Upregulating CD4+CD25+FOXP3+ Regulatory T Cells in Pancreatic Lymph Nodes in Diabetic NOD Mice by Adjuvant Immunotherapy

Department of Surgery and Diabetes Institute for Immunology and Transplantation, University of Minnesota, Minneapolis, MN 55455, USA.
Transplantation (Impact Factor: 3.83). 02/2009; 87(2):198-206. DOI: 10.1097/TP.0b013e3181933261
Source: PubMed


Immunotherapy with Complete Freund's adjuvant (CFA) is effective in ameliorating autoimmunity in diabetic nonobese diabetic (NOD) mice. We investigated whether CFA treatment up-regulates CD4+CD25+Foxp3+ regulatory T cells and increases transforming growth factor (TGF)-beta1 production in diabetic NOD mice.
New-onset diabetic NOD mice were treated with CFA and exendin-4, a potent analog of glucagon-like peptide-1. Reversal of diabetes was determined by monitoring blood glucose level. Ameliorating autoimmunity through immunoregulation was assessed by adoptive transfer. Regulatory T cells in the peripheral blood, spleen, thymus, and pancreatic nodes were measured. TGF-beta1 in plasma and the insulin content in the pancreas were also measured. Immunostainings for insulin and BrdU were performed.
New-onset diabetes could be reversed in 38% of NOD mice treated with CFA alone and in 86% of NOD mice treated with both CFA and exendin-4. Diabetes adoptive transfer by splenocytes from CFA-treated NOD mice was delayed. The percentage of CD4+CD25+Foxp3+ regulatory T cells in the pancreatic lymph nodes of CFA-treated NOD mice was significantly increased at 1, 5, and 15 to 17 weeks after treatment. TGF-beta1 in the plasma of CFA-treated NOD mice was also significantly increased. Combining CFA with exendin-4 treatment significantly increased the insulin content and the numbers of insulin and BrdU double-labeled beta cells in the islets.
Our results demonstrated that CFA treatment ameliorates autoimmunity in diabetic NOD mice by up-regulating CD4=CD25+Foxp3+ regulatory T cells and increasing TGF-beta1 production. Exendin-4 enhanced the effect of CFA on reversing diabetes in NOD mice by stimulating beta-cell replication.

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    • "The protective effects of CFA were suggested to be mediated through the downregulation of autoreactive CTLs and the stimulation of NK cells. Most recent data however, demonstrate that CFA treatment ameliorates autoimmunity in NOD mice by up-regulating CD4 + CD25 + Foxp3 + regulatory T-cells in pancreatic lymph nodes and for increasing TGF-í µí»½1 production [32], in spite of the fact that altered frequencies of peripheral CD4 + CD25 + Foxp3 + regulatory Tcells were not yet shown to be specifically associated with type 1 diabetes [33]. Molecular mechanisms involved in autoantigen-and adjuvant-dependant immune suppression detected in NOD mice remain to be further elucidated. "
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    Clinical and Developmental Immunology 11/2013; 2013(12):578786. DOI:10.1155/2013/578786 · 2.93 Impact Factor
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    • "The above feedback mechanism can explain the paradoxical finding that use of complete Freund's adjuvant (CFA) therapy raises inflammatory cytokines such as IFN-g and IL-17 while still being protective. This treatment also expands CD4 + CD25 + Foxp + T cells along inflammatory cytokines and establishes a new balance between regulatory and effector T cells with tolerogenic outcome (Tian and others 2009). We have shown the apoptosis of diabetogenic T cells in NOD mice by IFN-g and TNF-a as a mechanism of protection after mycobacterial adjuvant therapy (Qin and others 2004). "
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    • "Immunotherapy with CFA is effective in not only preventing spontaneous autoimmune diabetes, but also restoring self-tolerance to islet autoantigens. Recent studies conducted by Tian et al. (2009) demonstrated that CFA treatment ameliorates autoimmunity in diabetic NOD mice by up-regulating T regs and increasing TGF-β1 production. The percentage of T regs in the pancreatic lymph nodes of CFA-treated NOD mice was significantly increased at 1, 5, and 15 to 17 weeks after CFA treatment. "
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