Tran Khoi Q, Zhou Juhua, Durflinger Katherine H, Langhan Michelle M, Shelton Thomas E, Wunderlich John R, Robbins Paul F, Rosenberg Steven A, Dudley Mark E
Surgery Branch, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892-1201, USA.
J Immunother. 2008 Oct;31(8):742-51. doi: 10.1097/CJI.0b013e31818403d5.
Adoptive cell therapy (ACT) with tumor-reactive lymphocytes in patients with refractory melanoma can result in tumor regression and prolonged survival. Generating tumor-reactive lymphocyte cultures is technically difficult and resource intensive; these limitations have restricted the widespread application of ACT. Tumor-infiltrating lymphocytes (TIL) from melanoma contain tumor antigen-reactive cells. The "standard" method for producing TIL cultures for clinical administration requires extended in vitro expansion in interleukin-2, then identification of tumor-reactive cells by immunologic assays. We show here that limitations in reagents and methods during screening underrepresent the actual reactivity of TIL cultures. Furthermore, the extended culture times necessitated by the screening assays resulted in telomere shortening and reduced expression of CD27 and CD28 in the TIL cultures, properties that our prior studies showed are correlated with in vivo persistence and clinical response. We have thus developed an alternative "young" TIL method that demonstrated superior in vitro attributes compared with standard TIL. This approach uses the entire resected tumor to rapidly expand TIL for administration without in vitro testing for tumor recognition. Our observations suggest that younger TIL can have an undetermined but high level of antigen reactivity, and other advantageous attributes such as long telomeres and high levels of CD27 and CD28. We suggest that minimally cultured, unselected lymphocytes represent an alternative strategy for generating TIL cultures suitable for use in ACT that, if effective in vivo, may facilitate the widespread application of this approach to a broader population of patients with melanoma.
对难治性黑色素瘤患者采用肿瘤反应性淋巴细胞进行过继性细胞疗法（ACT）可导致肿瘤消退并延长生存期。生成肿瘤反应性淋巴细胞培养物在技术上具有挑战性且资源密集；这些限制阻碍了ACT的广泛应用。黑色素瘤的肿瘤浸润淋巴细胞（TIL）含有肿瘤抗原反应性细胞。用于临床给药的TIL培养物的“标准”生产方法需要在白细胞介素-2中进行长时间的体外扩增，然后通过免疫测定法鉴定肿瘤反应性细胞。我们在此表明，筛选过程中试剂和方法的局限性导致TIL培养物的实际反应性未得到充分体现。此外，筛选测定所需的延长培养时间导致TIL培养物中端粒缩短以及CD27和CD28表达降低，我们之前的研究表明这些特性与体内持久性和临床反应相关。因此，我们开发了一种替代性的“年轻”TIL方法，与标准TIL相比，该方法在体外表现出更优越的特性。这种方法使用整个切除的肿瘤快速扩增TIL以便给药，而无需进行体外肿瘤识别测试。我们的观察结果表明，较年轻的TIL可能具有未确定但高水平的抗原反应性，以及其他有利特性，如长端粒和高水平的CD27和CD28。我们认为，最少培养、未筛选的淋巴细胞代表了一种生成适用于ACT的TIL培养物的替代策略，如果在体内有效，可能有助于将这种方法更广泛地应用于更多黑色素瘤患者群体。
