Multivariate discovery and replication of five novel loci associated with Immunoglobulin G <i>N</i>-glycosylation

Multivariate analysis methods can uncover the relationship between phenotypic measures characterised by modern omic techniques. Here the authors conduct a multivariate GWAS on IgG N-glycosylation phenotypes and identify 5 novel loci enriched in immune system genes

Changes to serum sample tube and processing methodology does not cause inter-individual variation in automated whole serum N-glycan profiling in health and disease

Serum N-glycans have been identified as putative biomarkers for numerous diseases. The impact of different serum sample tubes and processing methods on N-glycan analysis has received relatively little attention. This study aimed to determine the effect of different sample tubes and processing methods on the whole serum N-glycan profile in both health and disease. A secondary objective was to describe a robot automated N-glycan release, labeling and cleanup process for use in a biomarker discovery system.25 patients with active and quiescent inflammatory bowel disease and controls had three different serum sample tubes taken at the same draw. Two different processing methods were used for three types of tube (with and without gel-separation medium). Samples were randomised and processed in a blinded fashion. Whole serum N-glycan release, 2-aminobenzamide labeling and cleanup was automated using a Hamilton Microlab STARlet Liquid Handling robot. Samples were analysed using a hydrophilic interaction liquid chromatography/ethylene bridged hybrid(BEH) column on an ultra-high performance liquid chromatography instrument. Data were analysed quantitatively by pairwise correlation and hierarchical clustering using the area under each chromatogram peak. Qualitatively, a blinded assessor attempted to match chromatograms to each individual.There was small intra-individual variation in serum N-glycan profiles from samples collected using different sample processing methods. Intra-individual correlation coefficients were between 0.99 and 1. Unsupervised hierarchical clustering and principal coordinate analyses accurately matched samples from the same individual. Qualitative analysis demonstrated good chromatogram overlay and a blinded assessor was able to accurately match individuals based on chromatogram profile, regardless of disease status.The three different serum sample tubes processed using the described methods cause minimal inter-individual variation in serum whole N-glycan profile when processed using an automated workstream. This has important implications for N-glycan biomarker discovery studies using different serum processing standard operating procedures

Comparative Analysis of the Effects of Anti-IL-6 Receptor mAb and Anti-TNF mAb Treatment on CD4(+) T-cell Responses in Murine Colitis

Background: The efficacy of anti-tumor necrosis factor monoclonal antibody (anti-TNF mAb) for Crohn&apos;s disease (CD) is well established, and anti-interleukin-6 receptor (anti-IL-6R) mAb has also been reported to be effective in CD. It is, however, unclear if the efficacy and mechanisms of both agents are different in CD therapy. Methods: Using an adoptive transfer colitis model, we compared the efficacy of anti-IL-6R mAb, anti-TNF mAb, and TNF receptor-Fc fusion protein (TNFR-Fc), and their modes of action on CD4(+) T cells. We also investigated the role of Th1 and Th17 cells in colitis using the same model. Results: The histological scores for the anti-IL-6R mAb and anti-TNF mAb groups but not for TNFR-Fc group were much lower than that for the control group, and the score was the lowest for the anti-IL-6R mAb group. The frequency of proliferating CD4(+) T cells was reduced in anti-IL-6R mAb and anti-TNF mAb groups, but not in the TNFR-Fc group, whereas the frequency of apoptotic CD4(+) T cells was similar in all groups. Anti-IL-6R mAb suppressed the induction of Th17 cells and increased the frequency of lamina propria regulatory T cells, whereas anti-TNF mAb exerted no influence on CD4(+) T-cell differentiation. A deficiency in interferon-gamma and/or IL-17 in CD4(+) T cells reduced the severity of colitis. Conclusions: Our findings suggest that suppression of the proliferation of pathogenic CD4(+) T cells is the major mode of action of biological agents for colitis therapy. Anti-IL-6R mAb might have benefits in CD patients with Th17 dominance and impaired Treg frequency.119sciescopu

Supplementary Material for: Narrow Band Imaging with Magnifying Endoscopy for Peyer's Patches in Patients with Inflammatory Bowel Disease

<b><i>Background/Aims:</i></b> Peyer's patches (PPs) play a major role in mucosal immunity, but little is known about their alterations in patients with inflammatory bowel disease (IBD). We aimed to evaluate endoscopic changes of PPs in IBD patients using narrow band imaging with magnifying endoscopy (NBI-ME). <b><i>Methods:</i></b> Images of PPs using NBI-ME by ileocolonoscopy were consecutively collected. Existence of branch-like structures and the vessel occupancy in the dome lesions of PPs were analyzed. Appearance of the surrounding villi of the domes in PPs was evaluated using a ‘villi index' consisting of irregular formation, hyperemia, and altered vascular network pattern. Vascularity of PPs was immunohistologically analyzed by anti-CD34 antibody. <b><i>Results:</i></b> 17 patients with Crohn's disease (CD), 43 with ulcerative colitis (UC), and 23 healthy control subjects (HC) were analyzed. Both CD and UC patients had a high prevalence of having branch-like structures and significantly higher vascularity in the dome lesions than HC. The villi indices and vascular widths in the villi were significantly larger in CD and UC patients than in HC. <b><i>Conclusions:</i></b> Precise examination with NBI-ME characterized alteration of vascular structure in the dome and surrounding villi lesions of PPs not only in CD but also in UC patients