Cytokines Levels and Salivary Microbiome Play A Potential Role in Oral Lichen Planus Diagnosis

Maria Fernanda Marques Silva De Carvalho,Talita Gomes Baeta Lourenço,Denise Cavalieri,Denise Da Cunha Pasqualin,Danielle Viana Ribeiro Ramos,Fernanda Agostini Rocha,Sabrina Do Nascimento,Luciana Marti,Débora Heller,Leandro Aurélio Liporoni Martins

doi:10.1038/s41598-019-54615-y

Abstract

Oral lichen planus (OLP) is a chronic Th1-mediated inflammatory mucocutaneous disease of the skin and oral mucosa that can have various clinical presentations. Lesions are usually bilateral and often painful. While cutaneous Lichen Planus (LP) lesions are self-limiting, the oral lesions are chronic and rarely remissive. The diagnosis of oral lichen planus (OLP) is often challenging, and confirmation by histopathological criterion is generally advised. The aim of our study was to identify the cytokines present in OLP-suggestive lesions and in non-specific inflammatory lesions (NSIL) used as controls. Moreover, assess cytokines protein levels and oral microbiota composition in whole saliva samples. Histopathological analysis, immunohistochemistry and gene expression were used as techniques to analyze the oral mucosal tissue samples. ELISA was conducted to analyze salivary cytokine levels and 16S rRNA sequencing was used to determine the salivary microbiome. As a result we observed larger number of infiltrated lymphocytes (p = 0.025), as well, more T CD4 lymphocytes in the epithelial tissue (p = 0.006) in OLP samples compared to NSIL. In addition, the OLP samples displayed more apoptotic cells compared to NSIL (p = 0.047). Regarding the cytokine analysis, IFN-γ and IL-33 were more expressed in OLP lesions than in NSIL samples (p < 0.001; p = 0.026). Furthermore, our results demonstrated higher levels of IFN-γ protein expression in the saliva of OLP group compared to controls (p = 0.0156). We also observed noted differences in the oral microbiota composition between OLP and NSIL saliva samples. In conclusion, OLP lesions presented larger numbers of apoptotic and inflammatory cells, higher levels of IFN-γ and IL-33 compared to NSIL, and these lesions also differ regarding oral microbiota composition. These results are consistent with the Th-1-mediated chronic inflammation nature of oral lichen planus investigated lesions and displayed unique features that could be used as a diagnostic tool.

Highlights

Oral lichen planus (OLP) is a chronic and inflammatory mucocutaneous disease of the skin and oral mucosa that can have various clinical features
In this study we found that OLP lesions in comparison to control lesions are characterized in the lamina propria by a more severe dense lymphocytic inflammatory infiltrate in which CD3+ cells predominate
Regarding the expression of cytokines in the lesions we found higher expression of IFN-γ and Interleukin 33 (IL-33) in OLP whereas no differences were found in IL-17 expression levels in comparison to control samples

Summary

Introduction

Oral lichen planus (OLP) is a chronic and inflammatory mucocutaneous disease of the skin and oral mucosa that can have various clinical features. A high proportion of the novel CD4+ T helper cells subset-Th17 cells were detected in OLP lesions These cells would release inflammatory cytokines leading to migration and activation of inflammatory cells and cytotoxic CD8 T cells with consequent damage to basal keratinocytes and their death by apoptosis[5,6,7,8,9,10]. Others studies have correlated Helicobacter pylori with OLP, but there are divergences between different sources[19,20] Even though, these events are not directly related to the OLP etiology, all these factors might influence the oral microbiome composition and as consequence alter the local immunity and favoring the OLP maintenance and progression. Non-specific inflammatory lesions (NSIL) of the oral mucosa were analyzed as controls

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Conclusion