RESUMO
BACKGROUND: Noninvasive sputum sampling has enabled the identification of biomarkers in asthmatic patients. Studies of discrete cell populations in sputum can enhance measurements compared with whole sputum in which changes in rare cells and cell-cell interactions can be masked. OBJECTIVE: We sought to enrich for sputum-derived human bronchial epithelial cells (sHBECs) and sputum-derived myeloid type 1 dendritic cells (sDCs) to describe transcriptional coexpression of targets associated with a type 2 immune response. METHODS: A case-control study was conducted with patients with mild asthma (asthmatic cases) and healthy control subjects. Induced sputum was obtained for simultaneous enrichment of sHBECs and sDCs by using flow cytometry. Quantitative PCR was used to measure mRNA for sHBEC thymic stromal lymphopoietin (TSLP), IL33, POSTN, and IL25 and downstream targets in sDCs (OX40 ligand [OX40L], CCL17, PPP1R14A, CD1E, CD1b, CD80, and CD86). RESULTS: Final analyses for the study sample were based on 11 control subjects and 13 asthmatic cases. Expression of TSLP, IL33, and POSTN mRNA was increased in sHBECs in asthmatic cases (P = .001, P = .05, and P = .04, respectively). Expression of sDC OX40L and CCL17 mRNA was increased in asthmatic cases (P = .003 and P = .0001, respectively). sHBEC TSLP mRNA expression was strongly associated with sDC OX40L mRNA expression (R = 0.65, P = .001) and less strongly with sDC CCL17 mRNA expression. sHBEC IL33 mRNA expression was associated with sDC OX40L mRNA expression (R = 0.42, P = .04) but not sDC CCL17 mRNA expression. CONCLUSIONS: Noninvasive sampling and enrichment of select cell populations from sputum can further our understanding of cell-cell interactions in asthmatic patients with the potential to enhance endotyping of asthmatic patients.
Assuntos
Asma/genética , Células Dendríticas/metabolismo , Células Epiteliais/metabolismo , Expressão Gênica , Transdução de Sinais/imunologia , Adulto , Antígenos CD1/genética , Antígenos CD1/imunologia , Asma/imunologia , Asma/patologia , Antígeno B7-1/genética , Antígeno B7-1/imunologia , Antígeno B7-2/genética , Antígeno B7-2/imunologia , Estudos de Casos e Controles , Comunicação Celular , Quimiocina CCL17/genética , Quimiocina CCL17/imunologia , Células Dendríticas/imunologia , Células Dendríticas/patologia , Células Epiteliais/imunologia , Células Epiteliais/patologia , Feminino , Perfilação da Expressão Gênica , Humanos , Interleucina-17/genética , Interleucina-17/imunologia , Interleucina-33/genética , Interleucina-33/imunologia , Peptídeos e Proteínas de Sinalização Intracelular , Masculino , Pessoa de Meia-Idade , Proteínas Musculares , Ligante OX40/genética , Ligante OX40/imunologia , Fosfoproteínas Fosfatases/genética , Fosfoproteínas Fosfatases/imunologia , Transdução de Sinais/genética , Escarro/citologiaRESUMO
Air pollution contributes to acute exacerbations of asthma and the development of asthma in children and adults. Airway epithelial cells interface innate and adaptive immune responses, and have been proposed to regulate much of the response to pollutants. Thymic stromal lymphopoietin (TSLP) is a pivotal cytokine linking innate and Th2 adaptive immune disorders, and is upregulated by environmental pollutants, including ambient particulate matter (PM) and diesel exhaust particles (DEP). We show that DEP and ambient fine PM upregulate TSLP mRNA and human microRNA (hsa-miR)-375 in primary human bronchial epithelial cells (pHBEC). Moreover, transfection of pHBEC with anti-hsa-miR-375 reduced TSLP mRNA in DEP but not TNF-α-treated cells. In silico pathway evaluation suggested the aryl hydrocarbon receptor (AhR) as one possible target of miR-375. DEP and ambient fine PM (3 µg/cm(2)) downregulated AhR mRNA. Transfection of mimic-hsa-miR-375 resulted in a small downregulation of AhR mRNA compared with resting AhR mRNA. AhR mRNA was increased in pHBEC treated with DEP after transfection with anti-hsa-miR-375. Our data show that two pollutants, DEP and ambient PM, upregulate TSLP in human bronchial epithelial cells by a mechanism that includes hsa-miR-375 with complex regulatory effects on AhR mRNA. The absence of this pathway in TNF-α-treated cells suggests multiple regulatory pathways for TSLP expression in these cells.
Assuntos
Citocinas/genética , Células Epiteliais/metabolismo , MicroRNAs/metabolismo , Material Particulado , Mucosa Respiratória/metabolismo , Emissões de Veículos , Células Cultivadas , Citocinas/metabolismo , Regulação da Expressão Gênica , Humanos , MicroRNAs/genética , Estabilidade de RNA , Receptores de Hidrocarboneto Arílico/genética , Receptores de Hidrocarboneto Arílico/metabolismo , Linfopoietina do Estroma do TimoRESUMO
To examine the effect of co-location of psychogeriatric and geriatric services on length of stay and changes in patients' psychosocial characteristics. A retrospective analysis of the performance indices of psychogeriatric patients aged > or =50 years admitted to a co-located psychogeriatric and geriatric unit at Bankstown-Lidcombe Hospital, New South Wales (NSW), Australia from April 2004 to June 2006. Comparisons were made between the performance of the Bankstown-Lidcombe's unit and the NSW state average (consisting of traditional solitary models of care) with respect to patients' length of stay (LOS) and changes in psychosocial indices. Bankstown's patients had a higher burden of psychosocial impairments. The mean LOS for psychogeriatric episodes was significantly shorter at Bankstown-Lidcombe Hospital than the NSW state average (28.3+/-19.6 days vs. 33.4+/-22.7 days, p<0.001). The overall improvement in aspects of mental state and social behaviors for psychogeriatric admissions at Bankstown-Lidcombe Hospital was significantly better than the NSW state average. Co-location of psychogeriatric and geriatric services reduced patients' LOS and improved psychosocial performance compared to traditional models of care. However, more robust studies are required to fully examine the benefits of this type of service.
