Enhanced SARS-CoV-2 Neutralization by Secretory IgA in vitro
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Abstract
SARS-CoV-2 primarily infects cells at mucosal surfaces. Serum neutralizing antibody responses are variable and generally low in individuals that suffer mild forms of the illness. Although potent IgG antibodies can neutralize the virus, less is known about secretory antibodies such as IgA that might impact the initial viral spread and transmissibility from the mucosa. Here we characterize the IgA response to SARS-CoV-2 in a cohort of 149 individuals. IgA responses in plasma generally correlate with IgG responses and clones of IgM, IgG and IgA producing B cells that are derived from common progenitors are evident. Plasma IgA monomers are 2-fold less potent than IgG equivalents. However, IgA dimers, the primary form in the nasopharynx, are on average 15 times more potent than IgA monomers. Thus, secretory IgA responses may be particularly valuable for protection against SARS-CoV-2 and for vaccine efficacy.
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SciScore for 10.1101/2020.09.09.288555: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Institutional Review Board Statement Consent: All participants provided written informed consent before participation in the study and the study was conducted in accordance with Good Clinical Practice and clinical data collection.
IRB: The study was performed in compliance with all relevant ethical regulations and the protocol was approved by the Institutional Review Board (IRB) of the Rockefeller University.Randomization not detected. Blinding not detected. Power Analysis not detected. Sex as a biological variable not detected. Cell Line Authentication Contamination: All cell lines have been tested negative for contamination with mycoplasma and parental cell lines were obtained from the ATCC. Table 2: …
SciScore for 10.1101/2020.09.09.288555: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Institutional Review Board Statement Consent: All participants provided written informed consent before participation in the study and the study was conducted in accordance with Good Clinical Practice and clinical data collection.
IRB: The study was performed in compliance with all relevant ethical regulations and the protocol was approved by the Institutional Review Board (IRB) of the Rockefeller University.Randomization not detected. Blinding not detected. Power Analysis not detected. Sex as a biological variable not detected. Cell Line Authentication Contamination: All cell lines have been tested negative for contamination with mycoplasma and parental cell lines were obtained from the ATCC. Table 2: Resources
Antibodies Sentences Resources Plates were washed 6 times with washing buffer and then incubated with anti-human IgG (Jackson Immuno Research 109-036-088) or anti-human IgA (Sigma A0295) secondary antibody conjugated to horseradish peroxidase (HRP) in blocking buffer at 1:5000 or 1:3000 dilution respectively. anti-human IgGsuggested: (Jackson ImmunoResearch Labs Cat# 109-036-088, RRID:AB_2337594)anti-human IgAsuggested: (Sigma-Aldrich Cat# A0295, RRID:AB_257876)The half-maximal and 90% inhibitory concentrations for purified plasma IgG or IgA or monoclonal antibodies (IC50 and IC90) were determined using 4-parameter nonlinear regression (GraphPad Prism). IgAsuggested: NoneIC90suggested: NoneThe antibody-virus-mix was then directly applied to VeroE6 cells (MOI of ∼0.1 PFU/cell; n=3) and incubated for 22h at 37°C. antibody-virus-mixsuggested: NoneGoat anti-rabbit AlexaFluor 594 (catalog no. A-11012; Life Technologies) was used as a secondary antibody at a dilution of 1:2,000. anti-rabbitsuggested: NoneThe frequency distributions of human V genes in anti-SARS-CoV-2 antibodies from this study was compared to 131,284,220 IgH and IgL sequences generated by (44) and downloaded from cAb-Rep (45), a database of human shared BCR clonotypes available at https://cab-rep.c2b2.columbia.edu/. anti-SARS-CoV-2suggested: NoneExperimental Models: Cell Lines Sentences Resources 14 cells (27), 293TAce2 cells (11) and VeroE6 kidney epithelial cells were cultured in Dulbecco’s modified Eagle medium (DMEM) supplemented with 10% FCS at 37°C and 5% CO2. 293TAce2suggested: RRID:CVCL_YZ65)In addition, medium for Ace2-overexpressing cell lines contained 5 µg/ml blasticidin and medium for VeroE6 cells was supplemented with 1 % nonessential amino acids. VeroE6suggested: JCRB Cat# JCRB1819, RRID:CVCL_YQ49)Pseudotyped virus neutralization assay: SARS-CoV-2 pseudotyped particles were produced by co-transfection of pSARS-CoV-2 Strunc and pNL4-3ΔEnv-nanoluc in 293T cells (11, 27). 293Tsuggested: NCBI_Iran Cat# C498, RRID:CVCL_0063)Software and Algorithms Sentences Resources The average of its signal was used for normalization of all the other values on the same plate with Excel software prior to calculating the area under the curve using Prism 8 (GraphPad). Excelsuggested: NoneThe half-maximal and 90% inhibitory concentrations for purified plasma IgG or IgA or monoclonal antibodies (IC50 and IC90) were determined using 4-parameter nonlinear regression (GraphPad Prism). GraphPadsuggested: (GraphPad Prism, RRID:SCR_002798)Sequence analysis was performed with MacVector. MacVectorsuggested: (MacVector, RRID:SCR_015700)Results from OddPub: Thank you for sharing your code and data.
Results from LimitationRecognizer: An explicit section about the limitations of the techniques employed in this study was not found. We encourage authors to address study limitations.Results from TrialIdentifier: No clinical trial numbers were referenced.
Results from Barzooka: We did not find any issues relating to the usage of bar graphs.
Results from JetFighter: We did not find any issues relating to colormaps.
Results from rtransparent:- Thank you for including a conflict of interest statement. Authors are encouraged to include this statement when submitting to a journal.
- Thank you for including a funding statement. Authors are encouraged to include this statement when submitting to a journal.
- No protocol registration statement was detected.
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