Establishment of murine animal model for acute graft-versus-host disease after allogeneic hematopoietic stem cell transplantation / 局解手术学杂志

ABSTRACT

Objective To establish the murine animal model for acute graft-versus-host disease after allogeneic hematopoietic stem cell transplantation (HSCT). Methods T cell-depleted (TCD) BM cells from allogeneic 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 cGy total body irradiation ( TBI) from a 137Cs source. Two to four hours later,the mice were injected intravenously ( i. v. ) 5 × 106 TCD-BM cells with 1. 25 × 106 or 2. 5 × 106 or 5 × 106 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 aGVHD 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 sur-vived on 10 days later in group 3. Conclusion BALB/c mice aGVHD model after allogeneic HSCT is successfully established by injecting i. v. 5 × 106 TCD-BM cells with 2. 5 × 106 splenic cells from allogeneic C57BL/6 mice.