CAJ | 학술논문

目的：应用B7-1和B7-2反义寡核苷酸(AS B7-1／AS B7-2 oligo)抑制CD80(B7-1)、CD86(B7-2)在供体小鼠骨髓树突状细胞(DC)上的表达，观察这类DC对同种异体小鼠心脏移植存活时间的影响并探讨其机理。方法：小鼠B7-1和B7-2反义寡核苷酸在Iipofectamine协助下分别转染供体鼠C57BL/10J(B10)小鼠骨髓DC，流式细胞仪检测其CD80／CD86的表达，证实为CD80low／CD86low。将各组DC经尾静脉输注到受体小鼠C3H/Hel(C3H)体内，1周后进行心脏移植术，观察存活时间；体外实验观察各组DC对同种异体T细胞的激活作用，包括混合淋巴细胞反应、细胞毒性效应、及IL-2的产生。结果：AS B7-1和AS B7-2分别显著抑制DC表达CD80／CD86；转输这些CD80low／CD86lowDC可使小鼠心脏移植物存活时间显著延长，分别为(18.6±O.89)d和(23.67±1O.73)d，与转输成熟骨髓DC(IL-4 DC)组和生理盐水注射组(6.22±0.97)d、(11.17±1.72)d比较，均有显著差异(P<0.O1；CD80low或CD86lowDC对异体T细胞激活作用较弱，表现为T细胞增殖能力、IL-2产生及细胞毒杀伤均明显低。结论：应用反义寡聚核苷酸转染供者DC，降低其CD8O或CD86的表达，可以抑制供者特异性的免疫应答，延长移植物存活时间。
목적：응용B7-1화B7-2반의과핵감산(AS B7-1／AS B7-2 oligo)억제CD80(B7-1)、CD86(B7-2)재공체소서골수수돌상세포(DC)상적표체，관찰저류DC대동충이체소서심장이식존활시간적영향병탐토기궤리。방법：소서B7-1화B7-2반의과핵감산재Iipofectamine협조하분별전염공체서C57BL/10J(B10)소서골수DC，류식세포의검측기CD80／CD86적표체，증실위CD80low／CD86low。장각조DC경미정맥수주도수체소서C3H/Hel(C3H)체내，1주후진행심장이식술，관찰존활시간；체외실험관찰각조DC대동충이체T세포적격활작용，포괄혼합림파세포반응、세포독성효응、급IL-2적산생。결과：AS B7-1화AS B7-2분별현저억제DC표체CD80／CD86；전수저사CD80low／CD86lowDC가사소서심장이식물존활시간현저연장，분별위(18.6±O.89)d화(23.67±1O.73)d，여전수성숙골수DC(IL-4 DC)조화생리염수주사조(6.22±0.97)d、(11.17±1.72)d비교，균유현저차이(P<0.O1；CD80low혹CD86lowDC대이체T세포격활작용교약，표현위T세포증식능력、IL-2산생급세포독살상균명현저。결론：응용반의과취핵감산전염공자DC，강저기CD8O혹CD86적표체，가이억제공자특이성적면역응답，연장이식물존활시간。
AIM:To investigate the effect of donor bone marrow derived dentritic cell (DC) treated with B7 - 1, B7 - 2 antisense oligonucleotide on mouse heart allografe survival time and its mechanism. METHODS: There were 7 groups of C57BL/10J (B10) mouse bone marrow DCs which were treated by 400 nM antisense oligonucleotide target to B7 -1, B7 -2 mRNA (AS B7- 1/2), B7- 1 mismatch oligo control ,B7- 2 mismatch control(mASB7- 1/2), lipofeetamine only and non-treatment, respectively. Each group of DC were named as ASB7- 1 DC, ASB7- 2 DC, mASB7 - 1 DC, mAS B7 - 2DC, and Lipo DC, respectively. RESULTS: Flow cytometer results shown that AS B7- 1/2 can inhibit B7- 1 (CD80)and B7- 2 (CD86) molecule express on DC surface, while control groups have no effects. To observe their tolerogenicity in mouse cardiac allograft model, B10→C3H heterotopic heart transplantation were performed. Recepients were received 2 x 106 of DC injection 7 days before transplantation. Results showed that both AS B7 - 1 DC and AS B7 - 2 DC can prolong mouse cardiac allograft survival time to (18.6 + 0.89) days and (23.67 + 10.73) days, respectively, compared with IL - 4 DC [ (6.22 + 0.97) days ( P < 0.01 ) ]. Two mismatch control groups can slightly prolong while oligo DC has no effect. For understanding its mechanism, each group of DC was used as stimulator to stimulated C3H spleen T cell. Results suggested that AS B7 - 1DC and AS B7 - 2 DC had less allo - stimulate function, including MLR and generation CTL and IL - 2 production than IL - 4 DC but control groups have no effect. CONCLUSION: Donor bone marrow derived DC treated with AS B7 - 1 oligo and AS B7 - 2 oligo expressed lower level of CD80 and CD86, respectively. These cells can induce allogeneic T cells anergy in vitro and markedly prolong mouse heart allograft survival time in vivo.