小鼠白细胞免疫球蛋白样受体2干扰RNA对树突状细胞表面CD11c、CD80和CD86的影响及意义

摘要

目的：探讨小鼠树突状细胞（DC）表面白细胞免疫球蛋白样受体2（LILRB2）改变对DC的CD11c、CD80和CD86表型的影响及意义。方法应用20 ng/ml重组小鼠粒细胞集落刺激因子（rGM-CSF）＋20 ng/ml IL-4刺激小鼠骨髓细胞生长，隔日进行细胞换液，第5天实验组转染LILRB2受体干扰RNA，空白对照组转染空质粒，对照组单纯换液。培养过程中观察细胞形态学变化，并于转染72 h后，光镜下观察DC形态，用流式细胞仪测定细胞表面CD11c、CD80和CD86分子表达。结果光镜下对照组与实验组均可见 DC 细胞形态，流式细胞仪测定实验组 CD11c （0.662±0.174）（n=12）与空白对照组 CD11c（0.675±0.237）和对照组（0.688±0.076）分子表达无明显差异（P＞0.05）。实验组 DC 细胞表面 CD80（0.626±0.060）较空白对照组 CD80（0.406±0.163）（n=12）和对照组CD80（0.409±0.100）表达均增加（P＜0.05）。实验组DC细胞表面CD86（0.730±0.102）较空白对照组CD86（0.497±0.278）和对照组CD86（0.368±0.073）表达均增加（P＜0.05）。结论应用LILRB2受体干扰RNA不影响DC细胞表面CD11c分子表达，而增加细胞表面CD80和CD86分子表达。细胞表面CD80和CD86分子作为免疫反应的重要协同刺激分子在抗原提呈细胞表面表达升高，则激活T细胞免疫反应的能力增强。%Objective To explore the efficacy of LILRB2 siRNA on the expression of phenotype of CD11c, CD80, CD86 and its underlying meaning on mouse borrow dendritic cells (DC). Methods On the base of the routine culture, both the test group and the two control groups which were subdivided into void control group and control group were treated with 20 ng/ml GM-CSF and 20 ng/ml IL-4 to stimulate the growth of DC cells extracted from mouse bone marrow. At the fifth day, the test group was further transfected with LILRB2-siRNA, the void control group was transfected with empty plasmid, and the control group was added nothing but fluid media. After transfection for 72 h, all groups were observed under microscope and CD11c, CD80, CD86 were measured with flow cytometry. Results At the eighth day, the DC cells were all found under microcopy. Flow cytometry demonstrated there was no significant difference of the expression of CD11c among test group (0.662±0.174), void control group (0.675±0.237) and control group (0.688±0.076) (P>0.05). The expression of CD80 in test group (0.626±0.060) was higher than its expression in void control group (0.406±0.163) and control group (0.409±0.100) (P<0.05). The expression of CD86 in test group (0.730±0.102) was also higher than its expression in void control group (0.497±0.278) and control group (0.368±0.073) (P<0.05). Conclusions The application of LILRB2-siRNA would not alter the expression of CD11c on DC cells but effectively promote the phenotype molecules expression of CD80 and CD86, this often represents the enhancement capability of DC cells to stimulate the activation of T cells as the expression of CD80, CD86 on the APC cells are thought as key co-stimulatory molecules to facilitate this process.