Reversal of autoimmune diabetes by restoration of antigen-specific tolerance using genetically modified Lactococcus lactis in mice
Tatiana Takiishi, … , Conny Gysemans, Chantal Mathieu
Tatiana Takiishi, … , Conny Gysemans, Chantal Mathieu
Published April 9, 2012
Citation Information: J Clin Invest. 2012;122(5):1717-1725. https://doi.org/10.1172/JCI60530.
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Research Article Autoimmunity

Reversal of autoimmune diabetes by restoration of antigen-specific tolerance using genetically modified Lactococcus lactis in mice

Abstract

Current interventions for arresting autoimmune diabetes have yet to strike the balance between sufficient efficacy, minimal side effects, and lack of generalized immunosuppression. Introduction of antigen via the gut represents an appealing method for induction of antigen-specific tolerance. Here, we developed a strategy for tolerance restoration using mucosal delivery in mice of biologically contained Lactococcus lactis genetically modified to secrete the whole proinsulin autoantigen along with the immunomodulatory cytokine IL-10. We show that combination therapy with low-dose systemic anti-CD3 stably reverted diabetes in NOD mice and increased frequencies of local Tregs, which not only accumulated in the pancreatic islets, but also suppressed immune response in an autoantigen-specific way. Cured mice remained responsive to disease-unrelated antigens, which argues against excessive immunosuppression. Application of this therapeutic tool achieved gut mucosal delivery of a diabetes-relevant autoantigen and a biologically active immunomodulatory cytokine, IL-10, and, when combined with a low dose of systemic anti-CD3, was well tolerated and induced autoantigen-specific long-term tolerance, allowing reversal of established autoimmune diabetes. Therefore, we believe this method could be an effective treatment strategy for type 1 diabetes in humans.

Authors

Tatiana Takiishi, Hannelie Korf, Tom L. Van Belle, Sofie Robert, Fabio A. Grieco, Silvia Caluwaerts, Letizia Galleri, Isabella Spagnuolo, Lothar Steidler, Karolien Van Huynegem, Pieter Demetter, Clive Wasserfall, Mark A. Atkinson, Francesco Dotta, Pieter Rottiers, Conny Gysemans, Chantal Mathieu

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Figure 2

CT preserves insulin content and limits insulitis.

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CT preserves insulin content and limits insulitis. (A) Evaluation by ELI...
(A) Evaluation by ELISA of insulin content in pancreatic extracts from cured (14 weeks) and control experimental groups, as indicated. Data are expressed as ng/mg of pancreas. Shown are the individual values (symbols) overlaid with mean (horizontal line), box (25th and 75th percentile), and Tukey whiskers. Statistical significance between 2 groups was calculated using Mann-Whitney U test; *P < 0.05, **P < 0.01. (B) Pancreatic sections were stained for insulin (green), Ki-67 (red), and DNA by Hoechst (blue) and analyzed with confocal microscopy at ×63 original magnification. Representative Ki-67/insulin microphotographs documenting the absence of proliferating β cells in the pancreas are shown. (C) Pancreatic sections of the indicated groups were stained with H&E, and insulitis was scored as indicated.