中华微生物学和免疫学杂志
中華微生物學和免疫學雜誌
중화미생물학화면역학잡지
CHINESE JOURNAL OF MICROBIOLOGY AND IMMUNOLOGY
2013年
10期
721-726
,共6页
HLA-G1%NK细胞%胞啃
HLA-G1%NK細胞%胞啃
HLA-G1%NK세포%포습
HLA-G1%NK cell%Trogocytosis
目的：自然杀伤细胞（ NK）不表达HLA-G抗原，将其与K562-G1细胞共培养，可检测到NK细胞表面表达HLA-G1蛋白，研究NK细胞表面获取HLA-G1抗原的机制。方法建立稳定表达HLA-G1抗原的K562细胞株；分别将K562-G1、K562、脱落的HLA-G1与NK-92MI细胞共培养，以及将HLA-G1与NK-92MI细胞上的HLA-G受体进行阻断后进行共培养，通过流式细胞术及荧光显微镜技术检测NK-92MI细胞对HLA-G1蛋白的获取情况。通过基于CD107a标记的流式细胞术，检测K562细胞表达HLA-G1前后对NK-92MI细胞杀伤活性的影响。结果 NK-92MI细胞能通过细胞间的相互接触转移快速获取K562-G1细胞表面的HLA-G1蛋白。 HLA-G特异性抗体87G阻断或其受体KIR2DL4特异性抗体＃33和ILT2受体特异性抗体阻断后，均不影响NK-92MI细胞对HLA-G1的获取。功能研究显示，HLA-G1能显著抑制NK细胞对K562细胞的杀伤活性（P＜0．01）。结论 NK细胞通过胞啃机制快速获取HLA-G1蛋白，其转移机制与受体配体间的亲和力、HLA-G1分子胞外结构域及非特异性的被动黏附作用无关。
目的：自然殺傷細胞（ NK）不錶達HLA-G抗原，將其與K562-G1細胞共培養，可檢測到NK細胞錶麵錶達HLA-G1蛋白，研究NK細胞錶麵穫取HLA-G1抗原的機製。方法建立穩定錶達HLA-G1抗原的K562細胞株；分彆將K562-G1、K562、脫落的HLA-G1與NK-92MI細胞共培養，以及將HLA-G1與NK-92MI細胞上的HLA-G受體進行阻斷後進行共培養，通過流式細胞術及熒光顯微鏡技術檢測NK-92MI細胞對HLA-G1蛋白的穫取情況。通過基于CD107a標記的流式細胞術，檢測K562細胞錶達HLA-G1前後對NK-92MI細胞殺傷活性的影響。結果 NK-92MI細胞能通過細胞間的相互接觸轉移快速穫取K562-G1細胞錶麵的HLA-G1蛋白。 HLA-G特異性抗體87G阻斷或其受體KIR2DL4特異性抗體＃33和ILT2受體特異性抗體阻斷後，均不影響NK-92MI細胞對HLA-G1的穫取。功能研究顯示，HLA-G1能顯著抑製NK細胞對K562細胞的殺傷活性（P＜0．01）。結論 NK細胞通過胞啃機製快速穫取HLA-G1蛋白，其轉移機製與受體配體間的親和力、HLA-G1分子胞外結構域及非特異性的被動黏附作用無關。
목적：자연살상세포（ NK）불표체HLA-G항원，장기여K562-G1세포공배양，가검측도NK세포표면표체HLA-G1단백，연구NK세포표면획취HLA-G1항원적궤제。방법건립은정표체HLA-G1항원적K562세포주；분별장K562-G1、K562、탈락적HLA-G1여NK-92MI세포공배양，이급장HLA-G1여NK-92MI세포상적HLA-G수체진행조단후진행공배양，통과류식세포술급형광현미경기술검측NK-92MI세포대HLA-G1단백적획취정황。통과기우CD107a표기적류식세포술，검측K562세포표체HLA-G1전후대NK-92MI세포살상활성적영향。결과 NK-92MI세포능통과세포간적상호접촉전이쾌속획취K562-G1세포표면적HLA-G1단백。 HLA-G특이성항체87G조단혹기수체KIR2DL4특이성항체＃33화ILT2수체특이성항체조단후，균불영향NK-92MI세포대HLA-G1적획취。공능연구현시，HLA-G1능현저억제NK세포대K562세포적살상활성（P＜0．01）。결론 NK세포통과포습궤제쾌속획취HLA-G1단백，기전이궤제여수체배체간적친화력、HLA-G1분자포외결구역급비특이성적피동점부작용무관。
Objective To investigate the mechanism of acquisition of HLA-G1 antigen by NK cells.Methods K562 cells stably expressing HLA-G1 antigen (K562-G1) were constructed.K562-G1 cells, K562 cells and shed HLA-G1 were respectively co-cultured with NK-92MI cells to observe the acquisi-tion of HLA-G by NK cells.To further investigate the mechanism , NK-92MI cells with blockage HLA-G re-ceptors were further co-cultured with K562-G1 cells and HLA-G1 proteins expressing on K 562-G1 cells were blocked and then co-cultured with NK-92MI cells. Acquisition of HLA-G 1 by NK-92MI cells was analyzed by flow cytometry and fluorescence microscopy .The effects of HLA-G1 expression on the cytotoxicity of NK-92MI cell were evaluated by flow cytometry analysis based on CD 107a labeling.R esults NK-92MI cells could quickly acquire HLA-G1 from K562-G1 cells in co-culture experiments .Blockade of HLA-G1 or its re-ceptors KIR2DL4 and ILT2 with specific mAbs did not affect the acquisition of HLA-G1 by NK-92MI cells. Moreover, HLA-G1 could significantly inhibit the cytotoxicity of NK cell ( P<0.01).Conclu sion NK-92MI cells acquire HLA-G1 from K562-G1 cells via trogocytosis , which is not associated with affinity be-tween receptor and ligand , extracellular domain of HLA-G1 or passive adhesion .
