HLA-B27 transgenic animals have significant alterations in gut microbiota by 16s rDNA sequencing.

<p>WT: wild-type Lewis rats; B27/B2M: (21–3×283–2)F1 HLA-B27/hβ2m transgenic Lewis rats; B2M: 283–2 hβ2m expressing transgenic rats; * designates p<0.05.</p

Genus level analysis differences from A, cecal lumen specimens, and B, cecal mucosal samples from cohort 3.

<p>Genus level analysis differences from A, cecal lumen specimens, and B, cecal mucosal samples from cohort 3.</p

Relative differences in <i>Bacteroides vulgatus</i> and <i>Akkermansia muciniphila</i> shown by A, quantitative PCR using the 16S rRNA gene as a reference, and B, by traditional PCR using whole cecum samples in cohort 1.

<p>Relative differences in <i>Bacteroides vulgatus</i> and <i>Akkermansia muciniphila</i> shown by A, quantitative PCR using the 16S rRNA gene as a reference, and B, by traditional PCR using whole cecum samples in cohort 1.</p

Rat cohorts used in experiments.

<p>WT: wild type; see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0105684#pone-0105684-t001" target="_blank">Table 1</a> for genotype designations.</p

Principal coordinates analysis from combined cecal lumen and cecal mucosa samples from cohort 3.

<p>Principal coordinates analysis from combined cecal lumen and cecal mucosa samples from cohort 3.</p

HLA-B27/hβ2m transgenic rat lines.

<p>Adapted from <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0105684#pone.0105684-Taurog2" target="_blank">[35]</a>.</p

16S rRNA gene sequencing principal coordinates analysis from non-co-housed rats shows significant differences in cecal microbiota between genotypes.

<p>A. Cecal lumen samples from cohort 3; B, Cecal mucosa samples from cohort 3; C, Phylum-level analysis from cecum lumen samples; D, Phylum-level analysis from cecum mucosa samples. WT: wild type rats; B2M: hβ2 microglobulin; B27/B2M: HLA-B27/hβ2 microglobulin transgenic rats.</p

BRISK analysis of whole cecum samples from cohort 1 shows microbial diversity differences on a species level between HLA-B27/hβ2m animals and co-housed wild type animals.

<p>A. Each line on the heat map represents a different bacterial species and the color code represents the relative abundance of that organism, with yellow indicating the highest abundance. B, Specific species differences are highlighted, with a black dot indicating statistically significant differences.</p

IgG4 Immunostaining and Its Implications in Orbital Inflammatory Disease

<div><p>Objective</p><p>IgG4-related disease is an emerging clinical entity which frequently involves tissue within the orbit. In order to appreciate the implications of IgG4 immunostaining, we analyzed gene expression and the prevalence of IgG4- immunostaining among subjects with orbital inflammatory diseases.</p><p>Methods</p><p>We organized an international consortium to collect orbital biopsies from 108 subjects including 22 with no known orbital disease, 42 with nonspecific orbital inflammatory disease (NSOI), 26 with thyroid eye disease (TED), 12 with sarcoidosis, and 6 with granulomatosis with polyangiitis (GPA). Lacrimal gland and orbital adipose tissue biopsies were immunostained for IgG4 or IgG secreting plasma cells. RNA transcripts were quantified by Affymetrix arrays.</p><p>Results</p><p>None of the healthy controls or subjects with TED had substantial IgG4 staining. Among the 63 others, the prevalence of significant IgG4-immunostaining ranged from 11 to 39% depending on the definition for significant. IgG4 staining was detectable in the majority of tissues from subjects with GPA and less commonly in tissue from subjects with sarcoidosis or NSOI. The detection of IgG4+ cells correlated with inflammation in the lacrimal gland based on histology. IgG4 staining tissue expressed an increase in transcripts associated with inflammation, especially B cell-related genes. Functional annotation analysis confirmed this.</p><p>Conclusion</p><p>IgG4+ plasma cells are common in orbital tissue from patients with sarcoidosis, GPA, or NSOI. Even using the low threshold of 10 IgG4+ cells/high powered field, IgG4 staining correlates with increased inflammation in the lacrimal gland based on histology and gene expression.</p></div

Case demographics.

<p>*Ages are not available for 4 subjects.</p><p>Case demographics.</p