Anti-Interferon Alpha Autoantibodies and Their Significance in Covid-19

During the last two years, treatment of patients with novel coronavirus infection COVID-19 remains an urgent health problem. Interferon proteins are known to play a significant role in antiviral immunity. Some pathological conditions are accompanied by production of neutralizing autologous immunoglobulins against own host interferons (autoIFN-Abs). There is evidence that autoantibodies against interferons alpha and omega are detected in patients with life-threatening course of COVID-19 pneumonia. The aim of our study was to analyze prevalence of autoantibodies against interferon alpha in patients with COVID-19 coronavirus infection and assess their impact on clinical course of the disease. We examined 70 patients with severe COVID-19, who received inpatient treatment at the intensive care units. Serum autoantibodies against interferon alpha were determined on day 8-50 after disease onset by using solid-phase enzyme immunoassay ( ELISA). Patients were divided into 2 groups: those with and without (group 2) autoantibodies against interferon alpha (group 1). Anti-COVID serum from 57 donors was used a control. Among patients, autoantibodies against interferon alpha were detected in 13 (18%) subjects, which level ranged from 26.8 to 1000 ng/ml. Among donors, autoIFN-Abs were detected in 5 (8.8%) subjects at trace concentrations (from 1.65 to 12.0 ng/ml). Respiratory failure developed significantly more often in patients with auto-IFN-Abs. While analyzing laboratory parameters, it was noted that the concentration of C-reactive protein was significantly higher in the group of patients with auto-IFN-Abs. Mortality rate of patients with high auto-IFN-Abs levels was 60%. In conclusion, it was found that serum autoantibodies against IFN alpha in COVID-19 patients caused lung damage that significantly more often required hardware respiratory support, so comparable by duration with it for patients without auto-IFN-Abs. High concentrations of auto-IFN-Abs (more than 100 ng/ml) in patients with COVID-19 can be considered as a predictor of unfavorable disease outcome.

Regulation of lymphocyte apoptosis in intensive care patients with COVID-19

Severity of a new coronavirus infection (COVID-19) caused by the SARS-COV-2 virus is largely due to abnormal immune condi-tion in these patients. Lymphopenia is observed in 85% of patients with severe COVID-19 that may be associated with enhanced apoptosis of lymphocytes. Objective. To analyze apoptotic death of lymphocytes and changes in proteins regulating apoptosis in patients with severe COVID-19. Material and methods. We analyzed 93 ICU patients. All patients were divided into three groups depending on severity and outcomes of disease: group 1 consisted of 53 patients with favorable course and outcomes of disease, group 2 included 26 patients with unfavorable course and favorable outcomes of disease, group 3 included 14 patients with unfavorable course and outcomes of disease. Blood sampling for analysis of apoptosis markers was carried out in 5-12 and 14-18 days after clinical manifestation of disease. Quantitative parameters of lymphocyte apoptosis were evaluated using flow cytometry. Regulatory proteins of apop-tosis (phosphorylated AKT, JNK, BAD, BCL-2, p-53, active caspase 8 and 9) were determined on the Luminex platform. We also assessed concentration of leukocytes, relative and absolute lymphocyte count, concentration of C-reactive protein (CRP), procal-citonin and lactate dehydrogenase. Results. Study groups significantly differed in NEWS score (p=0.001), SOFA score (p=0.001), CRP level (p=0.001), severity of lymph-openia (p=0.001) and level of CD14+HLA-DR+ monocytes (p=0.001). Quantitative parameters of lymphocyte apoptosis did not cor-relate with lymphopenia. The highest rates of lymphocyte apoptosis were observed in patients with favorable course and outcomes of disease. There was no correlation between concentration of lymphocytes in venous blood and level of proteins regulating apoptosis. Conclusion. Patients with severe COVID-19 are characterized by abnormal induction of lymphocyte apoptosis through external and internal activation pathways in response to viral aggression. In deceased patients, pro-apoptotic factors prevailed while activity of anti-apoptotic factors was decreased. © 2023, Media Sphera Publishing Group. All rights reserved.

Effect of vitamin D (25-OH D3 ) concentration on the course and outcomes of COVID-19 in intensive care patients

Objective. To study the effect of vitamin D concentration on the course and outcomes of COVID-19 in intensive care patients. Material and methods. A prospective study included 43 ICU patients admitted to the Sklifosovsky Research Institute for Emergency Care with a confirmed diagnosis of a novel coronavirus infection. Single blood sampling was performed in 1—2 days after admission for analysis of 25-OH D3 concentration. Patients were divided into 2 groups with normal and reduced level of vitamin D. We analyzed chest CT data and laboratory parameters at admission and at the peak of cytokine storm. Administration of IL-6 antagonists and IL-6 receptor blockers was compared. We also compared the need for respiratory support, period between the onset of disease and respiratory therapy, its total duration and type of respiratory support, incidence of complications (sepsis, multiple organ failure, nosocomial pneumonia, acute kidney injury, thrombosis and hemorrhagic events). We analyzed the outcomes of disease, length of ICU-stay and hospital-stay. Results. Patients with normal vitamin D level at admission were characterized by lower serum C-reactive protein (15.41±7.96 vs. 63.271±11.988;p=0.029), while lymphocyte count (1.69±0.49 vs. 0.921±0.0719;p=0.029), leukocyte count (7.88±1.33 vs. 5.305±0.468;p=0.041) and serum lactate dehydrogenase (454.50±102.5 vs. 271.315±16.867;p=0.024) were higher. Minimum daily saturation at the peak of cytokine storm was higher in patients with normal vitamin D level (95.2±1.39 vs. 90.947±0.9585;p=0.044). Conclusion. Patients with vitamin D deficiency have a more pronounced inflammatory response in initial stages of disease and more severe lung damage at the stage of cytokine storm. However, this does not increase aggressiveness and duration of respiratory support, ICU-and hospital-stay. There was no worsening of the outcomes too. © S.S. PETRIKOV1, G.E. SAVKOV1, M.A. GODKOV1, N.V. BOROVKOVA1, A.M. KVASNIKOV1, D.A. LEBEDEV1, K.V. KISELEV2, K.A. POPUGAEV1.

ANTI-INTERFERON ALPHA AUTOANTIBODIES AND THEIR SIGNIFICANCE IN COVID-19

During the last two years, treatment of patients with novel coronavirus infection COVID-19 remains an urgent health problem. Interferon proteins are known to play a significant role in antiviral immunity. Some pathological conditions are accompanied by production of neutralizing autologous immunoglobulins against own host interferons (auto-IFN-Abs). There is evidence that autoantibodies against interferons alpha and omega are detected in patients with life-threatening course of COVID-19 pneumonia. The aim of our study was to analyze prevalence of autoantibodies against interferon alpha in patients with COVID-19 coronavirus infection and assess their impact on clinical course of the disease. We examined 70 patients with severe COVID-19, who received inpatient treatment at the intensive care units. Serum autoantibodies against interferon alpha were determined on day 8–50 after disease onset by using solid-phase enzyme immunoassay (ELISA). Patients were divided into 2 groups: those with and without (group 2) autoantibodies against interferon alpha (group 1). Anti-COVID serum from 57 donors was used a control. Among patients, autoantibodies against interferon alpha were detected in 13 (18%) subjects, which level ranged from 26.8 to 1000 ng/ml. Among donors, auto-IFN-Abs were detected in 5 (8.8%) subjects at trace concentrations (from 1.65 to 12.0 ng/ml). Respiratory failure developed significantly more often in patients with auto-IFN-Abs. While analyzing laboratory parameters, it was noted that the concentration of C-reactive protein was significantly higher in the group of patients with auto-IFN-Abs. Mortality rate of patients with high auto-IFN-Abs levels was 60%. In conclusion, it was found that serum autoantibodies against IFN alpha in COVID-19 patients caused lung damage that significantly more often required hardware respiratory support, so comparable by duration with it for patients without auto-IFN-Abs. High concentrations of auto-IFN-Abs (more than 100 ng/ml) in patients with COVID-19 can be considered as a predictor of unfavorable disease outcome.

Hyperbaric oxygen therapy in patients with COVID-19

The aim of the study is to evaluate the efficacy of hyperbaric oxygen therapy and its effect on oxidative stress and apoptosis in patients with new coronavirus infection COVID-19. Materials and methods. 90 patients diagnosed with new coronavirus infection caused by SARS-CoV-2 virus were examined. Hyperbaric oxygen therapy sessions were conducted in 57 patients (38 in severe condition (CT 3–4), 19 in moderate condition (CT 1–2)). The procedures were performed in 1.4–1.6 ATA mode for 40 min-utes, 247 sessions in total were performed. The effect of hyperbaric oxygenation was assessed by measuring the level of oxygen saturation, the severity of oxidative stress and apoptosis of blood lymphocytes. Results. In all examined patients with new coronavirus infection caused by SARS-CoV-2, positive changes such as dyspnea reduction and improvement of general well-being were registered after hyperbaric oxygen therapy sessions. The level of oxygen saturation after the end of the hyperbaric oxygen therapy course was 95.0±1.6% (before the course — 91.3±5.9%), which allowed to return almost all patients to spontaneous res-piration without the need for further oxygenation therapy. Hyperbaric oxygen therapy did not reduce the total antioxidant activity, however, it was associated with a decrease in the blood malone dialdehyde from 4.34±0.52 µmol/l to 3.98±0.48 µmol/l and a decrease in open circuit potential of platinum electrode from-22.78±24.58 mV to-37.69±17.4 mV. Besides, the positive effect of hyperbaric oxygen therapy was manifested in normalization of blood cell apoptosis. Conclusion. Hyperbaric oxygen therapy in patients with new coronavirus infection caused by the SARS-CoV-2 virus is an effective treatment method with multiple effects resulting in improvement of subjective indicators of the patients' condition, increase of hemoglobin oxygen saturation, decrease of lipid peroxidation intensity, activation of antioxidant system, restoration of pro-and antioxidant balance and apoptosis normalization. © 2020, V.A. Negovsky Research Institute of General Reanimatology. All rights reserved.

Formation of a cohort of anticovid plasma donors with high-titer antibodies neutralizing SARS-CoV-2

Background. The application of convalescent plasma (CP) is currently seen as a feasible therapeutic approach in the treatment of COVID-19.Aim. To analyze the experience of recruiting a donor cohort from COVID-19 convalescents for banking of CP as part of a pilot project at the Moscow Healthcare Department.Materials and methods. A retrospective research included 493 COVID-19 convalescents as potential CP donors, all examined at the Sklifosovsky Research Institute for Emergency Medicine. CP was banked using the plasmapheresis method. Only those donors with a documented medical history of COVID-19, which was confi rmed by polymerase chain reaction of SARS-CoV-2 RNA in pharyngeal swabs, and no sooner than 14 days after complete recovery were eligible for donation. Viral neutralizing activity (VNA) was chosen as the key characteristic of the immunological viability of CP. All the donors having VNA titers were characterized in terms of gender, age, time interval since the disease onset, regression of clinical symptoms and clinical features of the COVID 19 course.Results. Effective (1:160 or more) and acceptable (1:80) VNA titers were found in 21.1 % and 24.75 % of donors, respectively. Signifi cant predictors for a donor having a high VNA titer included: male sex, age over 36 years and verifi ed viral pneumonia. The absence of a signifi cant body temperature response (38.5 °С) can be considered as a negative marker of a potential donor. Введение. Трансфузии плазмы антиковидной патогенредуцированной (ПАП) рассматривается как один из методов лечения COVID-19. Цель: проанализировать опыт формирования донорского резерва из реконвалесцентов COVID-19 для заготовки ПАП.Материалы и методы. В ретроспективное исследование включено 493 реконвалесцента COVID-19, проходивших обследование в ГБУЗ «НИИ скорой помощи им. Н. В. Склифосовского ДЗМ» в качестве потенциальных доноров ПАП. Заготовку ПАП осуществляли методом плазмафереза. К процедуре плазмодачи допускали доноров, у которых в анамнезе была перенесенная инфекция COVID-19, подтвержденная выявлением методом полимеразной цепной реакции РНК SARS-CoV-2 в мазках из глотки, наличие медицинской документации, срок с момента разрешения заболевания не менее 14 суток. В качестве характеристики иммунологической состоятельности ПАП был выбран титр вируснейтрализующих антител (ВНА). Сопоставлены характеристики доноров (пол, возраст, давность с момента начала заболевания и регресса клинической симптоматики, особенности течения COVID-19) с титром ВНА.Результаты. Эффективный титр ВНА (1:160 и более) выявлен у 21,10 % доноров, допустимый (1:80) - у 24,75 %. Значимыми предикторами высокого титра ВНА оказались: мужской пол донора, возраст старше 36 лет, наличие верифицированной вирусной пневмонии. Отсутствие значимой температурной реакции (38,5 °C) может рассматриваться как отрицательный маркер для привлечения потенциального донора. Заключение. Для получения ПАП с высокими титрами ВНА оптимально привлечение в качестве доноров-реконвалесцентов мужчин, переболевших COVID-19 с клинической картиной вирусной пневмонии и значимой температурной реакцией.