Demonstration of heat-labile and heat-stable epitopes of Rickettsia japonica on ultrathin sections

T. Uchiyama, T. Uchida, J. W. Wen, David Walker

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

BACKGROUND: Spotted fever group rickettsiae including Rickettsia japonica have two major polypeptides and lipopolysaccharide (LPS)-like antigen on the surface. The major part of the antibodies to these polypeptides generated by immunization with live rickettsiae are reactive with heat-labile and conformation-dependent epitopes. EXPERIMENTAL DESIGN: To demonstrate and precisely localize the heat-labile and heat-stable epitopes on the major surface polypeptides and LPS-like antigen of R. japonica, the immunocolloidal gold method was performed on ultrathin sections with polyclonal and monoclonal antibodies. The antigenicity and fine structure were preserved by using periodate-lysine-paraformaldehyde as fixative, followed by embedding with the resin Lowicryl K4M at a polymerization temperature of -35° C. RESULTS: Heat-labile and heat-stable epitopes on the two major surface polypeptides together with LPS-like antigen of R. japonica were demonstrated at the same location. These antigens were rather broadly distributed on the cell wall apart from the slime layer of the organism. The number of gold particles corresponding to the LPS-like antigen was much larger than that corresponding to the polypeptide antigen. CONCLUSIONS: Heat-labile epitopes of rickettsiae could be preserved and demonstrated by using the procedure cited here. Moreover, the precise localization of the major surface polypeptides and LPS-like antigen could be achieved in the cell wall by using ultrathin sections of infected cells instead of whole rickettsial cells. The LPS-like antigen was quantitatively predominant as judged by immunoelectron microscopy.

Original languageEnglish (US)
Pages (from-to)432-437
Number of pages6
JournalLaboratory Investigation
Volume71
Issue number3
StatePublished - 1994
Externally publishedYes

Fingerprint

Rickettsia
Epitopes
Hot Temperature
Lipopolysaccharides
Antigens
Peptides
Gold
Cell Wall
Fixatives
Immunoelectron Microscopy
Surface Antigens
Polymerization
Immunization
Fever
Monoclonal Antibodies
Temperature
Antibodies

Keywords

  • Immunocolloidal gold method
  • Immunoelectron microscopy
  • Lipopolysaccharide
  • Major surface polypeptides
  • Spotted fever group rickettsiae

ASJC Scopus subject areas

  • Pathology and Forensic Medicine

Cite this

Demonstration of heat-labile and heat-stable epitopes of Rickettsia japonica on ultrathin sections. / Uchiyama, T.; Uchida, T.; Wen, J. W.; Walker, David.

In: Laboratory Investigation, Vol. 71, No. 3, 1994, p. 432-437.

Research output: Contribution to journalArticle

@article{1ec739e66da049c680e1b704a2e43d95,
title = "Demonstration of heat-labile and heat-stable epitopes of Rickettsia japonica on ultrathin sections",
abstract = "BACKGROUND: Spotted fever group rickettsiae including Rickettsia japonica have two major polypeptides and lipopolysaccharide (LPS)-like antigen on the surface. The major part of the antibodies to these polypeptides generated by immunization with live rickettsiae are reactive with heat-labile and conformation-dependent epitopes. EXPERIMENTAL DESIGN: To demonstrate and precisely localize the heat-labile and heat-stable epitopes on the major surface polypeptides and LPS-like antigen of R. japonica, the immunocolloidal gold method was performed on ultrathin sections with polyclonal and monoclonal antibodies. The antigenicity and fine structure were preserved by using periodate-lysine-paraformaldehyde as fixative, followed by embedding with the resin Lowicryl K4M at a polymerization temperature of -35° C. RESULTS: Heat-labile and heat-stable epitopes on the two major surface polypeptides together with LPS-like antigen of R. japonica were demonstrated at the same location. These antigens were rather broadly distributed on the cell wall apart from the slime layer of the organism. The number of gold particles corresponding to the LPS-like antigen was much larger than that corresponding to the polypeptide antigen. CONCLUSIONS: Heat-labile epitopes of rickettsiae could be preserved and demonstrated by using the procedure cited here. Moreover, the precise localization of the major surface polypeptides and LPS-like antigen could be achieved in the cell wall by using ultrathin sections of infected cells instead of whole rickettsial cells. The LPS-like antigen was quantitatively predominant as judged by immunoelectron microscopy.",
keywords = "Immunocolloidal gold method, Immunoelectron microscopy, Lipopolysaccharide, Major surface polypeptides, Spotted fever group rickettsiae",
author = "T. Uchiyama and T. Uchida and Wen, {J. W.} and David Walker",
year = "1994",
language = "English (US)",
volume = "71",
pages = "432--437",
journal = "Laboratory Investigation",
issn = "0023-6837",
publisher = "Nature Publishing Group",
number = "3",

}

TY - JOUR

T1 - Demonstration of heat-labile and heat-stable epitopes of Rickettsia japonica on ultrathin sections

AU - Uchiyama, T.

AU - Uchida, T.

AU - Wen, J. W.

AU - Walker, David

PY - 1994

Y1 - 1994

N2 - BACKGROUND: Spotted fever group rickettsiae including Rickettsia japonica have two major polypeptides and lipopolysaccharide (LPS)-like antigen on the surface. The major part of the antibodies to these polypeptides generated by immunization with live rickettsiae are reactive with heat-labile and conformation-dependent epitopes. EXPERIMENTAL DESIGN: To demonstrate and precisely localize the heat-labile and heat-stable epitopes on the major surface polypeptides and LPS-like antigen of R. japonica, the immunocolloidal gold method was performed on ultrathin sections with polyclonal and monoclonal antibodies. The antigenicity and fine structure were preserved by using periodate-lysine-paraformaldehyde as fixative, followed by embedding with the resin Lowicryl K4M at a polymerization temperature of -35° C. RESULTS: Heat-labile and heat-stable epitopes on the two major surface polypeptides together with LPS-like antigen of R. japonica were demonstrated at the same location. These antigens were rather broadly distributed on the cell wall apart from the slime layer of the organism. The number of gold particles corresponding to the LPS-like antigen was much larger than that corresponding to the polypeptide antigen. CONCLUSIONS: Heat-labile epitopes of rickettsiae could be preserved and demonstrated by using the procedure cited here. Moreover, the precise localization of the major surface polypeptides and LPS-like antigen could be achieved in the cell wall by using ultrathin sections of infected cells instead of whole rickettsial cells. The LPS-like antigen was quantitatively predominant as judged by immunoelectron microscopy.

AB - BACKGROUND: Spotted fever group rickettsiae including Rickettsia japonica have two major polypeptides and lipopolysaccharide (LPS)-like antigen on the surface. The major part of the antibodies to these polypeptides generated by immunization with live rickettsiae are reactive with heat-labile and conformation-dependent epitopes. EXPERIMENTAL DESIGN: To demonstrate and precisely localize the heat-labile and heat-stable epitopes on the major surface polypeptides and LPS-like antigen of R. japonica, the immunocolloidal gold method was performed on ultrathin sections with polyclonal and monoclonal antibodies. The antigenicity and fine structure were preserved by using periodate-lysine-paraformaldehyde as fixative, followed by embedding with the resin Lowicryl K4M at a polymerization temperature of -35° C. RESULTS: Heat-labile and heat-stable epitopes on the two major surface polypeptides together with LPS-like antigen of R. japonica were demonstrated at the same location. These antigens were rather broadly distributed on the cell wall apart from the slime layer of the organism. The number of gold particles corresponding to the LPS-like antigen was much larger than that corresponding to the polypeptide antigen. CONCLUSIONS: Heat-labile epitopes of rickettsiae could be preserved and demonstrated by using the procedure cited here. Moreover, the precise localization of the major surface polypeptides and LPS-like antigen could be achieved in the cell wall by using ultrathin sections of infected cells instead of whole rickettsial cells. The LPS-like antigen was quantitatively predominant as judged by immunoelectron microscopy.

KW - Immunocolloidal gold method

KW - Immunoelectron microscopy

KW - Lipopolysaccharide

KW - Major surface polypeptides

KW - Spotted fever group rickettsiae

UR - http://www.scopus.com/inward/record.url?scp=0027984770&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0027984770&partnerID=8YFLogxK

M3 - Article

VL - 71

SP - 432

EP - 437

JO - Laboratory Investigation

JF - Laboratory Investigation

SN - 0023-6837

IS - 3

ER -