POS1429 ORAL DYSBIOSIS REFLECTS THE IMMUNOLOGICAL ALTERATION OF RA REGARDING TO HLA DRB1*SE, ACPA AND CIGARETTE SMOKING: NAGASAKI ISLAND STUDY

Abstract

Background:Anti-citrullinated protein antibody (ACPA) production is observed in several organs even prior to the onset of rheumatoid arthritis (RA), and oral mucosa is considered to be one of the important tissues. Saliva is considered to reflect the oral microbiota(oralMB) including periodontal disease. A gene-environment interaction between cigarette smoking and shared epitope genes in HLA-DRB1*shared epitope (SE) provides a high risk of ACPA-positive RA. However, the interaction of HLA-DRB1*SE, ACPA, cigarette smoking and oralMB of RA patients remains to be elucidated.Objectives:We investigated that the difference of oralMB among RA patients and healthy subjects(HS) regarding to ACPA, HLA-DRB1*SE and cigarette smoking.Methods:The Nagasaki Island Study, which had started in 2014 collaborating with Goto City, Nagasaki Prefecture, Japan, is intended for research of the preclinical stage of RA, including ACPA, HLA genotype screening, oralMB and lifestyle habit. Both of blood and salivary samples were obtained from 1422 subjects out of 4276 participants in this study from 2016 to 2018. ACPA positivity was 1.7 % in total 4276 subjects. At this point, we selected 291 subjects, who were ACPA positive non-RA HS(n=22) and patients with RA (n=33, 11 subjects were ACPA positive and 22 ACPA negative, respectively) as the case, age and gender matched ACPA negative non-RA HS (n=236) as the control. In RA subjects, current smoker was n=1(3.0%) and ever smoker was n=8(24.2%). In HS, current smoker was n=29(11.2%) and ever smoker was n=55(21.3%). ACPA was measured by ELISA, and HLA genotyping was quantified by next-generation sequencing (Ref.1). The operational taxonomic unit (OTU) analysis using 16S rRNA gene sequencing were performed. The richness of microbial diversity within subject (α-diversity) was scaled via Shannon entropy. The dissimilarity between microbial community composition was calculated using Bray-Curtis distance as a scale, and differences between groups (β-diversity) were tested by permutational multivariate analysis of variance (PERMANOVA). In addition, UniFrac distance calculated in consideration of the distance on the phylogenetic tree were performed.Results:Median age 71 y.o., % Female 58.4 %. Among RA and non-RA subjects, not α-diversity but β-diversity was statistically smaller significantly in RA (p=0.022). In the HS, there was no decrease in α-diversity between the ACPA-positive and HLA-DRB1*SE-positive groups, but in the ACPA-positive group, there was a decrease in α-diversity in the HLA-DRB1*SE-positive group. When we compared α-diversity stratified by the presence or absence of three factors (RA, ACPA, and HLA-DRB1*SE), the RA group with ACPA and HLA-DRB1*SE positive tended to have the lowest diversity (Figure 1 lower right). RA subjects, presence of HLA-DRB1*SE did not show the difference but the tendency of lower α-diversity (p=0.29).Conclusion:HS with ACPA-positive HLA-DRB1*SE tended to show lower α-diversity than ACPA-positive HS and HLA-DRB1*SE positive HS. Furthermore, RA subjects with ACPA-positive HLA-DRB1*SE showed lower α-diversity than HS with ACPA-positive HLA-DRB1*SE.Our study suggested that the oral dysbiosis may reflect the immunological status of patients with RA. Because of the small number of ACPA-positive patients, stratification by smoking history was difficult. Further examination is needed to clarify the gene-environment interaction and microbiome.