噬菌体颗粒抗原与MHC-Ⅱ类分子的交叉递呈位于内质网囊泡  

Cross-presentation of phage particle antigen in MHC class Ⅱ and endoplasmic reticulum marker-positive compartments

作　　者：郭晟[1] 万瑛[1] 周镜然[1] 邹丽云[1] 刘娜[1] 吴玉章[1] 

机构地区：[1]第三军医大学基础医学部全军免疫学研究所,重庆400038

出　　处：《第三军医大学学报》2005年第20期2031-2034,共4页Journal of Third Military Medical University

摘　　要：目的研究噬菌体呈现颗粒交叉递呈过程中,MHCⅠ抗原肽复合物在细胞内的形成部位,及其与MHCⅡ类分子阳性区域和内质网标志物阳性区域的关系。方法构建呈现OVA257264表位的噬菌体颗粒,大量制备此重组噬菌体颗粒。巨噬细胞与重组噬菌体颗粒共孵育后,使用抗MHCⅠ抗原肽复合物特异性单克隆抗体,观察巨噬细胞中MHCⅠ抗原肽复合物形成的部位。使用抗MHCⅡ类分子、内吞相关区域标志蛋白的单克隆抗体以及能显示内质网的特异性荧光染料,观察巨噬细胞内MHCⅠ抗原肽复合物和内吞相关区域及内质网的关系。结果成功构建了呈现OVA257264表位的噬菌体颗粒,纯化并鉴定了此噬菌体颗粒表达OVA257264表位的情况。共聚焦显微镜观察到MHCⅠ抗原肽复合物和MHCⅡ类分子阳性区域共聚,进一步染色发现此复合物位于转铁蛋白受体和LAMP1(晚期内体/溶酶体的表面标志物)阳性区域中,同时此区域呈现内质网标志蛋白阳性。结论重组噬菌体颗粒能在MHCⅡ类分子阳性、内体内质网标志物阳性的区域内完成交叉递呈。Objective To investigate the formation and intracellular location of MHC- Ⅰ -peptide complexes and discover the relationship between MHC class Ⅰ -peptide complexes and MHC class Ⅱ molecules and endoplasmic reticulum marker-positive compartments during the cross-presentation of phage particle antigens. Methods The hybrid filamentous phage particles were constructed, which displayed the OVA257-264 epitope. OVA257-264/Kb complexes and MHC class Ⅱ molecules or markers that distinguish early from late endocytic compartments and lysosomes or markers of ER were co-stained to further define the nature of the compartments stained with OVA257-264/Kb complexes. Results The phage particles displaying the OVA257-264 epitope were successfully constructed. We observed that the MHC class Ⅰ -peptide complexes co-localized with MHC class Ⅱ molecules and endocytic system makers or ER membrane markers during cross-presentation of phage particle antigens. Conclusion Endocytosed phage particles may be processed and cross-presented in organelles positive for phagosome and endoplasmic reticulum （ER） markers.

关 键 词：交叉递呈 CTL MHC 

分 类 号：R329.25[医药卫生—人体解剖和组织胚胎学] R373.9[医药卫生—基础医学]