T-cell proliferation assay for the detection of SARS-CoV-2-specific T-cells

• Published in: Clinica chimica acta 2022-07, Vol.532, p.130-136
• Main Authors: Chu, Chang, Schönbrunn, Anne, Elitok, Saban, Kern, Florian, Schnatbaum, Karsten, Wenschuh, Holger, Klemm, Kristin, von Baehr, Volker, Krämer, Bernhard K., Hocher, Berthold
• Format: Article
• Language: English
• Subjects: Antibodies, Viral; Cell Proliferation; COVID-19 - diagnosis; Humans; Humoral and cellular immune responses; Lymphocyte transformation test; Peptides; SARS-CoV-2; Spike Glycoprotein, Coronavirus; T-cell response; T-Lymphocytes; Vaccination
• ISSN: 0009-8981; 1873-3492
• DOI: 10.1016/j.cca.2022.05.025

•The present study shows that the novel SARS-CoV-2 lymphocyte transformation test (LTT) may improve the identification of those with infection or vaccine-induced immunity to the coronavirus and our data suggests that this new approach can be rolled out to a range of clinical situations.•The SARS-CoV-2 LTT can not only differentiate between acquired immunity due to natural infection or vaccination, but it may also help us examine and understand the complexity of the cellular immune response at an individual level. Both infection with and vaccination against SARS-CoV-2 trigger a complex B-cell and T-cell response. Methods for the analysis of the B-cell response are now well established. However, reliable methods for measuring the T-cell response are less well established and their usefulness in clinical settings still needs to be proven. Here, we have developed and validated a T-cell proliferation assay based on 3H thymidine incorporation. The assay is using SARS-CoV-2 derived peptide pools that cover the spike (S), the nucleocapsid (N) and the membrane (M) protein for stimulation. We have compared this novel SARS-CoV-2 lymphocyte transformation test (SARS-CoV-2 LTT) to an established ELISA assay detecting Immunoglobulin G (IgG) antibodies to the S1 subunit of the SARS-CoV-2 spike protein. The study was carried out using blood samples from both vaccinated and infected health care workers as well as from a non-infected control group. Our novel SARS-CoV-2 LTT shows excellent discrimination of infected and/or vaccinated individuals versus unexposed controls, with the ROC analysis showing an area under the curve (AUC) of > 0.95. No false positives were recorded as all unexposed controls had a negative LTT result. When using peptide pools not only representing the S protein (found in all currently approved vaccines) but also the N and M proteins (not contained in the vast majority of vaccines), the novel SARS-CoV-2 LTT can also discriminate T-cell responses resulting from vaccination against those induced by infection.