多种类型趋化性细胞因子在小鼠胸腺基质细胞系中的表达

来源 :中国医学科学院学报 | 被引量 : 0次 | 上传用户:wzcc1125
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目的分析趋化性细胞因子 SDF- 1α、 IP- 10、 KC、 MCP- 1和 RANTES在小鼠胸腺基质细胞系 MTEC1、 MTDC、 D2SC、 MTECB、 TEC 1C8、 TNC及原代培养胸腺基质细胞的半定量表达情况,并检测重组的趋化性细胞因子纯品 SDF- 1α、 IP- 10、 MCP- 1和 RANTES对小鼠胸腺细胞的趋化活性。方法以β- actin为内对照,用 RT- PCR方法,对上述 5种趋化性细胞因子进行 30个循环 PCR扩增,用凝胶成像系统观察扩增结果 ,并对各电泳带进行积分光密度扫描分析。用 Boyden小室法检测重组趋化性细胞因子 SDF- 1α、 IP- 10、 MCP- 1和 RANTES对全胸腺细胞的趋化活性,并计算其趋化指数。结果 SDF- 1α、 IP- 10、 KC、 MCP- 1和 RANTES在被检测胸腺基质细胞系的表达谱不同,表达强度亦有差异。 SDF- 1α和 MCP- 1在 D2SC和 TEC 1C8中未出现可见扩增条带;在 TEC 1C8中也未见 KC的扩增条带。重组趋化性细胞因子 SDF- 1α、 IP- 10、 MCP- 1和 RANTES对胸腺细胞的趋化指数分别为 3.7、 4.5、 6.2和 2.6。结论不同的胸腺基质细胞系,其趋化性细胞因子的表达谱和表达强度不同, 4种重组趋化性细胞因子对胸腺细胞表现出不同的趋化活性。 OBJECTIVE: To investigate the effects of chemotactic cytokines SDF-1α, IP-10, KC, MCP-1 and RANTES on the proliferation of mouse thymic stromal cell lines MTEC1, MTDC, D2SC, MTECB, TEC 1C8, TNC and primary culture of thymus stromal cells The chemotactic activity of recombinant chemotactic cytokines, pure SDF-1α, IP-10, MCP-1 and RANTES, on the thymocytes of mice was detected. Methods β-actin was used as an internal control. The RT-PCR method was used to amplify the above 5 chemotactic cytokines by 30 cycles. The amplification results were observed by gel imaging system. The integrated light Density scanning analysis. The chemotactic activity of recombinant chemotactic cytokines SDF-1α, IP-10, MCP-1 and RANTES on whole thymocytes was detected by Boyden chamber assay and the chemotactic index was calculated. Results The expression profiles of SDF-1α, IP-10, KC, MCP-1 and RANTES were different in the tested thymic stromal cell lines. No visible amplification bands were found for SDF-1α and MCP-1 in D2SC and TEC 1C8; no amplification band for KC was found in TEC 1C8. The chemotactic indexes of recombinant chemotactic cytokines SDF-1α, IP-10, MCP-1 and RANTES on thymocytes were 3.7, 4.5, 6.2 and 2.6, respectively. CONCLUSION: The expression and expression intensity of chemotactic cytokines in different thymic stromal cell lines are different. The four recombinant chemotactic cytokines show different chemotactic activities on thymocytes.
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