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Natalie Heinen, Corinna Marheinecke, Clara Bessen, Arturo Blazquez-Navarro, Toralf Roch, Ulrik Stervbo-Kristensen, Moritz Anft, Carlos Plaza Sirvent, Sandra Buße, Mara Klöhn, Jil Alexandra Schrader, Elena Vidal Blanco, Doris Urlaub, Carsten Watzl, Markus Hoffmann, Stefan Pöhlmann, Matthias Tenbusch, Eike Steinmann, Daniel Matthias Todt, Carsten Hagenbeck, Gert Zimmer, Wolfgang E. Schmidt, Daniel Robert Quast, Nina Babel, Ingo Schmitz, Stephanie Pfänder

• With the emergence of novel Severe Acute Respiratory Syndrome Coronavirus-2 (SARS-CoV-2) Variants of Concern (VOCs), vaccination studies that elucidate the efficiency and effectiveness of a vaccination campaign are critical to assess the durability and the protective immunity provided by vaccines. SARS-CoV-2 vaccines have been found to induce robust humoral and cell-mediated immunity in individuals vaccinated with homologous vaccination regimens. Recent studies also suggest improved immune response against SARS-CoV-2 when heterologous vaccination strategies are employed. Yet, few data exist on the extent to which heterologous prime-boost-boost vaccinations with two different vaccine platforms have an impact on the T cell-mediated immune responses with a special emphasis on the currently dominantly circulating Omicron strain. In this study, we collected serum and peripheral blood mononuclear cells (PBMCs) from 57 study participants of median 35-year old’s working in the health care field, who have received different vaccination regimens. Neutralization assays revealed robust but decreased neutralization of Omicron VOC, including BA.1 and BA.4/5, compared to WT SARS-CoV-2 in all vaccine groups and increased WT SARS-CoV-2 binding and neutralizing antibodies titers in homologous mRNA prime-boost-boost study participants. By investigating cytokine production, we found that homologous and heterologous prime-boost-boost-vaccination induces a robust cytokine response of \(CD4^{+}\) and \(CD8^{+}\) T cells. Collectively, our results indicate robust humoral and T cell mediated immunity against Omicron in homologous and heterologous prime-boost-boost vaccinated study participants, which might serve as a guide for policy decisions.