Up-regulated cytokine-inducible SH2-containing protein expression in allergen-stimulated T cells from hen's egg-allergic patients.
BACKGROUND
Although changes in the fine balance of allergen-specific T cells are crucial in the pathogenesis of allergic diseases, their roles in the allergic reaction to hen's eggs (HE) have not yet been fully analysed.


OBJECTIVE
Using microarray technology, allergen-stimulated T cells from HE-allergic children were analysed to identify genes that are specifically up-regulated in these cells.


METHODS
RNA from CD4(+) CD14(-) cells, fractionated from allergen-stimulated peripheral mononuclear cells, was analysed using a whole-genome microarray and real-time RT-PCR. The protein expression of selected genes was ascertained by flow cytometry.


RESULTS
In microarray analyses of allergen-stimulated T cells, 43 genes were up-regulated in HE-allergic children but not in non-HE-allergic children. Among these, up-regulation of three genes, cytokine -inducible SH2-containing protein (CISH), nuclear factor of kappa light polypeptide gene enhancer in B-cell inhibitor Z (NFKBIZ) and B-cell CLL/lymphoma 2 (BCL2), was confirmed by real-time quantitative RT-PCR. CISH, but not NFKBIZ or BCL2, showed a significantly higher ratio of antigen-stimulated cell transcription over unstimulated cells in HE-allergic than in non-HE-allergic children (P<0.01). Flow-cytometric analysis revealed that the percentage of CD25(+)CISH(+) cells in CD4(+) cells from patients with HE allergy was significantly higher than that in controls (P<0.01). The expression level of CISH was significantly higher in IL-4(+) Th2 cells than in IFN-gamma(+) Th1 cells.


CONCLUSIONS
We noted that CISH expression in allergen-stimulated CD4(+) T cells from HE-allergic patients was significantly increased in both mRNA and protein levels compared with that from non-HE-allergic children.
Nakajima
Y
Y
Department of Pediatrics, School of Medicine, Fujita Health University, Aichi-Ken, Japan. ynakaji@fujita-hu.ac.jp
Tsuge
I
I
Kondo
Y
Y
Komatsubara
R
R
Hirata
N
N
Kakami
M
M
Kato
M
M
Kurahashi
H
H
Urisu
A
A
Asano
Y
Y
D048868:Adaptor Proteins, Signal Transducing; D000485:Allergens / Q000276:immunology*; D000818:Animals; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology / Q000378:metabolism*; D002645:Chickens; D002675:Child, Preschool; D021181:Egg Hypersensitivity / Q000276:immunology / Q000378:metabolism*; D005260:Female; D006801:Humans; D020672:I-kappa B Proteins; D007223:Infant; D008297:Male; D009687:Nuclear Proteins / Q000378:metabolism; D020411:Oligonucleotide Array Sequence Analysis; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D050826:Suppressor of Cytokine Signaling Proteins / Q000378:metabolism*; D015854:Up-Regulation; D018909:src Homology Domains
D048868:Adaptor Proteins, Signal Transducing; D000485:Allergens; D020672:I-kappa B Proteins; D009687:Nuclear Proteins; D019253:Proto-Oncogene Proteins c-bcl-2; D012333:RNA, Messenger; D050826:Suppressor of Cytokine Signaling Proteins; C417234:NFKBIZ protein, human