Beta B1-crystallin

Identification of a candidate ciliary body uveitis antigen

David Stempel, Hallie Sandusky, Kirsten Lampi, Marianne Cilluffo, Joe Horwitz, Jonathan Braun, Lee Goodglick, Lynn K. Gordon

Research output: Contribution to journalArticle

17 Citations (Scopus)

Abstract

PURPOSE. Perineuclear anti-neutrophil cytoplasmic antibody (pANCA), a marker antibody present in 12% of patients with anterior uveitis, recognizes cytoplasmic antigens in the nonpigmented ciliary body epithelium, a probable site of immunologic reactivity in this inflammatory disease. In this study, a recombinantly isolated pANCA monoclonal antibody was used to identify the corresponding antigenic target(s) in the ciliary body. METHODS. Proteins from microdissected eye bank ocular ciliary body tissue were used to identify the corresponding ANCA antigen. Parallel two-dimensional protein gels were used for simultaneous identification of candidate antigenic protein spots by Western blot analysis and as a source of material for proteomic analysis. Multiple independent methods including Western blot analysis, confocal microscopy, and RT-PCR were used to provide additional characterization of the candidate protein. RESULTS. Proteomic analysis suggested that beta B1 (βB1)-crystallin is the primary ciliary body antigen. The presence of βB1-crystallin in the human ciliary body was confirmed by Western blot with a βB1 specific anti-peptide antibody. Confocal microscopy revealed colocalization of the antigenic reactivity of both anti-βB1 antibody and monoclonal pANCA. RT-PCR confirmed the presence of βB1-crystallin RNA in the ciliary body tissues. CONCLUSIONS. This study identified βB1-crystallin as a new cytoplasmic ciliary body antigenic target of a marker autoantibody associated with uveitis. This characterization of βB1-crystallin outside the lens raises questions about its extralenticular expression, intracellular role, and potential target of inflammation in uveitis.

Original languageEnglish (US)
Pages (from-to)203-209
Number of pages7
JournalInvestigative Ophthalmology and Visual Science
Volume44
Issue number1
DOIs
StatePublished - Jan 1 2003

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beta-Crystallins
Ciliary Body
Uveitis
Crystallins
Antineutrophil Cytoplasmic Antibodies
Antigens
Western Blotting
Confocal Microscopy
Proteomics
Anti-Idiotypic Antibodies
Proteins
Eye Banks
Anterior Uveitis
Polymerase Chain Reaction
Human Body
Autoantibodies
Lenses
Epithelium
Gels
Monoclonal Antibodies

ASJC Scopus subject areas

  • Ophthalmology

Cite this

Beta B1-crystallin : Identification of a candidate ciliary body uveitis antigen. / Stempel, David; Sandusky, Hallie; Lampi, Kirsten; Cilluffo, Marianne; Horwitz, Joe; Braun, Jonathan; Goodglick, Lee; Gordon, Lynn K.

In: Investigative Ophthalmology and Visual Science, Vol. 44, No. 1, 01.01.2003, p. 203-209.

Research output: Contribution to journalArticle

Stempel, D, Sandusky, H, Lampi, K, Cilluffo, M, Horwitz, J, Braun, J, Goodglick, L & Gordon, LK 2003, 'Beta B1-crystallin: Identification of a candidate ciliary body uveitis antigen', Investigative Ophthalmology and Visual Science, vol. 44, no. 1, pp. 203-209. https://doi.org/10.1167/iovs.01-1261
Stempel, David ; Sandusky, Hallie ; Lampi, Kirsten ; Cilluffo, Marianne ; Horwitz, Joe ; Braun, Jonathan ; Goodglick, Lee ; Gordon, Lynn K. / Beta B1-crystallin : Identification of a candidate ciliary body uveitis antigen. In: Investigative Ophthalmology and Visual Science. 2003 ; Vol. 44, No. 1. pp. 203-209.
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AU - Lampi, Kirsten

AU - Cilluffo, Marianne

AU - Horwitz, Joe

AU - Braun, Jonathan

AU - Goodglick, Lee

AU - Gordon, Lynn K.

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N2 - PURPOSE. Perineuclear anti-neutrophil cytoplasmic antibody (pANCA), a marker antibody present in 12% of patients with anterior uveitis, recognizes cytoplasmic antigens in the nonpigmented ciliary body epithelium, a probable site of immunologic reactivity in this inflammatory disease. In this study, a recombinantly isolated pANCA monoclonal antibody was used to identify the corresponding antigenic target(s) in the ciliary body. METHODS. Proteins from microdissected eye bank ocular ciliary body tissue were used to identify the corresponding ANCA antigen. Parallel two-dimensional protein gels were used for simultaneous identification of candidate antigenic protein spots by Western blot analysis and as a source of material for proteomic analysis. Multiple independent methods including Western blot analysis, confocal microscopy, and RT-PCR were used to provide additional characterization of the candidate protein. RESULTS. Proteomic analysis suggested that beta B1 (βB1)-crystallin is the primary ciliary body antigen. The presence of βB1-crystallin in the human ciliary body was confirmed by Western blot with a βB1 specific anti-peptide antibody. Confocal microscopy revealed colocalization of the antigenic reactivity of both anti-βB1 antibody and monoclonal pANCA. RT-PCR confirmed the presence of βB1-crystallin RNA in the ciliary body tissues. CONCLUSIONS. This study identified βB1-crystallin as a new cytoplasmic ciliary body antigenic target of a marker autoantibody associated with uveitis. This characterization of βB1-crystallin outside the lens raises questions about its extralenticular expression, intracellular role, and potential target of inflammation in uveitis.

AB - PURPOSE. Perineuclear anti-neutrophil cytoplasmic antibody (pANCA), a marker antibody present in 12% of patients with anterior uveitis, recognizes cytoplasmic antigens in the nonpigmented ciliary body epithelium, a probable site of immunologic reactivity in this inflammatory disease. In this study, a recombinantly isolated pANCA monoclonal antibody was used to identify the corresponding antigenic target(s) in the ciliary body. METHODS. Proteins from microdissected eye bank ocular ciliary body tissue were used to identify the corresponding ANCA antigen. Parallel two-dimensional protein gels were used for simultaneous identification of candidate antigenic protein spots by Western blot analysis and as a source of material for proteomic analysis. Multiple independent methods including Western blot analysis, confocal microscopy, and RT-PCR were used to provide additional characterization of the candidate protein. RESULTS. Proteomic analysis suggested that beta B1 (βB1)-crystallin is the primary ciliary body antigen. The presence of βB1-crystallin in the human ciliary body was confirmed by Western blot with a βB1 specific anti-peptide antibody. Confocal microscopy revealed colocalization of the antigenic reactivity of both anti-βB1 antibody and monoclonal pANCA. RT-PCR confirmed the presence of βB1-crystallin RNA in the ciliary body tissues. CONCLUSIONS. This study identified βB1-crystallin as a new cytoplasmic ciliary body antigenic target of a marker autoantibody associated with uveitis. This characterization of βB1-crystallin outside the lens raises questions about its extralenticular expression, intracellular role, and potential target of inflammation in uveitis.

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