Oral insulin for diabetes prevention in NOD mice: potentiation by enhancing Th2 cytokine expression in the gut through bacterial adjuvant.
Oral administration of insulin suppresses the development of diabetes in nonobese diabetic (NOD) mice and deviates the cytokine balance in the islets of Langerhans from a Th1 to a Th2 type cytokine pattern. However, the effect of oral insulin is limited and disease suppression is limited to a narrow dose range. Therefore we tried to improve the outcome of suboptimal insulin dosing by bacterial adjuvant. Mice treated with a suboptimal dose of oral insulin showed no change in diabetes incidence although a shift from Th1 towards Th2 cytokine expression occurred in inflamed islets. Significant suppression of diabetes development was only seen in NOD mice receiving both, insulin and the bacterial preparation OM-89 as adjuvant. OM-89 is a protein extract of Escherichia coli, with nonspecific immunostimulatory properties. Potentiation of the effect of oral insulin by the adjuvant was associated with upregulation of interleukin (IL)-4 Th2 cells in infiltrated islets and sustained local IL-2 gene expression. RT PCR analyses of cytokine expression in the gut showed a clear deviation to Th2 type reactivity and downregulation of inducible nitric oxide (NO) synthase (iNOS) expression by the bacterial adjuvant but not by oral insulin alone. Since macrophages are the primary target cells of adjuvant action we tested its effect on mouse macrophages in vitro. Treatment with OM-89 induced transient release of tumour necrosis factor alpha and nitrite but rendered macrophages refractory to restimulation by the potent macrophage activator lipopolysaccharide. In conclusion, the protective effect of oral insulin can be potentiated by pretreatment with the bacterial adjuvant OM-89. This effect correlates with enhanced Th2 cytokine and decreased iNOS gene expression in the gut, probably due to the downregulation of proinflammatory mediators by exposure to the adjuvant.
Hartmann
B
B
Clinical Department, Diabetes Research Institute at the Heinrich-Heine-University of Düsseldorf, Germany.
Bellmann
K
K
Ghiea
I
I
Kleemann
R
R
Kolb
H
H
D000276:Adjuvants, Immunologic / Q000008:administration & dosage / Q000494:pharmacology / Q000627:therapeutic use*; D000284:Administration, Oral; D000818:Animals; D000942:Antigens, Bacterial / Q000008:administration & dosage / Q000494:pharmacology / Q000627:therapeutic use*; D002460:Cell Line; D015331:Cohort Studies; D016207:Cytokines / Q000096:biosynthesis* / Q000187:drug effects / Q000235:genetics; D003922:Diabetes Mellitus, Type 1 / Q000453:epidemiology / Q000517:prevention & control*; D004195:Disease Models, Animal; D004357:Drug Synergism; D004926:Escherichia coli; D005260:Female; D005786:Gene Expression Regulation / Q000187:drug effects / Q000235:genetics; D007004:Hypoglycemic Agents / Q000008:administration & dosage / Q000494:pharmacology / Q000627:therapeutic use*; D007328:Insulin / Q000008:administration & dosage / Q000494:pharmacology / Q000627:therapeutic use*; D007515:Islets of Langerhans / Q000187:drug effects / Q000276:immunology / Q000473:pathology; D008264:Macrophages / Q000187:drug effects / Q000276:immunology; D051379:Mice; D016688:Mice, Inbred NOD; D019001:Nitric Oxide Synthase / Q000235:genetics; D011897:Random Allocation; D018418:Th2 Cells / Q000276:immunology*; D013997:Time Factors
D000276:Adjuvants, Immunologic; D000942:Antigens, Bacterial; D016207:Cytokines; D007004:Hypoglycemic Agents; D007328:Insulin; C050501:OM 89; D019001:Nitric Oxide Synthase