Use this URL to cite or link to this record in EThOS: http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.553905
Title: Identification of human hair follicle antigens targeted in the presumptive autoimmune hair follicle disorder alopecia areata and their potential functional relevance in vitro : methods development for isolation and identification of alopecia areata-relevant human hair follicle antigens using a proteomics approach and their functional assessment using an ex vivo hair follicle organ culture model
Author: Leung, Man Ching
Awarding Body: University of Bradford
Current Institution: University of Bradford
Date of Award: 2008
Availability of Full Text:
Access from EThOS:
Access from Institution:
Abstract:
Alopecia areata (AA) is a putative autoimmune hair loss disorder. It mainly affects the scalp hair but can also involve body hair, and can also affect the nail and the eye. While there are may be several lines of evidence to support the autoimmune basis of AA, there is still very little information on the hair follicle autoantigen(s) involved in its pathogenesis. In this project, serum antibodies (AA=10, control=10) were used to immunoprecipitate AA-relevant target antigens from normal human scalp hair follicle extracts. These immunoprecipitates were analysed by LC-MALDI-TOF/TOF mass spectrometry for target protein identification. This part of the project involved substantial methods development. Trichohyalin was immunoprecipitated by all AA sera, but by only 5 normal sera. Importantly, the mean Mascot scores of the AA group was significantly higher than the normal group (p=0.005). Keratin 16 was also identified from immunoprecipitates as another potential AA-relevant target antigen. Functional studies by ex vivo whole hair follicle organ culture using commercial antibodies to trichohyalin and keratin 16 significantly inhibited hair fibre elongation compared to controls. Indirect immunofluorescence studies revealed that AA sera contained higher immunoreactivity against normal human scalp anagen hair follicles compared to normal sera. Immunoreactivities were mainly in the outer root sheath and inner root sheath, and less so to the medulla and hair bulb matrix. Double immunofluorescence studies of AA and normal serum with anti-trichohyalin antibody (AE15) revealed co-localisation of 9 of the AA sera antibodies with trichohyalin in the inner root sheath (mostly in Henle's, less in Huxley's/inner root sheath cuticle), but only weakly in 3 normal sera. This study supports the involvement of an antibody response to anagen-specific hair follicles antigens in AA. Moreover, there may be some evidence that these antibodies may have a pathogenic role.
Supervisor: Tobin, Desmond J. Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID: uk.bl.ethos.553905  DOI: Not available
Keywords: Alopecia areata ; Hair follicle ; Trichohyalin ; Autoantigens ; Proteomics ; Tissue culture ; Immunoprecipitation ; 2D gel electrophoresis ; LC-MALDI-TOF/TOF Mass Spectrometry ; Immunohistochemistry
Share: