Carbohydrate Antibodies Research Articles

First-in-human Study With LIS1, a Next-generation Porcine Low Immunogenicity Antilymphocyte Immunoglobulin in Kidney Transplantation.

Polyclonal rabbit antithymocyte globulins (ATGs) are commonly used in organ transplantation as induction. Anti- N -glycolylneuraminic acid carbohydrate antibodies which develop in response to rabbit carbohydrate antigens might lead to unwanted systemic inflammation. LIS1, the first new generation of antilymphocyte globulins (ALGs) derived from double knockout swine, lacking carbohydrate xenoantigens was already tested in nonhuman primates and rodent models. This open-label, single-site, dose escalation, first-in-human, phase 1 study evaluated the safety, T cell depletion, pharmacokinetics, and pharmacodynamics of LIS1. In an ascending dose cohort (n = 5), a primary kidney transplant recipient at low immunologic risk (panel reactive antibody [PRA] < 20%), received LIS1 for 5 d at either 0.6, 1, 3, 6, or 8 mg/kg. After each patient completed treatment, the data safety monitoring board approved respective dose escalation. In the therapeutic dose cohort (n = 5) in patients with PRA <50% without donor specific antibodies, 2 patients received 8 mg/kg and 3 patients 10 mg/kg. CD3 + T cell depletion <100/mm 3 at day 2 was observed in all patients who received 6, 8, and 10 mg/kg of LIS1. The terminal half-life of LIS1 was 33.7 d with linearity in its disposition. Lymphocyte repopulation was fast and pretransplant lymphocyte subpopulation counts recovered within 2-4 wk. LIS1 was well tolerated, neither cytokine release syndrome nor severe thrombocytopenia or leukopenia were noticed. Antibodies to LIS1 were not detected. In this first-in-human trial, genome-edited swine-derived polyclonal LIS1 ALG was well tolerated, did not elicit antidrug antibodies, and caused time-limited T cell depletion in low- and medium-risk kidney transplant recipients.

Altered properties and structures of root exudate polysaccharides in a root hairless mutant of barley.

Root exudates and rhizosheaths of attached soil are important features of growing roots. To elucidate factors involved in rhizosheath formation, wild-type (WT) barley (Hordeum vulgare L. cv. Pallas) and a root hairless mutant, bald root barley (brb), were investigated with a combination of physiological, biochemical, and immunochemical assays. When grown in soil, WT barley roots bound ∼5-fold more soil than brb per unit root length. High molecular weight (HMW) polysaccharide exudates of brb roots had less soil-binding capacity than those of WT root exudates. Carbohydrate and glycan monoclonal antibody analyses of HMW polysaccharide exudates indicated differing glycan profiles. Relative to WT plants, root exudates of brb had reduced signals for arabinogalactan-protein (AGP), extensin, and heteroxylan epitopes. In contrast, the root exudate of 2-week-old brb plants contained ∼25-fold more detectable xyloglucan epitope relative to WT. Root system immunoprints confirmed the higher levels of release of the xyloglucan epitope from brb root apices and root axes relative to WT. Epitope detection with anion-exchange chromatography indicated that the increased detection of xyloglucan in brb exudates was due to enhanced abundance of a neutral polymer. Conversely, brb root exudates contained decreased amounts of an acidic polymer, with soil-binding properties, containing the xyloglucan epitope and glycoprotein and heteroxylan epitopes relative to WT. We, therefore, propose that, in addition to physically structuring soil particles, root hairs facilitate rhizosheath formation by releasing a soil-binding polysaccharide complex.

Association between Neu5Gc carbohydrate and serum antibodies against it provides the molecular link to cancer: French NutriNet-Sant\xe9 study

BackgroundHigh consumption of red and processed meat is commonly associated with increased cancer risk, particularly colorectal cancer. Antibodies against the red meat-derived carbohydrate N-glycolylneuraminic acid (Neu5Gc) exacerbate cancer in “human-like” mice. Human anti-Neu5Gc IgG and red meat are both independently proposed to increase cancer risk, yet how diet affects these antibodies is largely unknown.MethodsWe used world global data to demonstrate that colorectal cancer incidence and mortality are associated with increased national meat consumption. In a well-defined large cohort, we used glycomics to measure daily Neu5Gc intake from red meat and dairy, and investigated serum as well as affinity-purified anti-Neu5Gc antibodies. Based on 24-h dietary records, daily Neu5Gc intake was calculated for 19,621 subjects aged ≥ 18 years of the NutriNet-Santé study. Serum and affinity-purified anti-Neu5Gc antibodies were evaluated by ELISA and glycan microarrays in representative 120 individuals, each with at least eighteen 24-h dietary records (aged 45–60, Q1–Q4; aged > 60, Q1 and Q4; 10 men/women per quartile).ResultsWe found that high-Neu5Gc diet, gender, and age affect the specificity, levels, and repertoires of anti-Neu5Gc IgG immune responses, but not their affinity. Men consumed more Neu5Gc than women, mostly from red meat (p = 0.0015), and exhibited higher overall serum anti-Neu5Gc IgG levels by ELISA (3.94 ng/μl versus 2.22 ng/μl, respectively; p = 0.039). Detailed glycan microarray analysis against 56 different glycans revealed high Neu5Gc-specificity with increased anti-Neu5Gc IgG and altered repertoires, associated with higher consumption of Neu5Gc from red meat and cow dairy. Affinity purification of serum anti-Neu5Gc antibodies revealed increased levels and biased array repertoire patterns, without an increase in antibody affinity, in individuals consuming higher Neu5Gc levels. Furthermore, in a high-meat diet, antibody diversity patterns on glycan microarrays shifted towards Neu5Gcα3-linked glycans, increasing the α3/α6-glycans ratio score.ConclusionsWe found a clear link between the levels and repertoire of serum anti-Neu5Gc IgG and Neu5Gc intake from red meat and dairy. These precise rational methodologies allowed to develop a Gcemic index to simplify the assessment of Neu5Gc in foods that could potentially be adapted for dietary recommendations to reduce cancer risk.

Identification and validation of novel circulating biomarkers for early diagnosis of lung cancer

ObjectivesThis study aimed to identify novel circulating biomarkers in lung cancer. Materials and methodsExpression of 1000 secreted proteins in 15 early lung cancer patients and 10 healthy controls’ plasma were examined by RayBiotech antibody array. Candidate biomarkers were identified by SPSS analysis (p < 0.05 between lung cancer and healthy controls was considered significant) and validated by ELISA in 371 lung cancer patients and 388 healthy controls. Receive operating characteristic curve and binary logistic regression were performed to evaluate the diagnosis efficacy and to establish diagnostic models. Results and conclusionTwenty-two molecules expressed aberrantly in early lung cancer patients compared to healthy controls. Large sample validation showed significantly up-regulated levels of brain angiogenesis inhibitor 1, E-Cadherin, integrin-binding sialoprotein, and down-regulated expression of thrombospondin-1 (all p < 0.0001) in lung cancer patients. Receive operating characteristic curve analysis indicated E-Cadherin, brain angiogenesis inhibitor 1 and thrombospondin-1 had higher sensitivity and specificity than classical biomarkers carcinoenbryonic antigen, carbohydrate antibody 19-9 and cytokeratin 19 fragments. The high sensitivity and specificity of E-Cadherin brain angiogenesis inhibitor 1 and thrombospondin-1 were also confirmed in early lung cancer analysis. Combination analysis showed brain angiogenesis inhibitor 1, E-Cadherin and thrombospondin-1 had better diagnostic efficacy than classic lung cancer biomarkers. Our findings demonstrated the potential status of BAI-1, E-Cadherin and TSP-1 in diagnosis of lung cancer.

A Hybrid Approach Enabling Large-Scale Glycomic Analysis of Post-Golgi Vesicles Reveals a Transport Route for Polysaccharides.

The plant endomembrane system facilitates the transport of polysaccharides, associated enzymes, and glycoproteins through its dynamic pathways. Although enzymes involved in cell wall biosynthesis have been identified, little is known about the endomembrane-based transport of glycan components. This is partially attributed to technical challenges in biochemically determining polysaccharide cargo in specific vesicles. Here, we introduce a hybrid approach addressing this limitation. By combining vesicle isolation with a large-scale carbohydrate antibody arraying technique, we charted an initial large-scale map describing the glycome profile of the SYNTAXIN OF PLANTS61 (SYP61) trans-Golgi network compartment in Arabidopsis (Arabidopsis thaliana). A library of antibodies recognizing specific noncellulosic carbohydrate epitopes allowed us to identify a range of diverse glycans, including pectins, xyloglucans (XyGs), and arabinogalactan proteins in isolated vesicles. Changes in XyG- and pectin-specific epitopes in the cell wall of an Arabidopsis SYP61 mutant corroborate our findings. Our data provide evidence that SYP61 vesicles are involved in the transport and deposition of structural polysaccharides and glycoproteins. Adaptation of our methodology can enable studies characterizing the glycome profiles of various vesicle populations in plant and animal systems and their respective roles in glycan transport defined by subcellular markers, developmental stages, or environmental stimuli.

Starch Consumption May Modify Antiglycan Antibodies and Fecal Fungal Composition in Patients With Ileo-Anal Pouch.

Inflammatory bowel diseases (IBDs) are characterized by serologic responses to glycans. Patients with ulcerative colitis (UC) after proctocolectomy with ileo-anal anastomosis (pouch surgery) may develop inflammation (pouchitis) that resembles Crohn's disease (CD). We hypothesized that patients' serologic responses were affected by their consumption of dietary sugars. This study analyzed the correlations between antiglycan antibody expression and dietary sugar consumption in patients with UC pouch and the evolution in antibody levels over time. Patients were followed prospectively for 2 consecutive visits. The following antiglycan carbohydrate antibodies were detected by enzyme-linked immunosorbent assay: antichitobioside (ACCA), antilaminaribioside (ALCA), antimannobioside (AMCA), and anti-Saccharomyces cerevisiae (ASCA) antibodies. Patients completed a food frequency questionnaire. The fungal community in patients' fecal samples was analyzed by sequencing the internal transcribed spacer 2 (ITS2) region of nuclear ribosomal DNA. We included 75 UC pouch patients aged 45.2 ± 14 years who underwent pouch surgery 9.8 ± 6.7 years previously. Of these patients, 34.7% (n = 26) showed seropositivity for antiglycan antibodies. Starch consumption was significantly higher in patients with positive serologic responses (P = 0.05). Higher starch consumption was associated with higher AMCA and ACCA titers, which increased by 4.08% (0.8%-7.4%; P = 0.014) and 4.8% (0.7%-9.1%; P = 0.007), respectively, for each 10-g increase of dietary starch. The per-patient change in the relative abundance of Candida albicans in fecal samples correlated positively with changes in starch consumption (Spearman's r = 0.72; P = 0.012). Starch consumption correlated with positive antiglycan serology (ACCA and AMCA), suggesting that increased dietary starch intake may promote a specific immune response in patients with IBD.

Diagnostic utility of serological biomarkers in patients with Crohn's disease: A case-control study.

The aim of this study was to examine the expression of serological markers in patients with inflammatory bowel disease in China, and determine the diagnostic utility of serological markers, individually and in combination, for the diagnosis and differential diagnosis of Crohn's disease (CD).Serum samples were obtained from 160 participants in Eastern China. Among the participants, 98 were diagnosed with CD, 33 had ulcerative colitis (UC), and 29 were healthy controls (HC). The serum samples were tested for the presence of antibodies against outer membrane porin C (anti-OmpC), Pseudomonas fluorescens bacterial sequence I2 (anti-I2), anti-laminarin (anti-L), anti-chitin (anti-C), anti-chitobioside carbohydrate antibody (ACCA), anti-laminaribioside carbohydrate antibody (ALCA), anti-mannobioside carbohydrate antibody (AMCA), and anti-Saccharomyces cerevisiae antibody (ASCA) by indirect enzyme-linked immunosorbent assay (ELISA).Individually, anti-C, anti-L, ASCA-IgG, and ALCA lacked diagnostic value in the differentiation of CD. ASCA-IgA remained the most accurate marker for the diagnosis of CD, with an area under the curve (AUC) of 0.77; however, its sensitivity and specificity were both lower than 75%. Among the combinations of the 5 markers with significant diagnosing ability for CD, combinations with any 2 of the 3 markers, ASCA IgA, AMCA, and ACCA positive, provided the best accuracy in the diagnosis and differential diagnosis of CD (sensitivity and specificity both above 75%) and had the highest Youden index.Serological antibodies, when considered in combination, have remarkable value in the diagnosis and differential diagnosis of CD. Especially, the combination of any 2 of the 3 markers, ASCA-IgA, AMCA, ACCA positive, appears to be optimal.

Anticarbohydrate antibodies in patients with inflammatory bowel disease.

Evaluating antilaminaribioside carbohydrate antibodies (ALCA), antimannobioside carbohydrate antibodies (AMCA) and anti-Saccharomyces cerevisiae antibodies (ASCA), in patients with inflammatory bowel disease (IBD). Subjects and methods: 268 serum samples were used; 115 Crohn’s disease (CD), 83 ulcerative colitis, and 70 healthy control samples. All samples were evaluated using enzyme-linked immunosorbent assay for the following four anticarbohydrate antibodies: ACCA, ALCA, AMCA, and ASCA. Results: In patients with Crohn’s disease the prevalence of the anticarbohydrate antibodies was: ASCA 69%, AMCA 32%, ACCA 28% and ALCA 24% with the highest prevalence being for ASCA (P-value<0.0001) while in patients with ulcerative colitis the prevalence was: ACCA 46%, AMCA 35%, ALCA 23% and ASCA 15% with the highest prevalence being for ACCA (P- value<0.001). Conclusion: Anticarbohydrate antibodies are significantly present in patients with IBD. The use of a panel of anticarbohydrate antibodies may provide additional help in distinguishing IBD from non-IBD disease patterns and narrow the range of differential diagnosis in these patients.

Value of serological marker detection in hierarchical management of Crohn′s disease

Objective To explore the values of serological markers in the phenotype diagnosis of Crohn′s disease (CD). Methods From January 2015 to December 2016, 100 patients diagnosed as CD were enrolled. The levels of human anti-Saccharomyces cerevisiae antibody (ASCA) IgG, ASCA IgA, anti-mannobioside carbohydrate antibody (AMCA), anti-chitobioside carbohydrate antibody (ACCA), antibody against outer membrane porin C of Escherichia coli (anti-OmpC), antibody against pseudomonas fluorescens (anti-I2), flagella protein antibody, perinuclear anti-neutrophil cytoplasmic antibodies (pANCA) and antibodies to proteinase-3 (anti-PR3) were detected by indirect enzyme-linked immunosorbent assay (ELISA). Binary Logistic regression analysis was used to analyze the correlation between the serological markers and the phenotype of CD. The diagnosis values were evaluated by receiver operating characteristic (ROC) curve. Results The level of anti-I2 was significantly correlated with the narrow type of patients with CD (odd ratios (OR)=34.304, P=0.011); ACCA and flagella protein antibody were significantly correlated with the penetrating type of patients with CD (OR=1.024, P=0.027; OR=2.702, P=0.021). ASCA IgA and AMCA were significantly correlated with the lesion involved gastric, duodenum and small intestine in CD patients (OR=1.146, P=0.044; OR=1.035, P=0.013). In the diagnosis of narrow type CD, the diagnostic accuracy of anti-I2 was the highest (area under curve (AUC)=0.631, P=0.043), and according to this result, its best cutoff value was 0.650, and then the sensitivity increased to 28.6% (8/28). In the diagnosis of penetrating type CD, the diagnostic accuracy of flagella protein antibody was the highest (AUC=0.725, P<0.01). Anti-OmpC (AUC=0.677, P=0.006) and ACCA (AUC=0.644, P=0.025) also had higher diagnostic accuracy. According to these results, the best cutoff values of flagella protein antibody, anti-OmpC and ACCA were 0.395, 0.225 and 59.500, respectively, and then their sensitivity improved after correcting the cutoff values. The AUC of any one item positive in the combination of flagella protein antibody, anti-OmpC and ACCA in the diagnosis of penetrating type CD was 0.761 (P<0.01), which was significantly higher than the diagnostic accuracy of single antibody detection. In the diagnosis of the lesions involved gastric, duodenum and small intestine, the diagnostic accuracy of AMCA was the highest, and its AUC was 0.817 (P<0.01). ASCA IgA (AUC=0.772, P=0.001) and ACCA (AUC=0.752, P=0.001) also had higher diagnostic accuracy. The AUC of any one item positive of ASCA IgA and AMCA in the diagnosis of CD lesions involved gastric, duodenum and small intestine was 0.905 (P<0.01), which was significantly higher than the diagnostic accuracy of single antibody detection. The best cutoff values of ASCA IgA, AMCA and ACCA were 21.500, 55.000 and 29.500, respectively. And their sensitivity improved after correcting the cutoff values. The sensitivity of any one item positive in the three items also improved. Conclusion The detection of serological markers has great clinical values in the hierarchical management of phenotypes in Chinese CD population. Key words: Crohn disease; Enzyme-linked immunosorbent assay; Serological markers; CD phenotype

The value of serum antibodies in differentiating inflammatory bowel disease, predicting disease activity and disease course in the newly diagnosed patient

Background: Data on serum antibodies in untreated adult inflammatory bowel disease (IBD) patients at diagnosis are scarcely available, and results on the stability of antibody presence over time are inconsistent. Our aim was to investigate antibodies in newly diagnosed, untreated IBD patients in relation to disease phenotype and course. Furthermore, we analyzed antibody presence over time.Methods: Baseline anti-Saccharomyces cerevisiae antibodies (ASCA), anti-chitobioside carbohydrate antibodies (ACCA), anti-laminaribioside carbohydrate antibodies (ALCA) and anti-mannobioside carbohydrate antibodies (AMCA) were measured with enzyme-linked immunosorbent assays and perinuclear anti-neutrophilic cytoplasmic antibodies (pANCA) was measured by indirect immunofluorescence in serum of 120 untreated IBD patients at diagnosis and 19 healthy controls. Antibodies were related to disease outcomes. Serial measurements were available in 71 patients.Results: The combination of pANCA and ASCA enabled good discrimination between UC and CD (p = .004). Antibody presence was relatively stable over time, even though there were significant changes in concentrations. There was a trend towards larger fluctuations in concentration with immunosuppressive medication. Baseline pANCA in UC patients correlated with calprotectin values (rho = .545, p = .019) and change in pANCA status over time was associated with disease activity at that moment. No associations were found with antibodies at diagnosis and disease outcomes.Conclusion: Antibody profiles at diagnosis support the distinction between CD and UC. Anti-glycan antibodies are reasonably stable over time, but may fluctuate under the influence of immunosuppressive treatment which may explain the inconsistency in findings hitherto. The appearance or disappearance of pANCA antibodies during follow-up correlated with disease activity in UC and may be used in disease monitoring.

Humoral Immune Reconstitution Kinetics after Allogeneic Hematopoietic Stem Cell Transplantation in Children: A Maturation Block of IgM Memory B Cells May Lead to Impaired Antibody Immune Reconstitution

Although T cell immune reconstitution after allogeneic hematopoietic stem cell transplantation (allo-HSCT) has been well studied, long-term B cell immune reconstitution remains less characterized. We evaluated humoral immune reconstitution among 71 pediatric allo-HSCT recipients. Although tetanus toxoid antibody levels were normal at 1 year after allo-HSCT, antipolysaccharide carbohydrate antibodies remained persistently low for up to 5 years. While naive B cell counts normalized by 6 months, IgM memory B cell deficiency persisted for up to 2 years (P = .01); switched memory B cell deficiency normalized by 1 year after allo-HSCT. CD4+ T cell immune reconstitution correlated with that of switched memory B cells as early as 6 months after allo-HSCT (r = .55, P = .002) but did not correlate with IgM memory B cells at any time point after allo-HSCT. Taken together, this suggests that allo-HSCT recipients have impaired antibody immune reconstitution, mainly due to IgM memory B cell maturation block, compared with more prompt T cell–dependent switched memory cell immune reconstitution. We further explored other factors that might affect humoral immune reconstitution. The use of total body irradiation was associated with lower naive B cells counts at 6 months after HSCT (P = .04) and lower IgM (P = .008) and switched (P = .003) memory B cells up to 2 years. Allo-HSCT recipients with extensive chronic graft-versus-host disease had lower IgM memory B cell counts (P = .03) up to 2 years after allo-HSCT. The use of cord blood was associated with better naive (P = .01), IgM (P = .0005), and switched memory (P = .006) B cells immune reconstitution. These findings may inform future prophylaxis and treatment strategies regarding risk of overwhelming infection, graft-versus-host disease, and post-allogeneic HSCT revaccination.

Structural Insights into Reovirus σ1 Interactions with Two Neutralizing Antibodies.

Virus attachment proteins mediate binding to host cell receptors, serve critical functions in cell and tissue tropism, and are often targeted by the neutralizing antibody response. The structural investigation of antibody-antigen complexes can provide valuable information for understanding the molecular basis of virus neutralization. Studies with enveloped viruses, such as HIV and influenza virus, have helped to define sites of vulnerability and guide vaccination strategies. By comparison, less is known about antibody binding to nonenveloped viruses. Here, we structurally investigated two neutralizing antibodies that bind the attachment protein σ1 of reovirus. Furthermore, we characterized the neutralization efficiency, the binding affinity for σ1, and the effect of the antibodies on reovirus receptor engagement. Our analysis defines reovirus interactions with two neutralizing antibodies, allows us to propose a mechanism by which they block virus infection, and provides evidence for a conformational change in the σ1 protein during viral cell entry.

Latest developments in Semantic Web technologies applied to the glycosciences

Summary The Integrated Life Science Database Project of Japan funded a group of glycoscientists to carry out a project to integrate glycoscience databases using Semantic Web technologies. As a continuation of the previous project period, the Japan Consortium for Glycobiology and Glycotechnology Database (JCGGDB) developed several glycoscience-related databases. The GlycoProtDB database is among those being integrated, providing an important resource to understand protein glycosylation. Another database being integrated is GlycoEpitope, a comprehensive database of carbohydrate epitopes and antibodies. In the current project period, we started the development of GlyTouCan, the international glycan structure repository providing unique accession numbers to all glycan structures. Although such databases are sufficiently important in and of themselves, their integration with other—omics data such as the protein information in UniProt will be crucial to bring glycosciences to the forefront of life sciences. However, to integrate such disparate sets of data among different fields in a way such that future maintenance costs are minimal, standardized ontologies and formats must be established. Our latest project has attempted to define the minimal standards that are necessary to enable this integration. The technical challenges to integrate all these databases and the technologies to overcome these challenges will be described.

Characterization of Carbohydrate Surface Markers on Mouse Embryonic Stem Cells

Glycosylation of proteins and lipids on cell surface have been shown to be important in maintaining pluripotency and stem cell fate in embryonic stem cells. Lectins have been widely used to characterize carbohydrate modifications on cell surface of embryonic stem cells to determine pluripotency and stem cell fate. In the present study, a panel of 14 lectins and carbohydrate antibodies was used to characterize the carbohydrate surface markers of mouse Embryonic Stem (ES) Cells. SSEA-1-positive mouse ES cells were firstly enriched and the carbohydrate profile of the cells was determined by flow cytometry and immunocytochemistry. Enrichment of mouse ES cells yielded approximately 99.95 ± 0.87% of SSEA-1-positive mouse ES cells. A uniform and high percentage of binding was observed for PNA, DSL, JAC, GNL, PSA and LTL, with PNA, DSL, JAC and GNL having similar percentage of binding to SSEA-1 (99.9%), while PSA and LTL binding were approximately 95%-99%. Partial binding of WFL, SNA and AAL were observed in mouse ES cells which was also reflected by the respective immunocytochemistry images. A very low percentage of binding was observed for MAA and UEAI. The data showed that high expression of mannose, N-acetyllactosamine and galactose are present on the cell surface of mouse ES cells. Some reliable surface markers that can be used to determine pluripotency are PNA, DSL, JAC and GNL, which showed similar binding to SSEA-1, a well-established pluripotent marker. Taken together, the data has provided information on the cell surface carbohydrate profile of mouse ES cells.

Serology of Patients with Ulcerative Colitis After Pouch Surgery Is More Comparable with that of Patients with Crohn's Disease.

The serologic status of patients with ulcerative colitis (UC) who develop postoperative pouchitis was compared with that of patients with Crohn's disease (CD) and unoperated patients with UC. Pouch patients were stratified into normal pouch, acute/recurrent acute pouchitis, and chronic pouchitis/Crohn's-like disease of the pouch groups. Antibodies against glycans associated with CD (anti-Saccharomyces cerevisiae, anti-laminaribioside, anti-chitobioside, and anti-mannobioside carbohydrate antibodies [ASCA, ALCA, ACCA, and AMCA, respectively]) were detected and correlated with type of inflammatory bowel disease and pouch behavior. A total of 501 patients with inflammatory bowel diseases were recruited: 250 (50%) CD, 124 (24.7%) unoperated UC, and 127 (25.3%) UC-pouch. At least 1 positive antibody was detected in 77.6% CD, 52.0% UC-pouch and 33.1% unoperated UC (P < 0.0001 for all). ACCA and AMCA prevalence in CD, UC-pouch and unoperated patients with UC were 33.2%, 24.4%, and 16.9% (P = 0.003 for all) and 35.2%, 26.8%, and 7.3%, respectively (P < 0.0001 for all). ALCA and ASCA were more prevalent in patients with CD than unoperated UC and UC-pouch patients. A longer interval since pouch surgery was associated with inflammatory pouch behavior: 12.45, 11.39, and 8.5 years for acute/recurrent acute pouchitis, chronic pouchitis/Crohn's-like disease of the pouch, and normal pouch, respectively, P = 0.01 for all. The prevalence of the CD-associated anti-glycan antibodies ACCA and AMCA is significantly increased in UC-pouch patients, suggesting that pouch surgery may trigger differential immune responses to glycans. The finding that the serology of UC-pouch patients shares similarities with that of patients with CD supports the notion that those 2 inflammatory bowel diseases share a common pathogenic pathway.

Association of Anti-glycan Antibodies and Inflammatory Bowel Disease Course.

The usefulness of anti-glycan antibodies alone or combined with anti-Saccharomyces cerevisiae [ASCA] or perinuclear antineutrophil cytoplasmic [pANCA] antibodies for diagnosis of inflammatory bowel disease [IBD], differentiation between Crohn's disease [CD] and ulcerative colitis [UC], disease stratification including IBD phenotype, and also for determination of the course of the disease, remain unclear. A large panel of serological anti-glycan carbohydrate antibodies, including anti-mannobioside IgG antibodies [AMCA], anti-chitobioside IgA [ACCA], anti-laminaribioside IgG antibodies [ALCA], anti-laminarin [anti-L] and anti-chitine [anti-C] were measured in the serum from a cohort of 195 patients with IBD] [107 CD and 88 UC]. The respective accuracy of isolated or combined markers for diagnosis, disease differentiation, stratification disease phenotype, and severity of the disease course, defined by a wide panel of criteria obtained from the past medical history, was assessed. The positivity of at least one anti-glycan antibody was detected in a significant higher proportion of CD and UC compared with healthy controls [p < 0.0001 and p < 0.0007, respectively]. Whereas ASCA and ANCA antibody status had the highest efficacy to be associated with CD in comparison with UC (area under receiver operating characteristic curve [AUROC] = 0.70 for each], the adjunction of anti-laminarin antibody substantially improved the differentiation between CD and UC [AUROC = 0.77]. Titres of ACCA [> 51U/ml] and anti-laminarin [> 31U/ml] were significantly linked with a higher association with steroid dependency (odds ratio [OR] =2.0 [1.0-4.0], p = 0.03 and OR = 2.4 [1.1-5.2], p = 0.02, respectively]. We further defined the respective performance of anti-glycan antibodies to discriminate between patients with severe or not severe CD and UC course and determined the associated optimal cut-off values: severe CD course was significantly more likely in case of AMCA > 77U/ml [OR = 4.3; p = 0.002], ASCA > 63U/ml [OR = 3.5; p < 0.009] and at a lesser degree ACCA > 50U/ml [OR = 2.8; p < 0.02] and severe UC course was significantly associated with AMCA > 52U/ml [OR = 3.4; p = 0.04] and ACCA > 25U/ml [OR = 3.0; p < 0.04]. Anti-glycan antibodies are valuable serological markers, especially AMCA antibodies that may help clinicians to promptly classify patients into high risk for severe disease.

Present Status of ABO Incompatible Kidney Transplantation in Japan-Lessons From More Than 2,400 Cases to Date-.

Introduction: Since the first successful in January 1989, we have performed more than 2,400 ABO incompatible kidney transplantation(ABOIKT) to date.and it accounts for 30% of annual living kidney transplants in Japan. Purpose of this study: From the latest registry, we have reviewed the long term patient and graft survival, and analysed the factors which affected the outcome. Patients and Methods: 2,434 cases were enrolled in this study. Observation period was median 3 years 5 months( 1 day to 23 years 7 months) Average recipient age was 43.7 years. Blood type O accounts for 52%, type A for 25% and type B for 23%. HLA-AB and DR mismatches were 2.13±1.07 and 1.08± 0.65. Results: Overall patient survival rate at 1,5,10,15 and 20 years are 97, 93, 90, 83 and 73%, and graft survival rate are 94, 86, 71, 59, and 52%, respectively. Since 2001, the success rate has dramatically improved that patient survival rate at 1,5,10 years are 97, 95 and 93%, and graft survival rate are 93, 90 and 81%, respectively. 1,147 among 1,672 (68.6%)cases avoided splenectomy after 2001. Before 2000 era, graft survival rate of anti-A/B IgG antibody titer 64 folds and greater group were significantly worse than those with less than 32 folds, however, the difference disappered after 2001 era. Discussion: In the recent decade, we have elucidated the phenomenon of accommodation, and established the strategy of its induction by desensitization in ABOIKT. We have utilized the combination of CNI,MMF, steroids, anti-CD25 mAbs and rituximab which enabled to achieve the accommodation without splenectomy. In our study, the patient and graft outcome had dramatically improved even in patients wirh high preoperative anti A/B antibody titer groups after 2001 era, which was indicated as high risk group for AMR and poor graft outcome before 2000 era. We have really overcome the immunological barrier of ABO-histo-blood broup carbohydrate antigen and antibody in the field of kidney transplantation. Conclusion: ABO-IKTx has become accepted as a therapeutic alternative for end-stage kidney disease. The complete elucidation of the accommodation induction mechanism in near future will open the next door to the upcoming brand-new paradigm of the field of organ transplantation.

Study on joint detection of serological markers in the diagnosis of Crohn's disease

Objective To explore the values of detecting anti-Saccharomyces cerevisiae antibody (ASCA) IgA,ASCA IgG,anti laminaribioside carbohydrate antibody (ALCA),anti-chitobioside carbohydrate antibody (ACCA) and anti mannobioside carbohydrate antibody (AMCA) and antibody against outer membrane porin C of Escherichia coli (anti-OmpC) in the diagnosis of Crohn's disease (CD).Methods From October 10th 2012 to January 24th 2013,63 patients diagnosed as CD were selected.At the same period 30 patients diagnosed as ulcerative colitis (UC) were set as disease control group and 25 individuals without abnormal findings under endoscope and imaging examination were set as healthy control group.Fasting blood of all the subjects were drawn in the morning and serum was isolated after centrifuge.The level of ASCA IgG,ALCA,AMCA,ACCA,ASCA IgA,anti OmpC and I2 was detected by indirect enzyme linked immunosorbent assay.The sensitivity,specificity,positive predictive value and negative predictive value of each antibody were calculated and its value in the diagnosis was evaluated by receiver operating characteristic (ROC) curve.Results In the differential diagnosis of CD and UC by single antibody test,the accuracy of AMCA was the highest and the area under ROC curve was 0.739.In the screening of CD with single antibody examination,the diagnosis accuracy of I2 antibody was the highest and the area under ROC curve was 0.751.In multi antibodies joint examination,the accuracy of either ASCA IgA or AMCA positive was the highest in the differential diagnosis of CD and UC,also in the screening of CD and the area under ROC curve was 0.612 and 0.633,respectively.Conclusion In clinical practice,CD screening and the differential diagnosis of CD and UC may be achieved through detecting AMCA,ASCA IgA and I2 antibody. Key words: Crohn disease; Antibodies; Enzyme-linked immunosorbent assay; Serological markers; Diagnosis

Serum anti-glycan antibodies in paediatric-onset Crohn's disease: association with disease phenotype and diagnostic accuracy.

IntroductionAntibodies reacting with various microbial epitopes have been described in inflammatory bowel disease (IBD) and are associated with a specific diagnosis and clinical presentation.AimTo evaluate the profile of new anti-glycan antibodies, their potential association with disease phenotype and diagnostic accuracy in paediatric Crohn's disease (CD).Material and methodsBlood samples from 134 paediatric IBD patients (109 CD, 25 ulcerative colitis (UC)) and 67 controls were blindly analysed for anti-Saccharomyces cerevisiae (ASCA), anti-chitobioside carbohydrate (ACCA), anti-laminaribioside carbohydrate (ALCA), and anti-mannobioside carbohydrate (AMCA) antibodies using commercially available assays. The serological response to glycans was correlated with clinical disease characteristics.ResultsAt least one of the tested anti-glycan antibodies was present in 75% of CD patients. Despite the high frequency of reactivity to glycan epitopes, a limited overlap of serological markers was observed. In total, 49% of ASCA-negative patients presented with one of the following: ACCA, ALCA, or AMCA. The occurrence of one antibody from the anti-glycan panel was independently associated with complicated disease phenotype and ileocolonic disease location. A higher level of immune response as assessed by the quartile sum scores for ACCA, ALCA, and AMCA was linked with older age at diagnosis (10–17 years) and ileocolonic disease location. The ASCA had the greatest accuracy for diagnosis and differentiation of CD.ConclusionsQualitative and quantitative serologicalal response to glycan epitopes was associated with distinct clinical presentation in paediatric CD patients. This raises the possibility for the use of these markers to differentiate subgroups of CD patients with more sever clinical presentation. The ASCA was the most accurate serological marker for CD; however, testing for the new anti-glycan antibodies may constitute an adjunctive tool in a specific group of patients to aid in the differentiation of CD with absent ASCA from ulcerative colitis.

The Relation of the Level of Serum Anti-TF, -Tn and -Alpha-Gal IgG to Survival in Gastrointestinal Cancer Patients

Objective: To evaluate the relation of the level of serum anti-TF, -Tn and -αGal carbohydrate antibodies to survival in gastrointestinal cancer patients.Methods: The level of anti-TF (Thomsen-Friedenreich antigen), -Tn and -αGal IgG was analysed in the serum of patients with gastric (n = 83) and colorectal (n = 51) cancers in the long-term follow-up, using ELISA with polyacrylamide glycoconjugates. To evaluate overall survival and the risk of death, the Kaplan-Meier method and the Cox proportional hazards model were used in the univariate analysis of patients groups.Results: A significantly better survival was observed: (1) in patients with an increased level of anti-TF antibodies (all, stage III, T2-4, N1-2 and G3; P = 0.004-0.038, HR = 0.16-0.46); and (2) in patients with an increased level of anti-Tn antibodies (G1-2 tumors; P = 0.034-0.042, HR = 0.34-0.47). A significantly worse survival was observed in gastrointestinal, gastric and colorectal groups with an increased level of serum anti-αGal antibodies. This association depended on the patho-morphology of tumors (all, stages I-II, III, T2-4, N0, N1-2 and G1-2; P = 0.006-0.048, HR = 1.99-2.33). In the combined assessment of the anti-TF and -αGal antibodies level of the whole gastrointestinal group (n = 53), P = 0.002, HR = 0.25, 95% CI 0.094-0.655. In the follow-up, the survival time was shorter in patients whose level of anti-αGal antibodies rose (P = 0.009-0.040, HR = 2.18-4.27). The level of anti-TF antibodies inversely correlated with neutrophil/lymphocyte ratio (NLR, r = - 0.401, P = 0.004, n = 49). Patients with a higher level of anti-αGal antibodies and NLR values demonstrated a significantly worse survival (P = 0.009, HR = 2.98, n = 48).Conclusions: The preoperative levels of anti-TF, -Tn and -αGal antibodies and their dynamics are of prognostic significance. The method for the determination of circulating anti-carbohydrate antibodies may be a useful supplement in clinical outcome assessment.