Characterization of a human T cell line reactive to a 52 kDa islet protein

Douglas O. Sobel, Thomas Fleisher, Dennis G. Karounos

Research output: Contribution to journalArticlepeer-review

2 Scopus citations

Abstract

A 52 kDa islet protein has recently been identified as the target of autoantibodies in the NOD mouse model of IDDM and humans with IDDM. However, the presence of T cell immunity against the 52 kDa islet protein in IDDM has not been reported. We report the establishment and characterization of a T cell line (19KW) that reacts to purified 52 kDa islet protein (purified p52) from a subject with IDDM. The purified p52 induced a proliferative response as measured by thymidine incorporation in the 19KW T cell line with a stimulating index of up to 48. The proliferative responses were greater with increasing doses of purified p52 (0.1, 0.5, 2.0, and 6.0 μg/well). No reactivity was found to a liver fraction purified in the same manner as 52 kDa protein, BSA, ovalbumin extracts of rat muscle, fibroblast, adrenal, or pituitary tissue and to a rat exocrine cell tumor. Irradiated PBMC were required as antigen presenting cells (APC) for 19KW reactivity to the purified p52. The addition of anti-HLA DR or anti-HLA DQ antibodies significantly decreased the islet antigen-induced proliferative response. The addition of antibodies to HLA DP and class I MHC had no effect. Flow cytometric analysis revealed that the majority of T cells expressed CD4 and CD45RO molecules. T cell receptors Vβ6 and Vβ5.1 were found on 30 and 14% of the CD3+ (T cells) 19KW cells, respectively. In conclusion, a purified p52-reactive human T cell line predominantly consisting of TCR Vβ6+ and Vβ5.1+ cells has been established from a subject with IDDM. Reactivity to the purified p52 is antigen dose-dependent, tissue specific, requires irradiated PBMC as antigen presenting cells, and is HLA DR- and HLA DQ- restricted T cell lines specifically reactive to p52 may be useful for investigating further the role of this antigen in the pathogenesis of IDDM.

Original languageEnglish
Pages (from-to)387-394
Number of pages8
JournalJournal of Autoimmunity
Volume10
Issue number4
DOIs
StatePublished - Aug 1997

Bibliographical note

Funding Information:
This work was supported in part by the Diabetes Research and Education Program, Georgetown University, Veterans Affairs Merit Review WMU #92-00IV, and the Greenwall Foundation. We thank Mr Yang Xu, Behroux Ahvazi, Carter Hackney and S. L. Wilson for their excellent technical assistance and Mrs Barbara Runner for her secretarial support.

Copyright:
Copyright 2017 Elsevier B.V., All rights reserved.

Keywords

  • 52 kDa protein
  • Human
  • IDDM
  • Islet
  • T cell line

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology

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