Construction and modification of an antibody-mediated acute rejection model of renal graft in mice
Objective To construct and improve an acute antibody-mediated rejection(ABMR)model in allogeneic mouse kidney transplantation.Methods Totally,31 each of male C57BL/6 and BALB/c mice(6-8 weeks old,with complete mismatch of major histocompatibility complex(MHC)),ob-tained from Nanjing Medical University Medical Experimental Animal Center,were used as donors and re-cipients,respectively.The mice were randomly divided into 4 groups:ABMR group(10 mice):undergo-ing full-thickness skin transplantation from C57BL/6H2b to BALB/cH2d as a pre-sensitization procedure 5 days prior to allogeneic kidney transplantation using the end-to-side anastomosis method with renal artery patch;T-cell mediated rejection(TCMR)group(5 mice):undergoing C57BL/6H2b to BALB/cH2d kidney transplantation;syngeneic control(SYN)group(5 mice):undergoing BALB/cH2d to BALB/cH2d kidney transplantation;skin transplantation(ST)group(5 mice):undergoing only C57BL/6H2b to BALB/cH2d full-thickness skin transplantation.Recipient survival time was recorded.Renal blood flow velocity and distri-bution in transplanted kidneys were measured using a small animal Doppler ultrasound system.Renal pa-thology staining and Banff diagnosis were performed on days 3,5,and 7 after transplantation.CD4+T cells,complement fragment C3d deposition,donor-specific antibodies(DSA),peripheral blood IgG,ser-um creatinine(Cr),and blood urea nitrogen(BUN)levels were detected.Comparisons of means between groups were performed using T-tests,non-parametric tests,and survival analyses using the Kaplan-Meier method.P＜0.05 was considered statistically significant.Results On the 5th day post-operation,all mice in the SYN and TCMR groups survived,while the survival rate in the ABMR group was 66.7％,sig-nificantly lower than that in the SYN and TCMR groups(x2=17.02,P＜0.05).On day 5 post-transplan-tation,serum Cr in the ABMR group was higher than in the TCMR,SYN,and ST groups[(233.70± 22.13)μmol/L vs.(68.06±11.10),(20.08±2.42),(20.92±2.27)μmol/L,respectively,F=325.5,P＜0.01].Additionally,serum BUN in the ABMR group was also higher than in the TCMR,SYN,and ST groups[(120.20±13.46)μmol/L vs.(24.12±3.68),(20.48±5.39),(17.00± 2.55)µmol/L,respectively,F=216.4,P＜0.01).Color Doppler ultrasound showed no significant difference in renal blood flow distribution and peak systolic velocity in the SYN group compared to normal BALB/c mice at 7th day after transplantation[(373.40±6.44)mm/s vs.(387.90±6.90)mm/s,t=2.674,P＞0.05].Compared to the SYN group,pathological results in the ABMR group showed progres-sively worsening damage on days 3,5,and 7 post-operation,with the most typical damage observed on day 5.At day 5 post-transplantation,the number of CD4+T-cell infiltration in transplanted kidneys was greater in the TCMR group than in the ABMR group(20 310±5 079 vs.131 083±8 994,t=18.580,P＜0.01),and the peripheral intracellular blood IgG class DSA was lower in the TCMR group than in the ABMR group(426.9±37.4 vs.900.2±30.0,F=828.3,P＜0.01),consistent with the Banff criteria for ABMR diagnosis.Conclusion By improving the skin transplantation sensitization and vascular anasto-mosis methods,the allogeneic mouse kidney transplantation ABMR model was successfully modified and i-dentified,using C57BL/6H2b and BALB/cH2d mice as donors and recipients respectively,facilitating subse-quent gene knockout animal experiments.
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