A total of 18 healthy individuals (15 female) with no history of SARS-CoV-2 infection at baseline who worked in the microbiology laboratory (out of 50 potential participants) served as controls. in controls (17/18, 94.4%). A booster effect was documented in post-vaccination samples of nursing-home residents with prior COVID-19. Plasma antibody levels were higher (p?0.01) in recovered nursing-home residents (all 2500 IU/mL) than in individuals across the other two groups (median 1120 IU/mL in na?ve nursing-home residents and 2211 IU/ml in controls). A large percentage of nursing-home residents had SARS-CoV-2 S-reactive IFN- CD8+ (na?ve 31/49, 63.2%; recovered 8/10, 80%) or CD4+ T cells (na?ve 35/49, 71.4%; recovered 7/10, 70%) at baseline, in contrast to healthy controls (3/17, 17.6% and 5/17, 29%, respectively). SARS-CoV-2 IFN- CD8+ CPI-268456 and CD4+ T-cell responses were documented in 88% (15/17) and all control subjects after vaccination, respectively, but only in 65.5% (38/58) and 22.4% (13/58) of nursing-home residents. Overall, the median frequency of SARS-CoV-2 IFN- CD8+ and CD4+ T cells in nursing-home residents decreased in post-vaccination specimens, whereas it increased in controls. Conclusion The Comirnaty COVID-19 vaccine elicits robust SARS-CoV-2 S antibody responses in nursing-home residents. Nevertheless, the rate and frequency of detectable SARS-CoV-2 IFN- T-cell responses after vaccination was lower in nursing-home residents than in controls. Keywords: Comirnaty?COVID-19 vaccine, Nursing-home residents, SARS-CoV-2, SARS-CoV-2 S antibodies, SARS-CoV-2 S T cells Introduction The Comirnaty? (PfizerCBioNTech) coronavirus disease 2019 (COVID-19) vaccinea nucleoside-modified messenger RNA that encodes the full-length transmembrane S glycoprotein locked in its perfusion conformationelicits high levels of serum neutralizing antibodies (NtAbs), targeting mainly the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) receptor-binding domain (RBD), and Ednra strong TH1-skewed functional CD4+ and CD8+ T-cell responses in experimental models and humans [[1], [2], [3], [4]]. The efficacy of the vaccine has been shown to approach 95% in preventing COVID-19 across a wide range of age groups [5]. Real-world data have confirmed the efficacy of the vaccine in protecting against the occurrence of severe clinical forms of the disease [6,7]. Nevertheless, information on the immunogenicity and efficacy of this vaccine in elderly people with comorbidities and frailty is scarce [8,9]; such people have been prioritized for vaccination worldwide due to their increased risk of developing severe clinical forms of COVID-19 [10]. To gain further insight into this issue, we assess here SARS-CoV-2-S-targeted antibody and CPI-268456 functional T-cell responses after vaccination with Comirnaty in a cohort of nursing-home residents, most displaying one or more comorbidities, and either presumably SARS-CoV-2-na? ve or with documented prior SARS-CoV-2 infection. Material and methods Participants and study design CPI-268456 A total of 60 participants (44 female) in the study, selected from two nursing homes affiliated to the Clnico-Malvarrosa Health Department, Valencia, Spainwhich together provide care for 226 residentswere enrolled in the current study. A total of 18 healthy individuals (15 female) with no history of SARS-CoV-2 infection at baseline who worked in the microbiology laboratory (out of 50 potential participants) served as controls. Both patients and controls were randomly selected by creating an aleatory list using the Excel 2020 (Microsoft, Redmond, Washington, USA) INDEX function. Pre- and post-vaccination blood specimens from participants were collected in sodium heparin tubes (Beckton Dickinson, UK Ltd, UK). Informed consent was obtained from participants. The study was approved by the Hospital Clnico Universitario INCLIVA Research Ethics Committee (February, 2021). Antibody assays The following immunoassays were used in the current study. CPI-268456 (a) Roche Elecsys? Anti-SARS-CoV-2 S (Roche Diagnostics, Pleasanton, CA, USA), an electrochemiluminescence sandwich immunoassay (ECLIA) that quantifies total (IgG and IgM) antibodies directed against RBD. The assay is calibrated with the first WHO International Standard and Reference Panel for CPI-268456 anti-SARS-CoV-2 antibody [10]. (b) Elecsys? Anti-SARS-CoV-2 (Roche Diagnostics), a qualitative ECLIA detecting IgG and IgM antibodies against SARS-CoV-2 nucleoprotein. Both assays were run on cobas? e601 modular analyser (Roche Diagnostics, Rotkreuz, Switzerland). Plasma specimens were further diluted (1/10) for antibody quantification when appropriate. (c) LIAISON? SARS-CoV-2 TrimericS IgG assay (Diasorin S.p.A, Saluggia, Italy), run on a DiaSorin LIAISON platform (DiaSorin, Stillwater,.