New-onset IgG autoantibodies in hospitalized patients with COVID-19
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Abstract
COVID-19 is associated with a wide range of clinical manifestations, including autoimmune features and autoantibody production. Here we develop three protein arrays to measure IgG autoantibodies associated with connective tissue diseases, anti-cytokine antibodies, and anti-viral antibody responses in serum from 147 hospitalized COVID-19 patients. Autoantibodies are identified in approximately 50% of patients but in less than 15% of healthy controls. When present, autoantibodies largely target autoantigens associated with rare disorders such as myositis, systemic sclerosis and overlap syndromes. A subset of autoantibodies targeting traditional autoantigens or cytokines develop de novo following SARS-CoV-2 infection. Autoantibodies track with longitudinal development of IgG antibodies recognizing SARS-CoV-2 structural proteins and a subset of non-structural proteins, but not proteins from influenza, seasonal coronaviruses or other pathogenic viruses. We conclude that SARS-CoV-2 causes development of new-onset IgG autoantibodies in a significant proportion of hospitalized COVID-19 patients and are positively correlated with immune responses to SARS-CoV-2 proteins.
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SciScore for 10.1101/2021.01.27.21250559: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Institutional Review Board Statement not detected. Randomization Array probing: Serum or plasma samples were first heat inactivated at 56°C for one hour59 then tested at 1:100 dilution in 0.05% PBS-Tween supplemented with 1% (w/v) bovine serum albumin (BSA) and transferred into 96-well plates in a randomized layout. Blinding Samples were screened in a blinded fashion at a dilution of 1:80 with ultraviolet (UV) microscopy by clinical laboratory staff (A.G. and J.G.) who have extensive experience in the interpretation of ANA patterns. Power Analysis not detected. Sex as a biological variable not detected. Cell Line Authentication not detected. Table 2: Resources
Antibodies Sentences Resources In addition, … SciScore for 10.1101/2021.01.27.21250559: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Institutional Review Board Statement not detected. Randomization Array probing: Serum or plasma samples were first heat inactivated at 56°C for one hour59 then tested at 1:100 dilution in 0.05% PBS-Tween supplemented with 1% (w/v) bovine serum albumin (BSA) and transferred into 96-well plates in a randomized layout. Blinding Samples were screened in a blinded fashion at a dilution of 1:80 with ultraviolet (UV) microscopy by clinical laboratory staff (A.G. and J.G.) who have extensive experience in the interpretation of ANA patterns. Power Analysis not detected. Sex as a biological variable not detected. Cell Line Authentication not detected. Table 2: Resources
Antibodies Sentences Resources In addition, prototype human plasma samples derived from participants with autoimmune diseases with known reactivity patterns (e.g. ds-DNA, Scl-70, centromere, SSA, SSB, cardiolipin, whole histones, and RNP, all purchased from ImmunoVision; also from Stanford Autoimmune Diseases Biobank, and OMRF); APS-1, IPEX, PAP, or AMI associated with anti-IFN- γ blocking antibodies; as well as normal human sera (ImmunoVision, Product # HNP-0300, certified to be nonreactive to Hep-2 cell lysates at a titer of 1:100), were used for validation. SSBsuggested: Noneanti-IFN-suggested: (Novus Cat# NB100-78214, RRID:AB_1084710)Additional controls included samples from five healthy donors and three de-identified patients known to have clinically elevated PR3 and MPO antibody levels. MPOsuggested: NoneANAs were detected using a FITC- conjugated goat anti-human IgG antibody following vendor instructions. anti-human IgGsuggested: NoneGoat anti-human IgG-HRP (Cat# 109-035-008, Jackson ImmunoResearch Laboratories, West Grove, PA) was diluted 1:10,000 with sample dilution buffer and 50 μl of secondary antibody was added to each well. Goat anti-human IgG-HRPsuggested: (Santa Cruz Biotechnology Cat# sc-2907, RRID:AB_650497)anti-human IgG-HRPsuggested: (Santa Cruz Biotechnology Cat# sc-2769, RRID:AB_656966)Antibodies were considered “positive” if MFI was > 5 SD above the average MFI for HC for that antigen, and MFI was >3,000 units, a threshold which is more stringent than commonly published in related literature21. antigen,suggested: NoneELISA and antibody number data were visualized in GraphPad Prism v.9.0.0 (86). ELISAsuggested: NoneExperimental Models: Cell Lines Sentences Resources In addition, prototype human plasma samples derived from participants with autoimmune diseases with known reactivity patterns (e.g. ds-DNA, Scl-70, centromere, SSA, SSB, cardiolipin, whole histones, and RNP, all purchased from ImmunoVision; also from Stanford Autoimmune Diseases Biobank, and OMRF); APS-1, IPEX, PAP, or AMI associated with anti-IFN- γ blocking antibodies; as well as normal human sera (ImmunoVision, Product # HNP-0300, certified to be nonreactive to Hep-2 cell lysates at a titer of 1:100), were used for validation. Hep-2suggested: NoneSoftware and Algorithms Sentences Resources In addition, prototype human plasma samples derived from participants with autoimmune diseases with known reactivity patterns (e.g. ds-DNA, Scl-70, centromere, SSA, SSB, cardiolipin, whole histones, and RNP, all purchased from ImmunoVision; also from Stanford Autoimmune Diseases Biobank, and OMRF); APS-1, IPEX, PAP, or AMI associated with anti-IFN- γ blocking antibodies; as well as normal human sera (ImmunoVision, Product # HNP-0300, certified to be nonreactive to Hep-2 cell lysates at a titer of 1:100), were used for validation. ImmunoVisionsuggested: NoneImages were acquired with a Hamamatsu ORCA- ER B&W CCD Digital Camera controlled with Metamorph V7.10.3.390 software and 1×1 camera binning. Metamorphsuggested: NoneELISA and antibody number data were visualized in GraphPad Prism v.9.0.0 (86). GraphPad Prismsuggested: (GraphPad Prism, RRID:SCR_002798)Upon publication of this study in a peer-reviewed journal, deidentified array data will be uploaded to the Gene Expression Omnibus (GEO) database. Gene Expression Omnibussuggested: (Gene Expression Omnibus (GEO, RRID:SCR_005012)Results from OddPub: We did not detect open data. We also did not detect open code. Researchers are encouraged to share open data when possible (see Nature blog).
Results from LimitationRecognizer: We detected the following sentences addressing limitations in the study:Many studies of hospitalized COVID-19 patients, including our study, suffer from important limitations. First, confounding variables exist including heterogeneous demographics, medications at hospitalization, individualized treatment approaches, and, in some cases, unknown history of pre- existing medical or autoimmune conditions. Second, “Day 0” is not day 0 of infection but instead refers to a time point most proximate to hospitalization. Our viral array results (Figs. 4 and 5) confirm that the time between initial infection and sample acquisition was heterogeneous, potentially confounding interpretation of autoantibody and ACA results. Third, not all antigens (e.g., lipids, hydrophobic proteins and carbohydrates) are compatible with our screening methodology, and as a result we have certainly missed some reactivities. Fourth, we did not include patients who were asymptomatic, had mild COVID-19, were vaccinated for SARS-CoV-2, had other severe viral illnesses, or were children. Finally, our analysis was limited to hospitalized patients during acute illness, with follow up times of days rather than months or years. Although beyond the scope of these studies, our data generate many more questions that need to be addressed in the coming years – questions that can only be answered by generating large cohorts of prospectively enrolled subjects with new-onset viral syndromes, including patients with COVID-19, respiratory illnesses which resemble COVID-19, and subjects enrolled in...
Results from TrialIdentifier: No clinical trial numbers were referenced.
Results from Barzooka: We found bar graphs of continuous data. We recommend replacing bar graphs with more informative graphics, as many different datasets can lead to the same bar graph. The actual data may suggest different conclusions from the summary statistics. For more information, please see Weissgerber et al (2015).
Results from JetFighter: Please consider improving the rainbow (“jet”) colormap(s) used on page 75. At least one figure is not accessible to readers with colorblindness and/or is not true to the data, i.e. not perceptually uniform.
Results from scite Reference Check: We found no unreliable references.
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