SARS-CoV-2 specific T-cell humoral response assessment after COVID-19 vaccination using a rapid direct real-time PCR amplification
Author:
Cosma Chiara12, Galla Luisa12, Padoan Andrea123ORCID, Furlan Giulia23, Marchioro Lucio2, Zaninotto Martina12, Basso Daniela123, Plebani Mario23ORCID
Affiliation:
1. Laboratory Medicine Unit , University-Hospital of Padova , Padova , Italy 2. QI.LAB.MED., Spin-off of the University of Padova , Padova , Italy 3. Department of Medicine-DIMED , University-Hospital of Padova , Padova , Italy
Abstract
Abstract
Objectives
The SARS-CoV-2 immune response is mediated by both humoral and cellular immunity. In this study, SARS-CoV-2 specific cellular immunity was tested by a novel direct real-time PCR (dRT-PCR) assay, targeting mRNA of CXCL10, and compared with respect to an ELISA measuring interferon gamma (IFN-γ) release.
Methods
Whole blood (Li–He) and serum samples were collected from 92 healthcare workers (HCW), with three doses of homologous (Pfizer/BioNTech, n=74) or heterologous (Pfizer/BioNTech and Vaxzevria or Moderna, n=18) vaccinations. Li–He samples were incubated with SCV2 PANEL-1-T-ACTIVATION (Hyris srl, Lodi, Italy), or CoV-2 IGRA TUBE ELISA (Euroimmune, Lubeck, Germany). CXCL10 mRNA expression was analyzed by bCube/bApp (Hyris), while IFN-γ was evaluated by quant-T-Cell SARS-CoV-2 ELISA (Euroimmune). Anti-SARS-CoV-2 S-RBD IgG levels were measured in sera using a CLIA assay (Snibe, Shenzen, China).
Results
Imprecision of dRT-PCR assay was found to be satisfactory, and the two methods for measuring T cell immunity to SARS-CoV-2 peptides agreed in 82/87 (94.2%) of results. At qualitative dRT-PCR analyses, 81 subjects (93.2%) resulted as reactive to SARS-CoV-2 peptides, 3 (3.4%) were borderline and 3 were negative (3.4%). At univariate and multivariate analyses of quantitative dRT-PCR mRNA of CXCL10 and IFN-γ release results showed no difference between HCW with previous infection, homologous/heterologous vaccination, or demographical features. Anti-SARS-CoV-2 S-RBD IgG was associated with the previous infection and the time between the last vaccination or positivity.
Conclusions
Direct RT-PCR appeared accurate for determining the presence or absence of immunoreactivity of SARS-CoV-2 specific T cells, especially when rapid analyses are required, such as for organ transplantation.
Publisher
Walter de Gruyter GmbH
Subject
Biochemistry (medical),Clinical Biochemistry,General Medicine
Reference34 articles.
1. Ramasamy, MN, Minassian, AM, Ewer, KJ, Flaxman, AL, Folegatti, PM, Owens, DR, et al.. Safety and immunogenicity of ChAdOx1 nCoV-19 vaccine administered in a prime-boost regimen in young and old adults (COV002): a single-blind randomised, controlled, phase 2/3 trial. Lancet 2020;396:1979. https://doi.org/10.1016/S0140-6736(20)32466-1. 2. Polack, FP, Thomas, SJ, Kitchin, N, Absalon, J, Gurtman, A, Lockhart, S, et al.. Safety and efficacy of the BNT162b2 mRNA Covid-19 vaccine. NEJM 2020;383:2603. https://doi.org/10.1056/nejmoa2034577. 3. Baden, LR, Sahly, HME, Essink, B, Kotloff, K, Frey, S, Novak, R, et al.. Efficacy and safety of the mRNA-1273 SARS-CoV-2 vaccine. NEJM 2021;384:403. https://doi.org/10.1056/nejmoa2035389. 4. Sadoff, J, Gray, G, Vandebosch, A, Cárdenas, V, Shukarev, G, Grinsztejn, B, et al.. Safety and efficacy of single-dose Ad26.COV2.S vaccine against Covid-19. NEJM 2021;384:2187. https://doi.org/10.1056/nejmoa2101544. 5. Coronavirus (COVID-19) vaccinations. Available from: https://ourworldindata.org/covid-vaccinations [Accessed 2 Feb 2023].
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