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Protein microarray analysis as a tool for monitoring cellular autoreactivity in type 1 diabetes patients and their relatives
Author(s) -
Vrabelova Zuzana,
Kolouskova Stanislava,
Böhmova Kristyna,
Faresjö Maria Karlsson,
Sumnik Zdenek,
Pechova Marta,
Kverka Miloslav,
Chudoba Daniel,
Zacharovova Klara,
Stadlerova Gabriela,
Pithova Pavlina,
Hladikova Marie,
Stechova Katerina
Publication year - 2007
Publication title -
pediatric diabetes
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.678
H-Index - 75
eISSN - 1399-5448
pISSN - 1399-543X
DOI - 10.1111/j.1399-5448.2007.00308.x
Subject(s) - medicine , type 1 diabetes , diabetes mellitus , type 2 diabetes , microarray , microarray analysis techniques , computational biology , immunology , endocrinology , genetics , gene , gene expression , biology
Background: Autoreactive T cells have a crucial role in type 1 diabetes (T1D) pathogenesis. Objectives: The aim of our study was to monitor the in vitro production of cytokines by peripheral blood mononuclear cells (PBMCs) after stimulation with diabetogenic autoantigens. Subjects: Ten T1D patients (tested at the time of diagnosis and 6 and 12 months later), 10 first‐degree relatives of the T1D patients, and 10 controls underwent the study. Methods: PBMCs were stimulated with glutamic acid decarboxylase 65 (GAD65) amino acids (a.a.) 247–279, 509–528, and 524–543; proinsulin a.a. 9–23; and tyrosine phosphatase (islet antigen‐2)/R2 a.a. 853–872. Interleukin (IL)‐2, IL‐4, IL‐5, IL‐6, IL‐10, IL‐13, interferon (IFN)‐γ, tumor necrosis factor β, transforming growth factor β1, and granulocyte colony‐stimulating factor (GCSF) were analyzed by protein microarray. Results: Differences in cytokine(s) poststimulatory and mainly in basal production were observed in all groups. The most prominent findings were in controls, the higher basal levels of IL‐2, IL‐4, IL‐5, IL‐13, and GCSF were observed when compared with relatives (p < 0.05, for all). After stimulation in controls, there was a significant decrease in IL‐2, IL‐13, GCSF, and IFN‐γ (p < 0.05, for all). The group of relatives was the most variable in poststimulatory production. A strong correlation between cytokines production was found but groups differed in this aspect. Conclusion: By multiplex analysis, it may be possible, for example, to define the risk immunological response pattern among relatives or to monitor the immune response in patients on immune modulation therapy.