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Published on: May 8, 2016
[Multiple types of chemokines expressed in mouse thymic stromal cell lines]
1Department of Immunology, School of Basic Medical Sciences, Peking University, Beijing 100083, China.
Objective:
To investigate semi-quantitative mRNA expression of SDF-1 alpha, IP-10, KC, MCP-1, and RANTES in thymic stromal cell lines of MTEC1, MTDC, D2SC, MTECB, TEC 1C8, TNC as well as in the primary thymic stromal cell cultures. The chemotactic activities of recombinant SDF-1 alpha, IP-10, MCP-1, and RANTES to mouse thymocytes were detected.
Methods:
Using beta-actin as internal control, the mRNA of the chemokines listed above were amplified for 30 cycles with RT-PCR. The amplified products were observed by agarose electrophoresis, and each band was analyzed with integrated optical density. With the method of Boyden chamber assay, the chemotactic activities of recombinant SDF-1 alpha, IP-10, MCP-1, and RANTES were detected to thymocytes, and the chemotactic indices were calculated.
Results:
The expression intensity of SDF-1 alpha, IP-10, KC, MCP-1, and RANTES varied from each other in the stromal cell lines detected. The PCR products of SDF-1 alpha and MCP-1 were not seen in D2SC or TEC 1C8, nor was the band of KC observed in TEC 1C8 either. The chemotactic indices of recombinant SDF-1 alpha, IP-10, MCP-1, and RANTES to thymocytes were 3.7, 4.5, 6.2, and 2.6, respectively.
Conclusions:
Different thymic stromal cell lines could express different types of chemokines with different expression intensities. To thymocytes, recombinant SDF-1 alpha, MCP-1, and IP-10 showed strong chemotactic activities, while the chemotactic activity of RANTES was very weak.
