小鼠角膜移植排斥反应中植片浸润细胞类型及引流淋巴结细胞表型的变化

Characterization of cells infiltrating the graft and leukocyte phenotypes in draining lymph node in murine corneal allograft rejection

目的:探讨小鼠角膜移植排斥反应中植片浸润细胞类型及引流淋巴结细胞表型的变化。方法:建立小鼠角膜移植模型,术后不同时间点进行临床观察及角膜免疫荧光组织化学染色。采用三色流式细胞术分析移植术后3d颌下淋巴结(SMLN)、颈浅淋巴结(SCLN)的细胞表型变化。结果:角膜移植术后6h,中性粒细胞、树突状细胞、巨噬细胞开始聚集并逐渐浸润植片;24hCD4+和CD8+T细胞出现并侵入植片,2wk时达到高峰;术后12～21d所有植片均被排斥。移植后3dSMLN内CD4+T细胞比例较正常时明显下降(P=0.038),而SCLN各细胞比例无明显变化。结论:角膜移植术后植片迅速出现髓系及淋巴细胞浸润,伴有引流的SMLN内早期CD4+T细胞比例下降,二者间有一定关联。

AIM： To study the subpopulations of cells infiltrating the allograft and the phenotypes in draining lymph node in the murine corneal allograft rejection. METHODS： Using a mouse corneal graft model, eyes were evaluated by clinical observation and fluorescent immunohistochemistry at various times. The phenotype of the leukocytes in the submandibular lymph node （SM LN） and superfacial cervical lymph node （SC LN） were studied by three-color flow cytometric analysis （FACS） 3d after transplantation. RESULTS： 6h after corneal transplantation, neutrophils, dendritic cells, macrophages were present and gradually infiltrated the graft. CD4^＋Tand CD8^＋T cells appeared at 24h and reached peak at 2wk. All the allografts were rejected with in 12-21d after transplantation. 3d after the transplantation, there was an early reduction of CD4^＋T cells in SM LN compared with the nontransplanted mice （P = 0.038）, whereas no obvious changes of leukocyte in the SC LN. CONCLUSION： After corneal allograft transplantation, there is an acute infiltration of myeloid and lymphoid cells in the graft, which is consistent with the early reduction of CD4^＋T cells in SM LN.