Functional effects of proinflammatory factors present in Sj\xf6gren\u2019s syndrome salivary microenvironment in an in vitro model of human salivary gland

Mayte Arce-Franco,José David Machado,Teresa Giraldez,Pablo Miranda,Federico Díaz-González,Diego Alvarez De La Rosa,Martina K Pec,Carlos Martínez-Gimeno,María Dominguez-Luis,José María García-Verdugo

doi:10.1038/s41598-017-12282-x

Mayte Arce-Franco, José David Machado + Show 8 more

Open Access

https://doi.org/10.1038/s41598-017-12282-x

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Abstract

Primary Sjögren’s syndrome (pSS) is an autoimmune exocrinopathy in which the role that the immune response plays in reducing exocrine gland function, including the glandular microenvironment of cytokines, has not been fully understood. Epithelial cells from biopsies of human parotid gland (HPG) were used to establish a model of human salivary gland in vitro. In this model, the functional consequences of several proinflammatory soluble factors present in the pSS glandular microenvironment were assessed. Stimulation with isoproterenol and calcium produced a significant increase in the basal activity of amylase in the HPG cell supernatants. Under these conditions, the presence of TNF-α and CXCL12 increased amylase mRNA cellular abundance, but reduced the amylase activity in the cell-free supernatant in a dose-dependent manner. IL-1β and IFN-γ, but not TGF-β, also diminished amylase secretion by HPG cells. These results suggest that the glandular microenvironment of cytokine, by acting post-transcriptionally, may be responsible, at least in part, for the reduced exocrine function observed in pSS patients. These data may help to a better understanding of the pathogenesis of SS, which in turn would facilitate the identification of new therapeutic targets for this disorder.

Highlights

Primary Sjögren’s Syndrome is a chronic autoimmune disorder of unknown etiology characterized by lymphocytic infiltration of exocrine glands, both salivary and lachrymal, that results in dry eyes and mouth[1]
Quantitative real-time PCR studies have demonstrated that CD4+ T lymphocytes obtained from salivary gland biopsies of Primary Sjögren’s syndrome (pSS) patients express higher amounts of IL-2, IFN-γ, and IL-10 mRNA than peripheral blood CD4+ T cells, and salivary gland epithelial cells produced more IL-1, IL-6, and TNF-α mRNA than epithelial cells in normal salivary glands[16]
Human parotid glands (HPG) primary cells were isolated from tissue samples and expanded in culture using expansion medium, as described in Methods section

Summary

Introduction

Primary Sjögren’s Syndrome (pSS) is a chronic autoimmune disorder of unknown etiology characterized by lymphocytic infiltration of exocrine glands, both salivary and lachrymal, that results in dry eyes and mouth[1]. Quantitative real-time PCR (qRT-PCR) studies have demonstrated that CD4+ T lymphocytes obtained from salivary gland biopsies of pSS patients express higher amounts of IL-2, IFN-γ, and IL-10 mRNA than peripheral blood CD4+ T cells, and salivary gland epithelial cells produced more IL-1, IL-6, and TNF-α mRNA than epithelial cells in normal salivary glands[16]. Quantification of these cytokines by ELISA in saliva of patients respect to controls showed an elevated concentration that correlated with the mRNA levels[16]. This data may lead to a better understanding of the pathogenesis of pSS, which in turn would facilitate the identification of new therapeutic targets for this disorder

Methods

Results

Conclusion