HLA-B27 in Transgenic Rats Forms Disulfide-Linked Heavy Chain Oligomers and Multimers That Bind to the Chaperone BiP

Tri Minh Tran, Nimman Satumtira, Martha L. Dorris, Ekkehard May, Andrew Wang, Eiichi Furuta, Joel D. Taurog

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Abstract

To test the hypothesis that HLA-B27 predisposes to disease by forming disulfide-linked homodimers, we examined rats transgenic for HLA-B27, mutant Cys67Ser HLA-B27, or HLA-B7. In splenic Con A blasts from high transgene copy B27 lines that develop inflammatory disease, the anti-H chain mAb HC10 precipitated four bands of molecular mass 78-105 kDa and additional higher molecular mass material, seen by nonreducing SDS-PAGE. Upon reduction, all except one 78-kDa band resolved to 44 kDa, the size of the H chain monomer. The 78-kDa band was found to be BiP/Grp78, and the other high molecular mass material was identified as B27 H chain. Analysis of a disease-resistant low copy B27 line showed qualitatively similar high molecular mass bands that were less abundant relative to H chain monomer. Disease-prone rats with a Cys 67Ser B27 mutant showed B27 H chain bands at 95 and 115 kDa and a BiP band at 78 kDa, whereas only scant high molecular mass bands were found in cells from control HLA-B7 rats. 125I-surface labeled B27 oligomers were immunoprecipitated with HC10, but not with a mAb to folded B27-β 2-microglobulin-peptide complexes. Immunoprecipitation of BiP with anti-BiP Abs coprecipitated B27 H chain multimers. Folding and maturation of B27 were slow compared with B7. These data indicate that disulfide-linked intracellular H chain complexes are more prone to form and bind BiP in disease-prone wild-type B27 and B27-C67S rats than in disease-resistant HLA-B7 rats. The data support the hypothesis that accumulation of misfolded B27 participates in the pathogenesis of B27-associated disease.

Original languageEnglish (US)
Pages (from-to)5110-5119
Number of pages10
JournalJournal of Immunology
Volume172
Issue number8
StatePublished - Apr 15 2004

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Transgenic Rats
HLA-B27 Antigen
Disulfides
HLA-B7 Antigen
Transgenes
Immunoprecipitation
Polyacrylamide Gel Electrophoresis
Peptides

ASJC Scopus subject areas

  • Immunology

Cite this

Tran, T. M., Satumtira, N., Dorris, M. L., May, E., Wang, A., Furuta, E., & Taurog, J. D. (2004). HLA-B27 in Transgenic Rats Forms Disulfide-Linked Heavy Chain Oligomers and Multimers That Bind to the Chaperone BiP. Journal of Immunology, 172(8), 5110-5119.

HLA-B27 in Transgenic Rats Forms Disulfide-Linked Heavy Chain Oligomers and Multimers That Bind to the Chaperone BiP. / Tran, Tri Minh; Satumtira, Nimman; Dorris, Martha L.; May, Ekkehard; Wang, Andrew; Furuta, Eiichi; Taurog, Joel D.

In: Journal of Immunology, Vol. 172, No. 8, 15.04.2004, p. 5110-5119.

Research output: Contribution to journalArticle

Tran, TM, Satumtira, N, Dorris, ML, May, E, Wang, A, Furuta, E & Taurog, JD 2004, 'HLA-B27 in Transgenic Rats Forms Disulfide-Linked Heavy Chain Oligomers and Multimers That Bind to the Chaperone BiP', Journal of Immunology, vol. 172, no. 8, pp. 5110-5119.
Tran TM, Satumtira N, Dorris ML, May E, Wang A, Furuta E et al. HLA-B27 in Transgenic Rats Forms Disulfide-Linked Heavy Chain Oligomers and Multimers That Bind to the Chaperone BiP. Journal of Immunology. 2004 Apr 15;172(8):5110-5119.
Tran, Tri Minh ; Satumtira, Nimman ; Dorris, Martha L. ; May, Ekkehard ; Wang, Andrew ; Furuta, Eiichi ; Taurog, Joel D. / HLA-B27 in Transgenic Rats Forms Disulfide-Linked Heavy Chain Oligomers and Multimers That Bind to the Chaperone BiP. In: Journal of Immunology. 2004 ; Vol. 172, No. 8. pp. 5110-5119.
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AU - Taurog, Joel D.

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