Immunotherapy of pancreatic ductal adenocarcinoma with human gammadelta T lymphocytes

循环和肿瘤浸润的人γδ T细胞以mhc非限制性方式杀死各种肿瘤细胞。人类Vγ9Vδ2 γδ T细胞的优势亚群识别异戊烯基焦磷酸（IPP），这是甲羟戊酸途径的中间产物，在肿瘤细胞中增加。转化细胞中IPP的扩增生产可以通过治疗性给予含氮双磷酸盐（n- bp）进一步增加。我们已经在体外和体内证明了人v γ γ 9v δ2 γδ T细胞在将γδ T细胞和n- bp /IL-2过继转移到SCID小鼠后对胰腺导管腺癌（PDAC）的抑制作用。其他人发表的几项初步研究报告了n- bp和低剂量IL-2治疗后，肾或前列腺癌患者γδ T细胞的选择性激活和肿瘤生长的部分减少。为了促进基于γδ T细胞的免疫治疗，需要解决以下问题：(i)优化肿瘤反应性γδ T细胞的体外扩增和细胞毒效应活性；（ii）表征对PDAC细胞系的细胞毒性，包括n- bp的直接作用，γδ T细胞和PDAC细胞相互作用中相互作用分子的调节和肿瘤逃逸机制；（iii）使用SCID-Beige小鼠过继转移模型进一步临床前表征pdac反应性γδ T细胞和n- bp的抗肿瘤作用；（iv）在n- bp +低剂量IL-2治疗晚期PDAC患者的原理验证研究中，γδ T细胞聚焦免疫监测；(v)建立γδ T细胞最终过继转移到患者体内的GMP方案。

Circulating as well as tumor-infiltrating human γδ T cells kill various tumor cells in a MHC-non-restricted fashion. The dominant subset of human Vγ9Vδ2 γδ T cells recognizes isopentenyl pyrophosphate (IPP), an intermediate of the mevalonate pathway, which is increased in tumor cells. The augmented production of IPP in transformed cells can be further increased by therapeutically administered nitrogen-containing-bisphosphonates (n-BPs). We have shown the efficacy of human Vγ9Vδ2 γδ T cells against pancreatic ductal adenocarcinoma (PDAC) in vitro as well as in vivo after adoptive transfer of γδ T cells and n-BPs/IL-2 into SCID mice. Several pilot studies published by others reported a selective activation of γδ T cells and a partial reduction of tumor growth in renal or prostate cancer patients after treatment with n-BPs and low dose IL-2. To promote γδ T cell-based immunotherapy, the following issues will be addressed: (i) optimization of in vitro expansion and cytotoxic effector activity of tumor-reactive γδ T cells; (ii) characterization of cytotoxicity against PDAC cell lines including direct effects of n-BPs, regulation of interacting molecules and tumor escape mechanisms in the interplay of γδ T cells and PDAC cells; (iii) use of an adoptive transfer model in SCID-Beige mice for further preclinical characterization of PDAC-reactive γδ T cells and anti-tumor effects of n-BPs; (iv) γδ T cell focused immune-monitoring in proof-of-principle studies in patients with advanced PDAC treated with n-BPs plus low dose IL-2; and (v) establishment of GMP protocols for ultimate adoptive transfer of γδ T cells into patients.