decreased CD147 receptor expression on T-lymphocytes. Similar patterns were also found while examining the immunogram in males exepting increased level of lymphocytes and lowered CD147 expression on both T- and B-lymphocytes. No changes in the parameters of the immune T-cell arm was found. The high efficacy of the vaccine was confirmed by development of SARS-CoV-2-specific class G antiviral antibodies in 97.5% and 92.3% of vaccinated females and males, respectively. The data obtained evidence that: 1) vaccination induces a specific humoral immune response determined three months post-vaccination, and 2) it caused no serious disturbances in the immune system functioning, which could be reflected in the peripheral blood lymphocyte subset composition. Thus, the data presented allow to conclude that Gam-COVID-Vac is effective vaccine against SARS-CoV-2 infection.Copyright © 2022 Saint Petersburg Pasteur Institute. All rights reserved.

POST-GAM-COVID-VAC COMBINED VECTOR VACCINE CELLULAR AND HUMORAL IMMUNE RESPONSE

Currently, as the SARS-CoV-2 pandemic evolves, there has been increasingly more attention paid to building natural and vaccine-induced immunity against SARS-CoV-2 and related disease known as COVID-19. Widespread preventive vaccination plays an important role in effectively protecting people from viral infections and can reduce national economic costs. Purpose - to study peripheral blood cell subset composition and magnitude of humoral response in vaccinated Gam-COVID-Vac subjects. The prospective study included 352 patients, of which 194 (119 women and 75 men) underwent an immunogram study and assessed level of anti-SARS-CoV-2 antibodies. In patients, the study of the lymphocyte subset composition and estimation of anti-SARS-CoV-2 antibodies was carried out at two time points - prior to vaccination and 90 days after inoculated component 1 of the Gam-COVID-Vac vaccine. In general, vaccination was well tolerated by patients, with no serious adverse events after immunization. The reaction to the vaccine (fever, malaise, headache, local reactions) was short-term (1-2 days) and more often noted after inoculated vaccine component 2. Comparatively analyzed immunogram parameters in females before and after vaccination revealed increased relative level of T-lymphocytes (CD3+), T-helper cell subset (CD3+CD4+), increased absolute and relative level of activated CD3+CD25+ T-lymphocytes, but decreased absolute and relative level of natural killer (CD3-CD56+CD16+) and natural killer T-cell (CD3+CD56+CD16+) cell subsets as well as decreased CD147 receptor expression on T-lymphocytes. Similar patterns were also found while examining the immunogram in males exepting increased level of lymphocytes and lowered CD147 expression on both T- and B-lymphocytes. No changes in the parameters of the immune T-cell arm was found. The high efficacy of the vaccine was confirmed by development of SARS-CoV-2-specific class G antiviral antibodies in 97.5% and 92.3% of vaccinated females and males, respectively. The data obtained evidence that: 1) vaccination induces a specific humoral immune response determined three months post-vaccination, and 2) it caused no serious disturbances in the immune system functioning, which could be reflected in the peripheral blood lymphocyte subset composition. Thus, the data presented allow to conclude that Gam-COVID-Vac is effective vaccine against SARS-CoV-2 infection.

POST-GAM-COVID-VAC COMBINED VECTOR VACCINE CELLULAR AND HUMORAL IMMUNE RESPONSE.

Currently, as the SARS-CoV-2 pandemic evolves, there has been increasingly more attention paid to building natural and vaccine-induced immunity against SARS-CoV-2 and related disease known as COVID-19. Widespread preventive vaccination plays an important role in effectively protecting people from viral infections and can reduce national economic costs. Purpose - to study peripheral blood cell subset composition and magnitude of humoral response in vaccinated Gam-COVID-Vac subjects. The prospective study included 352 patients, of which 194 (119 women and 75 men) underwent an immunogram study and assessed level of anti-SARS-CoV-2 antibodies. In patients, the study of the lymphocyte subset composition and estimation of anti-SARS-CoV-2 antibodies was carried out at two time points - prior to vaccination and 90 days after inoculated component 1 of the Gam-COVID-Vac vaccine. In general, vaccination was well tolerated by patients, with no serious adverse events after immunization. The reaction to the vaccine (fever, malaise, headache, local reactions) was short-term (1-2 days) and more often noted after inoculated vaccine component 2. Comparatively analyzed immunogram parameters in females before and after vaccination revealed increased relative level of T-lymphocytes (CD3+), T-helper cell subset (CD3+CD4+), increased absolute and relative level of activated CD3+CD25+ T-lymphocytes, but decreased absolute and relative level of natural killer (CD3-CD56+CD16+) and natural killer T-cell (CD3+CD56+CD16+) cell subsets as well as decreased CD147 receptor expression on T-lymphocytes. Similar patterns were also found while examining the immunogram in males exepting increased level of lymphocytes and lowered CD147 expression on both T- and B-lymphocytes. No changes in the parameters of the immune T-cell arm was found. The high efficacy of the vaccine was confirmed by development of SARS-CoV-2-specific class G antiviral antibodies in 97.5% and 92.3% of vaccinated females and males, respectively. The data obtained evidence that: 1) vaccination induces a specific humoral immune response determined three months post-vaccination, and 2) it caused no serious disturbances in the immune system functioning, which could be reflected in the peripheral blood lymphocyte subset composition. Thus, the data presented allow to conclude that Gam-COVID-Vac is effective vaccine against SARS-CoV-2 infection. Copyright © 2022 Saint Petersburg Pasteur Institute. All rights reserved.

EFFECT OF COVID-19 VACCINATION ON THE IMMUNE STATUS AND AUTOANTIBODY PROFILE IN WOMEN OF REPRODUCTIVE AGE.

In the context of the COVID-19 pandemic, scientific interest is growing in studying the impact of the proposed vaccination on women's reproductive health. As is known, alterations in the state of the immune system and activation of an autoimmune response can lead to reproductive failure in women and potential complications of subsequent pregnancy. Objective(s): to evaluate the effect of the "Gam-COVID-Vac" on the immune status parameters, the relationship of their changes and the specific immune response to vaccination with the dynamics of the level of autoantibodies in women of reproductive age. The prospective study included 120 women who were vaccinated against COVID-19 with the "Gam-COVID-Vac". The criteria for inclusion in the study were: the age from 18 to 49 years, the absence of COVID-19 in the anamnesis, a negative result of a study on SARS-CoV-2 by PCR and negative results of tests for antibodies of classes G and M to SARS-CoV-2 before vaccination, the absence of pregnancy and serious somatic diseases. The patients were examined twice: immediately before vaccination and 90-100 days after the introduction of the 1st component of the vaccine. The level of IgG antibodies to SARS-CoV-2 after vaccination was assessed using ELISA. Before and after vaccination, the levels of antiphospholipid, anti-nuclear, organ-specific and antihormonal autoantibodies were determined, peripheral blood lymphocytes were immunophenotyped to determine the main subpopulations (CD3, CD4, CD8, CD19, CD5, CD16, CD56), as well as the expression of activation markers of lymphocytes (HLA-DR, CD25, CD147) using monoclonal antibodies. The effectiveness and safety of the combined vector vaccine against COVID-19 were high. Specific IgG antibodies to SARS-CoV-2 were produced in 98.3% of vaccinated women, no serious adverse reactions were observed. After vaccination, there was an increase in the level of some autoantibodies within the reference ranges, only IgM antibodies to phosphatidylethanolamine (PE) and IgG antibodies to DNA increased above the reference values. However, this increase was transient. After vaccination, the following changes in the parameters of the immunogram were observed: an increase in the content of cells with CD3+CD25+, CD19+ phenotype in peripheral blood and a decrease in the content of cells with CD56+CD16+ phenotype within the reference ranges, a decrease in CD147+/CD3+. Weak correlations were noted between these changes in immunogram parameters and the levels of some autoantibodies. The specific antiviral immune response to vaccination did not correlate with the autoimmune response. Vaccination with "Gam-COVID-Vac" is effective and safe and does not lead to disorders in the immune system. The observed increase in the level of autoantibodies to PE and DNA is transient. Changes in the parameters of the immune status within the reference ranges may be due to vaccination and the development of a specific antiviral immune response. Copyright © 2022, SPb RAACI.

POST-GAM-COVID-VAC COMBINED VECTOR VACCINE CELLULAR AND HUMORAL IMMUNE RESPONSE

Currently, as the SARS-CoV-2 pandemic evolves, there has been increasingly more attention paid to building natural and vaccine-induced immunity against SARS-CoV-2 and related disease known as COVID-19. Widespread preventive vaccination plays an important role in effectively protecting people from viral infections and can reduce national economic costs. Purpose - to study peripheral blood cell subset composition and magnitude of humoral response in vaccinated Gam-COVID-Vac subjects. The prospective study included 352 patients, of which 194 (119 women and 75 men) underwent an immunogram study and assessed level of anti-SARS-CoV-2 antibodies. In patients, the study of the lymphocyte subset composition and estimation of anti-SARS-CoV-2 antibodies was carried out at two time points - prior to vaccination and 90 days after inoculated component 1 of the Gam-COVID-Vac vaccine. In general, vaccination was well tolerated by patients, with no serious adverse events after immunization. The reaction to the vaccine (fever, malaise, headache, local reactions) was short-term (1-2 days) and more often noted after inoculated vaccine component 2. Comparatively analyzed immunogram parameters in females before and after vaccination revealed increased relative level of T-lymphocytes (CD3+), T-helper cell subset (CD3+CD4+), increased absolute and relative level of activated CD3+CD25+ T-lymphocytes, but decreased absolute and relative level of natural killer (CD3-CD56+CD16+) and natural killer T-cell (CD3+CD56+CD16+) cell subsets as well as decreased CD147 receptor expression on T-lymphocytes. Similar patterns were also found while examining the immunogram in males exepting increased level of lymphocytes and lowered CD147 expression on both T- and B-lymphocytes. No changes in the parameters of the immune T-cell arm was found. The high efficacy of the vaccine was confirmed by development of SARS-CoV-2-specific class G antiviral antibodies in 97.5% and 92.3% of vaccinated females and males, respectively. The data obtained evidence that: 1) vaccination induces a specific humoral immune response determined three months post-vaccination, and 2) it caused no serious disturbances in the immune system functioning, which could be reflected in the peripheral blood lymphocyte subset composition. Thus, the data presented allow to conclude that Gam-COVID-Vac is effective vaccine against SARS-CoV-2 infection. Copyright © 2022 Saint Petersburg Pasteur Institute. All rights reserved.

EFFECT OF COVID-19 VACCINATION ON THE IMMUNE STATUS AND AUTOANTIBODY PROFILE IN WOMEN OF REPRODUCTIVE AGE

In the context of the COVID-19 pandemic, scientific interest is growing in studying the impact of the proposed vaccination on women's reproductive health. As is known, alterations in the state of the immune system and activation of an autoimmune response can lead to reproductive failure in women and potential complications of subsequent pregnancy. Objective: to evaluate the effect of the "Gam-COVID-Vac” on the immune status parameters, the relationship of their changes and the specific immune response to vaccination with the dynamics of the level of autoantibodies in women of reproductive age. The prospective study included 120 women who were vaccinated against COVID-19 with the "Gam-COVID-Vac”. The criteria for inclusion in the study were: the age from 18 to 49 years, the absence of COVID-19 in the anamnesis, a negative result of a study on SARS-CoV-2 by PCR and negative results of tests for antibodies of classes G and M to SARS-CoV-2 before vaccination, the absence of pregnancy and serious somatic diseases. The patients were examined twice: immediately before vaccination and 90-100 days after the introduction of the 1st component of the vaccine. The level of IgG antibodies to SARS-CoV-2 after vaccination was assessed using ELISA. Before and after vaccination, the levels of antiphospholipid, anti-nuclear, organ-specific and antihormonal autoantibodies were determined, peripheral blood lymphocytes were immunophenotyped to determine the main subpopulations (CD3, CD4, CD8, CD19, CD5, CD16, CD56), as well as the expression of activation markers of lymphocytes (HLA-DR, CD25, CD147) using monoclonal antibodies. The effectiveness and safety of the combined vector vaccine against COVID-19 were high. Specific IgG antibodies to SARS-CoV-2 were produced in 98.3% of vaccinated women, no serious adverse reactions were observed. After vaccination, there was an increase in the level of some autoantibodies within the reference ranges, only IgM antibodies to phosphatidylethanolamine (PE) and IgG antibodies to DNA increased above the reference values. However, this increase was transient. After vaccination, the following changes in the parameters of the immunogram were observed: an increase in the content of cells with CD3+CD25+, CD19+ phenotype in peripheral blood and a decrease in the content of cells with CD56+CD16+ phenotype within the reference ranges, a decrease in CD147+/CD3+. Weak correlations were noted between these changes in immunogram parameters and the levels of some autoantibodies. The specific antiviral immune response to vaccination did not correlate with the autoimmune response. Vaccination with "Gam-COVID-Vac” is effective and safe and does not lead to disorders in the immune system. The observed increase in the level of autoantibodies to PE and DNA is transient. Changes in the parameters of the immune status within the reference ranges may be due to vaccination and the development of a specific antiviral immune response. © 2022, SPb RAACI.

IMPACT OF THE NOVEL CORONAVIRUS INFECTION (COVID-19) EXPERIENCED BY PREGNANT WOMEN ON THE HEALTH OF NEWBORNS

Background: Due to the high spread rate of SARS-CoV-2 and to the rapid increase in its incidence, including those among pregnant women, the novel coronavirus infection (COVID-19) has become a challenge in modern healthcare. Objective(s): To analyze the impact of the novel coronavirus infection experienced by pregnant women on the health of newborns in the early neonatal period. Material(s) and Method(s): A retrospective analysis was carried out of the birth records of 400 women who had experienced the novel coronavirus infection during pregnancy and the neonatal records of their newborns (n=500) who received health care in the clinical units of the V.I. Kulakov National Medical Research Center of Obstetrics, Gynecology, and Perinatology, Ministry of Health of Russia (Center), in July 2020 to July 2021. A comparison group consisted of randomly selected birth records of 495 pregnant women who had not been infected with COVID-19 and the neonatal records of their babies (n=500) born at the same Center at the same time. Result(s): The vast majority of women who had been infected with COVID-19 during pregnancy were found to have familial obstetric/gynecological and/or somatic histories. Among the factors aggravating pregnancy in the presence of COVID-19, chronic hypertension, hereditary thrombophilia, fat metabolism disorders, urogenital infections, and anemia are more common than those in the control group (p<0.05). This female group also tended to have miscarriage;however, no statistically significant differences could be detected (p=0.06). There were no statistically significant differences in the term and frequency of cesarean delivery in pregnant women in the study and control groups (p>0.05). Neonates born to women who had been infected with COVID-19 in the first trimester had its statistically significantly higher morbidity rates (p<0.05). The frequency of perinatal complications was higher in newborns whose mothers had experienced the novel coronavirus infection in the first trimester. Neonatal infants borns from women who had a new coronavirus infection in the third trimester, rhinitis and otitis media are statistically significantly more common in the early neonatal period. Among the factors leading to disruption of early neonatal adaptation of children whose mothers had a new coronavirus infection during pregnancy, the following were statistically significantly more common: infectious and inflammatory diseases (rhinitis, otitis media), hemorrhagic syndrome, and hypoglycemia (p<0.05). Neonates born to women who had been infected with COVID-19 in the first trimester were observed to have statistically significantly higher morbidity rates (p<0.05). The incidence of perinatal complications was higher in newborns whose mothers had experienced the novel coronavirus infection in the first trimester. Neonatal infants born to women who had the novel coronavirus infection in the third trimester were statistically significantly more commonly recorded to have rhinitis and otitis media in the early neonatal period. Among the factors leading to failure of early neonatal adaptation of babies whose mothers had the novel coronavirus infection during pregnancy, there were statistically significantly more often infectious and inflammatory diseases (rhinitis, otitis media), hemorrhagic syndrome, and hypoglycemia (p<0.05). Conclusion(s): The incidence of perinatal complications in babies born to women who had been infected with COVID-19 depended on their gestational age and was higher than that in newborns whose mothers had experienced the novel coronavirus infection in the first trimester. At the same time, the incidence of infectious and inflammatory diseases proved to be higher in infants whose mothers had a coronavirus infection in the third trimester. Failure of early neonatal adaptation of babies born to women who had an infection caused by SARS-CoV-2 during pregnancy may be due to both infectious and non-infectious factors that complicate the course of pregnancy and childbirth. Copyright © A group of authors, 2022.

SYSTEMIC OZONE THERAPY AND ITS IMMUNOMODULATORY EFFECTS IN COVID-19 PATIENTS

Objective: To evaluate the effect of systemic ozone therapy (OT) on the concentration of pro-inflammatory and anti-inflammatory cytokines in the blood in the complex treatment of COVID-19 patients. Material(s) and Method(s): The study included 65 patients with a confirmed diagnosis COVID-19 characterized by a moderate and severe course of the disease. The patients were admitted to the Infectious Disease Hospital of the V.I. Kulakov National Medical Research Centre for Obstetrics, Gynecology and Perinatology, Ministry of Health of Russia. The patients' age ranged from 29 to 78 years. All patients were treated in accordance with the Temporary Guidelines of the Ministry of Health of Russia "Prevention, Diagnosis and Treatment of Coronavirus Infection (COVID-19)". Two groups of patients were randomly formed. The first group included 35 patients whose complex therapy included OT: intravenous administration of 400 ml of ozonated saline solution with an ozone concentration of 4 mg/L;a total course consisting of 6 procedures performed every other day. The second group included 30 patients who did not have OT. Clinical and laboratory parameters were evaluated on admission to the Infectious Disease Hospital and after two weeks of complex treatment;clinical, laboratory, special, statistical research methods were used. The content of cytokines GM-CSF, IFN-gamma, TNF-alpha, IL-2, IL-4, IL-6, IL-8, IL-10 and their ratios were determined with a multiplex method on the Bioplex 200 analyzer (Bio-Rad, USA) using Bio-Plex Pro Human Cytokine 8-plex Panel (Bio-Rad, USA). Result(s): On admission to the Infectious Disease Hospital 47/65 (72,3%) patients had a moderate course of the disease and 18/65 (27.7%) patients had a severe one. The length of hospital stay in the group of patients with OT averaged 12.2 (2.7) (8-17) days, and it was 17.9 (4.2) (12-26) days in the group of patients who did not have OT. On admission, all patients had an increase in the level of C-reactive protein in their blood serum;the cytokine content in patients of the groups differed from the initial cytokine level and it was different between groups after two weeks of therapy. The medians of the pro-inflammatory cytokines IL-2, IL-6, IL-8 were particularly different. The content of these cytokines remained elevated in the second group of patients who did not have OT, compared with the baseline data and compared with the first group. The study of anti-inflammatory cytokines showed that the patients of the first group who had OT demonstrated a significantly higher IL-10 content compared to IL-10 content in the patients of the second group. Ratios of pro-and anti-inflammatory cytokines IL-2/IL-10, IL-2/ IL-4, IL-6/IL-10, IL-6/IL-4, IL-8/IL-10, IL-8/IL-4, TNF-alpha/IL-4 in the first group of patients with OT significantly decreased compared to the baseline indicator, which can be suggestive of a marked decrease in the activity of the inflammatory process. Conclusion(s): The positive effect of systemic OT on the clinical course of the disease has been revealed. Laboratory indicators in COVID-19 patients have shown a decrease in the severity of the inflammatory response. Besides having anti-inflammatory and immunomodulatory effect, OT helps to stop the process, improve the condition of patients and reduce the length of hospital stay. Systemic OT should be considered as an additional adjuvant method in the complex treatment of patients with SARS-CoV-2 infection. Copyright © A group of authors, 2022.

PROFILE OF ANTIPHOSPHOLIPID ANTIBODIES AND COMPLEMENT SYSTEM IN COVID-19 PATIENTS OF DIFFERENT SEVERITY

COVID-19, a severe acute respiratory syndrome caused by SARS-CoV-2, may predispose to thrombotic events, especially when combined with antiphospholipid antibodies (aPL). However, there are limited data on prevalence and antigenic specificity of aPL in COVID-19. Complement activation is assumed to play an important role in pathogenesis of COVID-19-associated coagulopathy. During the SARS-CoV-2 pandemic, it is necessary to identify important biomarkers for predicting severe course of COVID-19 and risk of thrombotic complications. Our objective was to evaluate the aPL profile, quantitative content and activity of complement and its components in COVID-19 patients graded by severity in the course of time. IgM and IgG antibodies to cardiolipin (CL), phosphatidylserine (PS), β2-glycoprotein-I (β2-GP-I), prothrombin (PT), annexin V (An V), as well as C1q complement component, content of its C3 and C4 components and total complement activity were determined in blood serum using ELISA approach. 141 patients with COVID-19 were included in the study. Group 1 consisted of 39 patients with mild form, group 2 (65 patients) presented with moderate form, and group 3 included 37 patients with severe form of COVID-19. Blood samples were obtained on day 3-7 of the disease (1st point) and after 14-28 days (2nd point). The results were as follows: aPL were detected in 29.1% of the total COVID-19 cohort, frequency of aPL detection by the severity grade did not differ (33.3%, 24.6% and 32.4%). In 8.5% of the patients, aPL were detected only at the 1st time point;in 14.2%, only at the 2nd point;and in 6.4% of the cases, at the both time points. Antibodies to PT (16.3%) and An V (11.3%) were revealed more frequently. The detection frequency of antibodies to PT was significantly higher than antibodies to CL and PS (7.1%), β2-GP-I (7.8%). The prevalence of aPL in groups 1 and 3 did not differ. At the 1st point in group 3, increased levels of C4 (89.2%) and C3 (24.3%) in blood, and a decrease in complement activity (35.1%) were more often observed than in group 1. At the 2nd time point in group 3, a decrease in complement activity was often detected (59.5%). The C3 levels exceeding 720 μg/ml were found to predict a 2.6-fold increased risk of severe COVID-19, and this risk became 3.3 times higher at C4 levels of > 740 μg/ml. The antibodies to PT and An V are often detected in COVID-19 patients, along with low prevalence of antibodies to CL and β2-GP-I. These antibodies can be involved in pathogenesis of COVID-19-associated coagulopathy, being detectable at the late stage of the disease, and they may trigger APS in predisposed patients and reconvalescents. Although presence of aPL antibodies is not associated with COVID-19 severity, their persistence over the period of convalescence may be an additional risk factor for thromboembolic complications. The COVID-19 patients are characterized by activation of the complement system, which increases in severe cases, and manifests with increased or decreased levels of C3 complement component, increased levels of C4 component in blood, and a decreased total complement activity. Quantitative determination of C3 and C4 complement components over the period of COVID-19 progression is of prognostic value, with respect to severity of the disease.

EFFECTS OF COVID-19 VECTOR VACCINE ON AUTOANTIBODY PROFILE IN REPRODUCTIVE AGE WOMEN

Autoimmune mechanisms have been implicated in the negative effects of vaccines on female reproductive health. This study evaluates the endogenous levels of self-reactive antibodies and ovarian reserve-associated hormones before and after immunization with the domestically developed Gam-COVID-Vac combined vector vaccine to check for possible reproductive sequelae. The prospective study enrolled 120 women aged 18–49, subject to vaccination with Gam-COVID-Vac. Ovarian reserve was assessed prior to vaccination and 90 days after the first component injection. Profiles of specific antibodies to self-antigens, including phospholipids, nuclear antigens, FSH, progesterone, and also thyroid, ovarian, trophoblast, and zona pellucida antigens, were assessed at the same time points by enzyme immunoassay. Overall, the vaccination had no effect on the levels of ovarian reserve-associated hormones and autoantibodies, apart from a transient increase in positivity for antiphosphatidylethanolamine IgM and anti-dsDNA IgG. Seroprevalence of elevated serum autoantibodies constituted 70.8% before and 75% after vaccination. According to the results, immunization with Gam-COVID-Vac does not affect ovarian reserve or autoimmune status, thus being safe for the female reproductive potential.

Immune status of covid-19 patients with different disease severity

Aim. To investigate the immune status and compare immunological parameters in COVID-19 patients with different disease severity. Materials and methods. The prospective study included 62 patients with COVID-19. The patients were stratified into three groups based on the disease severity, including mild (group 1, n=29), moderate (group 2, n=17), and severe (group 3, n=16) forms of COVID-19. On days 3–7 from the onset of the disease, peripheral blood lymphocytes were phenotyped by flow cytometry. Cytokine concentrations were measured using a multiplex immunoassay-standard 48-plex Bio-Plex Pro™ Human Cytokine Screening test system (Bio-Rad, USA) on a flow-based laser immuno-analyzer Bio-Plex 200. Results. Patients with severe COVID-19 had higher levels of leukocytes, neutrophils, CRP, and lower relative and absolute lymphocyte counts. There were low counts of CD3+, CD3+CD4+, CD3+CD8+, and T-lymphocytes expressing the activation marker HLA-DR (CD3+HLA-DR+), NK-cells, and PAN. In group 3, changes in 39 of the 48 investigated soluble factors were observed. Conclusion. High levels of leukocytes, neutrophils, CRP, neutrophilic-leukocyte index, low levels of absolute and relative lymphocyte counts, pronounced changes in immunological parameters, a systemic inflammatory reaction associated with the release of mediators called cytokines ("cytokine storm") predispose to a severe course of COVID-19. © A group of authors, 2021.

Gam-covid-vac (Sputnik v) vaccine has no adverse effect on ovarian reserve in reproductive-age women

Relevance. There is limited evidence on the effect of various vaccines on the human reproductive system. Potential adverse effects of vaccines on fertility are associated with autoimmune disorders, which might cause gonadal damage. Currently, several studies are underway investigating the impact of COVID-19 vaccines on human fertility. To our best knowledge, there is only one study published which demonstrated no adverse effects of the COVID-19 vaccine on assisted reproductive technology outcomes. Aim. To investigate the effect of the Gam-COVID-Vac (Sputnik V) vaccine on ovarian reserve and antiphospholipid antibody level in reproductive-age women. Materials and methods. The prospective study included 51 women vaccinated against COVID-19 with Gam-COVID-Vac (Sputnik V) vaccine. The inclusion criteria were age from 18 to 45, preserved menstrual function, normal ovarian reserve, no history of COVID-19, negative PCR test result for SARS-CoV-2 and negative SARS-CoV-2 IgG antibody test before vaccination, no pregnancy, and no history of serious illnesses. Clinical evaluation was carried out twice-immediately before immunization and 90 days after the first vaccine component administration. The antral follicle count was measured by pelvic ultrasound. Serum levels of AMH, FSH, TSH, estradiol, antiphospholipid antibodies (aPL) M and G isotypes against cardiolipin (aCL), β2-glycoprotein-1 (aβ2-GP-1), annexin V (aAn V), phosphatidylserine (aPS), and IgG antibodies against SARS-CoV-2 were measured by enzyme immunoassay. Results. There were no significant changes in hormones levels and antral follicle counts before and after vaccination, including in women of advanced reproductive age (≥37 years). After immunization, aPL antibody levels did not differ significantly from the baseline. There was no correlation between aPL antibody level dynamics and level of FSH and AMH, which indirectly demonstrates no possible autoimmune effect of vaccination on women's fertility. Conclusion. This is the first study investigating the effect of the Gam-COVID-Vac vaccine on ovarian reserve parameters and aPL antibody levels. The preliminary results prove that the Gam-COVID-Vac vaccine in women of reproductive age does not adversely impact ovarian reserve.

Gam-covid-vac (Sputnik v) vaccine has no adverse effect on spermatogenesis in men

Relevance. During the pandemic and large-scale vaccination against COVID-19, studies of the effect of various types of vaccines on human reproductive function have become particularly important. Vaccination may have a negative impact on the reproductive organs and tissues due to the development of an autoimmune process involving the gonads. Currently, there are some studies on the effect of vaccination against coronavirus infection caused by SARS-CoV2 on the male and female reproductive function. The absence of a negative effect of vaccination on spermatogenesis in men has already been demonstrated in the results of three studies. Objective. To evaluate the effect of the Gam-COVID-Vac (Sputnik V) vaccine on spermatogenesis and the level of antiphospholipid antibodies in men. Materials and methods. The prospective study included 45 men who were vaccinated against COVID-19 with Gam-COVID-Vac (Sputnik V). The criteria for inclusion in the study were the age from 18 to 55 years, the absence of a previous history of COVID-19, a negative RT-PCR result in SARS-CoV-2 testing, negative SARS-CoV-2 IgG results prior to vaccination, and the absence of severe somatic diseases. The patients were examined twice: immediately before vaccination and 90 days after the first dose was injected. We evaluated spermogram parameters, determined the levels of FSH, LH, TSH and total testosterone, and analyzed blood for IgM and IgG antiphospholipid antibodies (aPL) to cardiolipin (aCL), β2-glycoprotein-1 (aß2-GP1), annexin V (aAnV), phosphatidylserine (aPS), as well as IgG antibodies to SARS-CoV-2 using enzyme immunoassay. Results. There were no significant changes in the parameters of spermogram and hormone levels before and after vaccination;men with moderate impairment of spermatogenesis (oligoasteno-and/or teratozoospermia) did not show considerable changes either. The level of aPL after vaccination did not increase compared to the baseline level. Besides, there was no correlation between the dynamics of the level of antisperm antibodies, aPL and spermogram parameters. These findings indirectly indicate that there are no changes in the reproductive function associated with this autoimmune factor after vaccination in men. Conclusion. This is one of the first studies that has revealed the effect of the Gam-COVID-Vac vaccine on spermatogenesis, hormonal profile and aPL level in men. The obtained results demonstrate that the Gam-COVID-Vac vaccine does not have a negative effect on the reproductive function in men.

Plasma hemostatic system in patients with the novel coronavirus disease 2019 (COVID-19)

Aim. To investigate plasma hemostasis in patients with different levels of COVID-19 severity. Materials and methods. The study included 46 patients with confirmed COVID-19, who were stratified into four groups based on disease severity as mild (1), moderate (2), severe (3), and critically severe (4). Laboratory investigations included APTT, Quick's prothrombin time, concentration of D-dimer, fibrinogen, ATIII, and platelet count. Results. D-dimer concentration >450 ng/ml was found in 100% of patients increasing from 973 (545–1635) ng/ml in group 1 to 13513 (7627–22512) ng/ml in group 4 (p<0.05). Patients in group 2 had the highest fibrinogen level averaging 6.1(3.9–6.4) g/L. The highest and lowest platelet counts were in groups 1 (323x109) and 4 (155×109), respectively, (p <0.05). The ATIII levels in groups 1 and 2 were within the normal range, while in groups 3 and 4, they were decreased to 76% and 64%, respectively (p <0.05). All patients enrolled in the study had normal values of APTT and Quick's prothrombin time. Unfavorable outcomes were observed at a D-dimer concentration> 3633 ng/ml (specificity 92.3%, sensitivity 100%, AUC 0.94, p <0.001, positive predictive value 99.1%, and negative predictive value 100%) and ATIII <70.8% (specificity 70%, sensitivity 82.6%, AUC 0.85, p <0.001, positive predictive value 30.5%, and negative predictive value 96.1%). Conclusion. The levels of D-dimer, fibrinogen, and ATIII were found to be associated with the severity of COVID-19.

The impact of the immune status on COVID-19 severity

Aim. To investigate the impact of patient immune status on the severity of COVID-19. Materials and methods. The prospective study included 63 employees of the V.I. Kulakov NMRC for OG&P of Minzdrav of Russia with confirmed COVID-19. The patients were stratified into three groups based on the disease severity, including asymptomatic (group 1, n=17), mild (group 2, n=29), and moderate (group 3, n=17) form of COVID-19. On days 3–7 from the onset of the disease, peripheral venous blood samples were collected from the study subjects and tested for serum levels of anti-SARS-CoV-2 IgG antibodies and immune profile by ELISA. After day 20+, testing for serum levels of anti-SARS-CoV-2 IgG antibodies was repeated using ELISA. Results. Patients who had a higher BMI, blood group A(II), lower leukocyte and lymphocyte counts, higher relative monocyte count, changes in the immune profile in the form of a lower number of CD3+, CD3+ CD8+, СD19+, CD19+ CD5+, and phagocytic activity of neutrophils, developed more severe forms of COVID-19. They had severe clinical manifestations of the disease, and 100% of them developed antiviral immunity. Conclusion. This study identified several clinical, laboratory, and immune profile features that may be considered as predictive factors of severe COVID-19 and can be used in clinical practice to predict the clinical course of the disease.