ELECTROPORATION OF EXOGENOUS ANTIGEN INTO THE CYTOSOL FOR ANTIGEN PROCESSING AND CLASS-I MAJOR HISTOCOMPATIBILITY COMPLEX (MHC) PRESENTATION - WEAK BASE AMINES AND HYPOTHERMIA (18-DEGREES-C) INHIBIT THE CLASS-I MHC PROCESSING PATHWAY
ELECTROPORATION OF EXOGENOUS ANTIGEN INTO THE CYTOSOL FOR ANTIGEN PROCESSING AND CLASS-I MAJOR HISTOCOMPATIBILITY COMPLEX (MHC) PRESENTATION - WEAK BASE AMINES AND HYPOTHERMIA (18-DEGREES-C) INHIBIT THE CLASS-I MHC PROCESSING PATHWAY
复制标题
DOI：
10.1002/eji.1830220728
复制
发表时间：
1992-07-01
影响因子：
5.4
通讯作者：
HARDING, CV
中科院分区：
文献类型：
--
作者：
HARDING, CV
虽然内源性抗原是由I类主要组织相容性复合体(MHC)分子呈递的，但外源抗原通常需要在I类MHC呈递之前穿透到胞浆中进行加工。我们优化了电穿孔的条件，作为一种实验性地将外源抗原引入胞浆的手段，为解剖I类MHC加工途径提供了一种系统。通过I类或II类MHC限制性T杂交瘤细胞的反应来评估呈递。基本上，通过电穿孔瞬时递送抗原促进了I类途径的动力学分析，并研究了各种抑制剂或低温条件对I类MHC抗原处理的影响。这一途径被弱碱性胺(如氯喹和NH4Cl)、放线菌亚胺和低温(18摄氏度，抑制某些细胞内囊泡加工途径)抑制。电穿孔技术为分析I类MHC加工提供了一种简单、一致的快速胞浆抗原递送方法。
While endogenous antigens are presented by class I major histocompatibility complex (MHC) molecules, exogenous antigens generally require a means for penetration into the cytosol for processing prior to class I MHC presentation.We have optimized conditions for electroporation as a means to experimentally introduce exogenous antigens into the cytosol, providing a system with a number of advantages for dissecting the class I MHC processing pathway. Presentation was assessed by the response of class I or class II MHC-restricted T hybridoma cells. Essentially instantaneous antigen delivery by electroporation facilitated kinetic analysis of the class I pathway and investigation of the effects of various inhibitors or hypothermic conditions on class I MHC antigen processing. This pathway was inhibited by weak base amines (e.g. chloroquine and NH4Cl), cycloheximide, and hypothermia (18-degrees-C, which inhibits certain intracellular vesicular processing pathways). The electroporation technique provides a simple, consistent approach for rapid cytosolic antigen delivery for analysis of class I MHC processing.