| Summary: | Background and Objectives: Tumor-infiltrating regulatory T (TI-Treg) cells perform an important function in cancer immune escape. In this study, the third generation CAR construct was designed against human CD25 antigen as the cell surface biomarker of TI-Treg cells. Methods: At first, the construct of anti-CD25 CAR was designed. Using RNAfold web server, the RNA secondary structure was evaluated. Also, NK-92 cell line was transduced using lentiviral vectors. Then, the expression level of anti-CD25 CAR RNA, was assessed by qRT-PCR in NK-92 cells transduced with CAR and mock transfer vectors as well as untreated cells. Results: The RNA secondary structure was stable. Also, the expression level of anti-CD25 CAR RNA in the NK-92 cells transduced by pCDH-513B-1-anti-CD25 CAR transfer vector was significantly higher than NK-92 cells transduced by mock transfer vector and untreated cells (P˂0.0001). Conclusion: The present study on anti-CD25 CAR RNA showed that this type of CAR transcripts were stable and expressed at a high level. In fact, this type of CAR can be further studied in the future as a tool to remove the cancer immune escape in all types of solid and liquid cancers.
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