Despite improvement in insulin delivery, maintaining tight control of glucose homeostasis continues to be a challenge that results in bouts of severe hypo and hyperglycemia and serious long-term complications in many type 1 diabetes (T1D) patients. Therefore, developing an immunotherapy for the disease remains a major goal. Reaching this goal, however, requires deep knowledge of all facets of the diabetogenic process - which is generally believed to be initiated by an imbalance between pathogenic and regulatory mechanisms that allows diabetogenic T cells to infiltrate pancreatic islets and destroy insulin-producing ss-cells. Therefore, identifying and understanding roles of various molecules and cell types that tip the balance towards the immunopathogenic pathways in susceptible individuals and animal models is important for developing effective immunotherapy. This proposal investigates mechanisms that powerfully control ss-cell specific autoreactive T cells when Fas ligand (FasL), an apoptosis-inducing member of TNF family, is genetically or pharmacologically inactivated. Previously, the lack of appropriate models and efficacious FasL blocking mAb has severely hampered such investigation. In this application, we will use NOD mice that are haploinsufficient for FasL (NOD-gld/+ mouse) and a FasL- neutralizing mAb (MFL4 clone) to investigate the underlying mechanisms and therapeutic significance of FasL blockade using the MFL4 mAb. NOD-gld/+ mice are completely protected from T1D, immunocompetent, and have normal immune homeostasis. In addition, MFL4 mAb protects NOD-wt mice from diabetes without altering immune homeostasis and, more importantly, our preliminary data show it has promising efficacy in reversing hyperglycemia in new-onset cases. Based on our preliminary data generated using these model systems, we hypothesize that an IL-10-producing regulatory B cell subset that suppresses diabetogenic autoreactive T cells are negatively regulated by FasL. In NOD-wt mice, FasL-mediated apoptosis eliminates IL- 10-producing regulatory B cells thereby removing the brakes on autoreactive T-cells (tested Aim 1). We hypothesize that the MFL4 mAb can be used to reverse new-onset diabetes (tested in Aim 2). In NOD mice, haploinsufficiency for FasL (gld/+) or mAb blockade of FasL prevents IL-10-producing B cell elimination, leading to control of diabetogenic T cells and suppression of insulitis (tested in Aim 3). In this revised application, we will also assess relevance of our preclinical data to the human disease in samples from newly diagnosed patients at Hopkins and tissues provided by the JDRF sponsored nPOD project (Aim 3). Because the role of FasL in normal immune response and ss-cell death are dispensable, understanding how FasL modulates the diabetogenic process can lead to new mechanistic insights into the disease pathogenesis that could have important therapeutic implications.
Developing an immunotherapy for type 1 diabetes is a major goal. Understanding how Fas ligand potently prevents the disease in mouse model, focus of this application, may lead to designing new therapeutic approaches.
|Ramirez, Lourdes; Hamad, Abdel Rahim A (2015) From non-obese diabetic to Network for the Pancreatic Organ Donor with Diabetes: New heights in type 1 diabetes research. World J Diabetes 6:1309-11|
|Martina, Maria N; Noel, Sanjeev; Saxena, Ankit et al. (2015) Double negative (DN) Î±Î² T cells: misperception and overdue recognition. Immunol Cell Biol 93:305-10|
|Dai, Hong; Rahman, Ayesha; Saxena, Ankit et al. (2015) Syndecan-1 identifies and controls the frequency of IL-17-producing naÃ¯ve natural killer T (NKT17) cells in mice. Eur J Immunol 45:3045-51|
|Saxena, Ankit; Khosraviani, Sam; Noel, Sanjeev et al. (2015) Interleukin-10 paradox: A potent immunoregulatory cytokine that has been difficult to harness for immunotherapy. Cytokine 74:27-34|
|Noel, Sanjeev; Martina-Lingua, Maria N; Bandapalle, Samatha et al. (2014) Intestinal microbiota-kidney cross talk in acute kidney injury and chronic kidney disease. Nephron Clin Pract 127:139-43|
|Martina, Maria N; Bandapalle, Samantha; Rabb, Hamid et al. (2014) Isolation of double negative Î±Î² T cells from the kidney. J Vis Exp :|
|Martina, M N; Noel, S; Bandapalle, S et al. (2014) T lymphocytes and acute kidney injury: update. Nephron Clin Pract 127:51-5|
|Ramirez, Lourdes; Hamad, Abdel Rahim A (2013) Status of autoimmune diabetes 20-year after generation of BDC2.5-TCR transgenic non-obese diabetic mouse. World J Diabetes 4:88-91|
|Shao, Hanjuan; Scott, Sherri-Gae; Nakata, Chiaki et al. (2013) Extracellular matrix protein lumican promotes clearance and resolution of Pseudomonas aeruginosa keratitis in a mouse model. PLoS One 8:e54765|