DC-CIK细胞清除微小残留白血病的临床研究

Clinical study of dendritic cells and cytokine-induced killer cells for eliminating minimal residual leukemia

目的:观察异体树突状细胞(dendritic cell,DC)与细胞因子诱导的杀伤(cytokine-induced killer,CIK)细胞联合化疗清除微小残留白血病(minimal residual leukemia,MRL)的临床效果和安全性。方法:选择48例2009年1月至2011年6月石家庄平安医院收治的达到形态学完全缓解(complete remission,CR)但未达到分子学完全缓解(molecular CR,CRm)的急性白血病患者或MRL阳性白血病患者,依患者意愿分为联合组和化疗组,各24例,两组的一般资料和疾病程度相当;联合组应用DC-CIK细胞治疗和巩固化疗,化疗组仅应用巩固化疗。采集健康供者(患者的父母或子女)单个核细胞,制备DC-CIK细胞,每位患者输注4～6次,每次间隔15 d。Real-time PCR检测患者白血病特异基因或相关基因表达,流式细胞术检测患者MRL免疫表型组合、外周血淋巴细胞亚群的变化,记录治疗的不良反应发生情况。结果:随访至2012年6月。与化疗组相比,联合组CRm率显著提高[45.8%(11/24)vs 8.3%(2/24);χ2=8.55,P<0.01],四色流式细胞微小残留白血病免疫表型组合(four-color combination flow cytometric immunophenotype of minimal residual leukemia,CFIM)转阴率显著提高(66.7%vs 25.0%;χ2=8.39,P<0.01),MRL清除显著提高(66.7%vs 25.0%;χ2=8.39,P<0.01),3年以上持续完全缓解(continued completeremission,CCR)率也明显更高(79.2%vs 45.8%;χ2=5.69,P<0.05)。联合组治疗后患者淋巴细胞CD4+/CD8+比值较治疗前显著上升(1.3±0.4 vs 0.8±0.4,P<0.05)。DC-CIK细胞输注未见严重不良反应。结论:DC-CIK细胞联合化疗能够抑制白血病相关基因,促进CFIM转阴、提高MRL清除率、改善患者免疫功能、延长缓解期,DC-CIK输注无严重不良反应。

Objective：To evaluate the clinical efficacy and safety of allogenic dendritic cells （DCs） and cytokine-in- duced killer （CIK） cells combined with chemotherapy for eliminating minimal residual leukemia （MRL）. Methods： Forty-eight acute leukemia patients with morphological complete remission （CR） but molecular complete remission （CRm）, or patients with minimal residual leukemia （MRL） were selected from Ping＇ an Hospital of Shijiazhuang during Jan. 2009 to Jun. 2011. According to the patients＇ will, 48 patients were divided into combined group and chemotherapy group, with 24 each. All the patients were in the comparable general data and disease level. The combined group was treated with DC-CIK and consolidation chemotherapy, and the chemotherapy group was treated with consolidation chemo- therapy. PBMCs were collected from healthy donors （the patient＇ s parents or children） to prepare DC-CIK cells. DC-CIK ceils were intravenous injected into patients once every 15 days, a total of 4 -6 times infusion. Expression of leukemia specific and related genes were detected by Real-time PCR. Changes of peripheral lymphocyte subsets and MRL immuno- phentotype in patients were detected by flow cytometry. Adverse reactions were examined. Results： All the patients were followed up to Jun. 2012. Compared with the chemotherapy group, the CRm rate of combined group was significantly raised （45.8% [ 11/24 ] vs 8.3% [2/24 ] ; X2 = 8.55, P 〈 0.01 ） ; the four-color combination flow cytometric immuno- phenotype of minimal residual leukemia （CFIM） negative conversion rate of patients in the combined group was signifi- cantly raised （66.7% vs 25.0%, X2 = 8.39, P 〈 0. 01 ） ; the negative conversion rate of MRL was significant higher in the combined group （66.7% vs 25.0%, X^2 = 8.39, P 〈 0.01 ） ; the complete remission rate （CCR） of patients in the combined group after 3 years was significantly raised （79.2% vs 45.8% ; X^2 = 5.69, P 〈 0.05）. After treatment the ratio of CD4 ^＋/CD8 ^＋ lymphocyte was significant increased in the combined group （ 1.3 ± 0.4 vs O. 8 ± 0. 4, P 〈 O. 05 ）. No se- rious adverse reactions were observed after DC-CIK infusion. Conclusion： DC-CIK combined with chemotherapy can in- hibit leukemia related gene, promote the negative conversion of CFIM, facilitate the clear of MRL, improve immune func- tion and prolong remission of the patients. No serious adverse reactions are found in patients receiving DC-CIK infusion.