Maya Margalit

Glucocerebroside Ameliorates the Metabolic Syndrome in OB/OB Mice

Glucocerebroside (GC) is a naturally occurring glycolipid that may alter natural killer T (NKT) cell function. To determine the effect of GC on the metabolic derangements and immune profile in leptin-deficient mice, Ob/Ob mice were treated by daily injections of GC for 8 weeks and followed for various metabolic and immunological parameters. Marked amelioration of the metabolic alterations characteristic of leptin-deficient mice was observed in GC-treated animals compared with controls. A significant decrease in liver size and hepatic fat content were observed in GC-treated mice. Near-normalization of glucose tolerance and decreased serum triglyceride levels were observed. Fluorescence-activated cell sorting analysis of peripheral and intrahepatic lymphocytes revealed a 1.6-fold increase of the peripheral/intrahepatic NKT lymphocyte ratio. A 33% decrease of serum interferon-γ level and a 2.6-fold increase of serum interleukin 10 level were noted in GC-treated mice. Immune modulation by GC may have a role in the treatment of nonalcoholic steatohepatitis and other immune-mediated disorders.

Liver abscess in inflammatory bowel disease: Report of two cases and review of the literature

Hepatic abscesses are a rare complication of inflammatory bowel disease (IBD). Despite the fact that certain hepatobiliary complications of IBD, including cholelithiasis, primary sclerosing cholangitis (PSC) and cholangiocarcinoma predispose patients with IBD to ascending cholangitis, previously published data does not demonstrate that biliary infection is an important mechanism underlying liver abscess development in these patients. We describe two patients with inflammatory bowel disease, both with PSC, who developed multiple liver abscesses, and review the literature on liver abscesses in association with inflammatory bowel disease.

A double-blind clinical trial for treatment of Crohn's disease by oral administration of Alequel™, a mixture of autologous colon-extracted proteins : A patient-tailored approach

OBJECTIVES:In this study, we evaluated the safety and efficacy of a personalized mode of treatment for Crohns disease (CD) by oral administration of Alequel™, an extract of autologous colonic proteins.METHODS:Thirty-one patients with moderate to severe CD were enrolled in a 27-wk randomized, double-blind, placebo-controlled trial. Patients were randomized to receive either a placebo or the study drug prepared from autologous colonic extract.RESULTS:Oral administration of autologous colonic proteins resulted in clinical remission (58%vs 29%; 46.6%vs 26.6%, using an intention to treat analysis, P = NS), clinical response (67%vs 43%; 53.3%vs 40%, using an intention to treat analysis, P = NS) and improved quality of life (Inflammatory Bowel Disease Questionnaire score improvement 43%vs 12%) in the drug study group, compared to placebo group. No treatment-related adverse events were noted. Only in the study-drug-treated cohort who achieved clinical remission (DR), there was a decreased number of subject-specific, antigen-directed, IFNγ spot-forming colonies. DR subjects had a lower initial C-reactive protein level than DNOR or placebo subjects, an increased percentage of peripheral blood nature killer T cells, and an increased CD4+/CD8+ T-cell ratio throughout the period of drug administration.CONCLUSIONS:Oral administration of Alequel™ is a safe method for treatment of patients with moderate to severe CD, and its efficacy needs to be proven. Several markers may be applicable as surrogate markers for the clinical effect.

Suppression of hepatocellular carcinoma by transplantation of ex‐vivo immune‐modulated NKT lymphocytes

NKT cells are a regulatory subset of T lymphocytes with immune modulatory effects and an important role in anti‐tumor immunity. The feasibility of “ex‐vivo education” of NKT cells has recently been demonstrated. To evaluate the anti‐tumor effect of ex‐vivo immune‐modulated NKT lymphocytes in a murine model of hepatocellular carcinoma. Athymic Balb/C mice were sublethally irradiated and transplanted with human Hep3B HCC. NKT cells prepared from immunocompetent Balb/C mice were pulsed ex vivo with HCC‐derived antigens (Group A), Hep3B cells (group B) or BSA (group C), and adoptively transferred into HCC harboring mice (1 × 06 NKT cells per mouse). Group D mice did not undergo NKT cell transplantation. Group E mice were transplanted with 1 × 106 NKT cells from HBV‐immunized donors. Mice were followed for tumor size and weight. To determine the mechanism of the anti‐tumor effect, intrasplenic lymphocyte populations were analyzed by FACS for NKT, CD4+ and CD8+ lymphocyte subpopulations; STAT 1, 4 and 6 expression in splenocytes was assessed by Western blot, and serum cytokine levels were measured by ELISA. Adoptive transfer of NKT cells pulsed with HCC‐derived antigens (group A) and NKT cells from immunized donors (group E) resulted in complete disappearance of tumors within 4 weeks and attenuated weight loss (6.5% and 7% in groups A and E, respectively). In contrast, mice in groups B, C, and D developed large, necrotic tumors and severe weight loss (21%, 17% and 23% weight loss in groups B, C, and D, respectively). NKT/CD4 and CD8/CD4 ratios were significantly increased in groups A and E (12.3 and 17.6 in groups A and D, respectively, compared to 6.4, 4.8 and 5.6 in groups B, C and D, respectively, for the NKT/CD4 ratio; 41 and 19.8 in groups A and E, respectively, compared to 6.5, 11.8 and 3.2 in groups B, C, and D, respectively, for the CD8/CD4 ratio). Expression of the transcription factor STAT4 was evident in group A, but not in groups B‐D. Serum IFNγ, IL12 and IL4 levels were increased in groups A and E. Adoptive transfer of NKT lymphocytes exposed ex vivo by HCC‐derived antigens loaded on dendritic cells and NKT cells from immunized donors led to suppression of HCC in mice. NKT‐mediated anti‐tumor activity was associated increased NKT and CD8+ T lymphocyte numbers, increased expression of STAT4, a marker for IL‐12 activity and elevated serum levels of the proinflammatory cytokines IFNγ and IL12, and of IL4. Ex‐vivo modulation of NKT lymphocytes holds promise as a novel mode of immune therapy for HCC.

Amelioration of non-alcoholic steatohepatitis and glucose intolerance in ob/ob mice by oral immune regulation towards liver-extracted proteins is associated with elevated intrahepatic NKT lymphocytes and serum IL-10 levels.

Non‐alcoholic steatohepatitis (NASH) is a common cause of cryptogenic cirrhosis in the Western world. In an animal model of NASH, leptin‐deficient ob/ob mice present with alterations in number and function of hepatic NKT and peripheral CD4 lymphocytes. Oral immune regulation is a method to alter the immune response towards orally administered antigens. To determine the effect of oral immune regulation towards liver‐extracted proteins on the metabolic disorders in ob/ob mice, ob/ob mice and their lean littermates were orally administered liver extracts from wild‐type or ob/ob mice or bovine serum albumin for 1 month. The effect of treatment on hepatic fat content was measured by magnetic resonance imaging (MRI) and using a histological steatohepatitis grading scale. Glucose tolerance was measured by an oral glucose tolerance test (GTT). T lymphocyte subpopulations were assessed by flow cytometry analysis. Induction of immune regulation by oral presentation of liver‐extracted proteins resulted in a significant 18% reduction of the hepatic fat content in ob/ob mice fed with either wild‐type or ob/ob liver extracts for 1 month. The MRI signal intensity index in treated mice decreased to 0.48 and 0.51, respectively, compared with 0.62 in BSA‐fed controls (p = 0.037 and p = 0.019, respectively), while the histological steatohepatitis score decreased in both treated groups to 2.0, compared with 2.4 in BSA‐fed controls (p = 0.05). A significant improvement in GTT was noted in treated ob/ob mice. These changes were accompanied by a marked increase in the intrahepatic NKT lymphocyte population in mice fed with proteins extracted from both wild‐type and ob/ob mice (46.96% and 56.72%, respectively, compared with 26.21% in BSA‐fed controls; p < 0.05) and a significant elevation in serum IL‐10 levels. Oral immune regulation towards liver extracted proteins in leptin‐deficient mice resulted in a marked reduction in hepatic fat content and improved glucose tolerance. This effect was associated with a significant increase in the intrahepatic NKT lymphocyte population and serum IL‐10 levels, suggesting a Th1 to Th2 immune shift. Immune regulation towards disease‐associated antigens holds promise as a new mode of therapy for NASH. Copyright

Alleviation of acute and chronic graft-versus-host disease in a murine model is associated with glucocerebroside-enhanced natural killer T lymphocyte plasticity.

Background. Natural killer T (NKT) lymphocytes play a role in graft-versus-host disease GVHD (GVHD). Glucocerebroside (GC) is a naturally occurring glycolipid that plays a role in the immune modulation of NKT lymphocytes. This study aims to determine the effect of GC in murine models of semiallogeneic/acute and chronic GVHD. Methods. Acute and chronic GVHD were generated by fusion of splenocytes from C57BL/6 to (C57BL/6xBalb/c) F1 mice, and from B10.D2 donor mice into Balb/c, respectively. Recipients were treated daily with GC. Histological and immunological parameters of GVHD were assessed. Results. Treatment with GC significantly alleviated GVHD in both models The beneficial effect of GC was associated with an increase in the intrahepatic/peripheral NKT lymphocyte ratio in the semiallogeneic/acute model. This ratio was decreased in the chronic GVHD model. A significant increase in intrahepatic CD8+ lymphocyte trapping was noted in the semiallogeneic/acute model, whereas the opposite effect was observed in the chronic GVHD model. Administration of GC led to decreased serum interferon-&ggr; and increased serum interleukin-4 levels in the Th1-mediated model, whereas the opposite effect was observed in the Th2-mediated models. Conclusions. GC ameliorates GVHD in both Th1- and Th2-mediated murine models, suggesting it is able to differentially affect the immune system. GC may alleviate immunologically incongruous disorders, and may be associated with “fine tuning” of immune responses.

Induction of immune tolerance: a role for Natural killer T lymphocytes?

Abstract: The ability of the immune system to distinguish between harmful and harmless antigens is essential for mounting protective immune responses and preventing the induction of pathology. Tolerance is a mechanism that prevents or suppresses potentially injurious immune responses. Natural killer T (NKT) lymphocytes, a subset of regulatory T lymphocytes, can induce pro‐inflammatory or anti‐inflammatory immune responses. This subset of cells appears to be crucial for induction of tolerance by several immune‐modulatory interventions; these include immune manipulations in the setting of transplantation, induction of tolerance by introduction of antigen into immune‐privileged sites, and oral administration of disease‐associated‐antigen. The ability to predict whether tolerance or immunity will be generated in a given situation is essential for development of NKT lymphocyte‐based immune‐modulatory treatments. The role of NKT lymphocytes in these settings, and the requirements for development of tolerance, rather than immunity, are discussed.

W1091 The Impact of Concomitant Treatment with Immuno-Modulators and Antibiotics On the Outcome of C. difficile-Associated Inflammatory Bowel Disease Exacerbation: A European Multi-Center Retrospective Study

P129 The impact of concomitant treatment with immuno-modulators and antibiotics on the outcome of C. difficile-associated inflammatory bowel disease exacerbation: an ECCO multi-center retrospective study S. Ben-Horin1 *, M. Margalit2, P. Bossuyt3, J. Maul4, Y. Shapira5, D. Bojic6, I. Chermesh7, A. Al-Rifai8, A. Schoepfer9, M. Bosani10, M. Allez11, P.L. Lakatos12, F. Bossa13, A. Eser14, T. Stefanelli15, F. Carbonnel16, K. Katsanos17, D. Checchin18, I. Saenz de Miera19, Y. Chowers1, G.W. Moran20. 1Sheba Medical Center, Ramat-Gan, Israel, 2Haddassa Medical Center, Hebrew University, Jerusalem, Israel, 3University of Leuven Hospitals, Leuven, Belgium, 4Charite-Universitatsmedizin, Berlin, Germany, 5Tel-Aviv Medical Center, Tel-Aviv, Israel, 6Zvezdara University Medical Center, Belgrade, Serbia, 7Rambam Medical Center, Haifa, Israel, 8Salford NHS University Hospital, Manchester, United Kingdom, 9University Hospital of Bern, Bern, Switzerland, 10Sacco University Hospital, Milan, Italy, 11Saint-Louis Hospital, Paris, France, 12Semmelweis University, Budapest, Hungary, 13Casa Sollievo della Sofferenza Hospital, San Giovanni Rotondo, Italy, 14Medical University of Vienna, Vienna, Austria, 15Universita degli Studi di Milano, Milan, Italy, 16University hospital of Besancon, Besancon, France, 17Univeristy Hospital of Ioannina, Ioannina, Greece, 18University of Padua, Padua, Italy, 19Hospital General Yague, Burgos, Spain, 20University of Birmingham, Birmingham, United Kingdom

W1841 Esophageal Acid Reflux and Respiratory Symptoms: Disparate Relationship in Asthmatic and Non-Asthmatic Patients

(esophageal aperistalsis and hypotonic LES). Nine patients (60%) had endoscopic esophagitis, whereas all fifteen showed gastroesophageal reflux at the 24h pH-MII recording. Analyzing the type of reflux, 2 patients showed a pure acid reflux, five (33%) a non-acid reflux and the other eight (53%) had a mixed reflux, both acid and non acid. Regarding lung involvement, images of interstitial pneumonia were detected in 6 out 15 (40%). The proximal extent of liquid refluxate into the esophagus was documented in three patients, all of them with interstitial pneumonia (p>0.05 by Fishers exact test) Conclusions: the majority of SSc patients showed both acid and non acid GERD, and these findings might help for better tailoring medical treatment. Patients with proximal extent of liquid refluxate seem to be at risk of pulmonary involvement. These preliminary data need to be confirmed in a larger cohort of SSc patients.

Oral Tolerance for IBD

TO THE EDITOR: We read with great interest the article entitled “Reports of ‘Satisfactory Relief’ by IBS Patients Receiving Usual Medical Care Are Confounded by Baseline Symptom Severity and Do Not Accurately Reflect Symptom improvement,” by Whitehead et al. (1), and the accompanying editorial by Schoenfeld and Talley (2). We agree with the conclusions reached by Schoenfeld and Talley that binary end points are useful end points in IBS trials. In addition to the shortcomings in Whitehead et al. (1) described by Schoenfeld and Talley (2), we would like to correct several factual and interpretational errors.