Characteristics and Clinical Implications of Anti-IFN-alpha cytokine antibodies in partial Recombinase Activating Gene Deficiency patients before and during the COVID-19 Pandemic

Introduction: With the onset of the COVID-19 pandemic, there was increased attention on anti- IFN-alpha autoantibodies and its correlation with severe clinical outcomes in a large group of patients. However, this correlation has not been extensively investigated in patients with partial Recombinase Activating Gene Deficiency (pRD) who are known to have increased prevalence of anti- IFN-alpha autoantibodies. Therefore, there is a need to assess the presence of anti- IFN-alpha antibodies in pRD patients before and after the COVID-19 pandemic and explore the relationship between anti- IFN-alpha antibody presence and clinical outcomes. Method(s): Sera was collected from the whole blood after informed consent and Enzyme-Linked Immunosorbent Assay was conducted to confirm the presence of IgG-specific anti- IFN-alpha autoantibodies. Positive samples were determined as OD values above 3 standard deviations of the healthy donor OD mean. Result(s): Our cohort included both adult (n = 13) and pediatric (n = 9) patients with variants in RAG1 and RAG2. Eleven patients (50%) out of the 22 showed elevated anti- IFN-alpha autoantibodies levels. Five patients (23%) were defined as low positive for anti- IFN-alpha autoantibodies, and 6 patients had no autoantibody titers. Of the 22 patients, 16 were symptomatic with infectious and non-infectious complications including recurrent viral and/or bacterial infections, autoimmune cytopenias, and lymphoproliferation. Ten (63%) of the symptomatic patients demonstrated high anti-IFN-alpha autoantibodies titers. Of the 11 patients with no or low neutralizing anti- IFN-alpha autoantibodies levels, 5 were asymptomatic. In temporal comparison, 16 samples were collected pre-COVID-19 pandemic;8 samples were collected during the pandemic, 2 of which belonged to patients with samples collected before and during the pandemic. In the pre-pandemic cohort, 66% had anti- IFN-alpha autoantibodies. Conversely, during the COVID-19 pandemic, 89% had anti- IFN-alpha autoantibodies. Of note, one patient who had neutralizing anti- IFN-alpha autoantibodies remained positive both before and during the pandemic despite HSCT. Patient also had a SARS-CoV-2 infection in summer of 2022 with a mild clinical course. Conclusions & Next Steps: We observed persistence of anti-IFN-alpha autoantibodies in our cohort post-pandemic and even post-HSCT. It is unclear whether the presence of anti-cytokine antibodies are risk factor for severe COVID-19.Copyright © 2023 Elsevier Inc.

Anti-interferon-alpha autoantibodies in patients with inborn errors of Immunity and rheumatic diseases before and during the COVID-19 pandemic

Introduction: Type 1 interferon (IFN) autoantibodies, such as anti-IFNalpha, have pathogenic significance in life-threatening COVID-19 pneumonia. Ten to twenty percent of severe COVID cases are associated with type I IFN autoantibodies. These autoantibodies likely pre-exist while others arise de novo relative to SARS-CoV-2 infection. It is unclear to what extent type I anti-IFN autoantibodies are induced by SARS-CoV-2 infection and contribute to COVID-19 severity. We investigated these phenomena in those with inborn errors of immunity (IEI) and rheumatic disease (RHE). Aim(s): We aim to compare the prevalence and neutralization ability of anti-IFNalpha autoantibodies in IEI and RHE patients using archived blood samples before and after the COVID-19 pandemic began. Method(s): We determined the presence of autoantibodies against IFNalpha in plasma samples by enzyme linked immunosorbent assay in 453 patients with IEI or RHE who were testing either before or after the COVID-19 pandemic began in March 2020. Using flow cytometry, we determined the function of IFNalpha autoantibodies in plasma to block CD4T cell activation by inhibiting STAT-1 phosphorylation. Result(s): We found that 25 patients with IEI or RHE were positive for anti-IFNalpha autoantibodies. 10 out of 229 patient samples collected before the pandemic (4.2%) tested positive whereas 15 out of 224 patient samples collected after the pandemic began (7.0%) were positive. Seven of the 25 patients (28%) who tested positive had neutralizing antibodies in plasma, which prevented STAT-1 phosphorylation in CD4T cells;all of these patients had partial recombination activating gene deficiency (pRD) except for one patient with autoimmunity, leukemia and selective IgA deficiency. One pRD patient had anti-IFNalpha autoantibodies with neutralization capacity before the pandemic, which persisted after hematopoietic stem cell transplantation (HSCT) with full immune reconstitution. The patient was immunized for SARS-CoV-2 before and after HSCT and acquired COVID-19 infection a year after HSCT. The patient was symptomatic but never hospitalized and fully recovered despite having anti-IFNalpha autoantibodies. Conclusion(s): Anti-IFNalpha autoantibody levels were comparable before and after the start of the COVID-19 pandemic in IEI and RHE patients but only 28% of cases were neutralizing. The clinical implications of these autoantibodies are yet to be determined.Copyright © 2023 Elsevier Inc.

In silico and in vitro studies on inhibitors for SARS-CoV-2 non-structural proteins with dual herbal combination of Withania somnifera with five rasayana herbs.

Being highly transmissible, severe acute respiratory syndrome coronavirus (SARS-CoV-2) has affected millions of people causing devastating global impact and has also not slowed down even after vaccination. The emerges of new strains has made more concerns than the original one. We need a new therapeutic approach against the disease. Our comprehensive in silico study investigates dual herbal combinatorial methanolic extracts of W. somnifera (W) alone and with P. emblica (P) (W:P/1:4) , T. sinensis (T) (W:T/1:4), B. monnieri (B) (W:B/1:1), O. basilicum (O) (W:O/1:4), A. racemosus (A) (W:A/4:1) for potential four phytochemicals as ligands docked with eight COVID-19 Nonstructural proteins (nsp)-main protease (PDB ID:6LU7), papain-like protease (6WUU), helicase ADP (2XZL), N7-methyltransferase (5C8S), endoribonuclease (6WLC), 2'O-methyltransferase (6WVN), RNA dependent RNA polymerase (6M71), and 3Cprotease (6YNQ) along with Remdesivir and Hydroxychloroquine. Ligands from W:P/1:4 showed remarkable docking score (-9.01 kcal/mol) 6M71-(8E,11E,14E)-eicosa-8,11,14-trienoicacidmethylester (EIS) and (-9.99 kcal/mol) 6YNQ-N-[(E)-[4-[(2-methoxydibenzofuran-3-yl)amino]-4-oxobutan-2-ylidene]amino] 4nitrobenzamide (MET). Further, MD simulations were studied for 100 ns and showed the complexes were flexible, stable in the binding pockets of the receptors, and MM-PBSA analysis determined high binding energy of -129.673 ± 15.284 and -134.594 ± 7.085 for 6M71-EIS (Asn496, Lys577, Arg569) and 6YNQ-MET (Cys145, His41). Finally, in vitro JURKAT E6.1 cell lines treated with W:P/1:4 and W:O/1:4 methanolic extracts yielded 44.06 and 31.53 ng/mL levels for interferon alpha to counteract an external stimulus by establishing an antiviral state. Thus, nsp is targeted to design effective antiviral drugs for developing an effective therapeutic approach to combat viral RNA synthesis, processing, and suppression of host immunity.

Dynamics of Clinical Symptoms and Secretory Immunoglobulin a in Covid-19 Convalescent Patients

The post-COVID-19 recovery period is characterized by persistence of some symptoms, with immunological alterations being of great importance. Development of preventive measures to normalize mucosal immunity after a coronavirus infection determines the relevance of the current study. The aim was to study dynamics of clinical symptoms and level of secretory immunoglobulin A in individuals after a novel coronavirus infection as well as evaluate effectiveness of using IFN alpha-2b. Materials and methods. A study was conducted with patients aged 18 to 60 years old (n = 130), surveyed 1 to 9 months after post-infection, as well as in apparently healthy individuals lacking COVID-19 (n = 15). Previous novel coronavirus infection and post-COVID manifestations were verified based on medical documentation, complaints, anamnesis data, physical examination and questionnaires. The concentration of salivatory and nasopharyngeal mucosal sIgA was measured dynamically prior to and after administration of local therapy with IFN alpha-2b (gel applied intranasally twice a day for 30 days). Results. The acute period of COVID-19 was characterized by fever, anosmia, severe asthenia (fatigue and weakness), muscle and joint pain. Among the post-COVID manifestations at early period (1-3 months), pain in the joints and muscles (75.0%) as well as elevated body temperature (21.2%) were reliably detected, whereas in the long period (6-9 months) there were revealed dominance with the same frequency of shortness of breath, muscle and joint pain (75.8%, respectively). Based on examination data in healthy subjects, there was determined an arbitrary normal range of secretory IgA in saliva - 6.45 +/- 1.81 mg/ml and nasal swabs - 13.43 +/- 3.24 mg/ml. In the group of patients 1-3 months post-infection, therapy with IFN alpha-2b one month later resulted in significantly increased level of secretory IgA in saliva (from 1.84 +/- 0.28 to 5.78 +/- 1.96 mg/ml) and in nasal swabs (from 28.61 +/- 3.0 to 39.83 +/- 3.85 mg/ml) by more than 3-and 1.5-fold, respectively. In the group of patients without therapy was featured with stably sustained decline in sIgA level up to 9 months after COVID-19. In particular, the level of saliva sIgA ranged from 2.36 +/- 0.56 down to 2.16 +/- 0.66 mg/ml, and in nasal smears - from 15.66 +/- 1.32 to 10.23 +/- 1.07 mg/ml that differed insignificantly compared to baseline level. The rate of respiratory diseases prevailed in this group (27.6% of cases), which fully lacked in the group of topically administered IFN alpha-2b. Conclusion. In the post-COVID period, multiple organ disorders persist and reduced sIgA level is registered. Intranasally applied IFN alpha-2b made possible to normalize sIgA level and prevent accumulation of respiratory infectious pathologies.

Early antiviral therapy of abidol combined with lopinavir/ritonavir and recombinant interferon alpha-2b for patients with COVID-19 in Zhejiang: A multicenter prospective study.

Objective: To compare the efficacy of the combination of abidol, lopinavir/ritonavir plus recombinant interferon alpha-2b (rIFNalpha-2b) and the combination of lopinavir/ritonavir plus rIFNalpha-2b for patients with COVID-19 in Zhejiang province. Method(s): A multicenter prospective study was carried out to compare the efficacy of triple combination antiviral therapy and dual combination antiviral therapy in 15 medical institutions of Zhejiang province during January 22 to February 16, 2020. All patients were treated with rIFNalpha-2b (5 million U, 2 times/d) aerosol inhalation, in addition 196 patients were treated with abidol (200 mg, 3 times/d) + lopinavir/ritonavir (2 tablets, 1 time/12 h) (triple combination group) and 41 patients were treated with lopinavir/ritonavir (2 tablets, 1 time/12 h) (dual combination group). The patients who received triple combination antiviral therapy were further divided into three subgroups: <48 h, 3-5 d and >5 d according the time from the symptom onset to medication starting. The therapeutic efficacy was compared between triple combination group and dual combination group, and compared among 3 subgroups of patients receiving triple combination antiviral therapy. SPSS 17.0 software was used to analyze the data. Result(s): The virus nucleic acid-negative conversion time in respiratory tract specimens was (12.2+/-4.7) d in the triple combination group, which was shorter than that in the dual combination group [(15.0+/-5.0) d] (t=6.159, P<0.01). The length of hospital stay in the triple combination group [12.0 (9.0, 17.0) d] was also shorter than that in the dual combination group [15.0 (10.0, 18.0) d] (H=2.073, P<0.05). Compared with the antiviral treatment which was started within after the symptom onset of in the triple combination group, the time from the symptom onset to the viral negative conversion was 13.0 (10.0, 17.0), 17.0 (13.0, 22.0) and 21.0 (18.0, 24.0) d in subgroups of 48 h, 3-5 d and >5 d, respectively (Z=32.983, P<0.01), while the time from antiviral therapy to viral negative conversion was (11.8+/-3.9), (13.5+/-5.1) and (11.2+/-4.3) d, respectively(Z=6.722, P<0.05). Conclusion(s): The triple combination antiviral therapy of abidol, lopinavir/litonavir and rIFNalpha-2b shows shorter viral shedding time and shorter hospitalization time, compared with the dual combination antiviral therapy;and the earlier starting triple combination antiviral therapy will result in better antiviral efficacy.Copyright © 2020 by the Chinese Medical Association.

Risk signals mining of ribavirin and interferon-alpha based on the US FDA Adverse Event Reporting System database.

Objective: To identify which induced the symptoms/signs and laboratory abnormal findings occurred in patients with novel coronavirus pneumonia, by disease itself or by ribavirin and interferon-alpha treatments, through mining the adverse events (AEs) signals of the 2 antivirus agents. Method(s): According to the symptoms/signs and laboratory abnormal findings of novel coronavirus pneumonia mentioned in the literature and "Diagnosis and Treatment scheme of Novel Coronavirus Pneumonia (trial version 5)", AEs in this study were selected. Related data were collected from the U.S. FDA Adverse Events Reporting System (FARES) from Jan 1, 2004 to Dec 31, 2019, and the reporting odds ratio (ROR) method was used for signals detection for the above-mentioned 2 drugs. Result(s): A total of 7 582 463 AEs related to drugs were reported in the FAERS database, of which 31 775 related to ribavirin and 2 345 related to interferon-alpha. The results showed that AEs related to ribavirin in respiratory, thoracic, and mediastinal disorders were nasal congestion, cough, laryngeal pain, pharyngeal oedema, productive cough, and dyspnoea;AEs related to interferon-alpha were laryngeal pain and haemoptysis. In other system organ class, AEs related to above 2 drugs were pyrexia, feeling cold, pyrexia, nausea, vomiting, diarrhoea, headache, arthralgia, myalgia, and rash. AEs of laboratory abnormal results related to ribavirin were white blood cell/platelet count decrease and aspartate/alanine aminotransferase increase;AEs related to interferon-alpha were white blood cell/platelet count decrease, aspartate/alanine aminotransferase increase, and lymphocyte count decrease. Conclusion(s): Some AEs induced by ribavirin and interferon-alpha were similar to symptoms/signs and laboratory abnormal findings of novel coronavirus pneumonia, which should be distinguished in the clinical practice.Copyright © 2020 by the Chinese Medical Association.

DYNAMICS OF CLINICAL SYMPTOMS AND SECRETORY IMMUNOGLOBULIN A IN COVID-19 CONVALESCENT PATIENTS.

The post-COVID-19 recovery period is characterized by persistence of some symptoms, with immunological alterations being of great importance. Development of preventive measures to normalize mucosal immunity after a coronavirus infection determines the relevance of the current study. The aim was to study dynamics of clinical symptoms and level of secretory immunoglobulin A in individuals after a novel coronavirus infection as well as evaluate effectiveness of using IFNalpha-2b. Materials and methods. A study was conducted with patients aged 18 to 60 years old (n = 130), surveyed 1 to 9 months after post-infection, as well as in apparently healthy individuals lacking COVID-19 (n = 15). Previous novel coronavirus infection and post-COVID manifestations were verified based on medical documentation, complaints, anamnesis data, physical examination and questionnaires. The concentration of salivatory and nasopharyngeal mucosal sIgA was measured dynamically prior to and after administration of local therapy with IFNalpha-2b (gel applied intranasally twice a day for 30 days). Results. The acute period of COVID-19 was characterized by fever, anosmia, severe asthenia (fatigue and weakness), muscle and joint pain. Among the post-COVID manifestations at early period (1-3 months), pain in the joints and muscles (75.0%) as well as elevated body temperature (21.2%) were reliably detected, whereas in the long period (6-9 months) there were revealed dominance with the same frequency of shortness of breath, muscle and joint pain (75.8%, respectively). Based on examination data in healthy subjects, there was determined an arbitrary normal range of secretory IgA in saliva - 6.45+/-1.81 mg/ml and nasal swabs - 13.43+/-3.24 mg/ml. In the group of patients 1-3 months post-infection, therapy with IFNalpha-2b one month later resulted in significantly increased level of secretory IgA in saliva (from 1.84+/-0.28 to 5.78+/-1.96 mg/ml) and in nasal swabs (from 28.61+/-3.0 to 39.83+/-3.85 mg/ml) by more than 3- and 1.5-fold, respectively. In the group of patients without therapy was featured with stably sustained decline in sIgA level up to 9 months after COVID-19. In particular, the level of saliva sIgA ranged from 2.36+/-0.56 down to 2.16+/-0.66 mg/ml, and in nasal smears - from 15.66+/-1.32 to 10.23+/-1.07 mg/ml that differed insignificantly compared to baseline level. The rate of respiratory diseases prevailed in this group (27.6% of cases), which fully lacked in the group of topically administered IFNalpha-2b. Conclusion. In the post-COVID period, multiple organ disorders persist and reduced sIgA level is registered. Intranasally applied IFNalpha-2b made possible to normalize sIgA level and prevent accumulation of respiratory infectious pathologies.Copyright © 2022 Saint Petersburg Pasteur Institute. All rights reserved.

Features of secretory immunoglobulin A in patients after COVID-19 infection.

Secretory immunoglobulin A, as a marker of the immune response in the mucous membrane, is an available indicator for detecting changes in the local immunity of mucous patients who have undergone COVID-19. Objective. To evaluate the dynamics of changes in the level of sIgA in saliva samples and the effectiveness of the use of interferon alpha-2b in individuals after a coronavirus infection. Patients and methods. Patients aged 18 to 60 years after COVID-19 infection (group 1 on therapy, n = 65;group 2 without therapy, n = 65) and conditionally healthy individuals (control group, n = 15) were monitored. The material is saliva samples, where the sIgA level was determined initially and after a month. The drug - interferon alpha-2b, in the form of a gel for topical use (Viferon, dosage 36,000 IU/g) was administered intranasally 2 times a day, for 1 month. Results. In all groups of patients who underwent COVID-19, the level of saliva sIgA was lower compared to the conditional norm of healthy individuals (6,45 +/- 1,81 mg/ml). A month after the administration of interferon alpha-2b the best effect was observed in patients in the time interval of 1-3 months from the infection, where sIgA was noted a statistically significant increase from 1,84 +/- 0,28 to 5,78 +/- 1,96 mg/ml. In the groups of patients with later terms, a moderate increase in sIgA was determined (3-6 months: 2,83 +/- 0,71 to 3,33 +/- 1,78 mg/ml;6-9 months: 3,53 +/- 0,45 to 4,76 +/- 2,3 mg/ml) and the absence of infectious diseases during rehabilitation period. In the group without therapy, in all temporal aspects, a persistent decrease in sIgA indicators below normal values was revealed, and the frequency of incidence of respiratory viral infections was noted in 9,2% of cases. Conclusions. During the rehabilitation period, the greatest changes in sIgA in saliva were observed in patients in the first 3 months after the COVID infection. The administration of interferon alpha-2b to patients in the post-COVID period is accompanied by the normalization of sIgA and prevents the development of respiratory infections. In similar groups, after COVID-19 without therapy, the indicator tends to decrease, and this category of people is at a higher risk of developing other infectious pathologies.Copyright © 2022, Dynasty Publishing House. All rights reserved.

Epidemiological and clinical characteristics of 40 cases of COVID-19 outside Hubei Province, China

To explore characteristics of patients with pneumonia infected by 2019 Novel Coronavirus (COVID-19) in 2019 outside Hubei Province, China. 40 patients with pneumonia infected by COVID-19 which were confirmed by COVID-19 nucleic acid test were included. Procalcitonin (PCT), serum amyloid A (SAA), C-reactive protein (CRP) and computed tomography (CT) manifestations were analyzed. 40% of patients had clear contact history with Wuhan or other areas of Hubei Province. 60% of patients were clustered diseases and 40% were imported cases. 75% of patients had initial fever, 7.5% had cough, 5% had sore throat at first. 45% had decreased lymphocyte count, 72.5% and 55% patients had increased levels of SAA and CRP. 72.5% of the patients showed multiple ground glass lesions in one or two lungs on chest CT. 90% of the patients with pneumonia are of the common type, and a-interferon atomization inhalation combined with Lopinavir/Ritonavir tablets were given to patients during treatment. 62.5% of the patients were treated with antibiotics, and 15% with hormone. All patients improved after treatment, and 14 patients were cured and discharged. Family cluster infection and asymptomatic infection may be the main way of spreading of COVID-19 pneumonia outside Hubei Province in China.

Early antiviral therapy of abidol combined with lopinavir/ritonavir and recombinant interferon alpha-2b for patients with COVID-19 in Zhejiang: A multicenter prospective study.

Objective: To compare the efficacy of the combination of abidol, lopinavir/ritonavir plus recombinant interferon alpha-2b (rIFNalpha-2b) and the combination of lopinavir/ritonavir plus rIFNalpha-2b for patients with COVID-19 in Zhejiang province. Method(s): A multicenter prospective study was carried out to compare the efficacy of triple combination antiviral therapy and dual combination antiviral therapy in 15 medical institutions of Zhejiang province during January 22 to February 16, 2020. All patients were treated with rIFNalpha-2b (5 million U, 2 times/d) aerosol inhalation, in addition 196 patients were treated with abidol (200 mg, 3 times/d) + lopinavir/ritonavir (2 tablets, 1 time/12 h) (triple combination group) and 41 patients were treated with lopinavir/ritonavir (2 tablets, 1 time/12 h) (dual combination group). The patients who received triple combination antiviral therapy were further divided into three subgroups: <48 h, 3-5 d and >5 d according the time from the symptom onset to medication starting. The therapeutic efficacy was compared between triple combination group and dual combination group, and compared among 3 subgroups of patients receiving triple combination antiviral therapy. SPSS 17.0 software was used to analyze the data. Result(s): The virus nucleic acid-negative conversion time in respiratory tract specimens was (12.2+/-4.7) d in the triple combination group, which was shorter than that in the dual combination group [(15.0+/-5.0) d] (t=6.159, P<0.01). The length of hospital stay in the triple combination group [12.0 (9.0, 17.0) d] was also shorter than that in the dual combination group [15.0 (10.0, 18.0) d] (H=2.073, P<0.05). Compared with the antiviral treatment which was started within after the symptom onset of in the triple combination group, the time from the symptom onset to the viral negative conversion was 13.0 (10.0, 17.0), 17.0 (13.0, 22.0) and 21.0 (18.0, 24.0) d in subgroups of 48 h, 3-5 d and >5 d, respectively (Z=32.983, P<0.01), while the time from antiviral therapy to viral negative conversion was (11.8+/-3.9), (13.5+/-5.1) and (11.2+/-4.3) d, respectively(Z=6.722, P<0.05). Conclusion(s): The triple combination antiviral therapy of abidol, lopinavir/litonavir and rIFNalpha-2b shows shorter viral shedding time and shorter hospitalization time, compared with the dual combination antiviral therapy;and the earlier starting triple combination antiviral therapy will result in better antiviral efficacy.Copyright © 2020 by the Chinese Medical Association.

Immunopathogenetic Aspects and Effectiveness of Combination Therapy for Influenza A (H1N1)pdm2009 in Children.

Introduction. The continuing circulation of influenza A (H1N1)pdm2009 virus poses a threat of a new epidemic rise. It is known that influenza A (H1N1)pdm2009 is characterized by a severe course, development of life-threatening complications, and high mortality, which is associated not only with the biological features of the pathogen, but also with the induction of deep immunosuppression, especially the interferon system and the cellular-type immune response. The role of influenza in the development of severe forms of the new coronavirus infection COVID-19 has been revealed. The increase of the number of virus strains resistant to various classes of antiviral drugs is of unfavorable importance. This requires the development of new approaches to the treatment of influenza A (H1N1)pdm2009 with the combined use of drugs with complex antiviral and immunocorrective activity. Purpose. To substantiate the combination therapy of influenza A (H1N1)pdm2009 in children using oseltamivir (Tamiflu) and recombinant interferon-alpha2b (Viferon). Materials and methods. Clinical and laboratory examination of 85 children aged from 3 to 5 years with moderate (43) and severe forms (42) of influenza A (H1N1)pdm2009 was carried out. Results and discussion. In patients with severe forms of A(H1N1)pdm2009 influenza, a higher frequency of anamnestic risk groups (85.7%), frequent development of febrile fever (100%), severe intoxication symptoms (100%), symptoms of laryngitis (28.6%), tracheitis (57.1%), bronchitis (76.2%), dyspeptic (42.9%) and cerebral syndromes (62.9%), other complications (80.9%) were revealed. In these patients, more significant changes of the indicators of the cellular type of the immune response were found - the decrease of CD3, CD4, CD8, the humoral type of immune response - the increase of CD20, IgM, circulating immune complexes, the decrease of IgA and IgG, innate immunity factors - the decrease of the metabolic activity of neutrophils, moderate increase of CD16. The combined administration of recombinant interferon-alpha2b (Viferon) and oseltamivir (tamiflu) compared with oseltamivir (tamiflu) monotherapy reduced the duration of fever (Me 2, IQI 1-4 days and Me 3, IQI 2-4 days), intoxication (Me 3, IQI 2-4.5 days and Me 4.5, IQI 3-7 days), symptoms of rhinitis (Me 5, IQI 4-7 days and Me 6.5, IQI 4.5-7.5 days), pharyngitis (Me 5, IQI 4-7 days and Me 6.5, IQI 4.5-7.5 days), tracheitis (Me 2, IQI 1-3 days and Me 3.5, IQI 2-4 days), bronchitis (Me 3, IQI 2-5 days and Me 5, IQI 4-6 days). In this group, the complications developed less frequently (4.5% and 33.3%);there was the decrease of hospitalization time (Me 5, IQI 4-7 days and Me 6.5, IQI 5-7 days). There was the increase of the number of children, who (after 10 days from the start of therapy) had sanitation of the nasopharynx from the virus (90.9% and 61.9%). Conclusion. The high frequency of anamnestic risk groups and the induction of deep immunosuppression, especially the cellular component of immunity, are the cause of the formation of severe forms of influenza A (H1N1)pdm2009. This justified the appointment of combination therapy using the neuraminidase inhibitor oseltamivir (Tamiflu) and recombinant interferon-alpha2b (Viferon), which not only inhibits virus replication, but also has immunocorrective activity against the interferon system and cellular immunity. The high efficiency of the combined administration of recombinant interferon-alpha2b (Viferon) and oseltamivir (Tamiflu) lets to recommend the inclusion of these drugs in the treatment of severe forms of influenza A(H1N1)pdm2009 in children.Copyright © 2020, Professionalnye Izdaniya. All rights reserved.

Immunopathogenetic Aspects and Effectiveness of Combination Therapy for Influenza A (H1N1)pdm2009 in Children.

Introduction. The continuing circulation of influenza A (H1N1)pdm2009 virus poses a threat of a new epidemic rise. It is known that influenza A (H1N1)pdm2009 is characterized by a severe course, development of life-threatening complications, and high mortality, which is associated not only with the biological features of the pathogen, but also with the induction of deep immunosuppression, especially the interferon system and the cellular-type immune response. The role of influenza in the development of severe forms of the new coronavirus infection COVID-19 has been revealed. The increase of the number of virus strains resistant to various classes of antiviral drugs is of unfavorable importance. This requires the development of new approaches to the treatment of influenza A (H1N1)pdm2009 with the combined use of drugs with complex antiviral and immunocorrective activity. Purpose. To substantiate the combination therapy of influenza A (H1N1)pdm2009 in children using oseltamivir (Tamiflu) and recombinant interferon-alpha2b (Viferon). Materials and methods. Clinical and laboratory examination of 85 children aged from 3 to 5 years with moderate (43) and severe forms (42) of influenza A (H1N1)pdm2009 was carried out. Results and discussion. In patients with severe forms of A(H1N1)pdm2009 influenza, a higher frequency of anamnestic risk groups (85.7%), frequent development of febrile fever (100%), severe intoxication symptoms (100%), symptoms of laryngitis (28.6%), tracheitis (57.1%), bronchitis (76.2%), dyspeptic (42.9%) and cerebral syndromes (62.9%), other complications (80.9%) were revealed. In these patients, more significant changes of the indicators of the cellular type of the immune response were found - the decrease of CD3, CD4, CD8, the humoral type of immune response - the increase of CD20, IgM, circulating immune complexes, the decrease of IgA and IgG, innate immunity factors - the decrease of the metabolic activity of neutrophils, moderate increase of CD16. The combined administration of recombinant interferon-alpha2b (Viferon) and oseltamivir (tamiflu) compared with oseltamivir (tamiflu) monotherapy reduced the duration of fever (Me 2, IQI 1-4 days and Me 3, IQI 2-4 days), intoxication (Me 3, IQI 2-4.5 days and Me 4.5, IQI 3-7 days), symptoms of rhinitis (Me 5, IQI 4-7 days and Me 6.5, IQI 4.5-7.5 days), pharyngitis (Me 5, IQI 4-7 days and Me 6.5, IQI 4.5-7.5 days), tracheitis (Me 2, IQI 1-3 days and Me 3.5, IQI 2-4 days), bronchitis (Me 3, IQI 2-5 days and Me 5, IQI 4-6 days). In this group, the complications developed less frequently (4.5% and 33.3%);there was the decrease of hospitalization time (Me 5, IQI 4-7 days and Me 6.5, IQI 5-7 days). There was the increase of the number of children, who (after 10 days from the start of therapy) had sanitation of the nasopharynx from the virus (90.9% and 61.9%). Conclusion. The high frequency of anamnestic risk groups and the induction of deep immunosuppression, especially the cellular component of immunity, are the cause of the formation of severe forms of influenza A (H1N1)pdm2009. This justified the appointment of combination therapy using the neuraminidase inhibitor oseltamivir (Tamiflu) and recombinant interferon-alpha2b (Viferon), which not only inhibits virus replication, but also has immunocorrective activity against the interferon system and cellular immunity. The high efficiency of the combined administration of recombinant interferon-alpha2b (Viferon) and oseltamivir (Tamiflu) lets to recommend the inclusion of these drugs in the treatment of severe forms of influenza A(H1N1)pdm2009 in children.Copyright © 2020, Professionalnye Izdaniya. All rights reserved.

DYNAMICS OF CLINICAL SYMPTOMS AND SECRETORY IMMUNOGLOBULIN A IN COVID-19 CONVALESCENT PATIENTS.

The post-COVID-19 recovery period is characterized by persistence of some symptoms, with immunological alterations being of great importance. Development of preventive measures to normalize mucosal immunity after a coronavirus infection determines the relevance of the current study. The aim was to study dynamics of clinical symptoms and level of secretory immunoglobulin A in individuals after a novel coronavirus infection as well as evaluate effectiveness of using IFNalpha-2b. Materials and methods. A study was conducted with patients aged 18 to 60 years old (n = 130), surveyed 1 to 9 months after post-infection, as well as in apparently healthy individuals lacking COVID-19 (n = 15). Previous novel coronavirus infection and post-COVID manifestations were verified based on medical documentation, complaints, anamnesis data, physical examination and questionnaires. The concentration of salivatory and nasopharyngeal mucosal sIgA was measured dynamically prior to and after administration of local therapy with IFNalpha-2b (gel applied intranasally twice a day for 30 days). Results. The acute period of COVID-19 was characterized by fever, anosmia, severe asthenia (fatigue and weakness), muscle and joint pain. Among the post-COVID manifestations at early period (1-3 months), pain in the joints and muscles (75.0%) as well as elevated body temperature (21.2%) were reliably detected, whereas in the long period (6-9 months) there were revealed dominance with the same frequency of shortness of breath, muscle and joint pain (75.8%, respectively). Based on examination data in healthy subjects, there was determined an arbitrary normal range of secretory IgA in saliva - 6.45+/-1.81 mg/ml and nasal swabs - 13.43+/-3.24 mg/ml. In the group of patients 1-3 months post-infection, therapy with IFNalpha-2b one month later resulted in significantly increased level of secretory IgA in saliva (from 1.84+/-0.28 to 5.78+/-1.96 mg/ml) and in nasal swabs (from 28.61+/-3.0 to 39.83+/-3.85 mg/ml) by more than 3- and 1.5-fold, respectively. In the group of patients without therapy was featured with stably sustained decline in sIgA level up to 9 months after COVID-19. In particular, the level of saliva sIgA ranged from 2.36+/-0.56 down to 2.16+/-0.66 mg/ml, and in nasal smears - from 15.66+/-1.32 to 10.23+/-1.07 mg/ml that differed insignificantly compared to baseline level. The rate of respiratory diseases prevailed in this group (27.6% of cases), which fully lacked in the group of topically administered IFNalpha-2b. Conclusion. In the post-COVID period, multiple organ disorders persist and reduced sIgA level is registered. Intranasally applied IFNalpha-2b made possible to normalize sIgA level and prevent accumulation of respiratory infectious pathologies. Copyright © 2022 Saint Petersburg Pasteur Institute. All rights reserved.

[Anti-cytokine autoantibodies: Review of the literature]. / Anticorps anti-cytokines : quel impact clinique en pathologie humaine ?

Anti-cytokine antibodies (ACA) are an emerging cause of acquired immunodeficiency, especially in previously healthy adults. The most frequently reported are anti-IFN-γ responsible for disseminated non-tuberculous mycobacteria infections, and anti-GM-CSF mainly in mycobacteria, cryptococcosis and nocardiosis infections. The presence of anti-IFN-α in severe COVID-19 infections has recently been described. The search for and detection of these ACAs in an unusual infection situation makes it possible to set up specific therapies in addition to the anti-infective treatment. ACAs are also frequent in various autoimmune pathologies where, in addition to being indicators of the breakdown of immune tolerance, they can modulate the activity of the disease according to their cytokine target. In this review of the literature, we will focus on the epidemiology and the clinical impact of these ACAs in healthy subjects and in infectious or dysimmune diseases.

Features of secretory immunoglobulin A in patients after COVID-19 infection

Secretory immunoglobulin A, as a marker of the immune response in the mucous membrane, is an available indicator for detecting changes in the local immunity of mucous patients who have undergone COVID-19. Objective. To evaluate the dynamics of changes in the level of sIgA in saliva samples and the effectiveness of the use of interferon alpha-2b in individuals after a coronavirus infection. Patients and methods. Patients aged 18 to 60 years after COVID-19 infection (group 1 on therapy, n = 65;group 2 without therapy, n = 65) and conditionally healthy individuals (control group, n = 15) were monitored. The material is saliva samples, where the sIgA level was determined initially and after a month. The drug - interferon alpha-2b, in the form of a gel for topical use (Viferon, dosage 36,000 IU/g) was administered intranasally 2 times a day, for 1 month. Results. In all groups of patients who underwent COVID-19, the level of saliva sIgA was lower compared to the conditional norm of healthy individuals (6,45 +/- 1,81 mg/ml). A month after the administration of interferon alpha-2b the best effect was observed in patients in the time interval of 1-3 months from the infection, where sIgA was noted a statistically significant increase from 1,84 +/- 0,28 to 5,78 +/- 1,96 mg/ml. In the groups of patients with later terms, a moderate increase in sIgA was determined (3-6 months: 2,83 +/- 0,71 to 3,33 +/- 1,78 mg/ml;6-9 months: 3,53 +/- 0,45 to 4,76 +/- 2,3 mg/ml) and the absence of infectious diseases during rehabilitation period. In the group without therapy, in all temporal aspects, a persistent decrease in sIgA indicators below normal values was revealed, and the frequency of incidence of respiratory viral infections was noted in 9,2% of cases. Conclusions. During the rehabilitation period, the greatest changes in sIgA in saliva were observed in patients in the first 3 months after the COVID infection. The administration of interferon alpha-2b to patients in the post-COVID period is accompanied by the normalization of sIgA and prevents the development of respiratory infections. In similar groups, after COVID-19 without therapy, the indicator tends to decrease, and this category of people is at a higher risk of developing other infectious pathologies. Copyright © 2022, Dynasty Publishing House. All rights reserved.

Evaluation of the efficacy and safety of interferon alpha-2b in the prevention of acute respiratory infections (including COVID-19) among children in Uzbekistan

Objective. To study the preventive efficacy and safety of interferon alpha-2b (IFN-alpha2b) with antioxidants against acute respiratory viral infections (ARVI) among children in Uzbekistan during the COVID-19 pandemic. Patients and methods. An open-label prospective observational study included 300 first-grade pupils (age 6-7 years) from schools in Tashkent (Uzbekistan). Children were evenly divided into three groups: the first and second (study) groups from different schools received IFN-alpha2b (Viferon gel for external and local use, 36,000 IU/g);the third group (control) did not receive the medication. The main criteria for assessing the effectiveness of prevention were the incidence of ARVI within 3 months after inclusion in the study, duration and severity of clinical symptoms of the disease, and the frequency of adverse events associated with the use of the study medication. Results. Observation after 30 to 90 days from the beginning of treatment showed a statistically significant increase in the proportion of ARVI cases in the control group compared to the study groups where Viferon gel was used. The average duration of the febrile period in children with ARVI in the first group was the lowest and amounted to 3.1 +/- 0.4 days, in the second group - 3.8 +/- 0.5 days, in the third group - 5.4 +/- 1.1 days (p1-3 = 0.002 and p2-3 = 0.003, Mann-Whitney U test with adjustment to multiple comparisons). Also, the duration of intoxication symptoms and catarrhal phenomena in children with ARVI in the groups receiving IFN-alpha2b was significantly shorter than in the control group. No cases of COVID-19 were registered in the study groups during the entire period of observation, while in the control group, markers of infection were detected in 30 children. There were no adverse events associated with the administration of IFN-alpha2b (Viferon gel for external and local use 36,000 IU/g). Conclusion. In children who received IFN-alpha2b with antioxidants, a statistically significant decrease in ARVI episodes was found, as well as rapid relief of intoxication symptoms and catarrhal phenomena in the event of disease. The pronounced effectiveness and the absence of adverse events give reason to recommend the use of Viferon gel for the prevention of acute respiratory viral infections, as well as COVID-19 in children. Copyright © 2022, Dynasty Publishing House. All rights reserved.

Coordinated innate and T-cell immune responses in mild COVID-19 patients from household contacts of COVID-19 cases during the first pandemic wave.

Objective: To better define the immunopathogenesis of COVID-19, the present study aims to characterize the early immune responses to SARS-CoV-2 infection in household contacts of COVID-19 cases. In particular, innate, T- and B-cell specific responses were evaluated over time. Methods: Household contacts of COVID-19 cases screened for SARS-CoV-2 infection by nasopharyngeal swab for surveillance purposes were enrolled (T0, n=42). Of these, 28 subjects returned for a follow-up test (T1). The innate response was assessed by detecting a panel of soluble factors by multiplex-technology in plasma samples. Cell-mediated response was evaluated by measuring interferon (IFN)-γ levels by ELISA in plasma harvested from whole-blood stimulated with SARS-CoV-2 peptide pools, including spike (S), nucleocapsid (N) and membrane (M) proteins. The serological response was assessed by quantifying anti-Receptor-Binding-Domain (RBD), anti-Nucleocapsid (N), whole virus indirect immunofluorescence, and neutralizing antibodies. Results: At T0, higher levels of plasmatic IFN-α, IL-1ra, MCP-1 and IP-10, and lower levels of IL-1ß, IL-9, MIP-1ß and RANTES were observed in subjects with positive swab compared to individuals with a negative one (p<0.05). Plasmatic IFN-α was the only cytokine detectable in subjects with positive SARS-CoV-2 swabs with high accuracy for swab score positivity (0.93, p<0.0001). Among subjects with positive swabs, significant negative correlations were found among the RT-PCR cycle threshold values reported for genes S and N and IFN-α or IP-10 levels. At T0, the IFN-γ T-cell specific response was detected in 50% (5/10) of subjects with positive swab, while anti-RBD/anti-N antibodies showed a positivity rate of 10% (1/10). At T1, the IFN-γ T-cell specific response was detected in most of the confirmed-infection subjects (77.8%, 7/9), whereas the serological response was still observed in a minority of them (44.4%, 4/9). Overall, the swab test showed a moderate concordance with the T-cell response (78.6%, k=0.467), and a scarce concordance with the serological one (72.9%, k=0.194). Conclusions: Plasmatic IFN-α and the IFN-γ T-cell specific response appear early even in the absence of seroconversion, and show a greater positivity rate than the serological response in household contacts with positive swab.

Efficacy and safety of pegylated interferon-α2b in moderate COVID-19: a phase 3, randomized, comparator-controlled, open-label study.

OBJECTIVE: To evaluate the efficacy and safety of pegylated interferon alpha-2b (PEG IFN-α2b) administered in conjunction with the standard of care (SOC) in subjects with moderate coronavirus disease-19 (COVID-19). METHODS: In this study, adult subjects with confirmed moderate COVID-19 were randomized in a 1:1 ratio to receive either PEG IFN-α2b + SOC or SOC alone. The primary endpoint was a two-point improvement in clinical status on Day 11, measured by the World Health Organization's seven-point ordinal scale. RESULTS: Of 250 subjects, 120 were randomized to the PEG IFN-α2b + SOC arm and 130 were randomized to the SOC arm. The results for the PEG IFN + SOC arms vs the SOC arm for the proportion of subjects with a two-point improvement in the seven-point ordinal scale were 80.36% vs 68.18% (P=0.037) on Day 8, 91.60% vs 92.56% (P=0.781) on Day 11, and 94.12% vs 95.93% (P=0.515) on Day 15. There was a time-dependent decrease in the biomarkers in both arms, and no clinically significant changes in laboratory parameters. The safety profile was similar in both arms. CONCLUSION: PEG IFN-α2b induced early viral clearance, improved the clinical status, and decreased the duration of supplemental oxygen. It provides a viable treatment option and can limit the spread of severe acute respiratory syndrome coronavirus-2.

Antrodia cinnamomea Suppress Dengue Virus Infection through Enhancing the Secretion of Interferon-Alpha.

Dengue caused by dengue virus (DENV) is a mosquito-borne disease. Dengue exhibits a wide range of symptoms, ranging from asymptomatic to flu-like illness, and a few symptomatic cases may develop into severe dengue, leading to death. However, there are no effective and safe therapeutics for DENV infections. We have previously reported that cytokine expression, especially inflammatory cytokines, was altered in patients with different severities of dengue. Antrodia cinnamomea (A. cinnamomea) is a precious and endemic medical mushroom in Taiwan. It contains unique chemical components and exhibits biological activities, including suppressing effects on inflammation and viral infection-related diseases. According to previous studies, megakaryocytes can support DENV infection, and the number of megakaryocytes is positively correlated with the viral load in the serum of acute dengue patients. In the study, we investigated the anti-DENV effects of two ethanolic extracts (ACEs 1-2) and three isolated compounds (ACEs 3-5) from A. cinnamomea on DENV infection in Meg-01 cells. Our results not only demonstrated that ACE-3 and ACE-4 significantly suppressed DENV infection, but also reduced interleukin (IL)-6 and IL-8 levels. Moreover, the level of the antiviral cytokine interferon (IFN)-α was also increased by ACE-3 and ACE-4 in Meg-01 cells after DENV infection. Here, we provide new insights into the potential use of A. cinnamomea extracts as therapeutic agents against DENV infection. However, the detailed mechanisms underlying these processes require further investigation.

IFNAR2 relevance in the clinical outcome of individuals with severe COVID-19.

Interferons (IFNs) are a group of cytokines with antiviral, antiproliferative, antiangiogenic, and immunomodulatory activities. Type I IFNs amplify and propagate the antiviral response by interacting with their receptors, IFNAR1 and IFNAR2. In COVID-19, the IFNAR2 (interferon alpha and beta receptor subunit 2) gene has been associated with the severity of the disease, but the soluble receptor (sIFNAR2) levels have not been investigated. We aimed to evaluate the association of IFNAR2 variants (rs2236757, rs1051393, rs3153, rs2834158, and rs2229207) with COVID-19 mortality and to assess if there was a relation between the genetic variants and/or the clinical outcome, with the levels of sIFNAR2 in plasma samples from hospitalized individuals with severe COVID-19. We included 1,202 subjects with severe COVID-19. The genetic variants were determined by employing Taqman® assays. The levels of sIFNAR2 were determined with ELISA in plasma samples from a subgroup of 351 individuals. The rs2236757, rs3153, rs1051393, and rs2834158 variants were associated with mortality risk among patients with severe COVID-19. Higher levels of sIFNAR2 were observed in survivors of COVID-19 compared to the group of non-survivors, which was not related to the studied IFNAR2 genetic variants. IFNAR2, both gene, and soluble protein, are relevant in the clinical outcome of patients hospitalized with severe COVID-19.