Anti-GAD65 reactive peripheral blood mononuclear cells in the pathogenesis of cystic fibrosis related diabetes mellitus.
OBJECTIVE
A role of autoreactive T cells for type 1 diabetes pathogenesis is considered crucial. In our pilot study we addressed if autoreactive mononuclear cells are present also in peripheral blood of patients with other specific forms of diabetes as cystic fibrosis related diabetes (CFRD).


METHODS
Cellular immune responses to a known beta-cell autoantigen (GAD65 and GAD65 derived peptides) were analysed by ELISPOT (IFN-gamma) and by protein microarray analysis in four patients suffering from CFRD, in four cystic fibrosis (CF) patients without diabetes, in eight type 1 diabetes patients (without CF) and in four healthy controls.


RESULTS
Response to the autoantigen GAD65 (protein and peptides) was observed in 7/8 patients suffering from CF and in all type 1 diabetes patients. Post-stimulation production of Th1 cytokines (IFN-gamma, TNF-beta) was observed in 2/4 CFRD, 1/4 CF patients and in 7/8 type 1 diabetes patients. All these patients carry prodiabetogenic HLA-DQ genotype. Th2- and Th3 type of cytokine pattern was observed in 2/4 CF patients. Production of IL-8 was observed in the third CFRD as well as in the third CF patient and in 1/8 type 1 diabetes patient and borderline production of this chemokine was also observed in 2/4 healthy controls. No reaction was observed in the other 2/4 healthy controls and in the fourth CFRD patient who carried a strongly protective genotype and did not produce autoantibodies. The most potent peptide of GAD65 was amino acids 509-528.


CONCLUSIONS
We consider our observations as a sign of a reaction directed against the self-antigen GAD65 that are closely connected to type 1 diabetes. In CF patients who do not develop diabetes autoreactive mechanisms are very probably efficiently suppressed by immune self-tolerance mechanisms. CFRD patients are a heterogenic group. To disclose those who may display features of autoimmune diabetes could have an impact for their therapy and prognosis.
Stechova
Katerina
K
Department of Paediatrics, 2nd Medical Faculty of Charles University and University Hospital Motol in Prague, V Uvalu 84, Prague 5, Motol, 15006, Czech Republic. katerinastechova@seznam.cz
Kolouskova
Stanislava
S
Sumnik
Zdenek
Z
Cinek
Ondrej
O
Kverka
Miloslav
M
Faresjo
Maria Karlsson
MK
Chudoba
Daniel
D
Dovolilova
Eva
E
Pechova
Marta
M
Vrabelova
Zuzana
Z
Böhmova
Kristyna
K
Janecek
Lukas
L
Saudek
Frantisek
F
Vavrinec
Jan
J
D000293:Adolescent; D000328:Adult; D001323:Autoantibodies / Q000097:blood / Q000276:immunology*; D002648:Child; D003550:Cystic Fibrosis / Q000150:complications* / Q000276:immunology; D003920:Diabetes Mellitus / Q000209:etiology / Q000276:immunology*; D005260:Female; D005968:Glutamate Decarboxylase / Q000276:immunology*; D006683:HLA-DQ Antigens / Q000276:immunology; D006801:Humans; D007371:Interferon-gamma / Q000097:blood; D016209:Interleukin-8 / Q000097:blood; D007527:Isoenzymes / Q000276:immunology*; D007963:Leukocytes, Mononuclear / Q000276:immunology*; D008233:Lymphotoxin-alpha / Q000097:blood; D008297:Male; D010865:Pilot Projects; D040081:Protein Array Analysis
D001323:Autoantibodies; D006683:HLA-DQ Antigens; D016209:Interleukin-8; D007527:Isoenzymes; D008233:Lymphotoxin-alpha; D007371:Interferon-gamma; D005968:Glutamate Decarboxylase; C401141:glutamate decarboxylase 2