小鼠H-2单倍相合非清髓骨髓移植后细胞因子表达的动态研究

为了探讨H-2单倍相合小鼠非清髓骨髓移植后细胞因子表达变化在急性移植物抗宿主病（aGVHD）发生发展中的意义，建立了H-2单倍相合小鼠非清髓骨髓移植模型，环孢菌素A（CsA）联合霉酚酸酯（MMF）预防aGVHD，采用半定量逆转录聚合酶链反应（RT—PCR）方法检测移植后不同时间点上小鼠脾脏中IL-2、INF-γ、IL-4及IL—10的mRNA的表达。结果显示：仅接受非清髓性预处理小鼠的脾脏中各因子的mRNA表达较预处理前略有升高。未做aGVHD预防给药的小鼠（第2组）IL-2、IFN-γ的mRNA表达水平在移植后急剧升高，分别在21天、14天达峰值，之后显著下降，35天时基本恢复至移植前水平。接受GVHD预防给药的小鼠（第3组）IL-2、IFN-γ的mRNA表达的变化趋势与第2组相似，但峰值明显低于第2组。第2组和第3组小鼠IL-4的mRNA表达水平均在移植后14天达峰值，但第3组的峰值较第2组为高，且在此后的下降中较第2组缓和。IL—10的mRNA表达水平在移植后逐渐升高，在14天达到一个高峰，21天时下降，之后再次升高直至35天。结论：H-2单倍相合小鼠非清髓骨髓移植后脾脏内IL-2、IFN-γ、IL-4及IL—10均呈上调表达，应用CsA和MMF则可下调IL-2和IFN-γ的mRNA表达水平，从而降低了aGVHD的发生率和严重程度。

Kinetic Patterns of Cytokine Expressions in H-2 Haploidentical Nonmyeloablative Bone Marrow Transplantation in Mice

In order to explore what role relative cytokines play in acute GVHD (aGVHD) of mice transplanted with H-2 haploidentical nonmyeloablative bone marrow, a murine model was established by using C57BL/6J mouse as donor and (C57BL/6J x BALB/C) F1 mouse as the recipient. The recipient mice were given CsA and mycophenolate mofetile (MMF) for prophylaxis of aGVHD. The expression levels of IL-2, INFgamma, IL-4 and IL-10 in the spleen were detected by semi-quantitative RT-PCR at different time points after transplantation. The results showed that the expression levels of these cytokines increased slightly in the group only received nonmyeloablative conditioning. The expression levels of IL-2 and INF-gamma elevated significantly after transplantation in group 2 (without aGVHD prophylaxis), peaked at day 21 and day 14 respectively, then dropped rapidly, returned to the basal levels at day 35. The study on kinetic pattern of expression of IL-2 and INF-gamma in group 3 (with aGVHD prophylaxis) gave a similar result to group 2, but the peak value of cytokine expression in group 3 was much lower than that in group 2. The expression of IL-4 in Group 2 and group 3 all peaked at day 14, but the peak value in group 3 was higher than that in group 2, and decreased slowly in group 3. The expression of IL-10 increased gradually after transplantation, peaked at day 14, decreased after day 21, elevated again until day 35. It is concluded that the expression levels of these cytokines in the spleen all increase after H-2 haploidentical nonmyeloablative bone marrow transplantation. CsA and MMF can reduce the incidence and severity of aGVHD by down-regulating the expression levels of IL-2 and INF-gamma.