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目的 分析小鼠胸腺树突状细胞株 (MTDC)中化学趋化因子的类型。方法 以自行设计的引物 ,用逆转录聚合酶链式反应 (RT PCR)方法 ,从小鼠MTDC细胞株扩增SDF 1,IP 10 ,KC等几种化学趋化因子的cDNA片段 ,并对部份扩增产物进行核苷酸测序 ,或dotblotting杂交分析 ,以MTDC浓缩上清进行趋化实验。结果 自MTDC中扩增出了SDF 1、IP 10、KC、MCP 1和Lymphotactin等 5种化学趋化因子的cDNA片段 ,并对SDF 1、KC和Lymphotactin的PCR产物做了核苷酸测序鉴定证明 ,对MCP 1、IP 10和Lymphotactin亦进行dotblotting分析证实。MTDC浓缩上清对小鼠胸腺CD-4 CD-8细胞的趋化指数 (CI)为 3.0 4。结论 MTDC细胞株中存在着多种化学趋化因子。MTDC的浓缩培养上清对小鼠胸腺CD-4 、CD-8细胞具有中等强度的趋化活性 ,符合SDF 1的生物功能。
Objective To analyze the types of chemoattractant in murine thymic dendritic cell line (MTDC). Methods cDNA fragments of several chemokines such as SDF 1, IP 10 and KC were amplified from mouse MTDC cell line by reverse transcription-polymerase chain reaction (RT PCR) using primers designed by ourselves. Amplify products for nucleotide sequencing, or dotblotting hybridization analysis, MTDC concentrated supernatants for chemotaxis experiments. Results cDNA fragments of 5 chemokines such as SDF 1, IP 10, KC, MCP 1 and Lymphotactin were amplified from MTDC and nucleotide sequences of SDF 1, KC and Lymphotactin were identified , Dotblotting analysis of MCP 1, IP 10 and Lymphotactin was also confirmed. The chemotactic index (CI) of MTDC concentrated supernatant on mouse thymus CD-4 CD-8 cells was 3.04. Conclusion There are many chemotactic factors in MTDC cell line. MTDC concentrated culture supernatant of mouse thymus CD-4, CD-8 cells with moderate chemotactic activity, in line with the biological function of SDF1.