Department of Hematopoietic Stem Cell Transplantation, 307 Hospital of PLA, Academy of Military Medical Sciences, Beijing , China

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1 Med J Chin PLA, Vol. 41, No. 10, October 1, [ ] (DC) (CIK) 27 (PBMC) DC CIK DC CIK 3 27 DC-CIK 37% 85% % CD3 + CD4 + CD8 CD3 + CD4 CD8 + CD3 + CD19 CD3 CD19 + CD3 CD16 + CD56 + CD3 + CD16 + CD56 + CD3 + HLA-DR CD3 + HLA-DR + CD3 + CD28 + CD8 + Th2 (P 0.05) Th1 (P 0.05) CD3 + CD4 + CD25 + T ( T Treg ) (P 0.05) 27 DC-CIK [ ] T [ ] R [ ] A [ ] (2016) [DOI] /j.issn Autologous dendritic cells combined with cytokine-induced killer cells in the treatment of metastatic renal cell carcinoma ZHANG Jin-chao, YANG Yan-li, SUN Xue-dong, WU Qiong, ZHANG Xiao-yan, ZHAO Lai-wei, PAN Xin, JIANG Hao, DING Guo-liang, WANG Dan-hong *, CHEN Hu * Department of Hematopoietic Stem Cell Transplantation, 307 Hospital of PLA, Academy of Military Medical Sciences, Beijing , China * Corresponding author. WANG Dan-hong, wangdh307@sina.com; CHEN Hu, chenhu217@aliyun.com This work was supported by the Science and Technology Foundation of 12th Five-year Plan of China (2009zx09503) [Abstract] Objective To evaluate the clinical efficacy, the immune function and follow-up observation of autologous dendritic cells (DCs) combined with cytokine-induced killer (CIK) cells in the treatment of metastatic renal cell carcinoma. Methods Peripheral blood mononuclear cells (PBMCs) were collected from 27 patients with metastatic renal cell carcinoma, and cultured in vitro to produce DCs and CIK cells. After sterility test, phenotypic character ization by flow cytometry and cell count, the produced DCs and CIK cells were then returned to the patient. DCs were given subcutaneously on day 7, 9, 11 and 13 respectively, after PBMCs collection, and CIK cells were given intravenously on day 11 and 13 respectively. This treatment regimen was repeated at a 3 months interval until the disease progresses. Clinical outcomes and immune function were recorded during the treatment period. Results After DCs-CIK cells treatment, clinical efficacy showed an objective response rate (ORR) of 37%, a disease control rate (DCR) of 85% and 2 years overall survival rate of 81.5%. There were no significant changes of T cell subsets including CD3 + CD4 + CD8, CD3 + CD4 CD8 +, CD3 + CD19, CD3 CD19 +, CD3 CD16 + CD56 +, CD3 + CD16 + CD56 +, CD3 + HLA-DR, CD3 + HLA-DR +, CD3 + CD28 + CD8 + and Th2 cells except CD3 + CD4 + CD25 + T cells (Treg cells) and Th1 in peripheral blood between pre- and post-treatment. No serious adverse events were observed. Conclusion DCs-CIK cells immunotherapy provides a safe and effective treatment approach for patients with metastatic renal cell carcinoma, and may improve the immunosuppression status and [ ] (2009zx09503) [ ] [ ] ( ) [ ] wangdh307@sina.com chenhu217@aliyun.com

2 enhance the anti-tumor immunity without obvious adverse reaction. regulatory [Key words] dendritic cells; cytokine-induced killer cells; carcinoma, renal cell; lymphocyte subsets; T-lymphocytes, [1] 2%~3% 50~70 2:1 [2-3] 25%~57% [4] 20%~30% [5] 5 15% (OS) (dendritic cells DCs) (cytokineinduce killer cells CIK) [6-11] 27 DC-CIK DC-CIK DC-CIK ~81 57 Karnofsky(KPS) 70~90 ( ) DC-CIK 4 3 KPS D C - CI K A2 Thermo FACS Van-tage SE BD Olympus GT-T551 TaKaRa Baxter AG Cat LTS1077 rgm-csf ril ~60ml PBMC (8~12) 10 7 /ml 75cm min /ml IFN- 100ng/ml IL-2 500U/ml 2 50ng/ml Anti-CD3 (1~2) 10 6 /ml 11~13 CIK GM-CSF 1000U/ml IL-4 500U/ml 37 5% CO 2 4d TNF- 100ng/ ml 3d 6 survivin muc-1 DCs 7 DCs 1 27 Tab.1 General and clinical data of 27 patients with metastatic renal cell carcinoma Item Gender Number of patients Percentage (%) Male Female 4 15 Age(year) Pathologic type Clear-cell carcinoma Treatment before DC-CIK therapy Surgery 2 7 Surgery+Targeted 6 22 Surgery+IL-2/IFN 7 26 Surgery+IL-2/IFN+ Targeted 3 11 Combined treatment 9 34 Sites of metastatic Lung 9 33 Bone 2 7 Lung and bone 8 30 Other sites 8 30 Combined treatment means surgery, interleukin 2, IFN-, targeted drugs, local radiotherapy and interventional therapy 1.4 DCs CIK

3 Med J Chin PLA, Vol. 41, No. 10, October 1, DCs CIK 1.5 PBMC ml DCs (3~10) ml CIK (2~15) (complete respond CR) (partial response PR) (stable disease SD) (progressive disease PD) CR+PR (objective response rate ORR) CR+PR+SD (disease control rate DCR) 2 2 (overall survival OS) 1 CT 3 3~6 DC-CIK ORR DCR 2 WHO(1998) ( ) 2 m l CD 3 + C D 4 + C D 8 C D 3 + C D 4 C D 8 + C D 3 + C D 1 9 C D 3 C D C D 3 CD16 + CD56 + CD3 + CD16 + CD56 + CD3 + HLA-DR CD3 + HLA-D + CD3 + CD28 + CD8 + CD3 + CD4 + CD25 + T Th1 Th SPSS 19.0 x±s P 0.05 Kaplan-Meier DCs CIK DCs CIK DCs CIK DCs CD86 80% CD80 60% HL A-DR 85% CD11c 95% CD83 5% CCR7 3% CIK CD3 98% CD8 70% CD56 30% CD(3+56) 25% CD4 20% DC-CIK 1 CR(3.7%) 9 PR(33.3%) 13 SD(48.2%) 4 PD(14.8%) 37% 85% % ( 1) Overall survival 1 27 Fig.1 Overall survival of patients with metastatic renal cell carcinoma Time (month) 2 7 DC s (4.56~6.60) 10 7 CIK (4.12~6.05) 10 9 DCs CIK (P 0.05) 2. 3 Th1 Th2 DC-CIK Th1 (P 0.05) CD3 + CD4 + CD25 + T (Treg) (P 0.05) CD3 + CD4 + CD8 CD3 + CD4 CD8 + CD3 + CD19 CD3 CD19 + CD3 CD16 + CD56 + CD3 + CD16 + CD56 + CD3 + HLA-DR CD3 + HLA-DR + CD3 + CD28 + CD8 + T Th2 (P ) Th1, Th2 (%) 2 Th1 Th2 Fig.2 Changes of Th1 and Th2 pre- and post-treatment P=0.013 P=0.12 Pre-treatment Th1 Post-treatment Th1 Pre-treatment Th2 Post-treatment Th2

4 Tab. 2 Changes of T-lymphocyte subsets pre- and post- DC-CIK treatment in 27 patients with metastatic renal cell carcinoma T-lymphocyte subsets Pre-treatment Post treatment of DC-CIK After 1st course After 2nd course After 3rd course P value * CD3 + CD4 + CD CD3 + CD4 CD CD3 + CD CD3 CD CD3 CD16 + CD CD3 + CD16 + CD CD3 + HLA-DR CD3 + HLA-DR CD3 + CD28 + CD CD3 + CD4 + CD * In comparison with pre-treatment of DC-CIK IL -2 (TKI) IL-2 DC-CIK DC 1998 DC CIK CIK [9] DC-CIK 27 DC-CIK ORR DCR 2 27 DC-CIK CR 1 (3.7%) PR 9 (33.3%) SD 13 (48.2%) PD 4 (14.8%) 37% 85% % 27 DC CIK [11-15] Kim [16] DC OS 29 DC 2007 (PFS) 11 OS 26 [17] 2013 TIVO-1 24% PFS 9.1 OS 29.3 [18] OS DC CIK DC CIK DC-CIK [19] T B NK CD4 + CD25 + T (Treg ) ( B T ) CD4 + T [20] CD3 + T CD4 + T NK [21-22] Treg [23-24] Treg T

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