RESUMO
Members of the suppressor of cytokine signaling (SOCS) family are involved in the pathogenesis of many inflammatory diseases. SOCS-3 is predominantly expressed in T-helper type 2 (T(H)2) cells, but its role in T(H)2-related allergic diseases remains to be investigated. In this study we provide a strong correlation between SOCS-3 expression and the pathology of asthma and atopic dermatitis, as well as serum IgE levels in allergic human patients. SOCS-3 transgenic mice showed increased T(H)2 responses and multiple pathological features characteristic of asthma in an airway hypersensitivity model system. In contrast, dominant-negative mutant SOCS-3 transgenic mice, as well as mice with a heterozygous deletion of Socs3, had decreased T(H)2 development. These data indicate that SOCS-3 has an important role in regulating the onset and maintenance of T(H)2-mediated allergic immune disease, and suggest that SOCS-3 may be a new therapeutic target for the development of antiallergic drugs.
Assuntos
Asma/imunologia , Dermatite Atópica/imunologia , Hipersensibilidade/imunologia , Proteínas/metabolismo , Proteínas Repressoras , Células Th2/imunologia , Fatores de Transcrição , Animais , Hiper-Reatividade Brônquica/imunologia , Testes de Provocação Brônquica , Proteínas de Ligação a DNA/metabolismo , Humanos , Hipersensibilidade/metabolismo , Interleucina-12/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Proteínas/genética , Fator de Transcrição STAT4 , Transdução de Sinais/fisiologia , Proteína 3 Supressora da Sinalização de Citocinas , Proteínas Supressoras da Sinalização de Citocina , Células Th2/fisiologia , Transativadores/metabolismoRESUMO
The ability of glucocorticoids (GCs) to kill lymphoid cells via a process called apoptosis has led to their inclusion in essentially all chemotherapy protocols for lymphoid malignancies. Since GC receptor(GR) is a ligand-dependent transcription factor, there should be genes mediating apoptosis among the ones whose expression is induced by GC. This review summarizes recent advances related to the molecular basis of GC-induced apoptosis, focusing on microarray analysis. Various groups of genes have been identified as candidate target genes of GR including the ones encoding Bcl-2 family proteins and calcineurin inhibitors. Although further investigation is required to determine the genes causally involved in GC-induced apoptosis, the studies described here will hopefully lead to more efficient treatments of lymphoid malignancies.
Assuntos
Antineoplásicos/farmacologia , Glucocorticoides/farmacologia , Apoptose/efeitos dos fármacos , Humanos , Análise em Microsséries , Receptores de Glucocorticoides/genéticaRESUMO
We have analyzed transcription profiles in peripheral blood CD3(+) cells from patients with allergic diseases to better understand the genes that are involved. Transcription levels of the gene KIAA0603/AS160 in CD3(+) cells from patients with atopic dermatitis (AD) were significantly higher than in normal individuals. The KIAA0603 gene encodes a 1299 amino acid protein with two phosphotyrosine interaction domains at the N-terminal region and a TBC domain at the C-terminal region. The region containing the TBC domain has a 31% homology to human rab6 GTPase activating protein (GAP). When human primary CD3(+) cells were stimulated with anti-CD3 or calcium ionophore, the KIAA0603 transcript level was upregulated. The marked upregulation of KIAA0607 was accompanied by activation induced cell death of primary CD3+ cells. KIAA0603 is likely to be a Rab GAP that participates in the regulation of activated T cells, especially helper memory T cells. Expression of KIAA0603 in T cells may be involved in pathogenesis of AD.
Assuntos
Dermatite Atópica/metabolismo , Proteínas Ativadoras de GTPase/genética , Regulação da Expressão Gênica/genética , Linfócitos T/metabolismo , Transcrição Gênica/genética , Proteínas rab de Ligação ao GTP/genética , Sequência de Bases , Primers do DNA , Dermatite Atópica/imunologia , Proteínas Ativadoras de GTPase/metabolismo , Humanos , Reação em Cadeia da Polimerase/métodos , Valores de Referência , Subpopulações de Linfócitos T/imunologia , Linfócitos T/imunologia , Proteínas rab de Ligação ao GTP/metabolismoRESUMO
The analysis of genes preferentially expressed in the peripheral blood cells of atopic dermatitis patients may provide information on the molecular pathogenesis of the disease. We employed differential display PCR to clone a new gene (AB100163) with 99% homology to coiled-coil domain containing 132, transcript variant 1 (CCDC132) (NM_017667) (aliases, FLJ20097, FLJ23581, KIAA1861 and MGC176659). Full-length CCDC132 of approximately 4 kbp encodes mRNA expressed in almost all tissues, in particular brain tissue and skeletal muscle. A homologous gene has also been identified in mice. Using Western blot analysis, 111 kDa CCDC132 protein was detected in two human T-cell lines, MOLT-4 and Jurkat, and in the human cervical adenocarcinoma cell line HeLa. Quantitative RT-PCR revealed transcription levels of CCDC132 in the T cells of atopic dermatitis patients to be higher than in those of normal individuals. This suggests that changes in CCDC132 expression may be involved in the course of atopic dermatitis.
RESUMO
To determine the genes responsible for mediating the effects of glucocorticoids (GCs) on leukemic cells, transcriptional changes in GC-sensitive human pre-B leukemia 697 cells during GC-induced apoptosis were monitored using oligonucleotide microarrays. To circumvent the challenge of recovering mRNAs from dying cells, we compared the pattern of gene expression with that of 697 cells protected from apoptosis by transfection with bcl-2. Of the 12,000 genes examined for their response to GC, 93 genes were induced and 28 genes were repressed, many of which are known to be implicated in signal transduction, growth arrest, and transcription. These included the signal transduction-related genes encoding SOCS1, SOCS2, FKBP51, DSCR1, p56lck, and four protein kinase phosphatases. Growth arrest-related genes encoding p19(INK4d) and several Myc inhibitors were induced in response to the GC treatment. Anti-proliferative- or apoptosis-related genes encoding BTG1, BTG2, and granzyme A were also found to be transcriptionally up-regulated by GC. In addition, the regulation of genes encoding the glucocorticoid receptor and steroid receptor coactivator-1 suggested autoregulation of a GC-mediated signaling pathway.
Assuntos
Apoptose , Expressão Gênica , Glucocorticoides/metabolismo , Antineoplásicos Hormonais/farmacologia , Western Blotting , Sobrevivência Celular , DNA/metabolismo , Dexametasona/farmacologia , Regulação para Baixo , Citometria de Fluxo , Humanos , Neomicina/farmacologia , Análise de Sequência com Séries de Oligonucleotídeos/métodos , Inibidores da Síntese de Proteínas/farmacologia , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , RNA/metabolismo , RNA Mensageiro/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Transdução de Sinais , Fatores de Tempo , Células Tumorais Cultivadas , Regulação para CimaRESUMO
Analysis of patients with atopic dermatitis (AD) for differential expression of genes, as compared to normal individuals, will be useful for understanding the molecular pathogenesis of AD. We found that the expression of the gene ETEA in human peripheral blood CD3-positive cells from patients with atopic dermatitis was significantly higher than in normal individuals. Eosinophils from AD patients expressed ETEA at a significantly higher level than the healthy controls. The overall sequence of the 445 aa deduced polypeptide from the cloned ETEA cDNA showed homology to human Fas-associated factor 1 (FAF1), which is involved in Fas-mediated apoptosis. However, the interaction of ETEA with the Fas death domain was weaker than that of FAF1, as studied in yeast two-hybrid experiments. The ETEA-EGFP fusion protein was expressed in cytoplasm. During the course of activation-induced cell death of primary T cells, transcription levels of ETEA and FAF1 were upregulated with similar kinetics. The enhanced expression of ETEA may play a role in the regulating the resistance to apoptosis that is observed in T cells and eosinophils of AD patients.
Assuntos
Proteínas Sanguíneas/genética , Dermatite Atópica/sangue , Dermatite Atópica/genética , Sequência de Aminoácidos , Apoptose , Northern Blotting , Morte Celular , Linhagem Celular , Clonagem Molecular , Citoplasma/metabolismo , DNA Complementar/metabolismo , Eosinófilos/metabolismo , Perfilação da Expressão Gênica , Proteínas de Fluorescência Verde , Humanos , Proteínas Luminescentes/metabolismo , Proteínas de Membrana , Dados de Sequência Molecular , Ligação Proteica , Estrutura Terciária de Proteína , RNA/metabolismo , RNA Mensageiro/metabolismo , Proteínas Recombinantes de Fusão/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Homologia de Sequência de Aminoácidos , Fatores de Tempo , Distribuição Tecidual , Transcrição Gênica , Técnicas do Sistema de Duplo-HíbridoRESUMO
BACKGROUND: Monocytes, macrophages, and antigen-presenting cells (APCs) are key effectors of both innate and acquired immune responses. Such cells have been implicated in the pathogenesis of some inflammatory diseases. Differential gene expression in CD14-positive cells from patients with atomic dermatitis (AD) was studied using real-time quantitative RT-PCR to measure transcription levels of selected genes. METHODS: PBMCs were prepared by Ficoll gradient separation from 30 AD patients (the anti-mite-specific IgE RAST score: 0.75 to >100 UA/ml) and 10 healthy adult individuals (the RAST score: <0.34-0.37 UA/ml) and CD14-positive cells were selected. A total of 64 genes was selected for study from groups of genes with different molecular function. RESULTS: Genes involved in MHC class I antigen presentation, such as beta(2)-microglobulin, subunits of an immunoproteasome and ATP-binding cassette transporter TAP2 were expressed at higher levels in the AD patients than in the healthy controls. The genes for Toll-like receptors, CD36 and IFNgamma receptor were also upregulated in the AD patients. These genes are involved in MHC class I antigen presentation, recognition of bacterial pathogens and apoptotic cells. CONCLUSIONS: The upregulation of genes suggests that circulating monocytes in AD patients may be primed to differentiate into effector cells by conditions associated with AD. The upregulation of genes may prove to be a useful marker for AD.
Assuntos
Dermatite Atópica/genética , Regulação da Expressão Gênica/imunologia , Monócitos/fisiologia , Adolescente , Adulto , Antígenos CD36/genética , Antígenos CD36/imunologia , Criança , Pré-Escolar , Proteínas de Ligação a DNA/genética , Proteínas de Ligação a DNA/imunologia , Dermatite Atópica/imunologia , Dermatite Atópica/metabolismo , Genes MHC Classe I/genética , Genes MHC Classe I/imunologia , Genes MHC da Classe II/genética , Genes MHC da Classe II/imunologia , Humanos , Receptores de Lipopolissacarídeos/imunologia , Glicoproteínas de Membrana/genética , Glicoproteínas de Membrana/imunologia , Pessoa de Meia-Idade , Monócitos/imunologia , Análise de Sequência com Séries de Oligonucleotídeos , Receptores de Superfície Celular/genética , Receptores de Superfície Celular/imunologia , Receptores de Interferon/genética , Receptores de Interferon/imunologia , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Receptores Toll-Like , Receptor de Interferon gamaRESUMO
BACKGROUND: Analysis of genes that are differentially expressed in patients with atopic dermatitis (AD) and normal individuals will provide important information on the underlying molecular pathogenetic mechanisms of AD. METHODS: Transcript of freshly isolated peripheral blood T cells from 59 individuals were analyzed with a fluorescent differential display (FDD) method. Ninety-two differentially expressed genes were identified in this manner. Additionally, real-time quantitative RT-PCR was employed to investigate the expression of the FDD-selected genes and also genes related to T cell function. RESULTS: A number of genes, including CC chemokine receptor 4, T cell-specific tyrosine kinase (Emt/Itk), integrin beta1, integrin alpha6, IQGAP1 and MAR/SAR DNA-binding protein (SATB1), were shown to be more highly expressed in patients with moderate and/or severe AD than in controls or patients with mild AD. Because the products of these upregulated genes influence chemotaxis, adhesion, migration and Th2 polarization, it is suggested that in more severe AD, circulating T cells may function differently in this regard. Several other genes, the role of which in T cell function is currently unknown, were also found to be differentially expressed in AD. These included the heat shock protein 40 and vasopressin-activated calcium-mobilizing receptor 1. CONCLUSION: The upregulated genes identified in this work may serve as useful markers for moderate to severe AD as opposed to normal or mild AD and also as markers indicating progression to more severe AD. Further functional characterization will provide a better understanding of the pathophysiology of circulating T cells in AD.