Reversal of Diabetes in NOD Mice by Clinical-Grade Proinsulin and IL-10–SecretingLactococcus lactisin Combination With Low-Dose Anti-CD3 Depends on the Induction of Foxp3-Positive T Cells

Abstract

The introduction of β-cell autoantigens via the gut through Lactococcus lactis (L. lactis) has been demonstrated to be a promising approach for diabetes reversal in NOD mice. Here we show that a combination therapy of low-dose anti-CD3 with a clinical-grade self-containing L. lactis, appropriate for human application, secreting human proinsulin and interleukin-10, cured 66% of mice with new-onset diabetes, which is comparable to therapy results with plasmid-driven L. lactis Initial blood glucose concentrations (<350 mg/dL) and insulin autoantibody positivity were predictors of the stable reversal of hyperglycemia, and decline in insulin autoantibody positivity was an immune biomarker of therapeutic outcome. The assessment of the immune changes induced by the L. lactis-based therapy revealed elevated frequencies of CD4Foxp3 T cells in the pancreas-draining lymph nodes, pancreas, and peripheral blood of all treated mice, independent of metabolic outcome. Neutralization of cytotoxic T-lymphocyte antigen 4 and transforming growth factor-β partially abrogated the suppressive function of therapy-induced regulatory T cells (Tregs). Ablation or functional impairment of Foxp3 Tregs in vivo at the start or stop of therapy impaired immune tolerance, highlighting the dependence of the therapy-induced tolerance in mice with new-onset diabetes on the presence and functionality of CD4Foxp3 T cells. Biomarkers identified in this study can potentially be used in the future to tailor the L. lactis-based combination therapy for individual patients.

Immunotherapy with oral administration of humanized anti-CD3 monoclonal antibody: a novel gut-immune system-based therapy for metaflammation and NASH

“…This type of antigen-directed tolerance remains intact for long periods after treatment withdrawal. The mechanism has been shown to be associated with the generation of T regs [33]. The mechanism has been shown to be associated with the generation of T regs [33].…”

Reversal of Hyperglycemia and Suppression of Type 1 Diabetes in the NOD Mouse with Apoptotic DNA Immunotherapy™ (ADi™), ADi-100

“…However, there are very few published studies demonstrating that such ASIs (as monotherapies) can "reverse hyperglycemia" (i.e., Stage 2) in NOD mice [19]. This is in contrast to several non-specific immunomodulatory agents, such as anti-CD3 mAb, that have successfully reversed hyperglycemia in NOD mice, either alone or in combination with an ASI [13,[19][20][21] and have recently been effective at delaying insulin production loss in pre-diabetic (i.e., dysglycemia, Stage 2) subjects [22]. However, unlike ASIs, these non-specific therapies may not induce durable tolerance and thus would require long-term dosing with associated safety concerns.…”

From immunohistological to anatomical alterations of human pancreas in type 1 diabetes: New concepts on the stage

“…A limited number of B cells (CD20 + ) were observed in the early stages of insulitis, but their frequency increased alongside with the progression of β‐cell death . The presence of Natural Killer (NK) cells has been reported, and finally, FOXP3 + regulatory T‐cells were detected only in a single donor, whereas widely identified in a murine model of T1D (Non‐Obese Diabetic—NOD—mouse) . The authors observed that CD8 + T cells increased in number as β‐cells started to be destroyed and then declined dramatically once β‐cells were no longer present within the islets (Figure ), while macrophages have been described as the most prominent immune cell type infiltrating β‐cell‐depleted islets.…”