六种识别在大肠杆菌K-12中表达的痢疾志贺氏菌1型O抗原重复单元的单克隆抗体的表位作图。

Epitope mapping of six monoclonal antibodies recognizing the Shigella dysenteriae type 1 O-antigenic repeating unit expressed in Escherichia coli K-12.

作者信息

Fält I C, Lindberg A A

机构信息

Department of Clinical Bacteriology, Karolinska Institute, Huddinge Hospital, Sweden.

出版信息

Microb Pathog. 1994 Jan;16(1):27-41. doi: 10.1006/mpat.1994.1003.

DOI:10.1006/mpat.1994.1003

PMID:7520112

Abstract

Mouse and rat monoclonal antibodies were generated against lipopolysaccharides expressed in Escherichia coli K-12/Shigella dysenteriae type 1 hybrids and native Shigella dysenteriae type 1 bacteria. Six monoclonal antibodies, all IgM and reacting with the E. coli K-12/Shigella dysenteriae type 1 hybrids, were selected and characterized. The specificities were studied in inhibition experiments using purified native and synthetic saccharides, representing different parts of the S. dysenteriae type 1 tetrasaccharide repeating unit. Three patterns of specificities were seen: (i) MAEC-SD-1 and MAEC-SD-2 recognizing the terminal alpha-D-Galp-(1-->3)-beta-D-GlcpNAc disaccharide as present in an E. coli K-12 class I hybrid strain, (ii) MAEC-SD-11; a class IV specific mAb, recognizing the alpha-L-Rhap-(1-->2)-alpha-D-Galp-(1-->3)-beta-D-GlcpNAc-(1-->tris accharide, and (iii) MAEC-SD-21, MASD-1, MASD-2; all mAbs with class V specificity, recognizing the alpha-L-Rhap-(1-->3)-alpha-L-Rhap-(1-->2)-alpha-D-Galp-(1--> trisaccharide. Furthermore, the data strongly suggest that the native repeating unit for S. dysenteriae type 1 is the alpha-L-Rhap-(1-->3)-alpha-L-Rhap-(1-->2)-alpha-D-Galp-(1--> 3)-alpha-D-GlcpNAc tetrasaccharide.

摘要

针对大肠杆菌K - 12/痢疾志贺氏菌1型杂交菌及天然痢疾志贺氏菌1型中表达的脂多糖，制备了小鼠和大鼠单克隆抗体。筛选并鉴定了6种单克隆抗体，均为IgM，且与大肠杆菌K - 12/痢疾志贺氏菌1型杂交菌发生反应。利用纯化的天然和合成糖类进行抑制实验研究其特异性，这些糖类代表痢疾志贺氏菌1型四糖重复单元的不同部分。观察到三种特异性模式：(i) MAEC - SD - 1和MAEC - SD - 2识别存在于大肠杆菌K - 12 I类杂交菌株中的末端α - D - Galp-(1→3)-β - D - GlcpNAc二糖；(ii) MAEC - SD - 11，一种IV类特异性单克隆抗体，识别α - L - Rhap-(1→2)-α - D - Galp-(→3)-β - D - GlcpNAc-(1→三糖；(iii) MAEC - SD - 21、MASD - 1、MASD - 2，均为V类特异性单克隆抗体，识别α - L - Rhap-(1→3)-α - L - Rhap-(1→2)-α - D - Galp-(1→三糖。此外，数据强烈表明痢疾志贺氏菌1型的天然重复单元是α - L - Rhap-(1→3)-α - L - Rhap-(1→2)-α - D - Galp-(1→3)-α - D - GlcpNAc四糖。