Optimization of a Method to Detect Autoantigen-Specific T-Cell Responses in Type 1 Diabetes.

Autor:	Musthaffa Y; Department of Endocrinology and Diabetes, Queensland Children's Hospital, South Brisbane, QLD, Australia.; The University of Queensland Diamantina Institute, The University of Queensland, Brisbane, QLD, Australia., Nel HJ; The University of Queensland Diamantina Institute, The University of Queensland, Brisbane, QLD, Australia., Ramnoruth N; The University of Queensland Diamantina Institute, The University of Queensland, Brisbane, QLD, Australia., Patel S; The University of Queensland Diamantina Institute, The University of Queensland, Brisbane, QLD, Australia., Hamilton-Williams EE; The University of Queensland Diamantina Institute, The University of Queensland, Brisbane, QLD, Australia., Harris M; Department of Endocrinology and Diabetes, Queensland Children's Hospital, South Brisbane, QLD, Australia.; The University of Queensland Diamantina Institute, The University of Queensland, Brisbane, QLD, Australia., Thomas R; The University of Queensland Diamantina Institute, The University of Queensland, Brisbane, QLD, Australia.
Jazyk:	angličtina
Zdroj:	Frontiers in immunology [Front Immunol] 2020 Dec 07; Vol. 11, pp. 587469. Date of Electronic Publication: 2020 Dec 07 (Print Publication: 2020).
DOI:	10.3389/fimmu.2020.587469
Abstrakt:	The development of tolerizing therapies aiming to inactivate autoreactive effector T-cells is a promising therapeutic approach to control undesired autoimmune responses in human diseases such as Type 1 Diabetes (T1D). A critical issue is a lack of sensitive and reproducible methods to analyze antigen-specific T-cell responses, despite various attempts. We refined a proliferation assay using the fluorescent dye 5,6-carboxylfluorescein diacetate succinimidyl ester (CFSE) to detect responding T-cells, highlighting the fundamental issues to be taken into consideration to monitor antigen-specific responses in patients with T1D. The critical elements that maximize detection of antigen-specific responses in T1D are reduction of blood storage time, standardization of gating parameters, titration of CFSE concentration, selecting the optimal CFSE staining duration and the duration of T-cell stimulation, and freezing in medium containing human serum. Optimization of these elements enables robust, reproducible application to longitudinal cohort studies or clinical trial samples in which antigen-specific T-cell responses are relevant, and adaptation to other autoimmune diseases. Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. (Copyright © 2020 Musthaffa, Nel, Ramnoruth, Patel, Hamilton-Williams, Harris and Thomas.)
Databáze:	MEDLINE
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