细胞过继免疫治疗肿瘤的临床研究进展

细胞过继免疫治疗肿瘤的临床研究进展,张叔人中国医学科学院 北京协和医科大学肿瘤研究所,细胞免疫治疗肿瘤的发展,上世纪80年代初，Steven A. Rosenberg, Chief of Surgery at the National Cancer Institute，NIH，US LAK+IL-2 22例中： CR1，PR8（41%） LAK、TIL、基因修饰、 联合放、化疗,国内细胞免疫治疗研究热潮,80年代末90年代初LAK推动了国内细胞免疫治疗 研究的热潮。 LAK：自体、异体、胎儿脾、胸腺、脐血 90年代，我国药品监督管理局制定了相应法规，并 公布实施。此后又进行了多次修订。 首例申报CIK，北京人民医院（陆道培） DC疫苗，第二军医大学（曹雪涛） 上个世纪末CIK、DC-CIK、NK、 gdT、TIL 、 免疫杀手细胞等开展了多种免疫细胞抗肿瘤研究。,肿瘤的细胞免疫治疗经过体外培养加工,细胞过继免疫治疗(adoptive immunotherapy) LAK、TIL、CIK、DC-CIK、 NK、NKT、CTL 、 gdT 自体/异体，基因修饰 细胞瘤苗（tumor vaccine） 肿瘤细胞 树突状细胞 自体/异体、基因修饰 融合细胞,CIKCytokine induced killer,PBMCIFN-g anti-CD3, IL-2, IL-1a,2009，Germany，Prof. Dr. Schmidt-Wolf 建立了CIK临床研究登记网站 ，目前收集发表的文章39篇，中国人发表26篇(占2/3)。2011，J Cancer Res Clin Oncol. “Clinical trials on CIK cells: first report of the international registry on CIK cells (IRCC)”. PubMed the keywords CIK cells clinical trials，evaluated 11 studies（426人）. 并与作者联系核实涉及的杂志：Digestive and Liver Disease, Chinese Journal of Cancer, Anticancer Research, World Journal of Gastroenterology , Haematologica, British Journal of Cancer, Biology of Blood, and Marrow Transplantation. In some studies, abstracts were only available in Chinese.Unfortunately, these studies are often published only in Chinese. international standard,CIK 临床研究国际登记网站,Clinical trials on CIK cells: first report of the international registry on CIK cells (IRCC),C. Hontscha Y. Borck H. Zhou D. Messmer I. G. H. Schmidt-WolfJ Cancer Res Clin Oncol (2011) 137:305310,入组标准 排除标准,Table 2. Indications and contraindications of CIK immunotherapy in 11 clinical trials,Table 1 Tumor entity with number of respective patients, male-to-female ratio, andtherapeutic effects,Clinical trials on CIK cells: first report of the international registry on CIK cells (IRCC),脊髓发育不良,Cytokine Induced Killer Cells as Promising Immunotherapy for Solid Tumors Dario Sangiolo, Italy Journal of Cancer 2011; 2: 363-36,CIK治疗实体瘤的9个临床研究报道,Fig. 1. Disease-free survival curves of the three groups. KaplanMeier was employed to estimate the disease-free survival rates of the three groups. Logrank test shows a significantly higher rates in the CIK-I group (p = 0.001) and the CIK-II group (p = 0.004) than in the control group. No statistical significance was found between the CIK-I group and the CIK-II group (p = 0.345).,A randomized, controlled trial of postoperative adjuvant cytokine-induced killer cells immunotherapy after radical resection of hepatocellular carcinoma（127例）Hui Dong, Qiang Li , Jian Wang, Ti Zhang, Da-Lu Kong, Digestive and Liver Disease 41 (2009) 3641Department of Hepatobiliary Surgery, Cancer Hospital of Tianjin Medical University, Tianjin, Peoples Republic of China,无病生存期肝癌切除术后1个月开始治疗， CIK-I x3次CIK-II x6次对照,Fig. 2 The 1-, 3- and 5-year disease-free survival rates in patients receiving CIK cells (groups 1 and 2) when compared to no CIK cell treatment (127 patients); (trial 11). Group 1 was treated with 3 courses of CIK cells, group 2 was treated with 6 courses of CIK cells and the control group (group 3) received no CIK cell treatment for comparison,1-, 3- 和 5-年无病生存率,A randomized, controlled trial of postoperative adjuvant cytokine-induced killer cells immunotherapy after radical resection of hepatocellular carcinoma（127例）,A randomized, controlled trial of postoperative adjuvant cytokine-induced killer cells immunotherapy after radical resection of hepatocellular carcinoma,1-, 3- and 5-year overall survival rates,生存期,Figure 1 Survival rates of patients after cytokine-induced killer cells immunotherapy plus chemotherapy and chemotherapy alone. CIK: Cytokineinduced killer.,CIK治疗胃癌频率与疗效Increasing the frequency of CIK cells adoptive immunotherapy may decrease risk of death in gastric cancer patients (156例)Jing-Ting Jiang, . Xue-Guang Zhang 苏州大学World J Gastroenterol 2010 December 28; 16(48): 6155-6162,World J Gastroenterol 2010 December 28; 16(48): 6155-6162Jing-Ting Jiang, Yue-Ping Shen, Chang-Ping Wu, Yi-Bei Zhu, Wen-Xiang Wei, Lu-Jun Chen, Xiao Zheng,Jing Sun, Bin-Feng Lu, Xue-Guang Zhang 苏州大学,Increasing the frequency of CIK cells adoptive immunotherapy may decrease risk of death in gastric cancer patients,Cord blood-derived cytokine-induced killer cells biotherapy combined with second-line chemotherapy in the treatment of advanced solid malignancies (40例),Qi Niu, et.al., International Immunopharmacology 11 (2011) 449456Beijing GreatWall International Cancer Center, No. 309 PLA Hospital, Beijing, PR China,中位生存期 化疗 7.52个月化疗+CIK 11.17个月 差3.65个月,化疗联合脐血CIK治疗恶性肿瘤,Cord blood-derived cytokine-induced killer cells biotherapy combined with second-line chemotherapy in the treatment of advanced solid malignancies,中国肺癌杂志 2009, Vol.12, No. 9 昆明医学院第三附属医院赵光强 黄云超 叶联华 段林灿 周永春 杨凯云 马千里 雷玉洁 宋鑫 黄明 杨银山,全身化疗和支气管动脉灌注联合CIK过继免疫治疗期非小细胞肺癌的临床研究（113例）,3 CR, 1 PR, 6 SD，6PD / 16 CANCER BIOTHERAPY AND RADIOPHARMACEUTICALS 25% 62.5% Volume 25, Number 4, 2010,Immunotherapy with Cytokine-Induced KillerCells in Metastatic Renal Cell Carcinoma,Xiaosan Su，et al., The First Hospital of Kunming, Kunming, Peoples Republic of China.,目前对CIK治疗肿瘤的总体评价,相对安全材料、方法不一致评价标准不统一对血液系统来源的恶性肿瘤疗效好对实体瘤的疗效有待确定CIK应用前景较好；异体CIK克服同种异体排斥反应后是较好的治疗手段。,PBMC d0,粘附细胞,非粘附细胞,GM-CSF, IL-4,IFN-r Anti-CD3, IL-2, IL-1,Ag, TNF-a,CIK,DC,DC-CIK,Day5-9,DC-CIK,共培养的树突状细胞与CIK细胞治疗结肠癌血源性肺转移的实验研究,张嵩 王恩忠 白春学 徐永华中围科学院上海生命科学院生物化学与细胞生物学研究所，旦大学附属中山医院肿瘤 2003年 第23卷 6期,Lovo人结肠癌健康人血,共培养的树突状细胞与CIK细胞治疗结肠癌血源性肺转移的实验研究,裸鼠接种Lovo人结肠癌1x107 iv day 3,7, 10,P0.05,DC-CIK抑瘤效果优于CIK,提高急性白血病患者完全缓解病人的 无病生存期 陆道培教授领导的研究小组用自体CIK或自体DC-CIK治疗了111例经化疗或（和）自体造血干细胞移植（ASCT）治疗后完全缓解（CR）的急性白血病患者。 96例CR患者再接受CIK 或DC-CIK治疗后，5年无病生存率 （DFS）为66 %，明显高于在该医院接受单纯化疗或ASCT治疗的患者。 1415 例残留的白血病染色体和（或）基因标志均转阴。 无一例发生严重不良反应。主要不良反应为畏寒、寒战、发热、疲乏，均在24小时内消失。,该研究小组采用自CIK或自体DC-CIK治疗了24例白血病合并丙肝病毒感染的患者。 22例中有16例的血HCV-RNA转阴；在接受2疗程治疗的患者中，15例中有9例患者(60%) 血中HCV-RNA持续转阴。,使白血病合并丙肝病毒感染的患者 HCV-RNA持续转阴,Autologous cytokine-induced killer cell therapy in clinical trial phase I is safe in patients with primary hepatocellular carcinomaMing Shi, Fu-Sheng Wang, Division of Biological Engineering, Institute of Infectious Diseases, 302 Hospital of PLA, BeijingWorld J Gastroenterol 2004;10(8):1146-1151,CIK抗乙肝病毒作用,Autologous cytokine-induced killer cell therapy in clinical trial phase I is safe in patients with primary hepatocellular carcinoma,The growth of tumors in all patients became slow down, the tumor volume was decreased in 3 patients.,乙肝病毒测定,DC+CIK 联合化疗治疗晚期非小细胞肺癌的临床疗效评价（61对）,杨莉莉 曹水 李慧 于津浦 任宝柱 安秀梅 任秀宝天津市肿瘤防治重点实验室天津医科大学附属肿瘤医院研究所免疫室中国肿瘤临床 2009年第36卷第17期,DC+CIK 联合化疗治疗晚期非小细胞肺癌的临床疗效评价,Comparative study on antitumor immune response of autologous cytokineinduced killer (CIK) cells, dendritic cells CIK (DC-CIK), and semiallogeneic DC-CIK,QiJing Wang, .Jian-chuan Xia, State Key Laboratory of Oncology in Southern China, Guangzhou, Guangdong 510060, P. R. China 2 Department of Experimental Research, Sun Yatsen University Cancer Center, Guangzhou, Guangdong 510060, P. R. ChinaChinese Journal of Cancer2010 Vol. 29 Issue 7,Figure 2 Intracellular cytokine production of CIK, autologous DC-C IK, and semi-al logeneic DC-C IK cells A. IFN-g production of CIK cells, autologous DC-CI K cells, and semi-al logeneic DC-CI K cells from RCC patient; B. IL-4 production of CIK cells, autologous DC-CI K cells, and semi-al logeneic DC-CI K cells from RCC patient.,半相合DC-CIK增加产生IFN-g CIK降低产生IL-4 CIK,上颌窦鳞状细胞癌,Comparative study on antitumor immune response of autologous cytokineinduced killer (CIK) cells, dendritic cells CIK (DC-CIK), and semiallogeneic DC-CIK,NKNature Killer,Natural killer cell-based therapiesFranois Romagn and Eric Vivier, FranceF1000 Medicine Reports 2011, 3:9,NK cells sense interacting cells via their activating and inhibitory receptors. The density of ligands for these receptors dictates whether or not this interaction will lead to NK-cell activation and hence cytotoxicity and/or cytokine secretion. MHC, major histocompatibility complex; KIR, killer cell immunoglobulin-like receptor.,Fig 1. Protocol schema haplo-identical NK cell therapy combined with delayed autografting. Conditioning therapy comprised fludarabine (Flu, 25 mg/m2 on day )5 to day )2), dexamethasone (Dex, 40 mg/d on days )5 to )2), and melphalan (MEL, 140 mg/m2 IV on day )1). Flu and Dex were given to deplete patient lymphocytes in order to prevent rejection of allogeneic NK cells. MEL 140 mg/m2 was used for tumour reduction. NK cells from a haplo-identical KIR-ligand mismatched family donor were transfused on days 0 and +2. IL-2 was given daily to support the survival and expansion of NK cells in vivo. Autologous PBSCT was delayed to provide a window for donor NK cell product to kill residual myeloma cells,Infusion of haplo-identical killer immunoglobulin-like receptor ligand mismatched NK cells for relapsed myeloma in the setting of autologous stem cell transplantation,KIR错配半相合NK治疗复发骨髓瘤,Jumei Shi, et al., USA, British Journal of Haematology, 143, 641653，2008,氟达拉滨+地塞米松+美法轮,抗CD3磁珠去除T细胞，+IL-2活化扩增NK,Infusion of haplo-identical killer immunoglobulin-like receptor ligand mismatched NK cells for relapsed myeloma in the setting of autologous stem cell transplantation,CR 5+PR 1=60% ; MR 1, SD 1, PD 1,Fig 2. All donor NK products killed KIR-ligand mismatched MM cells. Donor NK cells lysed MM cell targets lacking inhibitory KIR-ligands including patient MM cells (when available), with the exception of donor 7, who did not have allo-reactive NK cells. K562, patient MM cells, U266 MM cell line (homozygous C group 1 and HLA-Bw4 negative), and patient PHA blasts were employed as targets in a standard 4-h 51Cr release assay, at E/T ratios of 10:1. Patient primary MM cells were available for patients 2, 3, 6, 7 and 8. Specific lysis percentage was calculated as (test release ) spontaneous release)/(maximal release ) spontaneous release) 100. All experiments were performed in triplicate wells and the mean SD were presented. One of three independent experiments was shown. All experiments were performed with the final NK cell product, which had been incubated overnight (products 14) or during cell processing with 300 IU/ml of IL-2 (products 510).,供者NK杀伤患者骨髓瘤细胞,Most of the donors were haploidentical to the corresponding recipient and at least one of the donors had a KIR mismatch.Donors PB by Ficoll-Hypaque density centrifugation, using standard procedures. CD56+ cells were isolated from PBMC using CliniMACS CD56 microbeads 10-5 M HC (氢化可的松), and 20 ng/mL IL-15,异体NK 治疗晚期非小细胞肺癌 A phase I trial of adoptive transfer of allogeneic natural killer cells in patients with advanced non-small cell lung cancer,Eleni G. et al., Greece, Cancer Immunol Immunother (2010) 59:17811789,异体NK 治疗晚期非小细胞肺癌 A phase I trial of adoptive transfer of allogeneic natural killer cells in patients with advanced non-small cell lung cancer,Fig. 1 a Percentages of CD56+CD3- cells among total CD56+ Gated cells isolated from donors peripheral blood on day 0 and after 1821 days of culture with 20 ng/mL IL-15 and 10-5 M HC(氢化可的松). b Expression of activating NK receptors (NKp30, NKp44, NKp46, NKG2D and 2B4) on donors CD56+ cells expanded for 1821 days of culture with 20 ng/mL IL-15 and 10-5M HC. Gray lines indicate isotype matched controls. Histograms from a representative donor. All donors gave similar results. c Cell growth (fold increase) and cytotoxic activity against K562 cells of donors CD56+cells after 2123 days of culture with 20 ng/mL IL-15 and 10-5M HC. Cytotoxicity assay was performed at an effector: target ratio 1:1. Horizontal bars indicate mean values,CD56+CD3- cells,Eleni G. et al., Greece, Cancer Immunol Immunother (2010) 59:17811789,2 PR 12.5%6 SD 50%8 PD,CD56+ + IL-15,异体NK 治疗晚期非小细胞肺癌 A phase I trial of adoptive transfer of allogeneic natural killer cells in patients with advanced non-small cell lung cancer,Fig. 2 Effect of number of infusions of allogeneic activated NK cells on progression-free survival (a) and overall survival (b) of all intended to treat non-small-cell lung cancer patients. c Overall survival （OS）of patients with stable or decreased pulmonary disease, receiving 23 and 4 NK cell infusions, respectively,异体NK 治疗晚期非小细胞肺癌 A phase I trial of adoptive transfer of allogeneic natural killer cells in patients with advanced non-small cell lung cancer,gdT,GDT: gd T cells, TTP: time to progression, OS: overall survival, PD: progressive disease, SD: stable disease, BSC: best supportive care, M: molecular targeted therapy, C: chemotherapy, R: radiotherapy, I: immunotherapy. a From the date of 1st injection. b Case #6, clinical response = SD, after 3rd injection.S, surgery; M, molecular targeted therapy; C, chemotherapy; R, radiotherapy; I, immunotherapy.,gd T 细胞治疗复发的非小细胞肺癌,A phase I study of adoptive immunotherapy for recurrent non-small-cell lung cancer patients with autologous gd T cellsJun Nakajima，Japan European Journal of Cardio-thoracic Surgery 37 (2010) 11911197Zoledronate（唑来膦酸 ） + IL-2.,无放化疗生存长？,SD 3 30%,Phase I/II study of adoptive transfer of T cells in combination with zoledronic acid and IL-2 to patients with advanced renal cell carcinoma,Hirohito Kobayashi， Japan， Cancer Immunol Immunother，26，April 2001,CR 1SD 5 54.5 %PD 5,唑来膦酸 + IL-2,CTLCytotoxic T LymphocyteTILTumor Infiltrating Lymphocyte,T细胞过激免疫治疗疫苗免疫-外周血、淋巴结T细胞体外扩增 TIL体外扩增,J. Clin. Invest. 117(6): 1466-1476 (2007),外周血淋巴结肿瘤组织,HLA-A2+ patients with metastatic melanoma received immunodepleting chemotherapy with cyclophosphamide and fludarabine（氟达拉滨） for 7 days before the adoptive transfer of highly selected tumor-reactive T cells and high-dose interleukin-2 (IL-2) therapy,patient 9 patient 10,Cancer Regression and Autoimmunity in Patients After Clonal Repopulation with Antitumor Lymphocytes， Steven A. Rosenberg SCIENCE 2002,298:850,抗肿瘤CTL克隆治疗转移性黑色素瘤,PR: 6/13 (46%) Mixed Respose: 4/13(30%),Patient demographics, treatments received, and clinical outcomes,Steven A. Rosenberg SCIENCE 2002,298:850,TIL与放、化疗联合应用治疗转移性黑色素瘤Adoptive Cell Therapy for Patients With Metastatic Melanoma: Evaluation of Intensive Myeloablative Chemoradiation Preparative Regimens Steven A. Rosenberg J Clin Oncol 26:5233-5239.2008,Fig 1. (A) Schedule of treatments administered for three lymphodepleting cell transfer regimens.Cy, cyclophosphamide; Flu, fludarabine; TBI, total-body irradiation; CD34, hematopoietic stem cells; TIL, tumor-infiltrating lymphocytes; IL-, interleukin.,化疗和全身放疗是去除抑制细胞,TIL与放、化疗联合应用治疗转移性黑色素瘤Adoptive Cell Therapy for Patients With Metastatic Melanoma: Evaluation of Intensive Myeloablative Chemoradiation Preparative Regimens Steven A. Rosenberg J Clin Oncol 26:5233-5239.2008,Objective clinical regressions in patients with metastatic melanoma treated with cell transfer therapy. (a) Regression of melanoma metastases in the heart (upper), adrenal (middle) and peritoneal cavity (lower) now ongoing at 34 months in a 53-year-old male. (b) Regression of multiple liver metastases now ongoing at 60 months in a 45-year-old male. (c) Rapid regression of multiple subcutaneous and nodal metastases now ongoing at 35 months in a 29-year-old male. (d) Regression of a large fungating scalp mass now ongoing at 34 months in a 40-year-old male.Steven A. Rosenberg， Adoptive cell therapy for the treatment of patients with metastatic melanoma，Current Opinion in Immunology 2009, 21:233240 ,TIL与放、化疗联合应用治疗转移性黑色素瘤,Survival of patients treated with cell transfer therapy in four consecutive clinical trials using increasing regimens of a lymphodepleting preparative regimen before adoptive cell transfer (NMA, non-myeloablative chemotherapy; TBI, total body irradiation). The number in parentheses is the number of patients in each trial.Current Opinion in Immunology 2009, 21:233240,Objective response rates using RECIST criteria in patients with metastatic melanoma treated in the Surgery Branch, NCI using different therapeutic strategies. Overall response rates in patients treated with vaccines is about 3% and with IL-2 or anti-CTLA4 is about 15%. With increasing levels of lymphodepletion, adding total body irradiation (TBI) and nonmyeloablative chemotherapy (NMA) to adoptive cell transfer (ACT) can achieve response rates as high as 72%.,TIL与放、化疗联合应用治疗转移性黑色素瘤,实体瘤的疗效评价标准Response Evaluation Criteria in Solid Tumors,ACT: 过继细胞输入NMA: 非清髓化疗TBI: 全身照射,Fig 2. Adoptive cell therapy after lymphodepletion. (A) Telomere length of infused tumor-infiltrating lymphocytes (TIL) correlates with clinical response. Flow-fluorescent in situ hybridization analysis was used to quantify the mean telomere length of infused TIL for patients on all three protocols.,Adoptive Cell Therapy for Patients With MetastaticMelanoma: Evaluation of Intensive MyeloablativeChemoradiation Preparative Regimens Mark E. Dudley,Steven A. Rosenberg, J Clinical Oncology, 2008,V26(32),5233-5239,过继免疫治疗效果与TIL端粒长短相关,改进过继免疫治疗的方法,Steven A Rosenberg ， Current Opinion in Immunology 2009, 21:233240,Suicide T cell therapy using adoptively transferred T cells. T cells can be