Howard Hughes Medical Institute---National Institutes of Health Research Scholars Program, Surgery Branch, National Cancer Institute, National Institutes of Health, Bethesda, Maryland 20892-1201, USA.
J Immunother. 2010 Sep;33(7):672-83. doi: 10.1097/CJI.0b013e3181e475cd.
Adoptive cell transfer using autologous tumor infiltrating lymphocytes or lymphocytes transduced with antitumor T-cell receptor (TCR) is an effective therapy for patients with metastatic melanoma. A limiting factor in the effectiveness of this treatment is the apoptosis of the transferred cells when Interleukin-2 (IL-2) administration is withdrawn. In an attempt to improve persistence of the transferred lymphocytes, we cotransduced human peripheral blood lymphocytes with retroviruses encoding Bcl-2 or Bcl-xL, antiapoptotic genes of the BCL2 family, and the MART-1 melanoma tumor antigen-specific TCR, DMF5. Lymphocytes were cotransduced with 38% to 64% cotransduction efficiency, and exhibited a marked delay in apoptosis after IL-2 withdrawal. Cotransduction with Bcl-2 or Bcl-xL did not affect cytokine secretion or lytic ability of the DMF5-transduced lymphocytes. After 5 days of IL-2 withdrawal, cotransduced lymphocytes produced similar levels of IFN-γ per cell as DMF5-alone transduced lymphocytes in response to tumor cells. Cotransduction did not alter the phenotype of lymphocytes with respect to a panel of T-cell differentiation markers. In a mouse model of melanoma, adoptively transferred T cells transduced with Bcl-2 persisted better in vivo at the site of tumor, 13 and 21 days after adoptive transfer (P=0.0064 and 0.041, respectively), with evidence of enrichment of the Bcl-2-transduced population over time (P<0.0001). Thus, by coexpressing Bcl-2 or Bcl-xL with a tumor-specific TCR, we have engineered a lymphocyte that resists apoptosis owing to IL-2 withdrawal without altering its tumor-specific function or phenotype, and thus may show improved antitumor effectiveness in vivo after cell transfer.
采用自体肿瘤浸润淋巴细胞或转导抗肿瘤 T 细胞受体（TCR）的淋巴细胞进行过继细胞转移是治疗转移性黑色素瘤患者的有效方法。这种治疗方法的有效性受到限制，因为当停止使用白细胞介素-2（IL-2）时，转移细胞会发生凋亡。为了提高转移淋巴细胞的持久性，我们使用逆转录病毒将 Bcl-2 或 Bcl-xL、BCL2 家族的抗凋亡基因和 MART-1 黑色素瘤肿瘤抗原特异性 TCR、DMF5 共转导到人外周血淋巴细胞中。淋巴细胞的共转导效率为 38%至 64%，并且在停止使用 IL-2 后凋亡明显延迟。共转导 Bcl-2 或 Bcl-xL 不会影响 DMF5 转导的淋巴细胞分泌细胞因子或裂解能力。在停止使用 IL-2 5 天后，共转导的淋巴细胞在对肿瘤细胞的反应中产生的 IFN-γ 量与 DMF5 单独转导的淋巴细胞相似。共转导不会改变 T 细胞分化标志物面板上的淋巴细胞表型。在黑色素瘤的小鼠模型中，过继转移的 T 细胞在接受 Bcl-2 转导后在肿瘤部位体内的存活时间更长，在过继转移后 13 天和 21 天（分别为 P=0.0064 和 0.041），随着时间的推移，Bcl-2 转导群体的富集得到证实（P<0.0001）。因此，通过与肿瘤特异性 TCR 共表达 Bcl-2 或 Bcl-xL，我们构建了一种淋巴细胞，由于 IL-2 的撤出而抵抗凋亡，而不会改变其肿瘤特异性功能或表型，因此在细胞转移后在体内可能显示出更好的抗肿瘤效果。
