RESUMO
Genetic studies on mouse models of asthma have identified interleukin-9 (IL9) as a determining factor in controlling bronchial hyperresponsiveness, a hallmark of the disease. Recently, the human IL9 receptor (hIL9R) gene locus has also been implicated in determining susceptibility to bronchial hyperresponsiveness and asthma. In order to evaluate the structure and function of the encoded product, we analyzed receptor transcripts derived from peripheral blood mononuclear cells of 50 unrelated donors. Sequence analysis of the entire coding region identified a splice variant that contains an in frame deletion of a single residue at codon 173 (DeltaQ). This variant could be permanently expressed in a cytokine-dependent murine T-cell line but lacked the ability to induce proliferation in response to human IL9. In situ analyses of cells expressing the wild-type and DeltaQ receptors found both forms to be expressed at the cell surface, but the DeltaQ receptor was unable to bind hIL9 and could not be recognized by N-terminal specific antibodies. These findings demonstrate that hIL9RDeltaQ presents an altered structure and function and suggests its potential role in down-regulating IL9 signaling in effector cells and associated biological processes.
Assuntos
Processamento Alternativo , Variação Genética , Interleucina-9/fisiologia , Ativação Linfocitária , Linfócitos/imunologia , Receptores de Interleucina/genética , Transcrição Gênica , Sequência de Aminoácidos , Animais , Células COS , Clonagem Molecular , Códon , Citometria de Fluxo , Humanos , Interleucina-9/farmacologia , Camundongos , Reação em Cadeia da Polimerase , Receptores de Interleucina/química , Receptores de Interleucina/fisiologia , Receptores de Interleucina-9 , Proteínas Recombinantes/biossíntese , Deleção de Sequência , Transdução de Sinais , Linfócitos T/imunologia , TransfecçãoRESUMO
Recent data have identified IL-9 as a key cytokine in determining susceptibility to asthma. These data are supported by the finding that allergen-exposed IL-9-transgenic mice exhibit many features that are characteristic of human asthma (airway eosinophilia, elevated serum IgE and bronchial hyperresponsiveness) as compared to the background strain. A striking feature of these animals is a robust peribronchial and perivascular eosinophilia after allergen challenge, suggesting that IL-9 is a potent factor in regulating this process. In an attempt to gain insights into the molecular mechanism governing IL-9 modulation of lung eosinophilia, we investigated the ability of this cytokine to induce the expression of CC-type chemokines in the lung because of their effect on stimulating eosinophil chemotaxis. Here we show that IL-9-transgenic mice in contrast to their congenic controls exhibit baseline lung eosinophilia that is associated with the up-regulation of CC-chemokine expression in the airway. This effect appears to be through a direct action of IL-9 because the addition of recombinant IL-9 to primary epithelial cultures and cell lines induced the expression of these chemokines in vitro. These data support a mechanism for IL-9 in regulating the expression of eosinophil chemotactic factors in lung epithelial cells.
Assuntos
Quimiocinas/biossíntese , Eosinofilia/etiologia , Interleucina-9/genética , Interleucina-9/fisiologia , Pulmão/imunologia , Alérgenos/administração & dosagem , Animais , Asma/etiologia , Asma/imunologia , Sequência de Bases , Linhagem Celular , Células Cultivadas , Quimiocinas/genética , Quimiotaxia de Leucócito , Primers do DNA/genética , Eosinofilia/imunologia , Células Epiteliais/imunologia , Humanos , Interleucina-9/farmacologia , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Dados de Sequência Molecular , Regulação para CimaRESUMO
Asthma is a complex heritable inflammatory disorder of the airways associated with clinical signs of atopy and bronchial hyperresponsiveness. Recent studies localized a major gene for asthma to chromosome 5q31-q33 in humans. Thus, this segment of the genome represents a candidate region for genes that determine susceptibility to bronchial hyperresponsiveness and atopy in animal models. Homologs of candidate genes on human chromosome 5q31-q33 are found in four regions in the mouse genome, two on chromosome 18, and one each on chromosomes 11 and 13. We assessed bronchial responsiveness as a quantitative trait in mice and found it linked to chromosome 13. Interleukin 9 (IL-9) is located in the linked region and was analyzed as a gene candidate. The expression of IL-9 was markedly reduced in bronchial hyporesponsive mice, and the level of expression was determined by sequences within the qualitative trait locus (QTL). These data suggest a role for IL-9 in the complex pathogenesis of bronchial hyperresponsiveness as a risk factor for asthma.
Assuntos
Asma/genética , Interleucina-9/genética , Animais , Células Cultivadas , Mapeamento Cromossômico , Feminino , Regulação da Expressão Gênica , Ligação Genética , Interleucina-9/metabolismo , Pulmão/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos , Fenótipo , Característica Quantitativa Herdável , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Especificidade da Espécie , Baço/citologia , Baço/metabolismoRESUMO
We examined the long arm XY pseudoautosomal region for linkage to asthma, serum IgE, and bronchial hyperresponsiveness. In 57 Caucasian families multipoint nonparametric analyses provide evidence for linkage between DXYS154 and bronchial hyperresponsiveness (P = 0.000057) or asthma (P = 0.00065). This genomic region is approximately 320 kb in size and contains the interleukin-9 receptor gene. These results suggest that a gene controlling asthma and bronchial hyperresponsiveness maybe located in this region and that the interleukin-9 receptor is a potential candidate.