Development of an in-house SARS-CoV-2 interferon-gamma ELISpot and plate reader-free spot detection method.
J Virol Methods
; 300: 114398, 2022 02.
Article
in English
| MEDLINE | ID: covidwho-1654867
ABSTRACT
Coronavirus disease 2019 (COVID-19) vaccination programs rolled out in an attempt to stop the COVID-19 pandemic. Besides neutralising antibodies, effective T cell responses are also crucial for protection against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection and COVID-19 disease severity. To assess SARS-CoV-2-specific T cell immunity, we developed an interferon-gamma (IFN-γ) enzyme-linked immunospot (ELISpot) that can be deployed in research and diagnostic settings. We optimised our ELISpot by testing multiple antigen concentrations to stimulate peripheral blood mononuclear cells of SARS-CoV-2-unexposed, COVID-19 convalescent and COVID-19 vaccinated volunteers. Also, we developed an ELISpot plate reader-free method to detect and quantify spots, which we compared to manual spot counting and automated analysis by an ELISpot plate reader. We observed strong SARS-CoV-2-reactive T cell responses in COVID-19 convalescent, and COVID-19 vaccinated volunteers but absent or only weak responses in unexposed volunteers. Overall, antigens with concentrations from 0.1 to 5.0 µg/mL per peptide elicited similar T cell responses. Also, our plate reader-free detection method reliably detected and quantified SARS-CoV-2-specific T cells, demonstrated by an excellent reliability when compared to manual analysis and automated analysis by an ELISpot plate reader.
Keywords
Full text:
Available
Collection:
International databases
Database:
MEDLINE
Main subject:
T-Lymphocytes
/
COVID-19
/
Immunity, Cellular
Type of study:
Diagnostic study
/
Prognostic study
Topics:
Vaccines
Limits:
Humans
Language:
English
Journal:
J Virol Methods
Year:
2022
Document Type:
Article
Affiliation country:
J.jviromet.2021.114398
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