CAJ | 학술논문

目的探讨从肝癌患者术中失血来源的单个核细胞中培养树突状细胞(DC)的可行性,为个体化的DC介导免疫治疗提供新的细胞来源.方法采集9例原发性肝细胞癌患者术中失血及8例脐血,分离其单个核细胞,其中贴壁的单个核细胞经重组人粒细胞巨噬细胞集落刺激因子(RHGM-CSF),重组人白细胞介素4(rhIL-4)诱导和负载癌细胞抗原,制成不同的DC瘤苗,悬浮的单个核细胞经细胞因子处理成为细胞因子诱导的杀伤细胞(CIK).采用二苯基溴化四氮唑蓝(MTT)法测定DC激活同源CIK的相对增殖率和CIK对肝癌细胞的杀伤效果.结果肝癌患者术中失血来源的单个核细胞在体外能够诱导分化为具有典型形态和表型的DC.术中失血来源的DC表面标志物表达水平低于脐血来源的DC,但二者均能有效地激活CIK,并增强其对肝癌细胞的杀伤活性.负载患者自身癌细胞抗原的术中失血IX:和脐血DC,其激活的CIK增殖率分别为(388.9±137.3)%和(315.1±44.5)%,对肿瘤细胞的杀伤率分别为(87.1±8.0)%和(90.0±5.1)%;而负载SMMC-7721抗原的术中失血DC和脐血DC,其激活的CIK增殖率分别为(239.9±48.7)%和(226.3±32.3)%,对肿瘤细胞的杀伤率分别为(76.4±7.9)%和(81.1 ±4.3)%.在激活CIK和增强对肝癌细胞杀伤能力方面,相同抗原负载的两种Dc差异无统计学意义,但负载自身抗原优于负载SMMC-7721抗原.结论肝癌患者术中失血来源的DC可有效激活CIK,并增强其对肝癌细胞的杀伤效应,为临床研究和应用DC瘤苗提供了一个新的来源.
목적 탐토종간암환자술중실혈래원적단개핵세포중배양수돌상세포(DC)적가행성,위개체화적DC개도면역치료제공신적세포래원.방법 채집9례원발성간세포암환자술중실혈급8례제혈,분리기단개핵세포,기중첩벽적단개핵세포경중조인립세포거서세포집락자격인자(RHGM-CSF),중조인백세포개소4(rhIL-4)유도화부재암세포항원,제성불동적DC류묘,현부적단개핵세포경세포인자처리성위세포인자유도적살상세포(CIK).채용이분기추화사담서람(MTT)법측정DC격활동원CIK적상대증식솔화CIK대간암세포적살상효과.결과 간암환자술중실혈래원적단개핵세포재체외능구유도분화위구유전형형태화표형적DC.술중실혈래원적DC표면표지물표체수평저우제혈래원적DC,단이자균능유효지격활CIK,병증강기대간암세포적살상활성.부재환자자신암세포항원적술중실혈IX:화제혈DC,기격활적CIK증식솔분별위(388.9±137.3)%화(315.1±44.5)%,대종류세포적살상솔분별위(87.1±8.0)%화(90.0±5.1)%;이부재SMMC-7721항원적술중실혈DC화제혈DC,기격활적CIK증식솔분별위(239.9±48.7)%화(226.3±32.3)%,대종류세포적살상솔분별위(76.4±7.9)%화(81.1 ±4.3)%.재격활CIK화증강대간암세포살상능력방면,상동항원부재적량충Dc차이무통계학의의,단부재자신항원우우부재SMMC-7721항원.결론 간암환자술중실혈래원적DC가유효격활CIK,병증강기대간암세포적살상효응,위림상연구화응용DC류묘제공료일개신적래원.
Objective To investigate the practical possibility of inducing dendritic cells (DCs) from mononuclear cells in the lost blood during operation of hepatocellular carcinoma (HCC) patients, and attempted to find a new source of precursor cells for the personalized immunothempy based on DCs. Methods Collected lost blood during hepatectomy from 9 HCC patients and human cord blood from 8 cases of healthy donors undergoing caesarean section. Their mononuclear cells were divided into monocytos and nonadherent lymphocytes. RhGM-CSF and rhIL-4 were administered to induce the monocytes differentiation into DCs, and then loaded with different antigens (lysate antigen of autolngous liver cancer cells and cell line SMMC-7721 cells). The lymphocytes were induced into cytokine-induced killer cells (CIK) with IL-2, CIY3-Ab, γ-IFN and PHA. MTr assay was performed to detect the proliferation rate of T lymphocytes mediated by DC and the cytotoxicity of CIK to liver cancer cells. Results DCs induced from monocytes of the intra-operative lost blood possessed typical morphology and phenotypos. Compared with the DCs from cord blood, the DCs from intra-operative lost blood expressed lower level of surface markers, but both could effectively induce proliferation of CIK and enhance the cytotoxicty of activated CIK against liver cancer cells at similar levels. When the DCs from lost blood and their counterpart from cord blood were both loaded with autologous tumor cell antigen, the proliferation rates of CIK were (388.9±137.3)% and (315.1±44.5)%, respectively, and the killing rates against tumor cells were (87.1±8.0)% and (90.0±5.1)%, respectively. When the two similar DC groups were loaded with lysate antigen of SMMC-7721 cells, the proliferation rates of CIK were (239.9±48.7) % and (226.3 ±32.3) %, respectively, and the killing rotes against tumor cells were (76.4±7.9) % and (81.1±4.3) %, respectively. There were no significant differences between those two DC groups. The data also showed that the proliferation and cytotoxicity of CIK induced by DCs loaded with autologous antigen were higher than that of DCs loaded with SMMC-7721 antigen. Conclusion Mononuclear cells separated from intra-operative lest blood of HCC patients can be induced into mature DCs, which can effectively activate CIK and significantly increase its killing effect on the fiver cancer cells, and may become a new source of DCs to study and develop vaccines for clinical application.