CAJ | 학술논문

目的探讨肝癌肝移植后应用活化供体脾脏来源的淋巴细胞,对受体实施过继性免疫治疗以预防肝癌复发的可行性.方法以培养黏附法体外分离培养Wistar大鼠骨髓树突状细胞(DCs)前体细胞,经诱导后与Wistar大鼠肝癌细胞CBRH-7919裂解物抗原共同孵育,形成负载癌抗原的DCs.以此DCs分别诱导受体(Wistar大鼠)、供体(SD大鼠)来源的脾淋巴细胞作为两个实验组C(受体活化组)、D(供体活化组),以未经抗原诱导的受、供体脾淋巴细胞作为对照组A(受体对照组)、B(供体对照组).对比活化、非活化以及供体来源、受体来源的脾淋巴细胞在体外对受体肿瘤细胞的杀伤活性.以上述不同来源和处理的脾淋巴细胞对SD→Wistar大鼠肝移植后肿瘤复发模型行过继性免疫治疗,以盐水治疗作为对照组E,每组6只大鼠,观察各组免疫治疗对于肿瘤浸润淋巴细胞、受体肝脏成瘤率和供肝的排斥反应的影响.结果 DCs诱导脾细胞过程中,培养上清液γ干扰素(IFN-γ)分泌C、D组较A、B组有明显的升高.C、D组对肝癌细胞的杀伤活性较A、B组显著增强,D组较C组显著增强.D组脾细胞回输体内后,在移植后大鼠体内观察到了肿瘤浸润淋巴细胞增多,肿瘤组织坏死和成瘤率下降.免疫治疗前后供肝未见严重排斥反应发生.结论活化的供体脾细胞较受体脾细胞有更强的肿瘤杀伤效果.应用供体脾淋巴细胞对肝移植受体进行过继性免疫治疗可以在不增加对移植肝排斥反应的同时,为预防肝癌肝移植术后复发、延长生存提供一种可能的方法.
목적 탐토간암간이식후응용활화공체비장래원적림파세포,대수체실시과계성면역치료이예방간암복발적가행성.방법 이배양점부법체외분리배양Wistar대서골수수돌상세포(DCs)전체세포,경유도후여Wistar대서간암세포CBRH-7919렬해물항원공동부육,형성부재암항원적DCs.이차DCs분별유도수체(Wistar대서)、공체(SD대서)래원적비림파세포작위량개실험조C(수체활화조)、D(공체활화조),이미경항원유도적수、공체비림파세포작위대조조A(수체대조조)、B(공체대조조).대비활화、비활화이급공체래원、수체래원적비림파세포재체외대수체종류세포적살상활성.이상술불동래원화처리적비림파세포대SD→Wistar대서간이식후종류복발모형행과계성면역치료,이염수치료작위대조조E,매조6지대서,관찰각조면역치료대우종류침윤림파세포、수체간장성류솔화공간적배척반응적영향.결과 DCs유도비세포과정중,배양상청액γ간우소(IFN-γ)분비C、D조교A、B조유명현적승고.C、D조대간암세포적살상활성교A、B조현저증강,D조교C조현저증강.D조비세포회수체내후,재이식후대서체내관찰도료종류침윤림파세포증다,종류조직배사화성류솔하강.면역치료전후공간미견엄중배척반응발생.결론 활화적공체비세포교수체비세포유경강적종류살상효과.응용공체비림파세포대간이식수체진행과계성면역치료가이재불증가대이식간배척반응적동시,위예방간암간이식술후복발、연장생존제공일충가능적방법.
Objective To explore the feasibility of using activated donor spleen lymphocytes to prevent liver cancer recurrence after liver transplantation.Methods The precursors of dendritic cells (DCs) were isolated from bone marrow of Wistar rats,and were then cultured and pulsed with CBRH7919 tumor cell antigens prepared with freeze-thawed tumor cell lysates. Spleen cells from donor (SD)/recipient (Wistar) rats were cocultured with these DCs respectively to induce CTL and to serve as the two experimental groups,C and D.Unactivited blank donor/recipient spleen cells were maintained to survive using low dose of rrIL-2.These served as the two control groups,A and B.Supernatant IFN-γ release were determined by ELISA,and the cytotoxicity of CTL was assayed by LDH release test.A post-transplantation HCC recurrence rat model was randomly assigned into five groups,6 rats each.Four groups were treated with adoptive lymphocyte infusion which came from the corresponding groups in vitro,and another group E was a saline infusion group to serve as a blank control.Tumorgenecity,tumor infiltrating-lymphocyte,and liver graft rejection were compared between the individual groups.Results The supernatant IFN-γ release level during DCs activation in group C and D were much higher than group A and B.Spleen cells in group C and D manifested much higher cytotoxicity to tumor cells than group A and B.The cytotoxicity of group D was significantly higher than group C.Aggregation of lymphocytes was detected within and near the tumor tissue with necrosis of tumor cells,and a decrease in tumorgenecity was observed in group D but without any evidence of liver graft rejection.Conclusion Activated donor spleen cells were more effective in preventing liver cancer recurrence after liver transplantation when compared with recipient spleen cells,and there was no impairment to the liver graft.