DNA MICROARRAY ANALYSIS OF TRANSFORMING GROWTH FACTOR-β AND RELATED TRANSCRIPTS IN NASAL BIOPSIES FROM PATIENTS WITH ALLERGIC RHINITIS

doi:10.1006/cyto.2002.1012

Cytokine

Volume 18, Issue 1, April 2002, Pages 20-25

https://doi.org/10.1006/cyto.2002.1012 Get rights and content

Abstract

Decreased activity of anti-inflammatory cytokines like transforming growth factor (TGF)-β may contribute to allergic inflammation. In vivo effects of TGF-β-effects are difficult to infer from local concentrations, since TGF-β-effects depend on a complex system of regulatory proteins and receptors. Instead the effects of TGF-β might be inferred by examining TGF-β-inducible transcripts. In this study DNA microarrays were used to examine local expression of TGF-β, TGF-β-regulatory and -inducible transcripts in nasal biopsies from patients with symptomatic allergic rhinitis and healthy controls. In addition, nasal fluids were analysed with cytological and immunological methods. Nasal fluid eosinophils, albumin, eosinophil granulae proteins and IgE, but not TGF-β, were higher in patients than in controls. DNA microarray analysis of nasal mucosa showed expression of transcripts encoding TGF-β, TGF-β-regulatory proteins and -receptors at variable levels in patients and controls. By comparison, analysis of 28 TGF-β-inducible transcripts indicated that 23 of these had lower measurement values in patients than in controls, while one was higher, and the remaining four were absent in both patients and controls. In summary, TGF-β and a complex system of regulatory genes and receptors are expressed in the nasal mucosa. Low expression of TGF-β-inducible transcripts may indicate decreased TGF-β activity in allergic rhinitis. DNA microarray analysis may be a way to study cytokine effects in vivo.

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Analyses of two profiling data sets from synovium from RA patients and healthy controls all showed significant activation of pathways with known pathogenic relevance, such as the Th1 pathway, the role of NFAT in regulation of the immune response, dendritic cell maturation, iCOS-iCOSL signaling in T helper cells, Fcγ receptor-mediated phagocytosis, interferon signaling, Cdc42 signaling, and cytotoxic T lymphocyte-mediated apoptosis. The most activated upstream regulators included TNF, an important drug target, as well as IFN-gamma and CD40LG, all of which are known to play important pathogenic roles in RA. The differentially expressed genes from synovium included several potential biomarkers, such as CCL5, CCL13, CCL18, CX3CL1, CXCL6, CXCL9, CXCL10, CXCL13, IL15, IL32, IL1RN, SPP1, and TNFSF11. By contrast, microarray studies of WBC, PBMC and CD4+ T cells showed variable pathways and limited pathway overlap with synovium. Similarly, scRNA-seq data from a mouse model of arthritis did not support that inflammatory responses in peripheral blood reflect those in the arthritic joints. These data showed pathway overlap between mouse joint cells and synovium from patients with RA, but not with cells in peripheral blood.
Our findings indicate a dichotomy between gene expression changes, pathways, upstream regulators and biomarkers in synovium and cell types in peripheral blood, which complicates identification of biomarkers in blood.
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TGF-β has anti-inflammatory effects. Decreased activity of the TGF-β pathway has previously been described in a DNA microarray study of allergic rhinitis.6 Moreover, TGF-β is released from TR1 cells, which are important regulators of peripheral tolerance.
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We sought to identify disease-associated pathways in the LPR using a network-based analysis.
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The DNA microarray analysis revealed several genes of known relevance to allergy. The eosinophil marker Charcot-Leyden crystal protein (CLC) that encodes Charcot-Leyden crystal protein differed most in expression. A network-based analysis showed upregulation of IL-4– and CCL4-dependent pathways and downregulation of a TGF-β–induced pathway. CCL4 is expressed by CD4⁺ T cells and chemotactic for eosinophils. We hypothesized that allergen induces release of CCL4 from T_H2 cells and that this contributes to influx of eosinophils. Further analysis showed increase of CCL4 protein in nasal fluids from allergic patients during the season. Allergen challenge of PBMCs resulted in proliferation of T_H2 cells and increased production of CCL4 in CD4⁺ T cells from allergic patients. An analysis of the DNA microarray data revealed a significant correlation between CCL4 and the eosinophil marker CLC.
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The nasal and bronchial mucosa present similarities and differences. Remodeling is defined as “model again or differently, reconstruct” and is present in the airways of most if not all asthmatic patients. Even though inflammation is similar in allergic rhinitis and asthma, the pathologic extent of nasal remodeling in patients with rhinitis seems to be far less extensive than that in the bronchi of asthmatic patients. Epithelial damage is only minimal, and the reticular basement membrane does not appear to be largely pseudothickened. Moreover, the demonstration of fibrogenic growth factors in the nasal mucosa of patients with allergic rhinitis is lacking because of the paucity of studies. The reasons why remodeling appears to be less extensive in the nasal mucosa than in the bronchial mucosa are still unclear, but 2 hypotheses can be put forward. On one hand, the cytokine production of smooth muscle cells might partly explain differences in remodeling of the 2 sites of the airways. On the other hand, the genes of the embryologic differentiation might persist in the nose and bronchi or might be re-expressed in asthma and rhinitis. Because the nose is of ectodermal origin and the bronchi of endodermal origin, these genes might also govern remodeling patterns. More studies are urgently required to better characterize nasal remodeling in patients with rhinitis. A better understanding of nasal and bronchial remodeling might help to identify new pathways and new therapeutic strategies to reduce long-term remodeling in asthma.
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^f1: Correspondence to: Mikael Benson, MD, PhD, Allergy Laboratory, Department of Oto-Rhino-Laryngology, Malmö University Hospital, S-205 02 Malmö, Sweden. Email: [email protected]

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Regular ArticlesDNA MICROARRAY ANALYSIS OF TRANSFORMING GROWTH FACTOR-β AND RELATED TRANSCRIPTS IN NASAL BIOPSIES FROM PATIENTS WITH ALLERGIC RHINITIS

Abstract

J Allergy Clin Immunol

Curr Opin Microbiol

FEBS Lett

Genomics

J Allergy Clin Immunol

Interleukin-10 and transforming growth factor-beta promoter polymorphisms in allergies and asthma

Am J Respir Crit Care Med

Transforming growth factor beta 1 gene expression in human airways

Thorax

Transforming growth factor beta abrogates the effects of hematopoietins on eosinophils and induces their apoptosis

J Exp Med

Eosinophil-associated TGF-beta1 mRNA expression and airways fibrosis in bronchial asthma

Am J Respir Cell Mol Biol

Expression of growth factors and remodelling of the airway wall in bronchial asthma

Thorax

Regular Articles
DNA MICROARRAY ANALYSIS OF TRANSFORMING GROWTH FACTOR-β AND RELATED TRANSCRIPTS IN NASAL BIOPSIES FROM PATIENTS WITH ALLERGIC RHINITIS