Cell surface localization of a 72 kilodalton heat shock protein in retroocular fibroblasts from patients with Graves' ophthalmopathy

A. E. Heufelder, B. E. Wenzel, R. S. Bahn

Research output: Contribution to journalArticle

64 Citations (Scopus)

Abstract

Heat shock proteins (HSPs) have been implicated in autoimmune disease, although they are generally considered to be intracellular in location. We demonstrate that, under certain circumstances, the inducible intracellular 72 kilodalton HSP can also be detected on the surface of cells. We used cultured retroocular fibroblasts derived from patients with severe Graves' ophthalmopathy (GO) and normal individuals in our studies. Sodium dodecylsulfate polyacrylamide-gel electrophoresis of immunoprecipitated cell lysates, derived from surface-radioiodinated GO cell monolayers, resulted in a single band of appropriate molecular weight on the autoradiogram. Further, a bright cell surface staining pattern was observed when indirect immunofluorescence was performed using the same anti-HSP 72 monoclonal antibody on parallel cell cultures. No cell-surface HSP 72 reactivity was detected in normal retroocular fibroblast monolayers by either method. These results are the first demonstration, by immunoprecipitation of surface- labeled proteins, of HSP expression on the surface of cells. This localization of HSP 72 on the surface of affected cells obtained from patients with an autoimmune disease may have implications concerning the role of this molecule in autoimmunity in general, and particularly in the immune process of GO.

Original languageEnglish (US)
Pages (from-to)732-736
Number of pages5
JournalJournal of Clinical Endocrinology and Metabolism
Volume74
Issue number4
DOIs
StatePublished - 1992

Fingerprint

HSP72 Heat-Shock Proteins
Graves Ophthalmopathy
Fibroblasts
Heat-Shock Proteins
Autoimmune Diseases
Monolayers
Membrane Proteins
Indirect Fluorescent Antibody Technique
Electrophoresis
Autoimmunity
Cell culture
Immunoprecipitation
Polyacrylamide Gel Electrophoresis
Demonstrations
Cell Culture Techniques
Molecular Weight
Sodium
Molecular weight
Monoclonal Antibodies
Staining and Labeling

ASJC Scopus subject areas

  • Biochemistry
  • Endocrinology, Diabetes and Metabolism

Cite this

Cell surface localization of a 72 kilodalton heat shock protein in retroocular fibroblasts from patients with Graves' ophthalmopathy. / Heufelder, A. E.; Wenzel, B. E.; Bahn, R. S.

In: Journal of Clinical Endocrinology and Metabolism, Vol. 74, No. 4, 1992, p. 732-736.

Research output: Contribution to journalArticle

@article{76f495418367467d8a9e526f7ba85b4e,
title = "Cell surface localization of a 72 kilodalton heat shock protein in retroocular fibroblasts from patients with Graves' ophthalmopathy",
abstract = "Heat shock proteins (HSPs) have been implicated in autoimmune disease, although they are generally considered to be intracellular in location. We demonstrate that, under certain circumstances, the inducible intracellular 72 kilodalton HSP can also be detected on the surface of cells. We used cultured retroocular fibroblasts derived from patients with severe Graves' ophthalmopathy (GO) and normal individuals in our studies. Sodium dodecylsulfate polyacrylamide-gel electrophoresis of immunoprecipitated cell lysates, derived from surface-radioiodinated GO cell monolayers, resulted in a single band of appropriate molecular weight on the autoradiogram. Further, a bright cell surface staining pattern was observed when indirect immunofluorescence was performed using the same anti-HSP 72 monoclonal antibody on parallel cell cultures. No cell-surface HSP 72 reactivity was detected in normal retroocular fibroblast monolayers by either method. These results are the first demonstration, by immunoprecipitation of surface- labeled proteins, of HSP expression on the surface of cells. This localization of HSP 72 on the surface of affected cells obtained from patients with an autoimmune disease may have implications concerning the role of this molecule in autoimmunity in general, and particularly in the immune process of GO.",
author = "Heufelder, {A. E.} and Wenzel, {B. E.} and Bahn, {R. S.}",
year = "1992",
doi = "10.1210/jc.74.4.732",
language = "English (US)",
volume = "74",
pages = "732--736",
journal = "Journal of Clinical Endocrinology and Metabolism",
issn = "0021-972X",
publisher = "The Endocrine Society",
number = "4",

}

TY - JOUR

T1 - Cell surface localization of a 72 kilodalton heat shock protein in retroocular fibroblasts from patients with Graves' ophthalmopathy

AU - Heufelder, A. E.

AU - Wenzel, B. E.

AU - Bahn, R. S.

PY - 1992

Y1 - 1992

N2 - Heat shock proteins (HSPs) have been implicated in autoimmune disease, although they are generally considered to be intracellular in location. We demonstrate that, under certain circumstances, the inducible intracellular 72 kilodalton HSP can also be detected on the surface of cells. We used cultured retroocular fibroblasts derived from patients with severe Graves' ophthalmopathy (GO) and normal individuals in our studies. Sodium dodecylsulfate polyacrylamide-gel electrophoresis of immunoprecipitated cell lysates, derived from surface-radioiodinated GO cell monolayers, resulted in a single band of appropriate molecular weight on the autoradiogram. Further, a bright cell surface staining pattern was observed when indirect immunofluorescence was performed using the same anti-HSP 72 monoclonal antibody on parallel cell cultures. No cell-surface HSP 72 reactivity was detected in normal retroocular fibroblast monolayers by either method. These results are the first demonstration, by immunoprecipitation of surface- labeled proteins, of HSP expression on the surface of cells. This localization of HSP 72 on the surface of affected cells obtained from patients with an autoimmune disease may have implications concerning the role of this molecule in autoimmunity in general, and particularly in the immune process of GO.

AB - Heat shock proteins (HSPs) have been implicated in autoimmune disease, although they are generally considered to be intracellular in location. We demonstrate that, under certain circumstances, the inducible intracellular 72 kilodalton HSP can also be detected on the surface of cells. We used cultured retroocular fibroblasts derived from patients with severe Graves' ophthalmopathy (GO) and normal individuals in our studies. Sodium dodecylsulfate polyacrylamide-gel electrophoresis of immunoprecipitated cell lysates, derived from surface-radioiodinated GO cell monolayers, resulted in a single band of appropriate molecular weight on the autoradiogram. Further, a bright cell surface staining pattern was observed when indirect immunofluorescence was performed using the same anti-HSP 72 monoclonal antibody on parallel cell cultures. No cell-surface HSP 72 reactivity was detected in normal retroocular fibroblast monolayers by either method. These results are the first demonstration, by immunoprecipitation of surface- labeled proteins, of HSP expression on the surface of cells. This localization of HSP 72 on the surface of affected cells obtained from patients with an autoimmune disease may have implications concerning the role of this molecule in autoimmunity in general, and particularly in the immune process of GO.

UR - http://www.scopus.com/inward/record.url?scp=0026535812&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0026535812&partnerID=8YFLogxK

U2 - 10.1210/jc.74.4.732

DO - 10.1210/jc.74.4.732

M3 - Article

C2 - 1548335

AN - SCOPUS:0026535812

VL - 74

SP - 732

EP - 736

JO - Journal of Clinical Endocrinology and Metabolism

JF - Journal of Clinical Endocrinology and Metabolism

SN - 0021-972X

IS - 4

ER -