The aim is to investigate the in vivo and in vitro killing effect of mesothelin chimeric antigen receptor T cells (MESO-CAR-T) in cervical squamous cell carcinoma. MESO-CAR-T cells were successfully constructed. In vitro verification of the killing effect of MESO-CAR-T cells was evaluated in the presence of SiHa cells by the lactate dehydrogenase release assay and cytokine release assay. The in vivo experiments were performed in immunodeficient NCG mice. After successful tumor formation with the subcutaneous implantation of SiHa cervical cancer cells, the injections of MESO-CAR-T cells into the tumors at different doses and frequencies were performed. Subsequently, the growth rate and size of the tumors in NCG mice were observed. A 17-fold increase in the number of MESO-CAR-T cells and a 16-fold increase in the number of Con-CAR-T cells were observed. The result of marker detection in the prepared MESO-CAR-T cells showed that CD3+ T lymphocytes accounted for 97.0 % of all cells, indicating successful preparation of MESO-CAR-T cells. Expression of the membrane protein MESO was detected in 12.8 % of SiHa cells. When the ratio of MESO-CAR-T cells to SiHa cells was 20:1, the lysis of target cells was most significant and was observed in 22 % of the cells. In the presence of SiHa cells, the secretion of IL-4、IL-2、IL-5、TNF-α and IFN-γ in MESO-CAR-T cells was higher than that in the control group. The effect of two consecutive intratumoral injections of MESO-CAR-T cells was more obvious than that of one injection. The pharmacological effect of the injection was observed within 1 week. Our finding identified the certain in vivo and in vitro killing activity of MESO-CAR-T cells.

中文翻译：

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通过特异性嵌合抗原受体修饰的 T 细胞杀死宫颈癌细胞。

目的是研究间皮素嵌合抗原受体T细胞（MESO-CAR-T）对宫颈鳞状细胞癌的体内外杀伤作用。MESO-CAR-T细胞构建成功。通过乳酸脱氢酶释放试验和细胞因子释放试验评估了在 SiHa 细胞存在下对 MESO-CAR-T 细胞杀伤效果的体外验证。体内实验是在免疫缺陷的 NCG 小鼠中进行的。在SiHa宫颈癌细胞皮下植入成功形成肿瘤后，将MESO-CAR-T细胞以不同的剂量和频率注射到肿瘤中。随后，观察 NCG 小鼠肿瘤的生长速度和大小。观察到MESO-CAR-T细胞数量增加17倍，Con-CAR-T细胞数量增加16倍。制备的MESO-CAR-T细胞标志物检测结果显示CD3+ T淋巴细胞占所有细胞的97.0%，表明MESO-CAR-T细胞制备成功。在 12.8% 的 SiHa 细胞中检测到膜蛋白 MESO 的表达。当 MESO-CAR-T 细胞与 SiHa 细胞的比例为 20:1 时，靶细胞的裂解最为显着，在 22% 的细胞中观察到。在SiHa细胞存在下，MESO-CAR-T细胞中IL-4、IL-2、IL-5、TNF-α和IFN-γ的分泌高于对照组。连续两次瘤内注射MESO-CAR-T细胞的效果比一次注射更明显。注射后1周内观察药理作用。我们的发现确定了 MESO-CAR-T 细胞的某些体内和体外杀伤活性。