Fleming M D, Bear H D, Lipshy K, Kostuchenko P J, Portocarero D, McFadden A W, Barrett S K
Department of Surgery, Medical College of Virginia, Virginia Commonwealth University, Richmond, USA.
J Immunother Emphasis Tumor Immunol. 1995 Oct;18(3):147-55. doi: 10.1097/00002371-199510000-00002.
Because the requirement for long-term cell culture can make adoptive cellular immunotherapy cumbersome, experiments were designed to determine whether smaller numbers of tumor-sensitized T cells activated briefly with bryostatin 1 and ionomycin (B/I) could be returned immediately to recipient mice without in vitro expansion and still have an anti-tumor effect in vivo. Popliteal tumor-draining lymph nodes (DLNs) from mice bearing progressive MCA-105 and MCA-203 footpad sarcomas were harvested and treated for 18 h with B/I. These cells were then washed and transferred immediately to naive C57B1/6 mice. In some experiments, these mice were irradiated (500 rads) before adoptive transfer and were given interleukin-2 (IL-2, 7,500 IU i.p., b.i.d. for 3 days) after receiving the activated lymphocytes. Recipient mice were challenged with sarcoma cells (4 x 10(5) i.v.) 6 to 32 days after receiving the activated lymphocytes. Mice receiving 10(6) B/I-activated lymphocytes before tumor challenge had significantly fewer metastases than did controls. This protective effect did not require exogenous IL-2 or host irradiation. Using Thy-1 congenic donors, it was shown that B/I-activated T cells expanded in recipients when IL-2 was also given, and these cells were a prominent component (15% of total cells) in the infiltrates found in the lungs of mice 7 days after i.v. tumor challenge. Combining these B/I-"pulsed" cells with cyclophosphamide (CYP) and IL-2 to treat mice with established (3-day) metastases resulted in significant reduction in pulmonary nodules, with complete regression in many of the treated mice, which was rarely seen with CYP alone or with CYP + IL-2. Thus, adoptive transfer of tumor-sensitized, B/I-activated DLN cells confers protection against i.v. tumor challenge, without prior in vitro expansion of the effector cells. Phenotyping studies demonstrate that donor cells activated with B/I do expand in recipient mice after adoptive transfer and can move to sites of tumor. Moreover, these cells can mediate a therapeutic effect on established tumor metastases, when combined with chemotherapy.
由于长期细胞培养的要求可能会使过继性细胞免疫疗法变得繁琐，因此设计了实验来确定用苔藓抑素1和离子霉素（B/I）短暂激活的数量较少的肿瘤致敏T细胞是否可以在不进行体外扩增的情况下立即回输到受体小鼠体内，并且在体内仍具有抗肿瘤作用。从小鼠的腘窝肿瘤引流淋巴结（DLN）中获取进行性MCA - 105和MCA - 203足垫肉瘤小鼠的淋巴结，并用B/I处理18小时。然后将这些细胞洗涤并立即转移到未接触过抗原的C57B1/6小鼠体内。在一些实验中，这些小鼠在过继性转移前接受照射（500拉德），并在接受活化淋巴细胞后给予白细胞介素-2（IL - 2，腹腔注射7500国际单位，每天两次，共3天）。受体小鼠在接受活化淋巴细胞后6至32天，静脉注射肉瘤细胞（4×10⁵个）进行攻击。在肿瘤攻击前接受10⁶个B/I活化淋巴细胞的小鼠的转移灶明显少于对照组。这种保护作用不需要外源性IL - 2或宿主照射。使用Thy - 1同基因供体，结果表明当也给予IL - 2时，B/I活化的T细胞在受体小鼠体内扩增，并且这些细胞是静脉注射肿瘤攻击7天后在小鼠肺中发现的浸润细胞中的主要成分（占总细胞的15%）。将这些B/I“脉冲”细胞与环磷酰胺（CYP）和IL - 2联合用于治疗已形成（3天）转移灶的小鼠，导致肺结节显著减少，许多治疗小鼠出现完全消退，这在单独使用CYP或CYP + IL - 2时很少见。因此，过继性转移肿瘤致敏的、B/I活化的DLN细胞可提供针对静脉注射肿瘤攻击的保护作用，而无需预先在体外扩增效应细胞。表型研究表明，用B/I活化的供体细胞在过继性转移后确实在受体小鼠体内扩增，并可迁移到肿瘤部位。此外，当与化疗联合使用时，这些细胞可对已形成的肿瘤转移灶产生治疗作用。
