蜕膜自然杀伤细胞趋化因子受体的表达及募集机制的研究  

The expression of chemokine receptors in CD56^(bright) CD16^- natural killer cells and the mechanism of their recruitment in decidua

作　　者：吴霞[1] 李大金[1] 袁敏敏[1] 朱影[1] 王明雁[1] 

机构地区：[1]复旦大学附属妇产科医院研究所,上海200011

出　　处：《中华医学杂志》2004年第12期1018-1023,共6页National Medical Journal of China

基　　金：复旦大学"985工程"基金资助项目 (985B3 6)

摘　　要：目的　研究早孕期人蜕膜自然杀伤 (NK)细胞趋化因子受体谱的表达及其特异性募集的机制。方法　收集早孕期蜕膜组织 ,免疫磁珠分选CD5 6 brightCD16 -NK细胞 ;逆转录 聚合酶链反应(RT PCR)检测CD5 6 brightCD16 -NK细胞中 18种趋化因子受体的转录水平 ,筛选出高表达的趋化因子受体 (即CXCR4及CXCR3)。原位杂交及免疫组织化学分析CXCR4特异性配体基质细胞衍生因子 (SDF 1)在早孕胎盘的定位表达 ;ELISA检测早孕滋养细胞培养上清液中SDF 1α的浓度水平。趋化试验研究重组人SDF 1α(rhSDF 1α)及滋养细胞培养上清液对蜕膜CD5 6 brightCD16 -NK细胞的趋化作用。结果　人蜕膜CD5 6 brightCD16 -NK细胞可转录多种趋化因子受体 ,其中CXCR4及CXCR3mRNA呈高水平表达。人早孕期滋养细胞表达趋化因子SDF 1;原代培养的滋养细胞可自发分泌SDF 1α,培养 6 0h后上清液中SDF 1α的浓度达 385ng/L± 91ng/L。rhSDF 1α及滋养细胞培养上清液对蜕膜CD5 6 brightCD16 -NK细胞具有显著趋化作用。当rhSDF 1α为 10 μg/L时趋化至Transwell下室中的细胞可达总细胞数的2 2 9%± 4 3%。结论　人早孕期蜕膜CD5 6 brightCD16 -NK细胞高水平转录CXCR4 ,而滋养细胞则表达并分泌CXCR4的配体SDF 1。SDF 1趋化、募集CD5 6ObjectiveTo investigate the transcriptions of 18 chemokine receptors in human decidual natural killer (NK) cells and explore the possible mechanisms of preferential accumulation of CD56 bright CD16 -NK cells in decidua during first-trimester pregnancy. MethodsVilli and decidual tissue were collected from normal pregnant women with 5～10 gestational weeks by artificial abortion. The decidual CD56 bright CD16 -NK cells were isolated by immune magnetic beads. The transcription levels of 18 chemokine receptors in the decidual CD56 bright CD16 -NK cells were assessed by reverse transcription-polymerase chain reaction (RT-PCR). After the high expression of CXCR4 and CXCR3 mRNA in these cells was found,the expression of stromal cell-derived factor-1 (SDF-1),the specific ligand of CXCR4,in first-trimester human placenta was detected by in situ hybridization and immunohistochemistry. The concentration of SDF-1α in the supernatant of culture of isolated trophoblast cells derived from the first-trimester human placentas was measured by ELISA. The chemotaxis of SDF-1 to decidual CD56 bright CD16 -NK cells was tested in Transwell,and the chemotactic activity was quantitatively examined. ResultsAmong the 18 chemokine receptors,CXCR4 and CXCR3 were found highly transcribed in decidual CD56 bright CD16 -NK cells. The concentration of SDF-1α in the supernatant was 385 ng/L±91 ng/L after trophoblast cells had been cultured for 60 hours. Both rhSDF-1α and supernatants in the culture of trophoblast cells exhibited chemotactic activity on decidual CD56 bright CD16 -NK cells. When the concentration of rhSDF-1α was 10 μg/L the number of cells that entered the lower chamber of Transwell accounted for 22.9%±4.3% of the total calls. ConclusionFirst-trimester human trophoblast cells produce SDF-1,which in turn endows the trophoblast cells with the capacity to attract decidual CD56 bright CD16 -NK cells highly expressing CXCR4. This activity contributes to the recruitment of decidual lymphocytes and may

关 键 词：蜕膜自然杀伤细胞 趋化因子受体 基因表达 NK 滋养细胞 基质细胞 

分 类 号：R392.1[医药卫生—免疫学]