Development of methods for the targeted delivery of antigens to antigen-presenting cells to detect SARS-CoV-2-specific T cell responses ex vivo

Abstract

Immunology is an attractive field due to its various applications such as immunotherapies. Monitoring of T cell responses plays a significant role in the development of immunotherapies and vaccines. There-fore, research has been directed towards the development of detection assays for the evaluation of modern vaccines. In course of evaluation of modern vaccines, a preliminary immunoassay has been developed in this work for the stimulation and monitoring of SARS-CoV-2-specific T cell responses in COVID-19 vaccinated patients. Intended for the stimulation assays, recombinant SARS-CoV-2 spike pro-tein was produced and purified. With the help of recombinant technology, DNA sequences encoding for SARS-CoV-2 antigens were inserted into an E. coli expression vector and overexpressed in E. coli BL21 (DE3) cells. With changing the expression conditions, the production of recombinant antigens was optimized. The recombinant SARS-CoV-2 spike protein was used to stimulate SARS-CoV-2-specific T cells in COVID-19 vaccinated patients. It was evidenced by e-Fluor proliferation assay that soluble recombinant SARS-CoV-2 spike protein induced CD8+ T cell proliferation COVID-19 vaccinated patients.