Comparison of anti-heat shock protein 70 (anti-hsp70) and anti-68-kDa inner ear protein in the sera of patients with Meniere's disease.
The 68-kDa antigen detected in the sera of patients with autoimmune inner ear disease is known to represent the highly inducible heat shock protein 70 (hsp70). To evaluate the existence of anti-hsp70 in the sera of patients with Meniere's disease and to develop a more reliable method to detect this antibody, the sera of patients and controls were examined. Bovine kidney (MDBK) cells were cultured and some of them were heat shocked. Proteins in the cells were separated using sodium dodecyl sulfate polyacrylamide gel electrophoresis. Sera were reacted simultaneously with the blots of non-heat-shocked cells and heat-shocked cells. The serum was considered positive if the band in the 70-kDa location was denser in the lane with heat-shocked cells relative to non-heat-shocked cells. Presence of the antibody against the 68-kDa protein was compared with the result of immunoblotting with MDBK cells. In immunoblotting with MDBK cells, 33.3% of patients with Meniere's disease had anti-hsp70, while in the control group, only 5% had this antibody. Of the 60 cases, 13 were positive against both hsp70 and the 68-kDa protein, whereas 7 were positive only against hsp70 and 6 only against the 68-kDa protein. These differences appeared to result from the greater sensitivity of the differential anti-hsp assay and from difficulties in interpreting the results in blots with bovine inner ear extracts because of faint, broad, or overlapping multiple bands. Quite a number of patients with Meniere's disease have anti-hsp70, and it may be indicative of an immune etiology of the disease. The Western blot using heat-shocked and non-heat-shocked cells could be a reliable method to detect this antibody.
Shin
S O
SO
Division of Otolaryngology--Head and Neck Surgery, University of California at San Diego, La Jolla 92093-0970, USA.
Billings
P B
PB
Keithley
E M
EM
Harris
J P
JP
D000328:Adult; D000368:Aged; D000818:Animals; D000906:Antibodies / Q000097:blood; D015153:Blotting, Western; D002417:Cattle; D002460:Cell Line; D007758:Ear, Inner / Q000276:immunology*; D005260:Female; D018840:HSP70 Heat-Shock Proteins / Q000276:immunology* / Q000378:metabolism; D006801:Humans; D015151:Immunoblotting; D007122:Immunoelectrophoresis; D007668:Kidney / Q000276:immunology; D008297:Male; D008575:Meniere Disease / Q000276:immunology* / Q000378:metabolism; D008875:Middle Aged
D000906:Antibodies; D018840:HSP70 Heat-Shock Proteins; C490786:HSPA1A protein, human