CAJ | 학술논문

目的探讨半相合基因骨髓细胞移植联合抗原免疫对移植物抗肿瘤效应的影响及其机制.方法 26只BALB/c青春期小鼠为受者,随机分为单纯骨髓细胞移植组(A组,10只)、联合抗原免疫移植组(B组,10只)、对照组(C组,6只).取BALB/c(H-2d)和C57BL/6(H-2b)交配后产生的F1代小鼠为供者.A、B组小鼠均行直线加速器全身照射(8 Gy),照射4～6 h内经尾静脉分别注射入供者小鼠的骨髓细胞(5×106/只),C组无处理.1周后3组小鼠均给予Renca肾癌细胞腋窝下注射(2.6×106/只),B组小鼠在荷瘤1 d后给予33 Gy照射的Renca肾癌细胞(5×105/只)皮下注射免疫,间隔1周,重复免疫1次.通过流式细胞术(FACS)测定移植后不同时间的嵌合体水平.断颈处死小鼠后,观察所有组别受者小鼠的肿瘤质量、体积增加量、肿瘤抑瘤率,取肿瘤组织做成石蜡切片HE染色后,观察肿瘤坏死面积比,采用原位末端标记法(TUNEL)检测各组小鼠肿瘤组织的凋亡指数(AI).结果 (1)FACS检测嵌合体水平:A、B组小鼠移植后14 d供体细胞嵌合体水平均＞90%,28 d后仍＞90%.(2)与C组和A组相比,B组的肿瘤生长速率明显减慢,肿瘤质量明显减小(P＜0.05),体积增加量明显降低(P＜0.01),抑瘤率高达60%,肿瘤坏死面积比明显增高(P＜0.05),AI明显增高(P＜0.01).结论抗原免疫能增强半相合基因骨髓细胞移植的抗肿瘤作用,其机制可能是促进肿瘤细胞坏死和细胞凋亡增加.
목적 탐토반상합기인골수세포이식연합항원면역대이식물항종류효응적영향급기궤제.방법 26지BALB/c청춘기소서위수자,수궤분위단순골수세포이식조(A조,10지)、연합항원면역이식조(B조,10지)、대조조(C조,6지).취BALB/c(H-2d)화C57BL/6(H-2b)교배후산생적F1대소서위공자.A、B조소서균행직선가속기전신조사(8 Gy),조사4～6 h내경미정맥분별주사입공자소서적골수세포(5×106/지),C조무처리.1주후3조소서균급여Renca신암세포액와하주사(2.6×106/지),B조소서재하류1 d후급여33 Gy조사적Renca신암세포(5×105/지)피하주사면역,간격1주,중복면역1차.통과류식세포술(FACS)측정이식후불동시간적감합체수평.단경처사소서후,관찰소유조별수자소서적종류질량、체적증가량、종류억류솔,취종류조직주성석사절편HE염색후,관찰종류배사면적비,채용원위말단표기법(TUNEL)검측각조소서종류조직적조망지수(AI).결과 (1)FACS검측감합체수평:A、B조소서이식후14 d공체세포감합체수평균＞90%,28 d후잉＞90%.(2)여C조화A조상비,B조적종류생장속솔명현감만,종류질량명현감소(P＜0.05),체적증가량명현강저(P＜0.01),억류솔고체60%,종류배사면적비명현증고(P＜0.05),AI명현증고(P＜0.01).결론 항원면역능증강반상합기인골수세포이식적항종류작용,기궤제가능시촉진종류세포배사화세포조망증가.
Objective To investigate whether whole tumor cell vaccination strategies in combination with bone marrow transplantation (BMT) can stimulate graft-versus-tumor effect (GVT). Methods Twentysix BALB/c mice were randomly divided into 3 groups: BMT group ( group A,n = 10) ,BMT + vaccination group (group B, n = 10), control group (group C, n=6). (BALB/c × C57BI/6) F1 mice [CB6F1, H-2K(b/d) ] were used as donors. BALB/c mice of group C were only inoculated with Renca cell (2. 6 × 106).Mice of group A and B were conditioned with 8 Gy irridation, followed by infusion by bone marrow cell of CB6F1 mice on day 1 ,then inoculated with Renca cell (2. 6 × 106) on day 8. All mice of group B were immunized subcutaneous on the back with 5 × 105 irradiated Renca tumor cells on day 9 and day 16. All mice of group C were inoculated with Renca cell (2. 6 × 106) on day 8. In group A and B, all mice were analyzed by fluorescence activated cell sorter (FACS) on day 14, and 28 day after BMT. Mice were killed on day 32after inoculation with tumor cell and collected blood sample. All tumors were taken out to be weighed and then fixed in 10% buffered formalin, embedded in paraffin, and cut into 5 μm slices. The slices were stained with HE and examined by TdT mediated-dUTP nick end labeling (TUNEL). Liver, skin, intestine ,and spleen were biopsied for histopathological examination. Results The results of chimera showed that engraftments of group A,B were full donor chimerism, and the chimerism of those remained above 90% and preserved even after 28 days. The tumor weight, tumor volume increment in the group B was lower than group A and C(P ＜0. 05). The tumor suppressing rates of the group A and B were 54%, 60% respectively. The area ratio of tumor necrosis and apoptosis index (AI) of the tumor in the group B were higher than group A and C(P ＜0. 05 ). Graft-versus-host disease was not observed in each group. Conclusion The mechanism of GVTafter haploidentical allogeneic bone marrow transplantation with tumor vaccination may be the promotion of tumor necrosis and apoptosis.