title: Islet-cell antigen-reactive T cells show different expansion rates and Th1/Th2 differentiation in type 1 diabetic patients and healthy controls.
abstract: The low frequency of islet-cell antigen-reactive T cells in type 1 diabetes makes their direct measurement difficult. Commonly used in vitro expansion could alter in vivo frequencies and Th1/Th2 differentiation states. Using IFN-gamma/IL-4 double color ELISPOT, we tested longitudinally the reactivity of PBMC from HLA-matched diabetic patients and healthy controls to GAD65, IA-2, and proinsulin peptides ex vivo and after in vitro culture. The peptide-reactive T cells showed IFN-gamma bias in the patients' PBMC in the primary assay. During in vitro culture, both IFN-gamma- and IL-4-producing cells were induced in controls, suggesting that the precursor cells were uncommitted naive T cells in vivo. In contrast, in diabetic patients, the ex vivo IFN-gamma response was conserved during culture, suggesting their Th1 commitment. Using CFSE-dye-dilution, we demonstrate that naive T cells expand in vitro at a faster rate than memory cells, which might account for the differences in expansion rates between diabetic patients and controls.
authors.affiliation: Department of Pathology, Case Western Reserve University, School of Medicine, Cleveland, OH 44106, USA.
authors.forename: Patrick A
authors.initials: PA
authors.lastname: Ott
authors.forename: Bernhard A
authors.initials: BA
authors.lastname: Herzog
authors.forename: Stefan
authors.initials: S
authors.lastname: Quast
authors.forename: Harald H
authors.initials: HH
authors.lastname: Hofstetter
authors.forename: Bernhard O
authors.initials: BO
authors.lastname: Boehm
authors.forename: Magdalena
authors.initials: M
authors.lastname: Tary-Lehmann
authors.forename: Ivana
authors.initials: I
authors.lastname: Durinovic-Bello
authors.forename: Beate R
authors.initials: BR
authors.lastname: Berner
authors.forename: Paul V
authors.initials: PV
authors.lastname: Lehmann
mesh_terms: D000328:Adult; D000595:Amino Acid Sequence; D001324:Autoantigens / Q000276:immunology; D002454:Cell Differentiation / Q000276:immunology; D003922:Diabetes Mellitus, Type 1 / Q000276:immunology*; D005434:Flow Cytometry; D005968:Glutamate Decarboxylase / Q000276:immunology; D006801:Humans; D007156:Immunologic Memory / Q000276:immunology; D007371:Interferon-gamma / Q000276:immunology; D015847:Interleukin-4 / Q000276:immunology; D007515:Islets of Langerhans / Q000276:immunology*; D007527:Isoenzymes / Q000276:immunology; D008213:Lymphocyte Activation; D008565:Membrane Proteins / Q000276:immunology; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000276:immunology; D011384:Proinsulin / Q000276:immunology; D054562:Protein Tyrosine Phosphatase, Non-Receptor Type 1; D017027:Protein Tyrosine Phosphatases / Q000276:immunology; D054635:Receptor-Like Protein Tyrosine Phosphatases, Class 8; D018417:Th1 Cells / Q000166:cytology / Q000276:immunology*; D018418:Th2 Cells / Q000166:cytology / Q000276:immunology*
chemical_list: D001324:Autoantigens; D007527:Isoenzymes; D008565:Membrane Proteins; D010446:Peptide Fragments; D015847:Interleukin-4; D007371:Interferon-gamma; D011384:Proinsulin; C516862:PTPRN protein, human; D054562:Protein Tyrosine Phosphatase, Non-Receptor Type 1; D017027:Protein Tyrosine Phosphatases; D054635:Receptor-Like Protein Tyrosine Phosphatases, Class 8; D005968:Glutamate Decarboxylase; C401141:glutamate decarboxylase 2