ABSTRACT
A lifelong gluten-free diet (GFD) is mandatory for celiac disease (CD) but has poor compliance, justifying novel strategies. We found that wheat flour transamidation inhibited IFN-γ secretion by intestinal T cells from CD patients. Herein, the primary endpoint was to evaluate the ability of transamidated gluten to maintain GFD CD patients in clinical remission. Secondary endpoints were efficacy in prevention of the inflammatory response and safety at the kidney level, where reaction products are metabolized. In a randomized single blinded, controlled 90-day trial, 47 GFD CD patients received 3.7 g/day of gluten from nontransamidated (12) or transamidated (35) flour. On day 15, 75% and 37% of patients in the control and experimental groups, respectively, showed clinical relapse (P = 0.04) whereas intestinal permeability was mainly altered in the control group (50% versus 20%, P = 0.06). On day 90, 0 controls and 14 patients in the experimental group completed the challenge with no variation of antitransglutaminase IgA (P = 0.63), Marsh-Oberhuber grading (P = 0.08), or intestinal IFN-γ mRNA (P > 0.05). Creatinine clearance did not vary after 90 days of treatment (P = 0.46). In conclusion, transamidated gluten reduced the number of clinical relapses in challenged patients with no changes of baseline values for serological/mucosal CD markers and an unaltered kidney function.
Subject(s)
Celiac Disease/diet therapy , Diet, Gluten-Free , Dietary Proteins/administration & dosage , Flour , Glutens/administration & dosage , Triticum/metabolism , Adolescent , Adult , Amidinotransferases/immunology , Autoantibodies/blood , Celiac Disease/immunology , Eating , Female , Humans , Interferon-gamma/metabolism , Intestinal Mucosa/immunology , Male , Middle Aged , Patient Compliance , Single-Blind Method , Transglutaminases/metabolism , Triticum/chemistry , Young AdultABSTRACT
BACKGROUND/AIMS: Cancer therapy with agonist anti-CD137 mAbs has been shown to induce immune-mediated tumor rejections in mice, and equivalent agents of this kind are currently being tested in cancer patients. Previous reports indicated that CD137 stimulation induced polyclonal infiltrates of T lymphocytes in the liver. This study characterizes the liver infiltrates and the target dependency of the phenomena and addresses the question of whether tumors nested in the liver are a more favorable target for CD137-based immunotherapy. METHODS: Liver infiltrates were studied with conventional histology and multiple color flow cytometry of total liver leukocytes. CD137(-/-) mice, mice with a single rearrangement of the TCR (OT-1 mice) and Rag(-/-) mice were used to clarify molecular requirements. Mice implanted with MC38 colon carcinomas either subcutaneously or inside the liver were used for comparative studies under treatment with agonist anti-CD137 mAbs. RESULTS: CD137 treatment caused mononuclear inflammation in the portal spaces of the liver, which gave rise to moderate increases in transaminases without signs of cholestasis. Marked increases in the numbers of CD8+ T cells were observed, including CD8+ T lymphocytes co-expressing CD11c. Infiltrates were absent in CD137(-/-) mice and mitigated in mice harboring a single transgenic TCR on their CD8 T cells. Despite the tumor-independent accumulation of T cells in the liver, immunotherapeutic effects were not more prominent against tumors located in this organ. CONCLUSIONS: Target-dependent effects of CD137 stimulation lead to liver infiltration with T cells, but lymphocyte enrichment in this organ does not privilege this site for immunotherapeutic effects against transplanted tumors.
Subject(s)
CD8-Positive T-Lymphocytes/metabolism , Colonic Neoplasms/immunology , Immunotherapy , Liver/pathology , Tumor Necrosis Factor Receptor Superfamily, Member 9/immunology , Amidinotransferases/immunology , Amidinotransferases/metabolism , Animals , Antibodies, Monoclonal/administration & dosage , CD8-Positive T-Lymphocytes/drug effects , CD8-Positive T-Lymphocytes/immunology , CD8-Positive T-Lymphocytes/pathology , Cell Count , Cell Line, Tumor , Cell Movement/drug effects , Colonic Neoplasms/pathology , Colonic Neoplasms/therapy , Liver/drug effects , Liver/immunology , Liver/metabolism , Lymphocytes, Tumor-Infiltrating/immunology , Mice , Mice, Inbred C57BL , Mice, Knockout , Mice, Transgenic , Neoplasm Transplantation , Organ Specificity , Receptors, Antigen, T-Cell, alpha-beta/genetics , Receptors, Antigen, T-Cell, alpha-beta/metabolism , Tumor Necrosis Factor Receptor Superfamily, Member 9/geneticsABSTRACT
Rat kidney L-arginine:glycine amidinotransferase (transamidinase) has been purified previously to homogeneity as two fractions, designated alpha and beta. No differences in the properties of these two fractions could be found. Two monoclonal antibodies (Tran/NS-1/1 and Tran/NS-1/3) to the purified alpha fraction of rat kidney transamidinase were produced, purified, and characterized. The results of competitive binding studies of the two monoclonal antibodies to alpha transamidinase were as follows: 1) Tran/NS-1/3 had no effect on 125I-Tran/NS-1/1 binding while Tran/NS-1/1 inhibited 125I-Tran/NS-1/1 binding; 2) Tran/NS-1/3 inhibited 125I-Tran/NS-1/3 binding while Tran/NS-1/1 had no effect on 125I-Tran/NS-1/3 binding. Therefore, Tran/NS-1/1 and Tran/NS-1/3 bound to different antigenic determinants on alpha transamidinase. 125I-Tran/NS-1/1 and 125I-Tran/NS-1/3 each had high avidity constants (approximately 10(7)-10(9)) for both alpha and beta rat kidney transamidinase. Tran/NS-1/1 and Tran/NS-1/3 bound to human kidney transamidinase in ELISA assays. A quantitative immunosorbent inhibition assay for rat kidney transamidinase was developed with 125I-Tran/NS-1/3. Approximately 30 ng of immunoreactive transamidinase could be detected by this immunosorbent inhibition assay. The amount of Tran/NS-1/3 immunoreactive species in rat lung and testicular tissue by the immunosorbent inhibition assay correlated well with the amount of transamidinase activity found in those tissues. The availability of the monoclonal antibodies, Tran/NS-1/1 and Tran/NS-1/3, should facilitate studies of rat and human transamidinase structure and regulation.