Aruga A, Aruga E, Cameron M J, Chang A E
Department of Surgery, University of Michigan, Ann Arbor 48109, USA.
J Leukoc Biol. 1997 Apr;61(4):507-16. doi: 10.1002/jlb.61.4.507.
We have previously demonstrated that the growth of weakly immunogenic murine sarcomas leads to the induction of immunologically specific pre-effector cells in tumor-draining lymph nodes (TDLN). The in vitro activation of TDLN cells with anti-CD3 monoclonal antibodies (mAbs) and interleukin-2 (IL-2) resulted in the acquisition of effector function as measured by tumor regression in the adoptive immunotherapy of pulmonary metastases. Further studies were performed to characterize the mechanisms associated with in vivo tumor reactivity mediated by activated TDLN cells. By positive selection, CD4+ and CD8+ T cells were purified and activated by the anti-CD3/IL-2 method. CD8+, but not CD4+, cells manifested tumor-specific granulocyte-macrophage colony-stimulating factor (GM-CSF) and interferon-gamma (IFN-gamma) release in vitro, and elicited tumor regression in vivo. By contrast, only activated CD4+ were found to release significant amounts of IL-2 in response to tumor antigen but did not mediate tumor regression in vivo. Mixing the two purified populations enhanced the antitumor activity of the CD8+ T cells. In culture, IL-2 was found to augment the relative amount of tumor-specific release of GM-CSF and IFN-gamma by activated TDLN cells. We found that the tumor-specific release of GM-CSF and IFN-gamma by activated lymphocytes was strongly associated with the in vivo therapeutic efficacy of these cells. Evidence in support of this included the following: (1) cytokine release of TDLN derived after different durations of tumor growth correlated with tumor reactivity in adoptive transfer studies, (2) cytokine release of T cells derived from different lymphoid organs corresponded with tumor reactivity in adoptive transfer, and (3) in vivo administration of neutralizing mAb to IFN-gamma and GM-CSF significantly inhibited the antitumor reactivity of TDLN cells. These studies document the contributory roles of IFN-gamma, GM-CSF, and IL-2 released by activated CD4+ and CD8+ T cells involved in tumor regression.
我们之前已经证明，弱免疫原性小鼠肉瘤的生长会导致在肿瘤引流淋巴结（TDLN）中诱导出免疫特异性的前效应细胞。用抗CD3单克隆抗体（mAb）和白细胞介素-2（IL-2）在体外激活TDLN细胞，导致其获得效应功能，这通过肺转移瘤过继免疫治疗中的肿瘤消退来衡量。我们进行了进一步研究，以阐明与活化的TDLN细胞介导的体内肿瘤反应性相关的机制。通过阳性选择，纯化CD4+和CD8+ T细胞，并通过抗CD3/IL-2方法进行激活。CD8+细胞（而非CD4+细胞）在体外表现出肿瘤特异性粒细胞-巨噬细胞集落刺激因子（GM-CSF）和干扰素-γ（IFN-γ）释放，并在体内引发肿瘤消退。相比之下，仅发现活化的CD4+细胞在响应肿瘤抗原时会释放大量IL-2，但在体内不介导肿瘤消退。将这两个纯化群体混合可增强CD8+ T细胞的抗肿瘤活性。在培养中，发现IL-2可增加活化的TDLN细胞肿瘤特异性释放GM-CSF和IFN-γ的相对量。我们发现，活化淋巴细胞肿瘤特异性释放GM-CSF和IFN-γ与这些细胞的体内治疗效果密切相关。支持这一点的证据包括：（1）肿瘤生长不同持续时间后获得的TDLN细胞因子释放与过继转移研究中的肿瘤反应性相关；（2）来自不同淋巴器官的T细胞因子释放在过继转移中与肿瘤反应性相对应；（3）在体内给予IFN-γ和GM-CSF的中和mAb可显著抑制TDLN细胞的抗肿瘤反应性。这些研究证明了活化的CD4+和CD8+ T细胞释放的IFN-γ、GM-CSF和IL-2在肿瘤消退中的作用。
