小鼠同种异体造血干细胞移植后急性移植物抗宿主病模型的构建

Establishment of murine animal model for acute graft-versus-host disease after allogeneic hematopoietic stem cell transplantation

目的 建立同种异体造血干细胞移植（HSCT）后的急性移植物抗宿主病（a GVHD）小鼠模型。方法 用C57BL/6小鼠胫、腓骨骨髓制备T细胞删除的（TCD）BM细胞,4~6周的同种异体BALB/c小鼠接受900 c Gy全身放疗2~4 h后,静脉注射供体5×10~6个TCD-BM细胞和不同剂量的脾细胞,分别以1.25×10~6、2.5×10~6、5×10~6个脾细胞作为实验组1、2、3,仅TCD-BM移植的作对照组,每组10只小鼠。通过临床GVHD评分、生存率和靶器官损害等检测模型建立情况。结果 实验组2于移植后7~14 d出现严重腹泻、拱背、活动性降低和毛皱褶等典型a GVHD临床表现,结肠、肺和肝等靶器官组织可见明显的GVHD病理学改变,45 d生存率为0。对照组和实验组1、3皆未见明显a GVHD临床表现。移植60 d后,实验组1生存率为80%,对照组为100%。实验组3移植后10 d,生存率为0。结论 用供体C57BL/6小鼠来源的5×10~6个TCD-BM细胞和2.5×10~6脾细胞静脉注射,成功构建HSCT后4~6周的BALB/c小鼠a GVHD模型。

Objective To establish the rnurine animal model for acute graft-versus-host disease after allogeneic hematopoietic stem cell transplantation （HSCT）. Methods T cell-depleted （TCD） BM cells from allogeneie C57BL/6 mice were harvested and prepared from the marrow of the femurs and tibias. Age of 4 to 6 weeks＇ BALB/c mice received 900 eGy total body irradiation （TBI） from a 137Cs source. Two to four hours later,the mice were injected intravenously （i. v. ） 5×10^6 TCD-BM cells with 1.25×10^6 or 2.5×10^6 or 5×10^6 splenic cells from allogeneic C57BL/6 mice,and respectively divided them into experimental group 1 ,experimental group 2 and experimental group 3. TCD-BM only was used as the negative control group of GVHD. Ten mice were used for each group. The establishment of aGVHD model was evaluated with a clinical GVHD scoring system,survival rate and target-organ damage. Results On 7 to 14 days after transplanta- tion, the typical clinical GVHD manifestation of severe diarrhea,hogback, activity reduced and ruffling were observed in experimental group 2. Furthermore,the aGVHD target organs of colon,lung and liver were harvested and made histological paraffin sections,then the obviously path- ological tissue damages of GVHD were detected under microscope. And the survival rate was lowed down to 0 on 45 days after transplantation in experimental group 2. On the contrary, no obviously clinical manifestation of aGYHD were observed in the control group, experimental group 1 and group 3. On 60 days later, the survival rate was 80% in experimental group 1 and 100% in control group. However,no mice was survived on 10 days later in group 3. Conclusion BALB/c mice aGVHD model after allogeneic HSCT is successfully established by injecting i.v. 5 ×10^6 TCD-BM cells with 2.5×10^6 splenic cells from allogeneic C57BL/6 mice.