异体手移植术后淋巴细胞亚群的早期监测

Early monitoring of lymphocyte subsets in patients with hand allotransplantation

目的 异体手移植术后患者外周血淋巴细胞亚群的变化和通过CD+ 3 细胞数量的监测指导抗胸腺细胞球蛋白 (antithymocyteglobulin ,ATG)的用量。方法 提取患者的外周血细胞 ,加入三荧光和双荧光标记的鼠抗人单克隆抗体CD3 /CD4 /CD8,CD3 /CD(16+ 56) ,CD3 /HLA DR ,用流式细胞分析仪测定。结果 术后 1周内CD+ 3 ,CD+ 4,CD+ 8T细胞 ,活化T细胞 [CD+ 3 /CD(16+ 56) + ,CD+ 3 /HLA DR+ ] ,静止性T细胞[CD+ 3 /CD(16+ 56) ,CD+ 3 /HLA DR-]明显下降 ,1周后除CD+ 8细胞比术前升高外 ,其余均恢复至术前水平 ;CD4 /CD8比值持续呈低水平 ;NK细胞 (CD-3 /CD(16+ 56) + )术后 1d内显著升高 ,但术后第 2天就直线下降并维持在低水平。结论 异体手移植中应用ATG的剂量以 ( 2mg·kg-1·d-1)较为适宜 ,其CD+ 3 细胞数量的监控范围为 0 .1× 10 9～ 0 .4× 10 9/L。说明T淋巴细胞亚群的监测有助于判断手的存活和观察排斥反应的发生。免疫抑制剂对移植术后的T淋巴细胞亚群有明显的影响。

Objective To investigate the changes of lymphocyte subsets in the peripheral blood of patients with hand allotransplantation and determine the dosage of antithymocyte globulin (ATG) by means of CD + 3 T cell count monitoring. Methods The lymphocytes gained from the peripheral blood of the patients in different intervals after hand allotransplantation were immunologically labeled by triple color fluorescent monoclonal antibodies CD 3/CD 4/CD 8 and dual color fluorescent monoclonal antibodies CD 3/CD (16+56) and CD 3/HLA DR. Flow cytometry (FCM) was used to determine CD 3, CD 4, CD 8, CD 3/CD (16+56) , and CD 3/HLA DR levels. Results The levels of CD + 3,CD + 4,CD + 8 T lymphocytes, the activated T lymphocyte [CD + 3/CD (16+56) +, CD + 3/HLA DR +], the silent T lymphocyte [CD + 3/CD (16+56) + , CD + 3/HLA DR -] decreased significantly within the first post transplantation week and then gradually returned to the pre transplantation level, except for CD + 8 T lymphocyte which increased to a level even higher than the pre transplantation level. The CD 4/CD 8 ratio remained at low level. Nature killer cell (CD - 3/CD (16+56) +)increased significantly at the first postoperative day and then dropped quickly and maintained at low level. Conclusions Administration of ATG at the dosage of 2 mg·kg -1 ·d -1 was sufficient to maintain the level of CD + 3 T cells at 0.1×10 9～0.4×10 9/L in patients with hand allotransplantation. Monitoring of T lymphocyte subsets is helpful for observation of survival and rejection in hand allotransplantation. The immunosuppressive agents have significant effects on the T lymphocyte subsets in the patients.