Enhanced induction of a 72 kDa heat shock protein in cultured retroocular fibroblasts

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

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

Fibroblasts in the retroocular connective tissue appear to play a central role in the pathogenesis of Graves' ophthalmopathy (GO). We hypothesize that specific attributes, differentiating normal from GO retroocular fibroblasts, may render the latter more vulnerable to the ongoing immunopathological process in GO. We investigated whether GO fibroblasts differ from normal fibroblasts with respect to their sensitivity and capacity to express the inducible 72 kDa heat shock protein (HSP) in response to stressful environmental stimuli. Cultured retroocular fibroblasts derived from patients with GO and normal individuals were exposed to various changes in the culture medium that may simulate conditions in the affected retroocular space in vivo. HSP 72 reactivity was determined using sodium dodecyl sulfate polyacrylamide-gel electrophoresis of cellular extracts, followed by immunoblotting with a mouse monoclonal anti-HSP 72 antibody and quantitative scanning densitometry. In addition, indirect immunofluorescence was performed on parallel fibroblast monolayers. Following exposure to heat and acidic pH, deprivation from nutrients, and high monolayer density, GO fibroblasts expressed HSP 72 with significantly greater sensitivity and in significantly higher abundance than did normal fibroblasts. These results demonstrate that changes in the physiological environment induce HSP 72 expression in cultured fibroblasts. The enhanced sensitivity and capacity of GO retroocular fibroblasts to express the inducible HSP 72 in response to stressful stimuli may play a role in the autoimmune process affecting the retroocular space in GO.

Original languageEnglish (US)
Pages (from-to)466-470
Number of pages5
JournalInvestigative Ophthalmology and Visual Science
Volume33
Issue number2
StatePublished - 1992

Fingerprint

HSP72 Heat-Shock Proteins
Graves Ophthalmopathy
Fibroblasts
Densitometry
Indirect Fluorescent Antibody Technique
Immunoblotting
Sodium Dodecyl Sulfate
Connective Tissue
Culture Media
Polyacrylamide Gel Electrophoresis
Hot Temperature

Keywords

  • autoimmunity
  • Graves' ophthalmopathy
  • heat shock protein
  • retroocular fibroblasts
  • stress response

ASJC Scopus subject areas

  • Ophthalmology

Cite this

Enhanced induction of a 72 kDa heat shock protein in cultured retroocular fibroblasts. / Heufelder, A. E.; Wenzel, B. E.; Bahn, R. S.

In: Investigative Ophthalmology and Visual Science, Vol. 33, No. 2, 1992, p. 466-470.

Research output: Contribution to journalArticle

Heufelder, A. E. ; Wenzel, B. E. ; Bahn, R. S. / Enhanced induction of a 72 kDa heat shock protein in cultured retroocular fibroblasts. In: Investigative Ophthalmology and Visual Science. 1992 ; Vol. 33, No. 2. pp. 466-470.
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