Protein microarray analysis as a tool for monitoring cellular autoreactivity in type 1 diabetes patients and their relatives
Article first published online: 10 SEP 2007
2007 Blackwell Munksgaard
Volume 8, Issue 5, pages 252–260, October 2007
How to Cite
Vrabelova, Z., Kolouskova, S., Böhmova, K., Faresjö, M. K., Sumnik, Z., Pechova, M., Kverka, M., Chudoba, D., Zacharovova, K., Stadlerova, G., Pithova, P., Hladikova, M. and Stechova, K. (2007), Protein microarray analysis as a tool for monitoring cellular autoreactivity in type 1 diabetes patients and their relatives. Pediatric Diabetes, 8: 252–260. doi: 10.1111/j.1399-5448.2007.00308.x
- Issue published online: 10 SEP 2007
- Article first published online: 10 SEP 2007
- Submitted 11 January 2007. Accepted for publication 16 July 2007
- autoreactive T cells;
- protein microarray;
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.