Furthermore, adoptive co-transfer experiments involving injection of Thy-1,2 recipients with diabetogenic T cells from syngeneic mice and T cells from congenic Thy-1,1 mice fed with CTB-insulin exhibited a selective recruitment of Thy-1,1 donor cells in the peripancreatic lymph nodes concomitant with reduced islet cell infiltration

Furthermore, adoptive co-transfer experiments involving injection of Thy-1,2 recipients with diabetogenic T cells from syngeneic mice and T cells from congenic Thy-1,1 mice fed with CTB-insulin exhibited a selective recruitment of Thy-1,1 donor cells in the peripancreatic lymph nodes concomitant with reduced islet cell infiltration. recipients with diabetogenic T cells from syngeneic mice and T cells from congenic Thy-1,1 mice fed with CTB-insulin exhibited a selective recruitment of Thy-1,1 donor cells in the peripancreatic lymph nodes concomitant with reduced islet cell infiltration. These results suggest that protection against autoimmune diabetes can be achieved by feeding minute amounts of a pancreas islet cell autoantigen linked to CTB and appears to involve the selective OGT2115 migration and retention of protective T cells into lymphoid tissues draining the site of organ injury. test for unpaired samples. RESULTS Oral Administration of CTB-Insulin Conjugate Suppresses Spontaneous Diabetes in NOD Mice. NOD females were fed once at 8 weeks of age with either 0.2 g, 2 g, or Rabbit polyclonal to Caspase 6 20 g of insulin conjugated to CTB or with 100 g of CTB, and animals were monitored for diabetes onset. As shown in Fig. ?Fig.1,1, onset of diabetes was delayed in animals fed a single dose of CTB-INS conjugate in comparison to control animals. For the whole period between 17 and 24 weeks of age the incidence of diabetes was significantly lower in the CTB-INS (20 g)-fed group than in the control group, which was fed CTB alone. Thus, at 23 weeks of age (i.e., 15 weeks after treatment), only 2 out of 17 (12%) mice fed insulin OGT2115 that was conjugated to CTB experienced developed diabetes, as compared with 7 out of 16 (44%) control mice that were fed CTB alone [protective efficacy (PE) = 73%; = 0.04]. In contrast, mice fed insulin (50 g) alone had only marginally lower disease incidence over the same time period, with 6 out of 16 animals (38%; PE = 14%; 0.10) having developed diabetes by week 23. Thereafter, this difference progressively vanished. Feeding mice with lower doses of CTB-INS experienced intermediate but not statistically significant effects on diabetes incidence; 2 out of 8 (25%) mice fed 2 g CTB-INS and 3 out of 8 (37.5%) mice fed 0.2 g developed diabetes by week 23 (data not shown). Open in a separate window Physique 1 Suppression of spontaneous autoimmune diabetes after oral treatment with CTB-insulin conjugate. NOD females were treated at 8 weeks of age with a single dental administration of 20 g of insulin conjugated to CTB (), insulin only (?), or CTB only (?). Data are mixed from two 3rd party tests. We also analyzed the result of repeated dental administrations of CTB-conjugated insulin on disease advancement. In the 1st experiment, just 2 out of 7 (28%) mice treated with 2 g CTB-INS, provided at 8, 10, 12, 14, and 16 weeks old, had created disease (PE = 60%; = 0.04) by 29 weeks, whereas 5 out of 7 (71%) mice given having a control (CTB-OVA) conjugate had developed diabetes. Nourishing mice five consecutive 10-g dosages of free of charge insulin (amounting to a complete of 50 g insulin) got no influence on disease advancement (data not demonstrated). Another experiment involving bigger numbers of pets from another NOD OGT2115 mouse colony yielded identical results. Thus, just 4 out of 35 (11%) mice given 40 g of CTB-conjugated insulin OGT2115 provided on five consecutive events had created diabetes by 20 weeks old (PE = 78%; 0.001) in comparison with 15 out of 30 (50%) sham-treated control mice (Desk ?(Desk1). 1). By 25 weeks old, 11 out of 35 (31%) pets given CTB-INS had created diabetes, in comparison with 20 out of 30 control pets (PE = 54%; 0.01). Repeated oral medication with unconjugated INS (5 100 g) got no appreciable influence on diabetes occurrence (Desk ?(Desk1). 1). Desk 1 Suppression of diabetes after repeated oral medication with low dosages of CTB-conjugated?insulin.