CAJ | 학술논문

目的研究甲泼尼龙对移植物细胞构成的影响及其对H-2半相合小鼠造血干细胞移植的作用.方法以C57BL/6J(H-2b)雄性小鼠为供鼠,C57BL/6J(H-2b)×BALB/c(H-2d)F1代雌性小鼠为受鼠,以常规动员方案(单用G-CSF)为对照,在此基础上加用3 d剂量分别为2 mg/kg(小剂量组)、10 mg/kg(中剂量组)及50 mg/kg(大剂量组)的甲泼尼龙动员造血干细胞,流式细胞术检测移植物中T细胞亚群、DCI(HLA-DR+CD11c+细胞)及CD34+细胞,干细胞回输后观察移植后的排斥、急性移植物抗宿主病(aGVHD)的发生及生存情况.结果与对照组比较,动员方案中加用不同剂量甲泼尼龙的各实验组移植物中CD3+T细胞比例均有显著下降(P<0.05),其中CD4+T细胞比CD8+T细胞下降更为明显,CD4/CD8+比值显著下降,而CD+CD25+T细胞比例则显著上升,DC1比例下降,CD34+细胞比例上升.CD3+T细胞、CD3+CD4+T细胞和CD34+细胞百分比在小剂量、中剂量和大剂量组间差异有统计学意义(P值均<0.05).对照组、小剂量组、中剂量组和大剂量组植活率分别为90%、100%、100%、80%.与对照组比较,各实验组HLA半相合移植受鼠aGVHD严重程度均明显下降(P<0.05),不同剂量的实验组间差异有统计学意义(P<0.05);移植后小鼠生存期均较对照组明显延长(P<0.05),中剂量组较小剂量组延长(P<0.05),但大剂量组反而较中剂量组缩短(P<0.05).结论动员方案中加用甲泼尼龙可选择性去除供鼠移植物中的T细胞,同时下调其中DC1比例,提升CD34+细胞比例,使H-2半相合小鼠移植后GVHD明显减轻,生存期延长.一定剂量的甲泼尼龙不会增加移植排斥风险.
목적 연구갑발니룡대이식물세포구성적영향급기대H-2반상합소서조혈간세포이식적작용.방법 이C57BL/6J(H-2b)웅성소서위공서,C57BL/6J(H-2b)×BALB/c(H-2d)F1대자성소서위수서,이상규동원방안(단용G-CSF)위대조,재차기출상가용3 d제량분별위2 mg/kg(소제량조)、10 mg/kg(중제량조)급50 mg/kg(대제량조)적갑발니룡동원조혈간세포,류식세포술검측이식물중T세포아군、DCI(HLA-DR+CD11c+세포)급CD34+세포,간세포회수후관찰이식후적배척、급성이식물항숙주병(aGVHD)적발생급생존정황.결과 여대조조비교,동원방안중가용불동제량갑발니룡적각실험조이식물중CD3+T세포비례균유현저하강(P<0.05),기중CD4+T세포비CD8+T세포하강경위명현,CD4/CD8+비치현저하강,이CD+CD25+T세포비례칙현저상승,DC1비례하강,CD34+세포비례상승.CD3+T세포、CD3+CD4+T세포화CD34+세포백분비재소제량、중제량화대제량조간차이유통계학의의(P치균<0.05).대조조、소제량조、중제량조화대제량조식활솔분별위90%、100%、100%、80%.여대조조비교,각실험조HLA반상합이식수서aGVHD엄중정도균명현하강(P<0.05),불동제량적실험조간차이유통계학의의(P<0.05);이식후소서생존기균교대조조명현연장(P<0.05),중제량조교소제량조연장(P<0.05),단대제량조반이교중제량조축단(P<0.05).결론 동원방안중가용갑발니룡가선택성거제공서이식물중적T세포,동시하조기중DC1비례,제승CD34+세포비례,사H-2반상합소서이식후GVHD명현감경,생존기연장.일정제량적갑발니룡불회증가이식배척풍험.
Objective To explore the influence of methylprednisolone (MP) on cellular component in donor graft and on H-2 haploidentical hematopoietic stem cell transplantation(HSCT) in mice. Methods A murine model of H-2 haploidentical HSCT was established by using of c57BL/6J male mouse as donor and (c57BL/6J × LB/C)F1 female mouse as recipient. The donor mouse received peripheral-blood(PB) progen-itor cells mobilization regimens consisted of recombinant human granulocyte colony-stimulating factor (rhG-CSF) alone (control group) or combined with MP in dose of 2 mg/kg daily [small-dose(SD) group], 10 mg/kg daily [middle-dose(MD) group], and 50 mg/kg daily [large-dose(LD) group] respectively. Per-centage of T cell subsets, DCI (HLA-DR+ CD11c+) and CD34+ cell in the grafts were detected by flow cy-tometry. Transplant rejection,severity of GVHD and survival time were observed. Results The percentages of CD3+ T cell in donor grafts in the three groups were significandy lower than that in control group (P < 0.05). The percentage of CD3+ CD4+ T cells decreased more significantly than that of CD3+ CD8+ T cells, and CD4/CD8 ratios decreased significantly. The percentage of CD4+ CD25+ T cells increased significantly, the percentage of DCI (HLA-DR+ CD11c+) decreased and the percentage of CD34+ cells increased in all the three groups than in control group. There were significant differences in the percentage of CD3+ T cells, CD3+ CD4+ T cells and CD34+ cells in donor grafts among SD group, MD group and LD group (P<0.05). The engraftment rates in control, SD, MD and LD groups were 90%, 100%, 100% and 80% respectively. Severity of aGVHD in each study group decreased significantly compared with that in control group (P < 0.05). There were statistical differences among different dosage groups (P<0.05). Survival time after trans-plantation in all study groups were significantly longer than that in control group (P<0.05), and in MD group was significantly longer than in SD group and LD groups (P<0.05). Conclusions Addition of meth-ylprednisolone to routine donor mice HSC mobilization regimen has a definite effect in alleviating aGVHD and prolonging survival time of mouse after H-2 haploidentical HSCT. With a suitable dosage addition of methyl-prednisolone to donor mice HSC mobilization regimen could avoid the increasing risk of graft rejection.