Animal models to study the pathogenesis and novel therapeutics of oral lichen planus

Oral lichen planus (OLP) is a prevalent oral mucosal disease characterized by an unknown etiology and a complex pathogenesis. Patients with OLP endure a chronic course marked by alternating non-erosive and erosive lesions, with no definitive cure currently available. Particularly challenging is the treatment of recalcitrant erosive OLP, highlighting an urgent need for therapies targeting specific pathogenic pathways. In diseases like OLP, where the etiopathogenesis is intricate and elusive, animal models are indispensable for hypothesis testing and elucidating disease mechanisms. To date, only three animal models for oral lichenoid lesions have been reported in the literature. This Perspective paper evaluates these existing models, along with a novel OLP mouse model introduced at the 3rd International Conference on Oral Mucosal Immunity and Microbiome. The validity of these models is critically assessed, and their potential future applications in advancing our understanding of OLP are discussed

The analysis of oral microbial communities of wild-type and toll-like receptor 2-deficient mice using a 454 GS FLX Titanium pyrosequencer

Background: Although mice have long served as an animal model for periodontitis, information on the composition of their indigenous oral microbiota is limited. The aim of the current study was to characterize mouse oral bacterial flora by applying extensive parallel pyrosequencing using the latest model pyrosequencer, a Roche/454 Genome Sequencer FLX Titanium. In addition, the effect of Toll-like receptor (TLR) 2 deficiency on oral microbiota was evaluated. Results: Eight oral bacterial communities of wild-type (n = 4) and TLR2 knock-out (n = 4) C57BL/6 mice were characterized by analyzing 80,046 reads of 16S rRNA genes obtained by pyrosequencing. Excluding the PCR primers, the average length of each sequencing product was 443 bp. The average species richness of the murine oral bacterial communities was estimated to be about 200, but the communities were dominated by only two main phyla and several species. Therefore, the bacterial communities were relatively simple. The bacterial composition of the murine oral microbiota was significantly different from that of humans, and the lack of TLR2 had a negligible effect on the murine oral microbiota. Conclusion: Pyrosequencing using the Roche/454 FLX Titanium successfully characterized mouse oral bacterial communities. The relatively simple oral bacterial communities of mice were not affected by TLR2 deficiency. These findings will provide a basis for future studies on the role of periodontal pathogens in the murine model of periodontitis.This study was supported by grants R13-2008-008-01003-0 from the Korea Science and Engineering Foundation.

Gut dysbiosis in autoimmune diseases: Association with mortality

To better understand the impact of gut dysbiosis on four autoimmune diseases [Sjögren’s syndrome (SS), systemic lupus erythematosus (SLE), rheumatoid arthritis (RA), and multiple sclerosis (MS)], this review investigated the altered gut bacteria in each disease and the shared ones among the four diseases. The enriched gut bacteria shared by three of the four autoimmune diseases were Streptococcus, Prevotella, and Eggerthella, which are associated with autoantibody production or activation of Th17 cells in immune-related diseases. On the other hand, Faecalibacterium comprises depleted gut bacteria shared by patients with SLE, MS, and SS, which is associated with various anti-inflammatory activities. The indexes of gut dysbiosis, defined as the number of altered gut bacterial taxa divided by the number of studies in SLE, MS, RA, and SS, were 1.7, 1.8, 0.7, and 1.3, respectively. Interestingly, these values presented a positive correlation trend with the standardized mortality rates —2.66, 2.89, 1.54, and 1.41, respectively. In addition, shared altered gut bacteria among the autoimmune diseases may correlate with the prevalence of polyautoimmunity in patients with SLE, SS, RA, and MS, that is, 41 percent, 32.6 percent, 14 percent, and 1–16.6 percent, respectively. Overall, this review suggests that gut dysbiosis in autoimmune diseases may be closely related to the failure of the gut immune system to maintain homeostasis

B cells activated in the presence of Th1 cytokines inhibit osteoclastogenesis.

Host immune response has been considered as an important disease-modifying factor of periodontitis, however, which immune cell(s) or factor(s) are involved in the destruction of periodontium remains unclear. Previously, we reported that osteoclastogenesis is enhanced by activated B cells but suppressed by activated CD8+ T cells. We present new data that B cells activated in the presence of Th1 cytokines inhibit osteoclastogenesis. Purified murine B cells were activated with anti-IgD mAb, IL-4, and anti-CD40 mAb, in the absence (BTh2) or presence of Th1 cytokines, either IL-2 (BIL-2) or IFN-gamma (BIFN-gamma). Each activated B cell population was co-cultured with RAW264.7 cells in the presence of soluble receptor activator of NF-kappaB ligand (sRANKL), and the effect on osteoclastic differentiation was evaluated. While BTh2 increased osteoclastogenesis, BIL-2 and BIFN-gamma suppressed it profoundly. To verify the mediating molecule(s), we analyzed cytokine profiles of the activated B cells. Compared to BTh2, BIL-2 expressed increased amount of IFN-gamma and BIFN-gamma expressed decreased amounts of IL-4, IL-5, and IL-10. IFN-gamma was a key negative regulator of osteoclastic differentiation, and mediated the inhibition by BIL-2. These results suggest that Th1 cytokines may have new important roles in resistance to periodontitis, acting directly on osteoclasts or indirectly through B cells.This study was supported by Korea Research Foundation grant 2000-041-F00276. The authors thank Dr. Jennifer M. Puck for the critical reading of manuscript

Phenotype and Tissue Residency of Lymphocytes in the Murine Oral Mucosa

The oral mucosa is a critical barrier tissue that harbors a series of distinct immune cell subsets. Immune surveillance in the oral mucosa is important for both local and systemic immunity because the oral cavity is a heavily utilized route of pathogen entry and also serves as site of pathogen propagation. Nonetheless, composition and phenotype of the lymphocyte pool in the oral mucosa have remained poorly characterized. Utilizing a newly established protocol for mucosal immune cell isolation, here, we report that the oral mucosa features a unique cellular composition of immune cells, which differed not only from secondary lymphoid organs but also from mucosal tissues in the gut and lung. We observed profound accumulation of CD11b(+)Ly6C(lo) monocytes in the oral mucosa that were maintained independently of T- and B-lymphocytes. Unlike the gut mucosa, the oral mucosa neither contained CD8αα T cells nor was it enriched for CD103(+)CD69(+) tissue-resident memory CD8 T cells. In fact, a major fraction of T cells circulated and trafficked through the mucosa as revealed by treatment with the S1P1 receptor antagonist, FTY720, a potent inhibitor of lymphocyte migration. Collectively, these results provide a comprehensive picture of immune cells in the oral mucosa as an active site of lymphocyte recruitment and surveillance

Ability of S100 proteins and matrix metalloproteinase-9 to identify periodontitis in a ligature-induced periodontitis dog model

Aims The present study aimed to monitor the levels of selected salivary biomarkers during the development and treatment of periodontitis and to evaluate their ability to identify periodontitis in dogs. Materials and methods A total of 15 beagle dogs were divided into a control group (no ligature), group 1 (ligature on six teeth), and group 2 (ligature on 12 teeth). The experimental periods consisted of 8 weeks of periodontitis induction and 4 weeks of treatment. Clinical measurements and the sampling of saliva were performed every 4 weeks. The levels of S100A8, S100A9, S100A8/A9, and matrix metalloproteinase (MMP)-9 were measured by enzyme-linked immunosorbent assay. Results All experimental animals and two control animals developed periodontitis, which was successfully treated. All salivary biomarkers were significantly increased in periodontitis with high diagnostic power (c-index >= 0.944) and were able to identify animals with periodontitis on a single tooth. Whereas the levels of salivary S100A8/A9 recovered to levels in health, those of S100A8, S100A9, and MMP-9 in periodontitis stability remained significantly higher than in health. Conclusion Salivary S100A8, S100A9, S100A8/A9, and MMP-9 may be used for the screening of periodontitis in dogs, but with caution of other conditions that can affect their levels in saliva.N

Associations of Anti-Aquaporin 5 Autoantibodies with Serologic and Histopathological Features of Sjogren's Syndrome

Biomarkers to stratify the complex and heterogeneous phenotypes of Sjogren's syndrome (SS) are needed. We aimed to validate the prevalence of anti-aquaporin 5 (AQP5) IgG in a non-Korean cohort and to optimize the method to screen the anti-AQP5 IgG. The study cohort included 111 primary SS and 43 non-SS Sjogren's International Collaborative Clinical Alliance (SICCA) controls that were obtained from the Sjogren's International Collaborative Clinical Alliance registry, in addition to 35 systemic lupus erythematosus (SLE) and 35 rheumatoid arthritis (RA) phenotypes. Anti-AQP5 IgG was screened by cell-based immunofluorescence cytochemistry (CB-IFC) assay in the absence or presence of epitope peptides, as well as by ELISA using the epitope peptides as coated antigens. Anti-AQP5 IgG specific to an E1 epitope was best at discriminating between SS and non-SS, and the two different methods (CB-IFC and ELISA) presented comparable performance in diagnostic accuracy (0.690 vs. 0.707). Notably, the SLE and RA groups had substantially lower levels of anti-AQP5 IgG than the SS group. In addition, the presence of anti-AQP5_E1 IgG was associated with serologic and histopathological features of SS. In conclusion, a similar prevalence of anti-AQP5 IgG was confirmed in a non-Korean cohort. Screening anti-AQP5 autoantibodies may help to form subgroups of SS for targeted therapy.Y

Prospective evaluation of clinical symptoms of chemotherapy-induced oral mucositis in adult patients with acute leukemia: A preliminary study

Objective The objective of this study was to prospectively evaluate the clinical features of chemotherapy-induced oral mucositis (CIOM) in adult patients with acute leukemia and the aggravating factors for such symptoms. Subjects Thirty-seven prospective patients aged >= 19 years with acute leukemia undergoing chemotherapy were enrolled. Oral and clinical investigations were performed at baseline and on Day 14 after starting chemotherapy. The presence and severity of cancer-induced oral mucositis were demonstrated using the World Health Organization (WHO) scoring system. Results On Day 14, we found that oral mucositis had developed in eight patients (21.6%). Hematopoietic stem cell transplantation (HSCT) in patients was a predictor of increased WHO scores (beta = 1.937, p < .001). Regarding oral sites, ventral tongue (beta = 1.670), soft palate (beta = 1.242), and buccal mucosa (beta = 0.593) were predictors for increased scores. In addition, the increase in WHO scores was positively correlated with the number of oral lesions (r = .521), the difficulty in eating (r = .250), and the overall oral health (r = .534; all p < .05). Conclusion The main factors affecting the severity of CIOM symptoms were the treatment with HSCT and the location of oral lesions. The incidence of CIOM and WHO scores were not significantly different between the subgroups of disease. Our findings will help clinicians investigate the oral findings after chemotherapy in adult patients with acute leukemia.Y

Immunohistochemical Characterization of the Human Sublingual Mucosa

The sublingual locus has recently received great attention as a delivery site for various immunotherapies, including those that induce allergen-specific tolerance, and for vaccines that generate protective immunity. To further understand the immune functions of the human sublingual mucosa, we characterized the distribution of various immunocytes therein by immunohistochemistry. We identified professional antigen presenting cells (APCs), including Langerhans cells (LCs) and macrophages. CD1a+ and langerin+ LCs were further found to be distributed in the basal and supra-basal layers of the epithelium, and macrophages were identified in the lamina propria. HLA-DR+ cells were observed in both the epithelium and the lamina propria, which mirrors the tissue distribution of LCs and macrophages within these tissues. CD3+, CD4+, and CD8+ T cells were found to be distributed along the basal layer of the epithelium and also in the lamina propria. Although B cells, plasma cells, and Foxp3+ regulatory T cells (Tregs) were only occasionally observed in the human sublingual mucosa in the absence of inflammation, they did show enrichment at inflammatory sites. Hence, we have further elucidated the immune cell component distribution in human sublingual mucosa

Mucosal and salivary microbiota associated with recurrent aphthous stomatitis

This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made.Abstract Background Recurrent aphthous stomatitis (RAS) is a common oral mucosal disorder of unclear etiopathogenesis. Although recent studies of the oral microbiota by high-throughput sequencing of 16S rRNA genes have suggested that imbalances in the oral microbiota may contribute to the etiopathogenesis of RAS, no specific bacterial species associated with RAS have been identified. The present study aimed to characterize the microbiota in the oral mucosa and saliva of RAS patients in comparison with control subjects at the species level. Results The bacterial communities of the oral mucosa and saliva from RAS patients with active lesions (RAS, n = 18 for mucosa and n = 8 for saliva) and control subjects (n = 18 for mucosa and n = 7 for saliva) were analyzed by pyrosequencing of the 16S rRNA genes. There were no significant differences in the alpha diversity between the controls and the RAS, but the mucosal microbiota of the RAS patients showed increased inter-subject variability. A comparison of the relative abundance of each taxon revealed decreases in the members of healthy core microbiota but increases of rare species in the mucosal and salivary microbiota of RAS patients. Particularly, decreased Streptococcus salivarius and increased Acinetobacter johnsonii in the mucosa were associated with RAS risk. A dysbiosis index, which was developed using the relative abundance of A. johnsonii and S. salivarius and the regression coefficients, correctly predicted 83 % of the total cases for the absence or presence of RAS. Interestingly, A. johnsonii substantially inhibited the proliferation of gingival epithelial cells and showed greater cytotoxicity against the gingival epithelial cells than S. salivarius. Conclusion RAS is associated with dysbiosis of the mucosal and salivary microbiota, and two species associated with RAS have been identified. This knowledge may provide a diagnostic tool and new targets for therapeutics for RAS