AUTHOR=Ishina Irina A. , Kurbatskaia Inna N. , Mamedov Azad E. , Shramova Elena I. , Deyev Sergey M. , Nurbaeva Kamila S. , Rubtsov Yury P. , Belogurov Alexey A. , Gabibov Alexander G. , Zakharova Maria Y. 

TITLE=Genetically engineered CD80–pMHC-harboring extracellular vesicles for antigen-specific CD4+ T-cell engagement

JOURNAL=Frontiers in Bioengineering and Biotechnology

VOLUME=Volume 11 - 2023

YEAR=2024

URL=https://www.frontiersin.org/journals/bioengineering-and-biotechnology/articles/10.3389/fbioe.2023.1341685

DOI=10.3389/fbioe.2023.1341685

ISSN=2296-4185

ABSTRACT=The identification of low-frequency antigen-specific CD4 + T cells is crucial for effective immunomonitoring across various diseases. However, this task still contains experimental challenges 2 This is a provisional file, not the final typeset article necessitating the implementation of enrichment procedures. While existing antigen-specific expansion technologies predominantly concentrate on the enrichment of CD8 + T cells, advancements in methods targeting CD4 + T cells have been limited. Here we report a technique, which harnesses antigenpresenting extracellular vesicles (EVs) for stimulation and expansion of antigen-specific CD4 + T cells. EVs are derived from a genetically modified HeLa cell line designed to emulate professional antigenpresenting cells (APCs) by expressing key costimulatory molecules CD80 and specific peptide-MHC-II complexes (pMHC). Our results demonstrate the beneficial potent stimulatory capacity of EVs for activating both immortalized and isolated from peripheral blood mononuclear cells (PBMC) human CD4 + T cells. Our technique successfully expands low-frequency Influenza-specific CD4 + T cells from healthy individuals. In summary, elaborated methodology represents a streamlined and efficient approach for the detection and expansion of antigen-specific CD4 + T cells, presenting a valuable alternative to existing antigen-specific T cell expansion protocols.