A sensitive method for detecting proliferation of rare autoantigen-specific human T cells

Mannering, SI; Morris, JS; Jensen, KP; Purcell, AW; Honeyman, MC; Van Endert, PM; Harrison, LC

Author(s): Mannering, SI; Morris, JS; Jensen, KP; Purcell, AW; Honeyman, MC; Van Endert, PM; Harrison, LC;
Details: Publication Year 2003-12,Volume 283,Issue #1-2,Page 173-183
Journal Title: JOURNAL OF IMMUNOLOGICAL METHODS
Publication Type: Journal Article
Abstract: The ability to measure proliferation of rare antigen-specific T cells among many bystanders is critical for the evaluation of cellular immune function in health and disease. T-cell proliferation in response to antigen has been measured almost exclusively by H-3-thymidine incorporation. This method does not directly identify the phenotype of the proliferating cells and is frequently not sufficiently sensitive to detect rare autoantigen-specific T cells. To overcome these problems, we developed a novel assay for antigen-specific human T-cell proliferation. Peripheral blood mononuclear cells (PBMC) were labelled with the fluorescent dye 5,6-carboxylfluorescein diacetate succinimidyl ester (CFSE) and cells that proliferated in response to antigen, with resultant reduction in CFSE intensity, were measured directly by flow cytometry. This assay was more sensitive than 3H-thymidine incorporation and detected the proliferation of rare antigen-specific CD4(+) T cells at 10-fold lower antigen concentrations. It also allowed the phenotype of the proliferating cells to be directly determined. Using the CFSE assay we were able to measure directly the proliferation of human CD4(+) T cells from healthy donors in response to the type I diabetes autoantigens glutamic acid decarboxylase (GAD) and proinsulin (PI). (C) 2003 Elsevier B.V. All rights reserved.
Publisher: ELSEVIER SCIENCE BV
Keywords: GLUTAMIC-ACID DECARBOXYLASE; FLOW-CYTOMETRY; APOPTOSIS; RESPONSES; INDIVIDUALS; PROINSULIN; TETRAMERS; WORKSHOP; CULTURE; ASSAYS
Publisher's Version: https://doi.org/10.1016/j.jim.2003.09.004
Terms of Use/Rights Notice: Refer to copyright notice on published article.

Creation Date: 2003-12-01 12:00:00