建立H-2单倍体相合造血干细胞移植（haplo-HSCT）与主要组织相容性复合体（MHC）不相合异基因造血干细胞移植（allo-HSCT）急性移植物抗宿主病（aGVHD）小鼠模型，观察造血干细胞移植后不同器官病理变化，为机制研究奠定基础。

通过给予受鼠8 Gy剂量的⁶⁰Coγ射线为预处理方案，以雄性C57BL /6 （H-2K^b）为供鼠，雌性BALB /C为受鼠，2 h内尾静脉回输200 μL骨髓细胞（5 × 10⁶/只）和脾脏单核细胞（1 × 10⁷/只）混悬液，构建MHC不相合allo- HSCT后aGVHD动物模型。相同预处理条件下，以父代雄性C57BL /6 （H-2K^b）为供鼠，雌性[C57BL/6 × BALB/C]F1 （H-2K^b/d）为受鼠的haplo-HSCT后aGVHD动物模型，2 h内尾静脉回输等量细胞；将两种受鼠分为TBI组（单纯全身照射组），全身照射+干细胞移植组（aGVHD模型组），分别设同周龄健康鼠作为对照。观察小鼠生存率、临床评分和体质量变化；HE染色观察器官（肺脏、肝脏、脾脏、肾脏、皮肤、肠）的病理学改变，两组间比较采用独立样本t检验，多组间比较采用单因素方差分析，组间两两比较采用LSD-t检验。

与对照组相比，两组aGVHD模型均出现生存率降低、临床评分[（6.18 ± 0.23比0）、（5.14 ± 0.38比0）分]增高、体重下降明显，器官病理损伤严重；两组单纯照射小鼠出现食欲下降且活动明显减少，但是未出现典型的aGVHD表现；且与TBI相比，aGVHD组临床评分[（6.18 ± 0.23）比（1.82 ± 0.11）、（5.14 ± 0.38）比（1.45 ± 0.23）分]升高，差异有统计学意义（P < 0.05）；与MHC不相合aGVHD组小鼠相比，haplo-HSCT后aGVHD组小鼠病理学观察发现器官损伤较轻，肾脏、皮肤病理评分[（3.67 ± 0.32）比（2.60 ± 0.25）、（7.50 ± 0.38）比（6.00 ± 0.32）分]升高，差异有统计学意义（P < 0.05）。肺脏、肝脏、脾脏、肠病理评分比较差异无统计学意义，但haplo-HSCT后aGVHD组小鼠生存时间延长。

研究表明，以TBI为预处理方案，以C57BL /6 （H-2^b）为供鼠，BALB/C与CB6F1 （H-2^b/d）为受鼠，均可稳定构建aGVHD模型，MHC不相合HSCT与haplo-HSCT相比，免疫排斥与器官损伤更为严重。

Establish the mouse models of acute graft-versus-host disease (aGVHD) after H-2 haploidentical (haplo-HSCT) and MHC incompatible allogeneic hematopoietic stem cell transplantation (allo-HSCT) and observe the pathological changes of different organs after HSCT, laying a foundation for mechanism research.

By administering the recipient mice with a dose of 8 Gy dose of ⁶⁰Coγ rays as the pretreatment protocol, using male C57BL/6 (H-2K^b) as the donor mice and female BALB /C (H-2K^d) as the recipient mice, 200 μL of suspension of bone marrow cells (5 × 10⁶/ mouse) and spleen monocytes (1 × 10⁷/ mouse) was reinfused via the tail vein within 2 hours to construct an animal model of aGVHD after allo-HSCT with MHC mismatch. Under the same pretreatment conditions, male C57BL/6 (H-2K^b) was used as the donor mouse and female[C57BL/6 × BALB/C]F1 (H-2K ^b/d) was used as the recipient mouse, and 200 μL of bone marrow cells (5 × 10⁶/ mouse) and spleen monocytes (1 × 10⁷/ mouse) were infused back into the tail vein within 2 hours to construct an animal model of aGVHD after haplo-HSCT. The two recipients were divided into TBI group, TBI plus allo-HSCT group (aGVHD model group). The two groups were given healthy mice of the same age as controls. The survival rate, clinical score and body mass changes of mice were observed, the pathological changes of organs (lungs, liver, spleen, kidneys, skin and intestines) were observed by HE staining. Data analysis was conducted using the independent samples t-test for inter-group comparisons. One-way analysis of variance was used for comparison between multiple groups, and LSD-t test was used for pairwise comparison between groups.

Compared with the healthy control group, the survival rate and body weight in both two aGVHD groups were decreased significantly, while the clinical score were increased (6.18 ± 0.23 vs 0, 5.14 ± 0.38 vs 0), and the pathological damage of organs was severe (P < 0.01). The mice in two TBI groups showed a decreased in appetite and activity, but did not have typical aGVHD manifestations. Compared with the TBI group, the clinical score of the aGVHD group was significantly higher (6.18 ± 0.23 vs 1.82 ± 0.11, 5.14 ± 0.38 vs 1.45 ± 0.23). Compared with the mice in the MHC-incompatible aGVHD group, the pathological observation of mice in the aGVHD group after haplo-HSCT showed that the organ damage was mild, the difference in kidney and skin were statistically significant (3.67 ± 0.32 vs 2.60 ± 0.25, 7.50 ± 0.38 vs 6.00 ± 0.32), while the difference in lung, liver, spleen and intestines was not statistically significant, and the survival time of mice in the aGVHD group was prolonged after haplo-HSCT.

Using TBI as the pretreatment regimen, C57BL/6 (H-2^b) as the donor mouse, BALB/C and CB6F1 (H-2^b/d) as the recipient mouse, the aGVHD model could be stably constructed, and the immune rejection and organ damage were more severe in MHC incompatible HSCT compared with haplo-HSCT.