小鼠皮肤移植耐受诱导及微嵌合体检测

Transplantation tolerance induction to mice skin allografts and detection of microchimerism

目的 探求简便易行、安全有效、有临床应用前景的移植耐受诱导方法，研究微嵌合体的形成情况及其与移植耐受的关系。方法 雄性ＢＡＬＢ／ｃ 和雌性Ｃ５７ＢＬ／６ 小鼠作为皮肤移植的供体和受体，移植前对受体小鼠单独或联合应用５Ｇｙ（５００ｒａｄ ，１Ｇｙ ＝ １００ｒａｄ） 全身照射，０ ．１ ｍｌ 抗淋巴细胞血清注射，３ ×１０７ 供体骨髓细胞静脉输注，１５０ｍｇ／ｋｇ 体重环磷酰胺腹腔注射。应用多聚酶链式反应（ＰＣＲ） 检测微嵌合体的形成。结果 单纯照射不能延长移植物的存活，也没有微嵌合体形成，而其它各实验组的皮肤移植物存活时间均较对照组明显延长，混合淋巴细胞反应和迟发型超敏反应也表现为供者特异性的降低，多种组织的微嵌合体检查均呈阳性。结论 采用比较温和的非照射的预处理方法，也可获得一定程度的免疫耐受，达到延长移植物存活的目的；移植前输注供体骨髓细胞能够促进微嵌合体的形成及移植物的存活。

Objective To establish a transplantation tolerance induction protocol which can be applied in clinical conditions and investigate the microchimerism formation in the tolerant mice. Methods Male BALB/c (H 2 d) and female C57BL/6 (B6, H 2 b) mice were used as skin transplant donors and recipients, respectively. Before operation B6 mice were conditioned with 500rad whole body irradiation (WBI), 0.1ml ALS injection for three times, 3×10 7 donor bone marrow cells transfusion (BMT), or 150mg/kg cyclophosphamide (CP) injection two days after BMT alone or combinatively. Microchimerism was detected with PCR method. Results When WBI was used combinatively with BMT and CP, the allograft survival was significantly prolonged, 4 of 7 mice kept their skin allografts more than 100 days. If ALS was introduced instead of WBI, the allograft survival was also prolonged, mean survival time was 23.57 day, just as the result of WBI+BMT induction. Microchimerism could be detected in all the four groups of mice which received BMT before transplantation. Conclusion Transplantation tolerance can also be induced by moderate non irradiate pretreatment. BMT before transplantation can promote the microchimerism.