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J. Mol. Sci.","IJMS"],"abstract":"<jats:p>Although SARS-CoV-2 immunity has been widely investigated, the relationship between antibody levels and cellular immune responses following different routes of exposure to SARS-CoV-2 remains not fully revealed. In this study, cellular immunity was assessed using a newly developed whole-blood interferon-gamma release assay (nano-IGRA) based on gold nanoparticle (AuNP)-conjugated SARS-CoV-2 antigens. AuNP-conjugated and soluble antigens were compared regarding interferon gamma (IFN-\u03b3) responses and intra- and interassay reproducibility in 112 individuals with immunity acquired through infection, vaccination, or both. Humoral immunity was evaluated by enzyme-linked immunosorbent assay (ELISA). Overall, 88% of participants were seropositive. AuNP-conjugated Spike (S) and Nucleocapsid (N) SARS-CoV-2 antigens induced significantly higher IFN-\u03b3 responses than soluble antigens in all exposure groups (p &lt; 0.001). AuNP conjugation reduced intraassay variability by 62.4% and 61.4% for S and N, respectively, and interassay variability by 71.7% and 68.3%, respectively. Antibody levels correlated significantly with IFN-\u03b3 responses to S (\u03c1 = 0.358, p &lt; 0.001) and N (\u03c1 = 0.309, p &lt; 0.001). An interesting finding is that cellular responses were also detected in some seronegative individuals. Nano-IGRA, based on a novel AuNP-based approach, provides antigen-specific cellular immune assessment, combining enhanced IFN-\u03b3 responses with improved reproducibility. Further studies should establish diagnostic cut-offs, sensitivity, specificity, and clinical performance.<\/jats:p>","DOI":"10.3390\/ijms27198664","type":"journal-article","created":{"date-parts":[[2026,9,28]],"date-time":"2026-09-28T16:50:54Z","timestamp":1790614254000},"page":"8664","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Integrated Assessment of Humoral and Cellular SARS-CoV-2 Immunity Using ELISA and a Gold Nanoparticle-Based IGRA"],"prefix":"10.3390","volume":"27","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-2225-8988","authenticated-orcid":false,"given":"Aleksandra","family":"Todorovi\u0107","sequence":"first","affiliation":[{"name":"Institute for the Application of Nuclear Energy INEP, University of Belgrade, Banatska 31b, 11080 Belgrade, Serbia"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"conceptualization","vocabulary":"credit"},{"role":"methodology","vocabulary":"credit"},{"role":"data-curation","vocabulary":"credit"},{"role":"writing-original-draft","vocabulary":"credit"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6145-3850","authenticated-orcid":false,"given":"Danica","family":"\u0106uji\u0107","sequence":"additional","affiliation":[{"name":"Institute for the Application of Nuclear Energy INEP, University of Belgrade, Banatska 31b, 11080 Belgrade, Serbia"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"methodology","vocabulary":"credit"},{"role":"data-curation","vocabulary":"credit"},{"role":"writing-review-editing","vocabulary":"credit"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3009-1698","authenticated-orcid":false,"given":"Zorica","family":"Kne\u017eevi\u0107 Jugovi\u0107","sequence":"additional","affiliation":[{"name":"Faculty of Technology and Metallurgy, University of Belgrade, Karnegijeva 4, 11000 Belgrade, Serbia"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"conceptualization","vocabulary":"credit"},{"role":"writing-original-draft","vocabulary":"credit"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9543-1732","authenticated-orcid":false,"given":"Petar","family":"Uskokovi\u0107","sequence":"additional","affiliation":[{"name":"Faculty of Technology and Metallurgy, University of Belgrade, Karnegijeva 4, 11000 Belgrade, Serbia"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"investigation","vocabulary":"credit"},{"role":"writing-review-editing","vocabulary":"credit"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6308-7586","authenticated-orcid":false,"given":"Du\u0161ica","family":"Stojanovi\u0107","sequence":"additional","affiliation":[{"name":"Faculty of Technology and Metallurgy, University of Belgrade, Karnegijeva 4, 11000 Belgrade, Serbia"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"methodology","vocabulary":"credit"},{"role":"writing-review-editing","vocabulary":"credit"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8233-1636","authenticated-orcid":false,"given":"Marija","family":"Gnjatovi\u0107","sequence":"additional","affiliation":[{"name":"Institute for the Application of Nuclear Energy INEP, University of Belgrade, Banatska 31b, 11080 Belgrade, Serbia"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"conceptualization","vocabulary":"credit"},{"role":"investigation","vocabulary":"credit"},{"role":"writing-original-draft","vocabulary":"credit"},{"role":"supervision","vocabulary":"credit"}]}],"member":"1968","published-online":{"date-parts":[[2026,9,28]]},"reference":[{"key":"ref_1","unstructured":"(2026, July 07). 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