Assessment of SARS-CoV-2 Specific CD4(+) and CD8 (+) T Cell Responses Using MHC Class I and II Tetramers
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Abstract
The success of SARS-CoV-2 (CoV-2) vaccines is measured by their ability to mount immune memory responses that are long-lasting. To achieve this goal, it is important to identify surrogates of immune protection, namely, CoV-2 MHC Class I and II immunodominant pieces/epitopes and methodologies to measure them. Here, we present results of flow cytometry-based MHC Class I and II QuickSwitch™ platforms for assessing SARS-CoV-2 peptide binding affinities to various human alleles as well as the H-2 Kb mouse allele. Multiple SARS-CoV-2 potential MHC binders were screened and validated by QuickSwitch testing. While several predicted peptides with acceptable theoretical Kd showed poor MHC occupancies, fourteen MHC class II and a few MHC class I peptides showed promiscuity in that they bind with multiple MHC molecule types. With the peptide exchange generated MHC tetramers, scientists can assess CD4+ and CD8+ immune responses to these different MHC/peptide complexes. Results obtained with several SARS-CoV-2 MHC class I and II peptides are included and discussed.
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SciScore for 10.1101/2020.07.08.194209: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Institutional Review Board Statement not detected. Randomization not detected. Blinding not detected. Power Analysis not detected. Sex as a biological variable not detected. Table 2: Resources
Antibodies Sentences Resources Briefly, 20 µL of magnetic beads linked with an anti HLA-ABC antibody (provided in the kit) were added into sample wells of a V-bottom 96-well microtiter plate with 5 µL of the Tetramer peptide exchange solution (from the 52 µL total volume). anti HLA-ABCsuggested: NoneFollowing a 150 µL wash with Peptide Exchange Assay Buffer #2 (provided in the kit, must be diluted 10x before use) the exchanged MHC Class II molecules were stained with 25 µL of Exiting Peptide Antibody-FITC (provided in the … SciScore for 10.1101/2020.07.08.194209: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Institutional Review Board Statement not detected. Randomization not detected. Blinding not detected. Power Analysis not detected. Sex as a biological variable not detected. Table 2: Resources
Antibodies Sentences Resources Briefly, 20 µL of magnetic beads linked with an anti HLA-ABC antibody (provided in the kit) were added into sample wells of a V-bottom 96-well microtiter plate with 5 µL of the Tetramer peptide exchange solution (from the 52 µL total volume). anti HLA-ABCsuggested: NoneFollowing a 150 µL wash with Peptide Exchange Assay Buffer #2 (provided in the kit, must be diluted 10x before use) the exchanged MHC Class II molecules were stained with 25 µL of Exiting Peptide Antibody-FITC (provided in the kit). Antibody-FITCsuggested: (Alomone Labs Cat# AAR-007-F, RRID:AB_2756537)Software and Algorithms Sentences Resources In Silico prediction of peptide binding affinities for MHC molecules: All of the theoretical binding affinities were derived from the Immune Epitope Database (IEDB) web resource funded by NIAID. NIAIDsuggested: (NIAID, RRID:SCR_016598)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: 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.
- No funding statement was detected.
- No protocol registration statement was detected.
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