MART-1-Specific Melanoma Tumor-Infiltrating Lymphocytes Maintaining CD28 Expression Have Improved Survival and Expansion Capability Following Antigenic Restimulation In Vitro
MART-1-Specific Melanoma Tumor-Infiltrating Lymphocytes Maintaining CD28 Expression Have Improved Survival and Expansion Capability Following Antigenic Restimulation In Vitro
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DOI：
10.4049/jimmunol.0901101
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发表时间：
2010-01-01
影响因子：
4.4
通讯作者：
Radvanyi, Laszlo
中科院分区：
文献类型：
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作者：
Li, Yufeng;Liu, Shujuan;Radvanyi, Laszlo
我们确定了CD 8（+）黑色素瘤肿瘤浸润淋巴细胞（TIL）是如何从两个不同的扩增阶段分离出来的。制备对黑色素瘤Ag再刺激应答的过继性T细胞治疗。我们发现，在快速扩增方案（REP）阶段后分离的TIL（用于产生最终的患者TIL输注产物）对MART-1肽脉冲树突状细胞的再刺激反应迟钝，许多CD 8（+）T细胞发生凋亡。端粒长度较短后REP，但足够的长度，以支持进一步的细胞分裂。表型分析显示REP后TILs细胞表面CD 28表达显著降低，而CD 27水平保持不变。通过CFSE稀释追踪REP后TIL增殖，以及对REP后CD 8（+）CD 28（+）和CD 8（+）CD 28（-）亚群进行分选，揭示REP后剩余的少数CD 28（+）TIL具有上级存活能力，并且在用MART-1肽再刺激后增殖。REP期间对不同支持性细胞因子混合物的分析发现，IL-15和IL-21的组合促进了与IL-2相当的CD 8（+）TIL扩增，但阻止了CD 28表达的丧失，并提高了REP后对抗原再刺激的反应性。这些结果表明，目前使用IL-2进行黑色素瘤过继性T细胞治疗的扩增方案主要产生CD 8（+）T细胞在Ag接触后不能在体内持续存在和分裂。剩下的少数CD 8（+）CD 28（+）T细胞可能是最终存活以重新填充宿主并介导长期肿瘤控制的唯一CD 8（+）TIL。使用IL-15和IL-21的REP方案可以大大增加能够长期持续的CD 28（+）TIL的数量。免疫学杂志，2010，184：452-465.
We determined how CD8(+) melanoma tumor-infiltrating lymphocytes (TILs) isolated from two distinct phases of expansion in. preparation for adoptive T cell therapy respond to melanoma Ag restimulation. We found that TILs isolated after the rapid expansion protocol (REP) phase, used to generate the final patient TIL infusion product, were hyporesponsive to restimulation with MART-1 peptide-pulsed dendritic cells, with many CD8(+) T cells undergoing apoptosis. Telomere length was shorter post-REP, but of sufficient length to support further cell division. Phenotypic analysis revealed that cell-surface CD28 expression was significantly reduced in post-REP TILs, whereas CD27 levels remained unchanged. Tracking post-REP TIL proliferation by CFSE dilution, as well as sorting for CD8(+)CD28(+) and CD8(+)CD28(-) post-REP subsets, revealed that the few CD28(+) TILs remaining post-REP had superior survival capacity and proliferated after restimulation with MART-1 peptide. An analysis of different supportive cytokine mixtures during the REP found that a combination of IL-15 and IL-21 facilitated comparable expansion of CD8(+) TILs as IL-2, but prevented the loss of CD28 expression with improved responsiveness to antigenic restimulation post-REP. These results suggest that current expansion protocols using IL-2 for melanoma adoptive T cell therapy yields largely CD8(+) T cells unable to persist and divide in vivo following Ag contact. The few CD8(+)CD28(+) T cells that remain may be the only CD8(+) TILs that ultimately survive to repopulate the host and mediate long-term tumor control. A REP protocol using IL-15 and IL-21 may greatly increase the number of CD28(+) TILs capable of long-term persistence. The Journal of Immunology, 2010,184: 452-465.