Validation of an interferon-gamma enzyme-linked immunosorbent spot assay to evaluate cell-mediated immunity against severe acute respiratory syndrome coronavirus 2
| Title: | Validation of an interferon-gamma enzyme-linked immunosorbent spot assay to evaluate cell-mediated immunity against severe acute respiratory syndrome coronavirus 2 |
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| Authors: | Yunhwa Kim, Eun-Jeong Jang, Ji-Young Hwang, Kyung-Min Lee, Sivilay Xayaheuang, Seok-Tae Choi, Hosun Park |
| Source: | Human Vaccines & Immunotherapeutics, Vol 20, Iss 1 (2024) |
| Publisher Information: | Taylor & Francis Group, 2024. |
| Publication Year: | 2024 |
| Collection: | LCC:Immunologic diseases. Allergy LCC:Therapeutics. Pharmacology |
| Subject Terms: | ELISpot assay, cell-mediated immunity, validation, SARS-CoV-2, COVID-19, Immunologic diseases. Allergy, RC581-607, Therapeutics. Pharmacology, RM1-950 |
| More Details: | The COVID-19 pandemic forced the rapid development of methods to measure humoral and cellular immunity against SARS-CoV-2. The lack of a global standardized protocol and the high variability of intra- and inter-assay precision of the T-cell response made it difficult to compare T-cell assay results with those of other laboratories. The interferon-gamma enzyme-linked immunosorbent spot (IFN-γ ELISpot) assay for immunogenicity evaluation was validated using naturally infected donor peripheral blood mononuclear cells, a commercially available IFN-γ ELISpot kit, and a SARS-CoV-2 specific peptide pool. Depending on anti-CD3 and peptide pool stimulation, the mean coefficients of variation (CVs) of the intra-assay precision were 19.0% and 13.4%, respectively. The mean CVs of the inter-assay precision were 26.1% and 25.4%, and the mean CVs for reproducibility were 6.7% and 15.9%, respectively. Linearity with an R-squared value between 0.98 and 0.99 was established, and the mean CVs between the lots were 17.6% and 6.6%, depending on the anti-CD3 and peptide pool stimulation, respectively. The limit of detection was 11 spot-forming counts per well. Taken together, we demonstrated that the IFN-γ ELISpot assay is feasible for evaluating SARS-CoV-2-specific cell-mediated immune function through validation based on standard operating procedures. |
| Document Type: | article |
| File Description: | electronic resource |
| Language: | English |
| ISSN: | 21645515 2164-554X 2164-5515 |
| Relation: | https://doaj.org/toc/2164-5515; https://doaj.org/toc/2164-554X |
| DOI: | 10.1080/21645515.2024.2428516 |
| Access URL: | https://doaj.org/article/af21b1388e1d42ecb577b4ce106501f3 |
| Accession Number: | edsdoj.f21b1388e1d42ecb577b4ce106501f3 |
| Database: | Directory of Open Access Journals |
| ISSN: | 21645515 2164554X |
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| DOI: | 10.1080/21645515.2024.2428516 |
| Published in: | Human Vaccines & Immunotherapeutics |
| Language: | English |