同种异基因骨髓细胞移植诱导嵌合体小鼠生成

Study on the Induction of Mouse Chimerism by Non-Myeloablative Allogeneic Bone Marrow Transplantation

采用C57BL/J和BALB/c两种小鼠进行骨髓细胞移植。移植前受体小鼠经 6 0Gy6 0 Coγ射线照射。用细胞染色法和流式细胞术分析了射线对小鼠淋巴细胞的损伤强度和照射后移植了骨髓细胞的小鼠在不同时间内的淋巴细胞亚群变化。结果表明 ,(1 ) 6 0Gyγ射线照射受体鼠 ,能杀伤 95 5 %的外周淋巴细胞 ,但是不影响自身H 2Kb 分子在淋巴细胞表面表达的百分数。残留细胞中CD3+ T细胞、NK1 1 + 淋巴细胞、CD3+ /NK1 1 + 淋巴细胞的阳性率分别由照射前的 40 44%增加到45 30 %、 1 1 3 %增加到 37 41 %和 1 0 7%增加到 5 86 % (P≤ 0 0 1 ) ;(2 )小鼠的淋巴细胞数量从照射后 30d开始恢复 ,到1 90d达到正常值 ;(3 )照射后进行同种异基因骨髓细胞移植后的第 60天 ,受体淋巴细胞表面自身H 2Kb 抗原表达下降 ,CD3+ /NK1 1 + 淋巴细胞百分数增加。随H 2Kb 的抗原表达的恢复 ,CD3+ /NK1 1 + 淋巴细胞百分数下降 ;(4)在细胞移植的第 1 90天 ,受体小鼠 (黑色 )表型呈现出供体鼠 (白色 )颜色特征。在嵌合体小鼠外周淋巴细胞中 ,有 2 5 %为供体H 2Kd 阳性细胞 ,CD3+ /NK1 1 + 淋巴细胞百分数为 1 88% ,明显高于正常值 (P≤ 0 0 1 )。以上结果表明 ,6 0Gy6 0 Coγ射线照射能诱导免疫耐受 。

C57BL/J and BALB/c mice were used for bone marrow transplantations in which the recipient mice were irradiated with 6 0 Gy 60 Co γ ray before transplantation The degree of damages on mouse lymphocytes after irradiation and the changes on lymphocyte subsets of mice in the different times after transplantation with bone marrow cells were determined with the cyto staining technique and flow cytometery analysis It was found that 4 5% of lymphocytes in the peripheral blood of the irradiated mice survived,of which most of the surviving lymphocytes were CD3 +T cells and NK1 1 + lymphocytes On the 60 days after allogeneic bone marrow transplantation,the expression of self H 2K b antigen of the recipient mice was lower,but the numbers of CD3 +/NK1 1 + NK T cells were increased On the 190 days after transplantation,the phenotypic character of the recipient mice (black color) converted to that of the donor mice (white color),and to become a chimeric mice. In the peripheral blood lymphocytes of the chimeric mice,2 5% of the lymphocytes were donor H 2K d positive cells,and the percentage of CD3 +/NK1 1 + cells was 1 88%,significantly higher than that of the normal value. These results indicate that irradiation with 6 0 Gy γ ray can induce tolerance,and the tolerance may be associated with increase of the CD3 +/NK1 1 + NKT cells