病毒受体拮抗性诱获配体H9的体外活性及其对人趋化因子受体CX_3CR1的作用  

The viral acceptor antagonist to entrapment ligand H9 in vitro activeness and the function to human acceptor CX3CR1

作　　者：张敏[1] 孙晗笑[1] 莫雪梅[1] 李秀英[1] 张光[1] 

机构地区：[1]暨南大学药学院基因组药物研究所,广东广州510632

出　　处：《细胞与分子免疫学杂志》2011年第6期597-601,共5页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家自然科学基金资助项目(3087221);广州市重大科技计划资助项目(2005Z1-E4081)

摘　　要：目的:来源于US28受体N末端的诱惑配体多肽H9,以阐明H9的体外活性及其对细胞表面趋化因子受体CX3CR1内化及调变研究,探讨H9对人趋化因子受体CX3CR1的作用及影响。方法:立足US28的广谱趋化因子结合活性,得到趋化因子受体拮抗性多肽H9。采用趋化抑制实验检测多肽对生理性趋化因子引起的细胞迁移活性的影响,流式细胞术方法(FCM)检测细胞内钙离子浓度变化,利用激光共聚焦显微镜和流式细胞仪分别定性、定量检测CX3CR1的内化,以阐明H9的生物活性及其对人源受体CX3CR1的作用及其机制。结果:多肽H9可以阻断受体结合生理性趋化因子形成的趋化作用,本身不引起趋化运动,不影响胞内信号转导和细胞自然活性;200 ng/mL H9在给药后的最初50min钟内能使细胞表面受体CX3CR1内化达到最大值,内化率约为70%,第100分钟,内化到细胞内的CX3CR1逐渐再循环到细胞表面。证明了H9能引起细胞表面受体CX3CR1内化,内化的受体部分再循环到细胞表面。结论:H9是一种趋化因子受体抑制短肽,影响人受体CX3CR1的内化,但内化后受体再循环到细胞表面,对人受体CX3CR1生理功能没有明显影响,可以作为特异性抗病毒多肽。AIM： To clarify the activeness of H9 in vitro and internalization and modulation of the surface chemokine receptor CX3CR1 induced by H9,To discuss the influence of H9 on the chemokine receptor CX3CR1.METHODS： Inhibition by chemotactic peptide on the physiological detection of chemokine induced cell migration activity.Flowcytometry examined the effection of H9 on intracellular calcium.Laser scanning confocal microscopy and flow cytometry were used to determine thequality and quantity of CX3CR1 internalization.RESULTS： H9 was able to block the migration induced by chemokine receptor.In the chemoattraction test,H9 was unable to induce the chemotactic movement,and it does not affect the signal transduction and activeness of cells.It was found that H9 could induce internalization with a maximal rate of 70%,at the concentration of 200 ng/mL.The internalized CX3CR1 molecules could recycled to the cell surface.CONCLUSION： H9 makes human CX3CR1 internalize.After internalizing,the CX3CR1 receptor recirculates the cell surface.It does not affect CX3CR1 physiology function.H9 could be used as a specificity anti-virus peptide.

关 键 词：病毒受体 趋化因子受体 趋化抑制 诱惑配体 趋化因子受体拮抗剂 受体内化 

分 类 号：R392.1[医药卫生—免疫学]