IL-2短期活化的HLA半相合的外周血单个核细胞治疗晚期恶性实体瘤初步观察

目的:初步探讨IL-2短期活化的HLA半相合的异体外周血单个核细胞(allo-mPBMCs)治疗晚期转移性/化疗抗性实体瘤的可行性。方法:11例晚期转移性/化疗抗性恶性实体瘤患者,供者为与患者HLA半相合直系亲属。供者经10μg/kg·dG-CSF动员3天后采集PBMC,体外用大剂量rhIL-2培养后回输患者。观察治疗后临床反应,采用流式细胞仪、LDH和ELISA试剂盒检测动员、活化前后表型,IL-2短期活化前后的非特异性杀瘤活性以及患者回输前后血清中各种细胞因子的含量变化。结果:11例患者回输的HLA半相合allo-mPBMCs总数达(2.5～4.3)×1010。体外经G-CSF动员后T细胞,尤其是CD4+T细胞显著下降,而NK细胞比例显著上升;经大剂量IL-2短期活化后,各种细胞亚群比例未见改变,但活化细胞(包括CD69+和CD25+细胞)亚群比例显著升高。体外实验中可观察到IL-2短期活化的allo-mPBMCs具有类似LAK样的较高的非特异性杀瘤活性。经过治疗,11例患者血清中Th1类细胞因子,如IL-2、IL-12、IFN-γ、TNF-α含量明显升高,而Th2类细胞因子,如TGF-β含量明显下降。治疗后8例观察到临床症状的缓解,或肿瘤标志物的降低,或影像学资料的改善,其中1例治疗后4个月达到部分缓解,而3例出现进展,其中2例死亡。结论:IL-2短期活化的HLA半相合的allo-mPBMCs具有明显的抗肿瘤效应,为治疗晚期转移性/化疗抗性实体瘤提供了新的希望。

Preliminary Observation on Antitumor Effect of IL-2 Activated HLA Haploidentical Allogenic Peripheral Blood Monocular Cells in Treatment of Metastatic or Chemo-resistant Solid Tumor

Objective: To study the feasibility of IL-2 activated HLA haploidentical mobilized al-logenic peripheral blood monocular cells (allo-mPBMCs) in treatment of metastatic or chemoresistant solid tumor patients. Methods: Eleven patients with malignant solid tumor were enrolled, with HLA haploidentical relatives as the donors. After mobilization with G-CSF for 3 days, allo-mPBMCs were collected and activated by high dose of rhIL-2. Phenotypes and lysis activity of allo- mPBMCs were compared using flow cytometer and LDH kit. Multiple cytokines secreted in serum were examined by ELISA kit. Clinical effects were evaluated by CT scan. Results: In the 11 patients, 2.5-4.3×10¹⁰ acti-vated haploidentical allo-mPBMCs were transfused. After mobilization, the proportion of T cells, espe-cially CD4⁺T cells, decreased significantly and the proportion of NK cells increased remarkably which showed a statistical significance. Whereas after activation with high dose of IL-2, except a dramatic ele-vation in the proportion of activated cells which highly expressed CD69 and CD25, no change was found in the proportion of the other cell subgroups. In LDH test, a potent LAK-like non-specific tumorlysis activity was observed against NK non-sensitive tumor cell line Raji. After treatment, the contents of Th1 type cytokines, such as IL-2, IL-12, IFN-2 and TNF-2, rose significantly while the contents ofTh2 type cytokines TGF-β dropped. Eight of the 11 patients showed relief in symptoms, tumor markersin serum or regression confirmed by CT scan after treatment, one of which reached PR 4 months post-treatment. Progression was observed in 3 cases, including 2 deaths. Conclusion: IL-2 activated hap-loidentical allo-mPBMCs possessed considerable antitumor effect, which could give a new light ontreatment of the metastatic or chemo-resistant solid tumor patients.