FIGURE 2.

Virus‐specific CAR T cells. To synthetize scFv fragment of chimeric antigen receptor (CAR), neutralizing antibodies against virus is produced in hybridoma cells, and their RNA is extracted to synthesis cDNA. Heavy and light chains variable regions are amplified by PCR, and PCR products are confirmed by DNA sequencing. PCR products are inserted into viral vectors containing an Ig‐based hinge region, TCR transmembrane domain, co‐signaling domain, and intracellular signaling domains (CD28 and CD3). The cloned viral vectors containing CAR sequences are amplified. Primary CD8⁺ T cells are isolated from healthy donor, and stimulated in vitro. Viral vectors are delivered to stimulated primary T cells and cultured in appropriate cell culture medium. Enriched CAR T cells are then confirmed by functional testing experiments