Transcription profiling of intraepithelial and peripheral CTL from patients with celiac disease or cytomegalovirus seropositive patients suggest that a stepwise transformation of CTLs into NK-like cells may underlie immunopathology in various chronic infectious and inflammatory diseases.

ABSTRACT: Celiac disease is an intestinal inflammatory disorder induced by dietary gluten in genetically susceptible individuals. The mechanisms underlying the massive expansion of interferon g–producing intraepithelial cytotoxic T lymphocytes (CTLs) and the destruction of the epithelial cells lining the small intestine of celiac patients have remained elusive. We report massive oligoclonal expansions of intraepithelial CTLs that exhibit a profound genetic reprogramming of natural killer (NK) functions. These CTLs aberrantly expressed cytolytic NK lineage receptors, such as NKG2C, NKp44, and NKp46, which associate with adaptor molecules bearing immunoreceptor tyrosine-based activation motifs and induce ZAP-70 phosphorylation, cytokine secretion, and proliferation independently of T cell receptor signaling. This NK transformation of CTLs may underlie both the self-perpetuating, gluten-independent tissue damage and the uncontrolled CTL expansion leading to malignant lymphomas in severe forms of celiac disease. Because similar changes were detected in a subset of CTLs from cytomegalovirus-seropositive patients, we suggest that a stepwise transformation of CTLs into NK-like cells may underlie immunopathology in various chronic infectious and inflammatory diseases. Experiment Overall Design: Using a standard GeneChip preparation protocol (Affymetrix, Santa Clara CA), gene array hybridizations and data collection on 5 paired NKG2C+ and NKG2C- IE and PB –CTL samples was preformed at the Functional Genomics Facility, University of Chicago. Briefly, 10ug of total RNA, extracted using the RNeasy Mini kit (Qiagen, Valencia, CA), was used to generate double-stranded cDNA using a T7 linked oligo dT primer and the Superscript II RT system (Invitrogen Carlsbad CA). Purified cDNA was then used to generate biotin-labeled cRNA using the BioArray High Yield RNA Transcript Labeling Kit (Enzo Diagnostics, Farmingdale, NY) according to the manufacturers protocol. This biotinylated-cRNA was then fragmented and hybridized to an Affymetrix GeneChip HG-U133. The arrays were then washed and stained with phycoerythrin-conjugated streptavidin according to the Affymetrix GeneChip protocol and scanned using the Affymetrix Agilent GeneArray Scanner (Affymetrix, Santa Clara CA). Data analyses were preformed using DNA-Chip Analyzer 1.3 (dChip) with the *.CEL files obtained from GeneChip Operating System 1.3 (GCOS 1.3). A PM-only model was used to estimate gene expression level and the invariant set approach was used for normalization. For comparison analyses of the paired NKG2C+ and NKG2C- populations, thresholds for selecting significant genes were set at a relative difference > 1.5 fold, absolute difference > 100 and a significant paired t-test for the signal intensity difference at p < 0.05. Duplicate genes and genes of unknown function were removed. The remaining genes were then grouped based on function and cluster analysis was preformed using dChip.

ORGANISM(S): Homo sapiens

DISEASE(S): cytomegalovirus seropositive

SUBMITTER: Bana Jabri 

PROVIDER: E-GEOD-4592 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress