| ObjectiveTo establish a suitable animal model of acute rejection after limb allograft,improve the pathologic diagnosis of graft's acute rejection,observe the influence of radiated donators on recipients before hindlimb allograft,search a new way to prevent acute rejection and to induce immunologic tolerance of limb transplantation in the adult rats.Methods1. Wistar rats were chosen as donators,SD rats as recipients. Sciatic nerve,femoral nerve,femoral artery and femoral vein were anastomosised in operations of hindlimb allograft. Postoperation all receptors were given Benzathine benzylpenicillin. Then the symptoms and physical signs of recipients were observed.2. Acute rejection occurred and grafts were cut at 1st day,3rd day,5th day,7th day and more days later. The pathology of grafts was observed3. The rats were radiated with x-ray of 5 Gy,hindlimb biopsy was taken on 0 day,1st day,3rd day,5th day,and 7th day after the operation. Leucocytes,lymphocyte,death leucocytes,CD80,and CD95 in bone marrow were labeled by monoclonal antibodies,and detected by the flow cytometer,then were compared with those of non-irradiation rats.4. The operations of hindlimb allograft were made on the 1st day after the irradiation on Wistar rats. Then the symptoms and physical signs of the recipients,the pathology of grafts were observed,and were compared with those of non-irradiation rats.Results1. Acute rejection occurred after one week or so postoperations. The survival period of grafts was about 13+2.4 days. The swelling and necrosis of grafts were gradually appeared,but the necrosis was not fatal to the hosts.2. There was rejection of different degrees at different time shown on the pathological sections of grafts.3. After the rats were radiated by x-ray of 5Gy,gentle radiation injuries in soft tissues of rats' hindlimb occurred at 0 day,1 day,3rd day,but mostly recovered at 5th day. The death rate of Leukocytes in bone marrow reached the peak at 0 day,at the same time,the percentage of lymphocytes was the lowest,CD80 was statistically insignificant,but CD95 at 1st day was statistically significant.4. The hindlimb transplantation of Wistar-SD rats were made on Ist day after the irradiation on donators with X-ray of 5 Gy. The acute rejection did not take place with in the first two weeks. The new hair began to grow on the grafts after about three weeks. The survival period of grafts was 24.7+8.1days in irradiation group. Therefore,there was significant difference in the survival period of grafts between the irradiation group and the non-irradiation group( P<0.05 ).Conclusions1. The acute rejection can be stably induced in the animal model,which is fitting to study the transplantation immunology and pathology. But proficient microsurgical technique required.2. The degree of acute rejection rank from strong to mild in skin,blood vessel,interstitial,muscularis,and cartilage,and bone tissue.3. After the rats are radiated by x-ray of 5Gy,gentle radiation injuries in soft tissues of rats' hindlimb can recover,and the death of Antigen presenting cell in hindlimb's bone marrow were mostly of apoptosis. The best time for transplantation is on the 1st day after the irradiation.4. The acute rejection can be prevented if the donors have been irradiated with X-ray of 5Gy before the operations,but the mechanism should be study deeply. |
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