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目的研究CD54和LFA-1的相互作用在6A8α甘露糖苷酶表达抑制的Jurkat细胞(AS)的黏附性增强中的作用。方法用细胞凝集试验确证AS细胞间黏附的增强,用细胞与细胞间黏附分子1人IgG的Fc片段(ICAM1Fc)的黏附试验和阻断性抗CD11a抗体的阻断试验研究CD54LFA1的作用,用单克隆抗体MEM148检测AS细胞LFA-1亲和力的变化,用单克隆抗体NKIL16检测AS细胞LFA1亲合力的变化,用鬼笔环肽染色细胞骨架,用JurkatRaji细胞间的作用作模型研究6A8α甘露糖苷酶表达抑制对T和B细胞间黏附的影响,用ConA结合试验检测细胞中蛋白质N糖基化的变化。结果(1)AS细胞间的黏附性增强主要与CD54及CD11a表达的增强相关,也与LFA-1亲和力的增高相关;(2)AS细胞的细胞骨架发生重排;(3)AS细胞与Raji细胞间的黏附也增强;(4)ConA与AS细胞的结合增强。结论CD54和LFA-1的相互作用在ASJurkatT细胞的黏附性增强中起重要作用。细胞骨架重排也可能起作用。AS细胞的蛋白质发生了N-糖基化修饰。
Aim To investigate the role of CD54 and LFA-1 interactions in the enhancement of adhesion of Jurkat cells (AS) with inhibition of 6A8α-mannosidase expression. Methods The effect of CD54LFA1 on the adhesion of AS cells was confirmed by cell agglutination test. The effect of CD54LFA1 on the adhesion of ICAM1Fc and the blockade of anti-CD11a antibody was studied. The changes of affinity of LFA-1 in AS cells were detected by the monoclonal antibody MEM148, the affinity of LFA1 in AS cells was detected by the monoclonal antibody NKIL16, the cytoskeleton was stained with phalloidin, and the expression of 6A8α-mannosidase was studied by the interaction between Jurkat-Raji cells Inhibition of T and B cell adhesion between cells using ConA binding assay of protein N glycosylation changes. Results (1) The enhancement of adhesion between AS cells was mainly associated with the enhanced expression of CD54 and CD11a, and also with the increased affinity of LFA-1. (2) The cytoskeleton of AS cells was rearranged. (3) Cell adhesion is also enhanced; (4) ConA and AS cells increased binding. Conclusion The interaction between CD54 and LFA-1 plays an important role in the adhesion enhancement of ASJurkat T cells. Cytoskeleton rearrangements may also work. N-glycosylation of AS cell proteins occurs.