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A Barcoded Flow Cytometric Assay to Explore the Antibody Responses Against SARS-CoV-2 Spike and Its Variants.
Vesper, Niklas; Ortiz, Yaneth; Bartels-Burgahn, Frauke; Yang, Jianying; de la Rosa, Kathrin; Tenbusch, Matthias; Schulz, Sebastian; Finzel, Stephanie; Jäck, Hans-Martin; Eibel, Hermann; Voll, Reinhard E; Reth, Michael.
  • Vesper N; Institute of Biology III, Faculty of Biology, University of Freiburg, Freiburg, Germany.
  • Ortiz Y; Research Centres Bioss, Centre for Biological signal studies, CIBSS, Centre for Integrative Biological Signalling Studies, University of Freiburg, Freiburg, Germany.
  • Bartels-Burgahn F; Institute of Biology III, Faculty of Biology, University of Freiburg, Freiburg, Germany.
  • Yang J; Research Centres Bioss, Centre for Biological signal studies, CIBSS, Centre for Integrative Biological Signalling Studies, University of Freiburg, Freiburg, Germany.
  • de la Rosa K; Institute of Biology III, Faculty of Biology, University of Freiburg, Freiburg, Germany.
  • Tenbusch M; Research Centres Bioss, Centre for Biological signal studies, CIBSS, Centre for Integrative Biological Signalling Studies, University of Freiburg, Freiburg, Germany.
  • Schulz S; Institute of Biology III, Faculty of Biology, University of Freiburg, Freiburg, Germany.
  • Finzel S; Research Centres Bioss, Centre for Biological signal studies, CIBSS, Centre for Integrative Biological Signalling Studies, University of Freiburg, Freiburg, Germany.
  • Jäck HM; Department of Cancer and Immunology, Max-Delbrück-Center for Molecular Medicine in the Helmholtz Association (MDC), Berlin, Germany.
  • Eibel H; Institute of Clinical and Molecular Virology, University Hospital Erlangen, Friedrich-Alexander University Erlangen-Nuremberg, Erlangen, Germany.
  • Voll RE; Division of Molecular Immunology, Internal Medicine III, Nikolaus-Fiebiger-Center of Molecular Medicine, Friedrich-Alexander University Erlangen-Nuremberg, Erlangen, Germany.
  • Reth M; Department of Rheumatology and Clinical Immunology, Medical Center - University of Freiburg, Faculty of Medicine, University of Freiburg, Freiburg, Germany.
Front Immunol ; 12: 730766, 2021.
Article in English | MEDLINE | ID: covidwho-1463473
ABSTRACT
The SARS-CoV-2 pandemic has spread to all parts of the world and can cause life-threatening pneumonia and other severe disease manifestations known as COVID-19. This health crisis has resulted in a significant effort to stop the spread of this new coronavirus. However, while propagating itself in the human population, the virus accumulates mutations and generates new variants with increased fitness and the ability to escape the human immune response. Here we describe a color-based barcoded spike flow cytometric assay (BSFA) that is particularly useful to evaluate and directly compare the humoral immune response directed against either wild type (WT) or mutant spike (S) proteins or the receptor-binding domains (RBD) of SARS-CoV-2. This assay employs the human B lymphoma cell line Ramos, transfected for stable expression of WT or mutant S proteins or a chimeric RBD-CD8 fusion protein. We find that the alpha and beta mutants are more stably expressed than the WT S protein on the Ramos B cell surface and/or bind with higher affinity to the viral entry receptor ACE2. However, we find a reduce expression of the chimeric RBD-CD8 carrying the point mutation N501Y and E484K characteristic for the alpha and beta variant, respectively. The comparison of the humoral immune response of 12 vaccinated probands with 12 COVID-19 patients shows that after the boost, the S-specific IgG class immune response in the vaccinated group is similar to that of the patient group. However, in comparison to WT the specific IgG serum antibodies bind less well to the alpha variant and only poorly to the beta variant S protein. This is in line with the notion that the beta variant is an immune escape variant of SARS-CoV-2. The IgA class immune response was more variable than the IgG response and higher in the COVID-19 patients than in the vaccinated group. In summary, we think that our BSFA represents a useful tool to evaluate the humoral immunity against emerging variants of SARS-CoV-2 and to analyze new vaccination protocols against these variants.
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Full text: Available Collection: International databases Database: MEDLINE Main subject: Cell Separation / Spike Glycoprotein, Coronavirus / Flow Cytometry / SARS-CoV-2 / COVID-19 Type of study: Experimental Studies Topics: Vaccines / Variants Limits: Female / Humans / Male / Middle aged Language: English Journal: Front Immunol Year: 2021 Document Type: Article Affiliation country: Fimmu.2021.730766

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Full text: Available Collection: International databases Database: MEDLINE Main subject: Cell Separation / Spike Glycoprotein, Coronavirus / Flow Cytometry / SARS-CoV-2 / COVID-19 Type of study: Experimental Studies Topics: Vaccines / Variants Limits: Female / Humans / Male / Middle aged Language: English Journal: Front Immunol Year: 2021 Document Type: Article Affiliation country: Fimmu.2021.730766