[Animal model of experimental autoimmune inner ear disease induced by P0 protein purified from inner ear].
OBJECTIVE
To purify P0 protein from guinea pig's inner ear by preparative SDS-PAGE and to study the possible role it may play in the etiology of autoimmune inner ear disease.


METHODS
A mixture of membraneous proteins of inner ear was separated by preparative SDS-PAGE. The corresponding band at 30,000 was cut and electrically eluted. The protein collected was identified by analytical SDS-PAGE and Western blot assay. A group of 20 guinea pigs were immunized with P0 protein emulsified in complete Freunds adjuvant, another 10 guinea pigs were immunized with complete Freunds adjuvant only as control. The guinea pigs' hearing thresholds, serum IgG level and morphological changes in the inner ear were investigated. The distribution of P0 protein in the cochlear was detected by immunohistochemical technique.


RESULTS
The purity of the protein was demonstrated by a single band at the 30000 site in SDS-PAGE, which was identified as P0 protein by western blot analysis assay. About 17.5% P0-immunized guinea pigs showed increased hearing thresholds, elevated IgG level (F = 6.48, P<0.01), as well as a decreased number of spiral ganglion cells and inflammatory cell infiltration in the cochlear nerve region. The P0 protein is distributed in the cochlear nerve and spiral ganglion only.


CONCLUSIONS
P0 protein from guinea pigs in ner ear can be successfully purified by preparative SDS-PAGE and an animal model of experimental autoimmune inner ear disease induced by P0 protein was successfully established.
Lin
Xi
X
Department of Otorhinolaryngology, the First Affiliated Hospital, Fujian Medical University, Fuzhou, 350005, China.
Cheng
Qing
Q
Wang
Jibao
J
Huang
Xiang
X
Zhang
Rong
R
D000818:Animals; D001327:Autoimmune Diseases / Q000276:immunology*; D004195:Disease Models, Animal; D006168:Guinea Pigs; D007759:Labyrinth Diseases / Q000276:immunology*; D018993:Myelin P0 Protein / Q000276:immunology* / Q000302:isolation & purification
D018993:Myelin P0 Protein