Surgery Branch, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892-1201, USA.
J Immunother. 2010 Jun;33(5):547-56. doi: 10.1097/CJI.0b013e3181d367bd.
Adoptive cell therapy (ACT) using tumor-infiltrating lymphocytes (TIL) for metastatic melanoma has shown objective response rates as high as 72%. The successful application of this therapy requires the selection of unique tumor-reactive lymphocyte cultures for each patient. This is a technically and logistically difficult undertaking, and patients who do not have tumor-reactive TIL are not considered eligible for treatment. To simplify the methods of TIL generation and extend TIL-based immunotherapy to additional patients, methods were developed to use unselected, minimally cultured ("young") TIL. Young TIL cultures contain a variable number of CD8(+), CD4(+), and CD3(-)CD56(+) natural killer cells. In this study we retrospectively investigated a role for these subsets in the clinical outcome of patients treated with TIL derived from selected microcultures. This analysis demonstrated a suggestive but nonsignificant association between the number of CD8(+) cells administered and tumor regression. We therefore investigated the feasibility of selecting CD8(+) cells from young TIL cultures for ACT therapy. The available methods for clinical scale CD8(+) enrichment proved inadequate for TIL, so an optimized CD8(+) enrichment method was developed and is reported here. We observed that CD8 (+)enrichment of some TIL cultures revealed in vitro tumor recognition that was not evident in bulk culture, and an improved in vitro recognition of tumor in other TIL cultures. In addition, the enriched CD8(+) young TIL expanded more reliably and predictably in rapid expansions than the bulk TIL. Thus, optimized CD8(+) selection combines the benefits of antigen-selected TIL and young TIL for generating lymphocyte cultures for ACT, and should be evaluated in cell transfer therapy protocols.
过继细胞疗法（ACT）使用肿瘤浸润淋巴细胞（TIL）治疗转移性黑色素瘤，客观反应率高达 72%。这种疗法的成功应用需要为每位患者选择独特的肿瘤反应性淋巴细胞培养物。这是一项技术和后勤上都具有挑战性的工作，没有肿瘤反应性 TIL 的患者不被认为有资格接受治疗。为了简化 TIL 生成方法并将基于 TIL 的免疫疗法扩展到更多的患者，开发了使用未经选择的、最小培养的（“年轻”）TIL 的方法。年轻的 TIL 培养物中含有数量不定的 CD8(+)、CD4(+)和 CD3(-)CD56(+)自然杀伤细胞。在这项研究中，我们回顾性地研究了这些亚群在接受源自选定微培养物的 TIL 治疗的患者的临床结果中的作用。该分析表明，给予的 CD8(+)细胞数量与肿瘤消退之间存在提示性但无统计学意义的关联。因此，我们研究了从年轻的 TIL 培养物中选择 CD8(+)细胞用于 ACT 治疗的可行性。用于临床规模 CD8(+)富集的现有方法对 TIL 不够有效，因此开发了一种优化的 CD8(+)富集方法，并在此报告。我们观察到，一些 TIL 培养物的 CD8(+)富集显示出体外肿瘤识别，而在批量培养中则不明显，而在其他 TIL 培养物中则提高了体外对肿瘤的识别。此外，与批量 TIL 相比，富集的 CD8(+)年轻 TIL 在快速扩增中更可靠且可预测地扩增。因此，优化的 CD8(+)选择结合了抗原选择的 TIL 和年轻 TIL 的优势，用于生成用于 ACT 的淋巴细胞培养物，并且应该在细胞转移治疗方案中进行评估。
