Anti-CD3 and nasal proinsulin combination therapy enhances remission from recent-onset autoimmune diabetes by inducing Tregs
Damien Bresson, … , Kevan C. Herold, Matthias von Herrath
Damien Bresson, … , Kevan C. Herold, Matthias von Herrath
Published May 1, 2006
Citation Information: J Clin Invest. 2006;116(5):1371-1381. https://doi.org/10.1172/JCI27191.
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Research Article Metabolism

Anti-CD3 and nasal proinsulin combination therapy enhances remission from recent-onset autoimmune diabetes by inducing Tregs

Abstract

Safe induction of autoantigen-specific long-term tolerance is the “holy grail” for the treatment of autoimmune diseases. In animal models of type 1 diabetes, oral or i.n. immunization with islet antigens induces Tregs that are capable of bystander suppression. However, such interventions are only effective early in the prediabetic phase. Here, we demonstrate that a novel combination treatment with anti-CD3ε–specific antibody and i.n. proinsulin peptide can reverse recent-onset diabetes in 2 murine diabetes models with much higher efficacy than with monotherapy with anti-CD3 or antigen alone. In vivo, expansion of CD25+Foxp3+ and insulin-specific Tregs producing IL-10, TGF-β, and IL-4 was strongly enhanced. These cells could transfer dominant tolerance to immunocompetent recent-onset diabetic recipients and suppressed heterologous autoaggressive CD8 responses. Thus, combining a systemic immune modulator with antigen-specific Treg induction is more efficacious in reverting diabetes. Since Tregs act site-specifically, this strategy should also be expected to reduce the potential for systemic side effects.

Authors

Damien Bresson, Lisa Togher, Evelyn Rodrigo, Yali Chen, Jeffrey A. Bluestone, Kevan C. Herold, Matthias von Herrath

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Figure 4

hpIIp–specific T cells from mice treated with anti-CD3 and i.

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hpIIp–specific T cells from mice treated with anti-CD3 and i. . proinsul...
. proinsulin peptide exhibit a regulatory cytokine profile after in vitro stimulation. (A) Splenocytes and PLNs from animals treated and cured by the anti-CD3 alone or in combination with the hpIIp were used in vitro in a cytokine secretion assay. Cells were stimulated with a mixture of anti-CD3 and anti-CD28 Abs, various peptides (hpIIp, insB9–23, and LCMV-specific GP61–80 and NP118–126) or remained nonstimulated. After 72 hours, the supernatants were analyzed by ELISA as well as using the Luminex 100 LabMAP System. The data represent the levels of cytokine above the background observed without stimulation (NM-anti-CD3 [IL-4: 17 pg/ml, IL-5: 430 pg/ml, IL-10: 10 pg/ml, TGF-β1: 530 pg/ml, and IFN-γ: 2,500 pg/ml] or combination therapy [IL-4: 41.5 pg/ml, IL-5: 380 pg/ml, IL-10: 200 pg/ml, TGF-β1: 580 pg/ml, and IFN-γ: 3,000 pg/ml]). Data for an average of 4 mice per group are shown. *P < 0.05 compared with the nonstimulated control. (B) To confirm the cytokine profile found in the supernatants, intracellular cytokine stainings were performed. The cells were stimulated specifically with the hpIIp or unspecifically with a mixture PMA/ionomycin or remained unstimulated (nonstimulated) for 6 hours in the presence of an inhibitor of intracellular protein transport (monensin). The cells were recovered and stained at the surface for CD4 and intracellularly for Foxp3 and the cytokines TGF-β, IL-10, and IL-4. The cytokine expression was analyzed in the CD4+Foxp3+ (upper panel) and CD4+Foxp3neg (lower panel) populations.