Clay T M, Custer M C, Spiess P J, Nishimura M I
National Cancer Institute, National Institutes of Health, Surgery Branch, Bethesda, MD 20892, USA.
Pathol Oncol Res. 1999;5(1):3-15. doi: 10.1053/paor.1999.0003.
The purpose of this review is to illustrate some of the technical and biological hurdles that need to be addressed when developing new gene therapy based clinical trials. Gene transfer approaches can be used to "mark" cells to monitor their persistence in vivo in patients, to protect cells from toxic chemotherapeutic agents, correct a genetic defect within the target cell, or to confer a novel function on the target cell. Selection of the most suitable vector for gene transfer depends upon a number of factors such as the target cell itself and whether gene expression needs to be sustained or transient. The TCR gene transfer approach described here represents one innovative strategy being pursued as a potential therapy for metastatic melanoma. Tumor reactive T cells can be isolated from the tumor infiltrating lymphocytes (TIL) of melanoma patients. A retroviral vector has been constructed containing the T cell receptor (TCR) alpha and beta chain genes from a MART-1-specific T cell clone (TIL 5). Jurkat cells transduced with this virus specifically release cytokine in response to MART-1 peptide pulsed T2 cells, showing that the virus can mediate expression of a functional TCR. HLA-A2 transgenic mice are being used to examine whether transduced bone marrow progenitor cells will differentiate in vivo into mature CD8+ T cells expressing the MART-1-specific TCR. Expression of the human TCR alpha and beta chain genes has been detected by RT-PCR in the peripheral blood of HLA-A2 transgenic mice reconstituted with transduced mouse bone marrow. Expression of the TIL 5 TCR genes in the peripheral blood of these mice was maintained for greater than 40 weeks after bone marrow reconstitution. TIL 5 TCR gene expression was also maintained following transfer of bone marrow from mice previously reconstituted with transduced bone marrow to secondary mouse recipients, suggesting that a pluripotent progenitor or lymphocyte progenitor cell has been transduced.
本综述的目的是阐述在开展基于新基因疗法的临床试验时需要克服的一些技术和生物学障碍。基因转移方法可用于“标记”细胞，以监测其在患者体内的持久性，保护细胞免受毒性化疗药物的影响，纠正靶细胞内的遗传缺陷，或赋予靶细胞新功能。选择最适合基因转移的载体取决于多种因素，如靶细胞本身以及基因表达需要持续还是短暂。本文所述的TCR基因转移方法是作为转移性黑色素瘤潜在治疗方法而正在探索的一种创新策略。肿瘤反应性T细胞可从黑色素瘤患者的肿瘤浸润淋巴细胞（TIL）中分离出来。构建了一种逆转录病毒载体，其包含来自MART-1特异性T细胞克隆（TIL 5）的T细胞受体（TCR）α和β链基因。用这种病毒转导的Jurkat细胞在接触MART-1肽脉冲处理的T2细胞时会特异性释放细胞因子，表明该病毒可介导功能性TCR的表达。HLA-A2转基因小鼠正用于研究转导的骨髓祖细胞在体内是否会分化为表达MART-1特异性TCR的成熟CD8 + T细胞。在用转导的小鼠骨髓重建的HLA-A2转基因小鼠外周血中，通过RT-PCR检测到了人TCRα和β链基因的表达。骨髓重建后，这些小鼠外周血中TIL 5 TCR基因的表达维持了超过40周。将先前用转导的骨髓重建的小鼠的骨髓转移到二级小鼠受体后，TIL 5 TCR基因表达也得以维持，这表明已转导了多能祖细胞或淋巴细胞祖细胞。
