Goedegebuure P S, Zuber M, Leonard-Vidal D L, Burger U L, Cusack J C, Chang M P, Douville L M, Eberlein T J
Brigham and Women's Hospital, Department of Surgery, Harvard Medical School, Boston, MA 02115.
Surg Oncol. 1994 Apr;3(2):79-89. doi: 10.1016/0960-7404(94)90003-5.
Previously we described the use of solid-phase anti-CD3 monoclonal antibody (mAb) to stimulate murine tumour-infiltrating lymphocytes (TIL) and their subsequent expansion in recombinant interleukin 2 (rIL-2). In a pulmonary metastases model using the methylcholanthrene-induced sarcoma MCA-105 anti-CD3 activated TIL were capable of eradicating disease similar to TIL cultured in rIL-2 only. Here we extend these observations by characterizing the biological effects of sequential solid-phase anti-CD3 activation. TIL from MCA-105 tumour activated with solid-phase anti-CD3 on day 1 were reactivated on day 14, or day 26, or both and compared to TIL grown in rIL-2 only or TIL activated with anti-CD3 once on day 1. Reactivation enhanced in vitro proliferation 1.8- to 4-fold compared to TIL activated once with anti-CD3 (P < 0.05). In addition, the total lytic capacity of the cultures was enhanced after reactivation without changing the phenotype of the TIL cultures. Reactivation resulted in a greater in vivo efficacy when the TIL were administered within 72 h of reactivation. In contrast, TIL activated with anti-CD3 on day 1 and day 14 were least effective of all TIL cultures (P < 0.05). This correlated with in vitro cytokine production. The most effective TIL cultures in vivo produced 4- to 100-fold higher amounts of cytokines, especially interferon gamma (IFN gamma) and granulocyte macrophage colony stimulating factor (GM-CSF), than the other cultures. On the other hand, the least effective in vivo TIL culture, TIL activated with anti-CD3 on day 1 and 14, produced little or no cytokines. These data suggest that in vitro production of cytokines is indicative of in vivo efficacy of anti-CD3 activated TIL.
此前我们描述了使用固相抗CD3单克隆抗体（mAb）刺激小鼠肿瘤浸润淋巴细胞（TIL）及其随后在重组白细胞介素2（rIL-2）中的扩增。在使用甲基胆蒽诱导的肉瘤MCA-105的肺转移模型中，抗CD3激活的TIL能够根除疾病，类似于仅在rIL-2中培养的TIL。在此，我们通过表征连续固相抗CD3激活的生物学效应来扩展这些观察结果。第1天用固相抗CD3激活的来自MCA-105肿瘤的TIL在第14天、第26天或这两天均再次激活，并与仅在rIL-2中生长的TIL或第1天用抗CD3激活一次的TIL进行比较。与用抗CD3激活一次的TIL相比，再次激活使体外增殖提高了1.8至4倍（P < 0.05）。此外，再次激活后培养物的总杀伤能力增强，而TIL培养物的表型未改变。当在再次激活后72小时内给予TIL时，再次激活导致更大的体内疗效。相比之下，在第1天和第14天用抗CD3激活的TIL在所有TIL培养物中效果最差（P < 0.05）。这与体外细胞因子产生相关。体内最有效的TIL培养物产生的细胞因子量比其他培养物高4至100倍，尤其是干扰素γ（IFNγ）和粒细胞巨噬细胞集落刺激因子（GM-CSF）。另一方面，体内效果最差的TIL培养物，即在第1天和第14天用抗CD3激活的TIL，几乎不产生或不产生细胞因子。这些数据表明，细胞因子的体外产生指示抗CD3激活的TIL的体内疗效。
