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 5

Tregs from CT-cured mice can suppress CD4+ T cell activation in an Ag-specific manner.

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Tregs from CT-cured mice can suppress CD4+ T cell activation in an Ag-sp...
Cell Proliferation Dye eFluor–labeled (eBioscience) CD4+CD25– Tresps, isolated from normoglycemic NOD mice that had received a prime-boost with human PINS or chicken OVA, were cocultured for 4 days with activated PINS- or OVA-pulsed APCs and the indicated ratios of CD4+CD25+ T cells, isolated from CT-cured mice at the end of the 6-week treatment. (A) Activation of Tresps was measured by flow cytometric analysis of CD44 and shown as MFI normalized to responder-only culture. (B) ELISA measurement of IFN-γ in the supernatant of Treg-Tresp cultures. (C) Percentage of IFN-γ–positive cells in CD4+ Tresps in Treg-Tresp cocultures stimulated with PINS or OVA as indicated. (D) Percentage of IL-10–producing cells in CD4+ Tregs in Treg-Tresp cocultures stimulated with PINS or OVA as indicated. Statistical significance was calculated using Mann-Whitney t test (*P < 0.05, **P < 0.01, ***P < 0.001).