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Background: The rapid emergence of the SARS-CoV-2 Omicron variant that evades many therapies illustrates the need for antiviral treatments with high genetic barriers to resistance. The small molecule PAV-104, identified through a moderate-throughput screen involving cell-free protein synthesis, was recently shown to target a subset of host protein assembly machinery in a manner specific to viral assembly with minimal host toxicity. The chemotype shows broad activity against respiratory viral pathogens, including Orthomyxoviridae, Paramyxoviridae, Adenoviridae, Herpesviridae, and Picornaviridae, with low susceptibility to evolutionary escape. Here, we investigated the capacity of PAV-104 to inhibit SARS-CoV-2 replication in human airway epithelial cells (AECs). Method(s): Dose-dependent cytotoxicity of PAV-104 in Calu-3 cells was determined by MTT assay. Calu-3 cells were infected with SARS-CoV-2 isolate USA-WA1/2020 (MOI=0.01). Primary AECs were isolated from healthy donor lung transplant tissue, cultured at air liquid interface (ALI), and infected with SARS-CoV-2 Gamma, Delta, and Omicron variants (MOI=0.1). SARS-CoV-2 replication was assessed by RT-PCR quantitation of the N gene, immunofluorescence assay (IFA) of nucleocapsid (N) protein, and titration of supernatant (TCID50). Transient co-expression of four SARS-CoV-2 structural proteins (N, M, S, E) to produce virus-like particles (VLPs) was used to study the effect of PAV-104 on viral assembly. Drug resin affinity chromatography was performed to study the interaction between PAV-104 and N. Glycerol gradient sedimentation was used to assess N oligomerization. Total RNA-seq and the REACTOME database were used to evaluate PAV-104 effects on the host transcriptome. Result(s): PAV-104 reached 50% cytotoxicity in Calu-3 cells at 3732 nM (Fig.1A). 50 nM PAV-104 inhibited >99% of SARS-CoV-2 infection in Calu-3 cells (p< 0.01) and in primary AECs (p< 0.01) (Fig.1B-E). PAV-104 specifically inhibited SARS-CoV-2 post entry, and suppressed production of SARS-CoV-2 VLPs without affecting viral protein synthesis. PAV-104 interacted with SARS-CoV-2 N and interfered with N oligomerization. Transcriptome analysis revealed that PAV-104 treatment reversed SARS-CoV-2 induction of the interferon and maturation of nucleoprotein signaling pathways. Conclusion(s): PAV-104 is a pan-respiratory virus small molecule inhibitor with promising activity against SARS-CoV-2 in human airway epithelial cells that should be explored in animal models and clinical studies.
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Background: One of the efforts to control SARS-CoV-2 infection in health workers is vaccination. In this study, the levels of SARS-CoV-2 neutralizing antibody (nAb) in health workers were measured with Ichroma and iFlash. Method(s): This study applied an observational analytic design with a prospective cohort and was conducted at Dr. Soetomo Regional Public Hospital, Surabaya, from January to November 2021. The population of this study included a total of 75 health workers after taking the second dose of the SARS-CoV-2 (Sinovac) vaccine. The Covid-19 NAb levels of the population were tested with Ichroma and iFlash on day 0 before vaccination, as well as days 14 and 28, and months 3 and 6 after vaccination. Result(s): The Friedman test indicated a significant difference in NAb levels according to the iFlash test on day 14, day 28, month 3, and month 6 compared to those before vaccination (p < 0.05). The Wilcoxon test revealed a significant difference in NAb levels on day 14, day 28, month 3, and month 6. The results of the Cochran test showed a significant difference in the positivity of NAb according to the Ichroma test on day 14, day 28, month 3, and month 6 compared to those before vaccination (p < 0.05). McNemar's test demonstrated that the COI at month 3 was not significantly different from that before vaccination;The COI at month 6 was not significantly different from those at days 14 and 28. The results of the Pearson correlation test and Bland-Altman plot indicated a moderate correlation between Ichroma and iFlash (r = 0.592, p = 0.002). Conclusion(s): Neutralizing antibodies for Covid-19 were formed after day 14 and started to increase on day 28 and started to decrease in months 3 and 6. The levels of NAb for Covid-19 were measured with Ichroma and iFlash in roughly the same pattern and had a moderate positive correlation.Copyright © 2023 Phcogj.Com.
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The 2019 novel coronavirus (SARS-CoV-2) causes severe pneumonia called COVID-19 and leads to severe acute respiratory syndrome with a high mortality rate. The SARS-CoV-2 virus in the human body leads to jumpstarting immune reactions and multi-organ inflammation, which has poorer outcomes in the presence of predisposing conditions, including hypertension, dyslipidemia, dysglycemia, abnormal adiposity, and even endothelial dysfunction via biomolecular mechanisms. In addition, leucopenia, hypoxemia, and high levels of both cytokines and chemokines in the acute phase of this disease, as well as some abnormalities in chest CT images, were reported in most patients. The spike protein in SARS-CoV-2, the primary cell surface protein, helps the virus anchor and enter the human host cells. Additionally, new mutations have mainly happened for spike protein, which has promoted the infection's transmissibility and severity, which may influence manufactured vaccines' efficacy. The exact mechanisms of the pathogenesis, besides molecular aspects of COVID-19 related to the disease stages, are not well known. The altered molecular functions in the case of immune responses, including T CD4+, CD8+, and NK cells, besides the overactivity in other components and outstanding factors in cytokines like interleukin-2, were involved in severe cases of SARS-CoV-2. Accordingly, it is highly needed to identify the SARS-CoV-2 bio-molecular characteristics to help identify the pathogenesis of COVID-19. This study aimed to investigate the bio-molecular aspects of SARS-CoV-2 infection, focusing on novel SARS-CoV-2 variants and their effects on vaccine efficacy.Copyright © 2022 NRITLD, National Research Institute of Tuberculosis and Lung Disease, Iran.
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Background: Immune check point inhibitors (ICPi) have become the first line treatment for most of the cancers and have shown promising results. However, they can provoke reactions, the most feared being immune related adverse events (irAE). Case presentation: We present a series of three cases, of patients recieving ICPi. All three patients developed AKI after administration of SARS-CoV-2 mRNA vaccine. Two patients had kidney-biopsy-proven acute interstitial nephritis (AIN) which responded to ICPi discontinuation and treatment with steroids. One had presumed AIN based on the high levels of CRP and urine retinol binding protein to creatinine ratio and responded to cessation of ICPi alone. Conclusion(s): These three cases demonstrate that a strong immune response from the SARS-CoV-2 mRNA vaccine combined with an uninhibited immune system under influence of ICPi led to an amplification of autoimmunity leading to AKI presenting as AIN.Copyright © The Author(s) 2022.
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Infectious bronchitis virus (IBV) initially establishes the infection in the respiratory tract and then spreads to other tissues depending on its virulence. During 2011-2018, the 4/91 IBV strain was isolated from poultry flocks affected by decreased egg production and quality in Eastern Canada. One of the Canadian 4/91 IBV isolates, IBV/Ck/Can/17-038913, was propagated in embryonated chicken eggs and molecularly characterized using whole genome sequencing. An in vivo study in laying hens was conducted to observe if IBV/Ck/Can/17-038913 isolate affects the egg production and quality. Hens were infected with IBV/Ck/Can/17-038913 isolate during the peak of egg lay, using a standard dose and routes maintaining uninfected controls. Oropharyngeal and cloacal swabs were collected at predetermined time points for the quantification of IBV genome loads. At 6 and 10 days post-infection, hens were euthanized to observe the lesions in various organs and collect blood and tissue samples for the quantification of antibody response and IBV genome loads, respectively. Egg production was not impacted during the first 10 days following infection. No gross lesions were observed in the tissues of the infected birds. The IBV genome was quantified in swabs, trachea, lung, proventriculus, cecal tonsils, kidney, and reproductive tissues. The serum antibody response against IBV was quantified in infected hens. In addition, histological changes, and recruitment of immune cells, such as macrophages and T cell subsets in kidney tissues, were measured. Overall, data show that IBV/Ck/Can/17-038913 isolate is not associated with egg production issues in laying hens infected at the peak of lay, while it demonstrates various tissue tropism, including kidney, where histopathological lesions and immune cell recruitments were evident.
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The aim of this paper was to prepare specific monoclonal antibody (mAb) against African swine fever virus (ASFV) p54 protein. The p54 protein was expressed in Escherichia coli expression system and used as the antigen in mAb production. The spleen cells from the immunized BALB/c mice were fused with myeloma cells SP2/0. To screen the positive hybridoma cells, the purified p54 protein was used as envelope antigen for indirect ELISA. After four times' subcloning, the supernatant of hybridoma cells were used to identify mAb subtype, ascites were prepared via in vivo induction method in mice and then the mAb was purified. The titer of the mAb was detected by indirect ELISA, and the specificity of the mAb was identified by cross reactivity assay, IFA and Western blot. According to the predicted secondary structure of p54 protein, using the stepwise truncation method identified the epitope region of mAbs, and labeled the region in tertiary structure of p54 protein. Results were as follows: six hybridoma cells secreting p54 monoclonal antibody were successfully screened and named 28G12-1, 31G7-1, 31G7-2, 35F10-1, 35F10-2, 38D3-1, respectively. The heavy chains of 28G12-1, 31G7-1, and 31G7-2 were IgG2a type, the heavy chains of 35F10-1, 35F10-2, 38D3-1 were IgG1 type, light chains were all kappa chains. The lowest titer of mAb was 1:25 600, and having no cross reaction with PRRSV, PRV, PEDV, PPV, SADS-CoV, PCV2, the specificity was strong. All six monoclonal antibodies could recognize the 127-146 aa on carboxyl end. In this study, ASFV p54 protein and p54 monoclonal antibody were successfully obtained, and the epitopes of six mAbs were identified, these experimental data laid a foundation for the functional research of p54 protein and the study of ASFV epitope vaccine. Copyright © 2023 Editorial Board, Institute of Animal Science of the Chinese Academy of Agricultural Sciences. All rights reserved.
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Background: In the COVID-19 pandemic, individuals may be asymptomatic, with mild or severe symptoms associated with respiratory tract infections. Patients with COVID-19 have IgG antibodies on average two weeks after infection and persist at stable levels for a few months. The influence of the levels of these antibodies and the time in circulation, protection, and severity of the disease in reinfections, has not yet been completely elucidated. The objective of this work was to evaluate the titers of anti-SARS-CoV-2 IgG antibodies in asymptomatic patients or with mild symptoms in a period of six months. Method(s): In this longitudinal study, we selected 62 individuals (median age 42.5 years;IQR 33.3-52.0;59.7% female) tested positive for SARS-CoV-2 Immunoglobulin G (IgG) antibodies using a Fluorescence Immunoassay (FIA) (iChroma II, BioSys + Kovalent) from July 2020 (peak of the first wave) to March 2021 (begging of the second wave associated with transmission of the Gamma variant) in the state of Sergipe, Northeast Brazil. All participants included in the present study had a history of asymptomatic or mild COVID-19, were not vaccinated against the disease, and were selected from the EpiSERGIPE Project conducted in the state of Sergipe. An in-house indirect Enzyme-Linked Immunosorbent assay (ELISA)-based method was used to evaluate the serum titers of anti-nucleocapsid SARS-CoV-2 antibodies in three moments: baseline (D0), 90 days (D90), and 180 days (D180). Briefly, 96-microwell plates (Nunc, Thermo Fisher Scientific). Result(s): In D0, 79% (49 of 62) of individuals had a positive result for the presence of anti-nucleocapsid SARS-CoV-2 antibodies. In D90 and D180, the percentage of positive results was 69.3% (43 of 62) and 53.2% (33 of 62), respectively. We found a progressive decline in the levels of anti-nucleocapsid SARS-CoV-2 antibodies over time, ranging from 26.2 (Interquartile Range [IQR] 12.4-37.7) in D0 to 11.7 (IQR 5.6-18.2) in D180 (p<0.001). In addition, we found that individuals over 40 years old had higher levels of IgG antibodies to the SARS-CoV-2 nucleocapsid in D90, regardless of sex. However, an influence of sex and age was not observed on D0 and D180. Discussion(s): Studies describe that IgG levels remain for only 3-4 months in the body of individuals who have had previous contact with SARS-CoV-2, however in this study it was observed that most individuals had a significant and gradual decrease in anti-nucleocapsid SARS-CoV-2 antibodies circulating, even before completing three months after exposure to the virus. Conclusion(s): This study showed a progressive decline in the levels of anti-nucleocapsid SARS-CoV-2 antibodies during the first six months after SARS-CoV-2 infection among individuals with asymptomatic or mild COVID-19. Approximately 50% of individuals have no detectable antibodies six months after infection. Moreover, we found a potential influence of age on the humoral response against SARS-CoV-2. Copyright © 2022
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Introduction: Viral infections have been implicated in the initiation of the autoimmune diseases. Recent reports suggest that a proportion of patients with COVID-19 develop severe disease with multiple organ injuries. We evaluated the relationship between COVID-19 severity, prevalence and persistence of antinuclear and other systemic and organ specific autoantibodies as well as SARS-CoV-2 infection specific anti-nucleocapsid (N) IgG antibodies and protective neutralizing antibody (Nab) levels. Methods: Samples from 119 COVID-19 patients categorized based on their level of care and 284 healthy subjects were tested for the presence and persistence of antinuclear and other systemic and organ specific autoantibodies as well as SARS-CoV-2 and neutralizing antibody levels. Results: The data shows significantly increased levels of anti RNP-A, anti-nucleocapsid and neutralizing antibody among patients receiving ICU care compared to non-ICU care. Furthermore, subjects receiving ICU care demonstrated significantly higher nucleocapsid IgG levels among the RNP-A positive cohort compared to RNP-A negative cohort. Notably, the expression of anti RNP-A antibodies is transient that reverts to non-reactive status between 20 and 60 days post symptom onset. Conclusions: COVID-19 patients in ICU care exhibit significantly higher levels of transient RNP-A autoantibodies, anti-nucleocapsid, and SARS-CoV-2 neutralizing antibodies compared to patients in non-ICU care.
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High-throughput and rapid screening testing is highly desirable to effectively combat the rapidly evolving COVID-19 pandemic co-presents with influenza and seasonal common cold epidemics. Here, we present a general workflow for iterative development and validation of an antibody-based microarray assay for the detection of a respiratory viral panel: (a) antibody screening to quickly identify optimal reagents and assay conditions, (b) immunofluorescence assay design including signal amplification for low viral titers, (c) assay characterization with recombinant proteins, inactivated viral samples and clinical samples, and (d) multiplexing to detect a panel of common respiratory viruses. Using RT-PCR-confirmed SARS-CoV-2 positive and negative pharyngeal swab samples, we demonstrated that the antibody microarray assay exhibited a clinical sensitivity and specificity of 77.2% and 100%, respectively, which are comparable to existing FDA-authorized antigen tests. Moreover, the microarray assay is correlated with RT-PCR cycle threshold (Ct) values and is particularly effective in identifying high viral titers. The multiplexed assay can selectively detect SARS-CoV-2 and influenza virus, which can be used to discriminate these viral infections that share similar symptoms. Such protein microarray technology is amenable for scale-up and automation and can be broadly applied as a both diagnostic and research tool.
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Background: During the COVID-19 pandemic, it remains a major concern whether patients with rheumatic musculoskeletal disease treated with conventional (cs) or biologic (b) disease modifying drugs (DMARDs) exhibit an adequate immune response to the currently available SARS-CoV2 vaccines. There remains an urgent need for more data on SARS-CoV-2 vaccine efficacy to inform healthcare providers on the efficiency of the applied vaccination, potential need of and period for booster and/or re-vaccination. Objectives: To assess and compare the efficacy of the SARS-CoV2 vaccines BNT162b2 vaccine (Pfzer/BioNTech) and mRNA-1273 vaccine (Moderna). (The vaccines were administered as part of the Danish vaccine roll out and offered each with two doses and approximately four weeks apart). Patients' SARS-CoV2 IgG serum level was used as proxy to determine vaccination response. Methods: We established the 'Detection of SARS-CoV2 antibodies in Danish Infammatory Rheumatic Outpatients' study (DECODIR) as a longitudinal prospective cohort study. Patients with rheumatoid arthritis (RA), spondyloarthrop-athies (SpA) or psoriatic arthritis (PsA) receiving their outpatient treatment and monitored in the Danish DANBIO registry at the Danish Hospital for Rheumatic Diseases (DG), Sonderborg were included (April-June 2021). Bloods, patient reported outcome measurements (PROMS), clinical data and treatment information (cs/bDMARD) were collected at baseline (prior to vaccination) and after six weeks and six months. SARS-CoV2 IgG levels in serum were assessed by ELISA (ThermoFischer), and manufacturer's cut-off (>=10 EliA U/mL) selected as defnition of sufficient IgG response. Associations between antibody response, age, gender, disease (RA/PsA/SpA), treatment with no or cs/bDMARDs and disease activity were tested using proportional odds regression and bootstrapped tests of medians. Results were reported using mean, median (IqR) and bootstrapped 95% confdence interval (CI) of the median. Results: A total of 243 patients were included at baseline and after six weeks;at six months' follow-up data were available for 233 patients. After six weeks, vaccination was followed by a signifcant increase in IgG levels (median of <0.7 EliA U/mL at baseline versus 36.5 EliA U/mL). Patients treated with a combination of both cDMARD and bDMARD had signifcantly lower IgG levels compared to patients without any DMARD treatment (8,2 EliA U/mL vs 19.5 EliA U/mL (p<0.001)). Patients treated with oral prednisolone (any dose) also showed signifcantly lower median IgG levels compared to patients without DMARD treatment (3,8 EliA U/mL vs 19.5 EliA U/mL (p<0.01)). The actual measurements six months after baseline demonstrated a signifcant decrease of IgG levels for the whole study population (median of 16 EliA U/mL at six month vs 36.5 EliA U/mL at six weeks, p < 0.001) (Figure 1). Similar to week 6, lowest response rates were found in patients treated with prednisolone or combination of csDMARD and bDMARD. After 6 months, the proportional odds model revealed signifcantly lower median IgG antibody level in patients who received Pfzer compared to Moderna (median 15 EliA U/mL (95%CI: 13-18) vs 44.5 EliA U/mL (95%CI: 36-83) (p<0.001). Conclusion: IgG levels decreased markedly six months after the initial double dose regimen. Patients treated with a combination of cs/bDMARD or oral pred-nisolone are at higher risk of inadequate vaccine response as measured by IgG level. Our results support the decision for the need of a third booster vaccine in patients with infammatory rheumatic diseases, especially in the case of cs/bDMARD combination treatment and prednisolone. The data may indicate a need for further revaccination in these patients.
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Background: The relevance of studying immune response after SARS-CoV-2 vaccination in patients with infammatory immune-mediated diseases (IMIDs) represents a deep concern regarding the risk estimation and management of patients with these diseases on immunomodulatory drugs. It is well known that certain treatments as anti CD20 therapies results in a diminished immunogenicity against common vaccines but it is a scarce data regarding the cellular protection obtained upon vaccination between patients with different IMID and between different treatments. Objectives: To compare a potential detriment on cellular and antibody-mediated protection upon SARS-CoV-2 vaccination in patients with IMIDs treated with immunosuppressive drugs. Methods: We recruited 73 patients with rheumatoid arthritis-RA-(n=49), spondy-larthritis-SpA-(n=19), infammatory bowel disease-IBD-(n=5), idiopathic juvenile arthritis-IJA-(n=2) and heterogenous group composed of sclerodermia, lupus, uveitis(n=6). They were treated mainly with rituximab (n=27), TNFi (n=37) or JAKi (n=3). We collected data of age,sex, csDMARDs, previous SARS-CoV-2 infection, last RTX infusion and prednisone use. After one month of vaccination, we assessed the humoral response performing the Thermo Scientific EliA SARS-CoV-2-Sp1 IgG Test (positivity cut-off >0.70 IU/ml) which was also compared with the data with of 35 healthy controls. In addition, in 40 patients who had serum antibody levels under 100UI/ml, we analysed the cellular response by the use of the QuantiFERON SARS-CoV-2 Starter Pack (Quiagen). A cut-off value of 0.15 IU/ml discriminate between positive or negative cell-mediated immune responses. We compared differences among the different IMIDs and between the different immu-nosuppressive treatments through non-parametric test (p<0.05) Results: Regarding demographic characteristics of patients, older patients (>56 years) and female sex were factors which were associated with low titles of serum antibodies. Anti-spike IgG antibodies were present in an 86% of the IMIDs patients and in 100% healthy controls with signifcant different IgG titre (median [IQR]): 51[11-184] vs 700[440-940];p<0.0001. The differences between (median [IQR]) serum antibody levels were statistically different between IMID type: 33[1-138] in RA vs 94[34-191] in SpA vs 204[187-204] in IBD vs 133[61-204] in IJA vs 13[1.5-31.8] in the rest;p=0.04. Remarkably, patients with IBD who had the highest antibodies titles were the youngest compared with the other patients. Target of the therapy played also an important role in serum antibody levels being these: 3.6 [0.7-51] in RTX patients vs 156 [45-204] in TNFi vs 40 [18-58] in JAKi patients;p<0.0001. In those patients who the last infusion of rituximab was, at least, one year before vaccination presented CD19+ B cells detected by fow cytometry and anti-spike IgG antibodies as well. Cell-mediated responses to SARS-CoV-2 were positive in 33% of IMIDs patients, indeterminated in 3% and negative in 65% of the patients. Strikingly, out of the 33% positive patients, 85% were treated with RTX. A 61% of the RTX patients had inducible cell-mediated responses vs 14% of the patients treated with TNFi;p<0.01. On the other hand, there were not differences in cell-mediated responses between positive and negative antibody patients. Conclusion: Titres of serum antibodies against spike protein of SARS-CoV-2 were lower in IMIDs patients than in controls. Patients with RTX had lower rates of positivity humoral response as well as lower serum titles than patients treated with other therapies regardless the patients 'age. Neverthless, in those patients in whom RTX infusion was delayed because of vaccination they conserved a humoral response. On the other hand, more patients treated with RTX had inducible cell-mediated responses compared with patients with TNFi.
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The novel coronavirus 2019 (COVID-19) was first discovered in Wuhan city (China) in patients with severe respiratory disorders and the disease which it cause named scientifically as Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2). Since, its discovery, more than 1,338,604 of COVID-19 case was confirmed between Iraqi peoples a while 30 June 2021 with nearly 17,156 death cases. In order to detect the association of COVID-19 with elevated IL-6 and others blood parametres as D-dimer, C-reactive protein, Ferritin and Lactate dehydrogenase and some other risk factors as age, gender, smoking and place of residence, the study conducted 70 people were devided into three groups. Under 30 year, between 30 to 40 year and above 40 year and also 20 healthy subject were taken as a control group. Levels of IL-6 and the other hematological parameters were estimeated using Automated fluorescent immunoassay system (AFIAS-6) technique. The result showed a significant increase in the level of IL-6 and the other hematological parameters studied in all age groups of COVID-19 patients in comparison with the healthy subjects group. The results also showed that patients with blood group type (o+) were the most frequent in the disease in the study by percent (35.71%) followed by patients with AB+, A+ and B+ and A- blood group type and amounted (25.71%, 18.57%, 14.9% and 5.72%), respectively. Results also showed that the age groups above 40 years were most frequent with the disease in comparison with group of between 30 to 40 and group of under 30 year. The disease was more common in males compared to females in this study and also was more common among non-smokers and between urban residents more than among city residents.
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Objective: To report a pediatric case of severe, treatment-resistant, COVID-19-associated acute longitudinally extensive transverse myelitis (LETM). Background: Since the COVID-19 global pandemic, there is evolving literature reporting the neurological manifestations of the novel coronavirus. COVID-19-associated acute LETM was first reported in an elderly Asian man with lower-extremity weakness <1-week after onset of fever and respiratory distress. In childhood, this is rarely reported with only few reports of COVID-19-associated acute LETM. Design/Methods: We reviewed clinical and radiographic reports of our patient. We searched PubMed for literature using terms “transverse myelitis & COVID-19” and “pediatric transverse myelitis&COVID-19.” Results: A 5-year-old previously healthy boy presented with altered mental status. Prior to admission, he was exposed to COVID-19 and had consumed an unknown quantity of sertraline and risperidone tablets. Thereafter, he stated that he could not feel his legs, fell, and hit his head. In the emergency department, he was intubated. EKG revealed QTc prolongation (486-ms). SARS-CoV-2-PCR positive. Thereafter, flaccid quadriparesis, bulbar dysfunction, left-sided numbness, and hyperreflexia were noted;he communicated by eye blinking. MRI-spine revealed C1-C4 hyperintensity (T2-weighted) consistent with LETM;DWI negative for acute stroke. CSF basic labs, viral and MS panels, and ACE unremarkable. Serum anti-aquaporin-4 and myelin-oligodendrocyte-glycoprotein antibodies negative. Serum West Nile-IgM-IgG negative. Mycoplasma pneumoniae IgM-reactive, IgG-positive;confirmatory IgM immunofluorescence assay-negative. He received IV-methylprednisolone ×5-days, plasmapheresis ×10-sessions, pulsed steroid ×3-days. Minimal neurological improvement was noted. Repeat MRI-spine 2-weeks later unchanged. Tracheostomy and gastric tube were placed. He was transferred to a neurology topic. Conclusions: COVID-19-associated acute LETM in childhood can have a rapid, devastating clinical course. Clinicians should maintain a high-index of suspicion for LETM in COVID-19 pediatric patients presenting with neurological manifestations and consider alternative strategies for severe, treatment-resistant cases.
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BACKGROUND AND AIMS: Vaccination against coronavirus disease 2019 (COVID- 19) can reduce disease incidence and severity. Dialysis patients demonstrate a delayed immunologic response to vaccines. We determined factors affecting the immunologic response to COVID-19 vaccines in hemodialysis patients. METHOD: All patients within a Swedish hemodialysis network, vaccinated with two doses of COVID-19 vaccine 2-8 weeks before inclusion, were eligible for this cross-sectional study. Severe adult respiratory syndrome coronavirus 2 (SARS-CoV-2) spike protein antibody levels were determined by the EliA SARS-CoV-2-Sp1 IgG test (Thermo Fisher Scientific, Phadia AB) and related to clinical and demographic parameters. Eighty-nine patients were included. RESULTS: Patients were vaccinated with two doses of Comirnaty (BNT162b2, 73%) or Spikevax (mRNA-1273, 23.6%). Three patients received combinations of different vaccines. Response rate (antibody titres >7 U/mL) was 89.9%, while 39.3% developed high antibody titres (>204 U/mL), 47 (43-50) days after the second dose. A previous COVID-19 infection associated with higher antibody titres [median (25th-75th percentile) 1558.5 (814.5-3763.8) U/mL versus 87 (26-268) U/mL;P = 0.002], while the time between vaccine doses did not differ between groups (P = 0.7). Increasing SARS-CoV-2 antibody titres were independently associated with increasing time between vaccine doses, decreasing serum calcium levels and previous COVID-19 (Table 1). CONCLUSION: In conclusion, a longer interval between COVID-19 mRNA vaccine doses, lower calcium and a previous COVID-19 infection were independently associated with a stronger immunologic vaccination response in hemodialysis patients. While the response rate was good, only a minority developed high antibody titres 47 (43-50) days after the second vaccine dose. (Table Presented).
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Background: Galectin-9 (Gal-9) is a β-galactoside-binding lectin involved in immune regulation and viral immunopathogenesis. Multiple recent reports demonstrate that plasma levels of Gal-9 are elevated in the setting of severe COVID-19 disease. However, a causal role of Gal-9 in SARS-CoV-2 pathology remains to be elucidated. Here, we determined the impact of Gal-9 on SARS-CoV-2 replication and pro-inflammatory signaling in immortalized and primary human airway epithelial cells (AECs). Methods: Dose-dependent cytotoxicity of recombinant human Gal-9 in the Calu-3 AEC line was determined by MTT assay. Calu-3 cells were infected with SARS-CoV-2 isolate USA-WA1/2020 (MOI=0.01). Primary AECs were isolated from healthy donor lung transplant tissue, cultured at air liquid interface (ALI), and infected with SARS-CoV-2 lineage P.1 (MOI=0.1). SARS-CoV-2 replication was assessed by RT-PCR quantitation of the nucleocapsid (N) gene, immunofluorescence assay (IFA) of N protein, and titration of supernatant (TCID50). Viral entry was measured using luciferase activity of VSV-SARS-CoV-2 S-ΔG-Luciferase reporter pseudovirus. ACE2 and TMPRSS2 cell-surface expression were measured by flow cytometry. Pro-inflammatory factors (IL-6, IL-8, and TNFα) were detected by RT-PCR. Total RNA-seq was used to evaluate Gal-9 effects on the host transcriptome. Groups were compared by Student's t-test, and differential expression analyses were performed using DESeq2. Results: Gal-9 reached 50% cytotoxicity in Calu-3 cells at 597 nM. Gal-9 significantly increased SARS-CoV-2 expression (8.1 to 25.5 fold;p<0.0001) and infectious virus release (1.9 to 17.8 fold;p<0.038) in a dose-dependent manner in Calu-3 cells. Pseudovirus entry into Calu-3 cells was enhanced by Gal-9 (2.4 to 5.6 fold;p<0.0016), and the enhanced entry was inhibited by anti-ACE2 antibody (p<0.0027). Cell surface ACE2 and TMPRSS2 expression were unaffected by Gal-9. Gal-9 treatment accelerated virus-induced expression of IL-6, IL-8, and TNFα (p<0.018) in Calu-3 cells. Gal-9 increased SARS-CoV-2 production (p=0.03) and pro-inflammatory factor expression (p<0.05) in primary AECs (N=5 donors). RNA-seq data revealed that Gal-9 significantly induced IL-17, EIF2, IL-8 and IL-6 signaling pathways in the setting of SARS-CoV-2 infection. Conclusion: Gal-9 facilitates SARS-CoV-2 entry, replication, and virus-induced pro-inflammatory signaling in AECs ex vivo. Our data suggest that pharmacologic manipulation of Gal-9 should be explored as a SARS-CoV-2 therapeutic strategy.
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Background:Most people with coronavirus infectious disease (covid-19) have mild to moderate symptoms and recover after appropriate medical interventions. However 15-32% develop severe or critical COVID-19 with case fatality rate of 1-15%.Treatment in intensive care unit has become a major challenge, early recognition of severe forms is absolutely essential for timely triaging of patients. Hyper inflammation is one of the character associated with disease progression of COVID-19 but the association between C - reactive protein (CRP) and COVID-19 is not clear. So this study aimed to evaluate the association between CRP & severity of COVID-19 confirmed cases. Methods:Clinically confirmed 100 COVID-19 cases referred to Covid Care Center, SSMC, Tumkur were included in the study. CRP was done by Fluorescence Immunoassay by Finecare as per manufacturer’s instructions soon after admission to covid care center. Results:Out of 100 confirmed covid-19 cases 65% were males and 35% were females, predominantly in the age group of 60-80years. CRP was >100mg/dl in 21 patients among this group there was male preponderance(15) with>60 years of age, who were severly ill and were in need of ICU admission, remaining 79 cases showed CRP levels of <100mg/dl were isolated in covid care center. Conclusions:Patients with severe COVID-19 have a higher level of CRP than those with a non-severe disease. Higher lev- els of CRP (>100mg/dl) at presentation might indicate impending clinical deterioration and the need for invasive me- chanical ventilation
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Because of the senescence of the immune system, antibody response to the COVID-19 vaccines may differ from older to younger adults. The study aim compares the titers of SARS-CoV-2 IgG antibody of patients ≥60 years who received three doses of CoronaVac vaccine and those who received two doses of CoronaVac+1 dose of Pfizer-BioNTech after 1 month of the last vaccination. Patients ≥60 years who received the CoronaVac vaccine between March 1, 2021, and April 30, 2021, who did not have COVID-19 disease before the first dose of vaccination and were negative for COVID-19 antibodies, whose antibodies were tested before the third dose of vaccination, and who did not have any COVID-19 disease during the follow-up were included. The demographic characteristics and comorbidities of patients were recorded. An immunofluorescence assay (IFA) fast test and a chemiluminescent microparticle immunoassay (Abbott) were used to measure SARS-CoV-2 quantitative antibody levels at the first month after the third-dose vaccine. Totally 81 patients, 41 patients in third dose of the CoronaVac group (female:male 18:23, mean age 69.4 ± 8.5), and 40 patients in third dose of the Pfizer-BioNTech group (female:male 15:25, mean age 69.9 ± 9.1) were included. The patients' comorbidities in the groups were similar. The titers of IgG antibodies to SARS-CoV-2 measured according to both IFA and Abbott Kit at first month the third dose vaccination was significantly higher in the Pfizer-BioNTech group (p ≥ 0.001, p = 0.012, respectively). The results report that the formed immunity in the first month after the two doses of CoronaVac+1 dose Pfizer-BioNTech vaccine was higher than three doses of CoronaVac vaccine in older adults.
Subject(s)
Antibodies, Viral/blood , COVID-19 Vaccines/immunology , SARS-CoV-2/immunology , Aged , COVID-19/blood , COVID-19/prevention & control , COVID-19 Vaccines/administration & dosage , Cross-Sectional Studies , Female , Humans , Immunogenicity, Vaccine , Immunoglobulin G/blood , Immunosenescence , Male , Middle Aged , Retrospective Studies , VaccinationABSTRACT
Introduction: Kidney biopsy (KB) tissues are usually dissected 0.5-1.0 cm for fresh sample immunofluorescence study (FSIF). Paraffin washed-out immunofluorescence (PWIF) technique doesn’t require segmentation of the kidney biopsy sample that may jeopardise the sample adequacy for histopathological examination (HPE). Hypothetically, it reduces biopsy passes and complications. We examine the safety and adequacy of native kidney biopsy (NKB) and transplant kidney biopsy (TKB) using the FSIF technique and PWIF technique in HPE preparation for immunofluorescent study. Methods: We retrospectively collected clinical history, blood results, and renal biopsy reports for all the patients who had undergone KB using electronic medical records at University Malaya Medical Centre, Kuala Lumpur, Malaysia from 1/1/2015 to 31/4/2021. We performed 1520 KBs (795 NKBs & 725 TKBs) and 1336 KBs with complete data analysed (670 NKBs & 666 TKBs). Sixty-one of the 670 (9.1%) NKBs and 48 of the 666 (7.2%) TKBs employed the PWIF method from 13/10/2020 to 31/4/2021 (6 months) because of the COVID-19 pandemic. We collected 62 NKBs and 65 TKBs from 13/10/2018 to 31/4/2019 (6 months) as the control group which used the FSIF method. The control is chosen as such to correlate with the same months of another year. Results: There were no statistically significant differences in the baseline characteristics between the two groups of each cohort respectively except for the ethnicity distribution in the TKB cohort (p=0.037) and INR in NKB and TKB cohorts (p=0.000 & p=0.002 respectively). PWIF group in NKB and TKB recorded lesser pass (p=0.000 for both), longer HPE core (p=0.001 & p=0.024 respectively), better HPE adequacy (p=0.006 & p=0.012 respectively). There were no statistical differences in the diagnostic yield, immediate pain, gross haematuria, haematoma, and admissions or prolong hospitalisation due to biopsy complications in both groups for NKB and TKB cohorts (Table 1 & 2). [Formula presented] Conclusions: Paraffin washed-out immunofluorescence technique increased the native and transplant kidney biopsy adequacy with a lesser pass and better biopsy sample adequacy for histopathological examination but no difference in the incidence of complications. However, a greater number of cases are required to assess the statistically significant difference in the incidence of complications. No conflict of interest
ABSTRACT
INTRODUCTION: Most adults receiving mRNA vaccines for SARS-CoV-2 (SCV2) exhibit IgG antibodies (Ab) targeting the S1 spike protein within a week of dose 2. However, correlates of protection are still not fully understood. The aim of this study was to better quantify the % of neutralizing Ab (nAb) that develop after dose 2 and also identify factors affecting the timing and degree of nAb production. METHODS: Using a fluorescence immunoassay to quantify the % of SCV2-Ab capable of blocking S1 at its receptor binding domain (for attaching to ACE-2 receptors), residents/ staff (n=70;ages 23-100 yrs) of an assisted living facility had blood samples measured on day 7 and 21 following dose 2 of the Pfizer-BioNTech mRNA vaccine. Based on existing research, %nAb < 30% is delineated as inadequate protection (“nAb negative”). RESULTS: Except for a 58 yo man taking daily prednisone (asthma) and a 55 yo man on levothyroxine, 100% of those < 70 yrs (n=33) were nAb+ (>30% nAb) on day 7 after dose 2. However, if >70 yo (n=37), the % of nAb+ findings diminished with age. Only half of those 71-80 yo, 33% of those 81-90 yo and 11% of those >90 yo were nAb+. Nonetheless, 2 weeks later, the %+ among those tested had increased to 83%, 71%, and 50% for those respective age groups. When examining the average of nAb% measurements within each of the various age group stratifications 1 week after dose 2, the averages ranged 96-100% for the 3 age groups < 50 years (ie, 23-30, 31-40 and 41-50), while the age group averages were borderline or inadequate for those >70 yo. However, 21 days after dose 2, the average %nAb measurement had become 91% for those 61 to 70 years of age, 75% for those 71-80, and 55% for those 81-90. For persons > 90 yo (n=8), the average %nAb was 35% but half of those persons (n=4) had no detectable nAb, either at day 7 or day 21. No persons had any significant declines in %nAb between Day 7 and 21 and the majority sustained or improved their %nAb. CONCLUSIONS: Escalating age and immunomodulating medications/conditions do impact the timing and degree of nAb developing after mRNA vaccination. Most persons < 90 yo are observed to be “positive” for protective levels of nAb by 3 weeks after dose 2. On-going investigations are addressing the duration and sustained degree of nAb+ findings as well as external validation of the tool used in this research.