再生障碍性贫血患者T淋巴细胞早期激活及可溶性肿瘤坏死因子受体的研究

Study on the T lymphocytes early activation and soluble tumor necrosis factor receptor in patients with aplastic anemia

目的研究再生障碍性贫血 (再障 )患者外周血CD4+ 、CD8+ T淋巴细胞早期激活标志CD6 9的表达及血清、骨髓中可溶性肿瘤坏死因子受体 1和 2 (sTNF R1和sTNF R2 )的水平及其意义。方法将外周血在 2 0 μg ml植物血凝素 (PHA)条件下进行全血细胞培养 ,于 0h和 4h分别用双色免疫荧光标记流式细胞术对CD4+ 、CD8+ T淋巴细胞CD6 9的表达进行分析。用ELISA法检测血清和骨髓中sTNF R1和sTNF R2的水平。结果PHA刺激前重型再障 (SAA)患者CD4+ 、CD8+ 细胞CD6 9的表达率增高 [分别为 (8.96± 7.2 3) %和 (10 .6 7± 7.5 8) % ],慢性再障 (CAA)患者CD8+ 细胞CD6 9的表达率增高[(7.36± 5 .4 9) % ];PHA刺激后再障患者CD4+ 、CD8+ 细胞表达CD6 9明显增强 [SAA为 (71.73±11 91) %和 (6 1.74± 13.4 4 ) % ;CAA为 (5 9.35± 10 .15 ) %和 (4 8.78± 8.2 5 ) % ],CD4+ 细胞CD6 9的表达率高于CD8+ 细胞。SAA患者两种sTNF R水平均明显升高 [sTNF R1血清为 (12 5 8.5 8± 385 .6 9)ng L ,骨髓为 (16 5 0 .6 0± 6 6 5 .0 1)ng L ;sTNF R2血清为 (12 5 7.10± 2 95 .4 9)ng L ,骨髓为 (2 0 73.5 7± 5 6 1.17)ng L]。CAA患者骨髓两种sTNF R水平升高 [sTNF R1为 (10 94 .2 1± 2 91.93)ng L ,sTNF R2为 (15 0 2 .4 8±385 .

ObjectiveTo investigate the expression of T cell early activation marker (CD 69) on peripheral CD 4 +and CD 8 +lymphocytes and serum levels of soluble tumor necrosis factor receptor 1 and 2 ( sTNF-R1 and sTNF-R2) in serum and bone marrow in patients with aplastic anemia (AA) and their pathophysiological significance. Methods In vitro activation of T lymphocytes was carried out by whole blood cell culture containing PHA (20 μg/ml). The CD 69 expressions on CD 4 + and CD 8 +lymphocytes at 0 h and 4 h after PHA exposure were analyzed by two-color flow cytometry. The levels of sTNF-R1 and sTNF-R2 in serum and bone marrow were measured by ELISA. Results The CD 69 expression rates of CD 4 +and of CD 8 + cells in SAA patients were (8.96±7.23)% and (10.67±7.58)%,respectively, and that of CD 8 +cells in CAA patients was (7.36±5.49)% before PHA stimulation. The CD 69 expression rates of CD 4 +and of CD 8 +cells in SAA patients were (71.73±11.91)% and (61.74±13.44)% and in CAA (59.35±10.15)% and (48.78±8.25)% respectively, and were significantly elevated after PHA stimulation.CD 69 expression on CD 4 +cells was much higher than that on CD 8 +cells after stimulation.The levels of the two sTNF-R (sTNF-R1 and sTNF-R2) in peripheral blood and bone marrow of SAA patients were elevated and in the bone marrow of CAA patients were also increased.The serum levels of sTNF-R2 were positively related to the CD 69 expression rates of CD 8 +cells before PHA stimulation. Conclusion Increased early activation and activated potentials of T lymphocytes,along with abnormally elevated immunologically active molecules might play a major role in the pathogenesis of AA.