CD8+ CD122+ PD-1− effector cells promote the development of diabetes in NOD mice

Abstract

It is well established that CD4 and CD8 T cells are required for the initiation of autoimmune diabetes in NOD mice. However, different subsets of CD4 or CD8 cells may play different roles in the initiation of insulitis. In this study, we evaluated the role of the previously described CD8(+) CD122(+) in this process. We found that prediabetic NOD mice have an almost 50% reduction of CD8(+) CD122(+) T cells in their secondary lymphoid organs compared with BL/6 or Balb/c mouse strains. This reduction is explained by the lack of the regulatory CD8(+) CD122(+) PD-1(+) cell population in the NOD mice, as we found that all CD8(+) CD122(+) T cells from prediabetic NOD mice lack PD-1 expression and regulatory function. Depletion of CD8(+) CD122(+) PD-1(-) cells through injection of anti-CD122 mAb in prediabetic female NOD mice reduced the infiltration of mononuclear cells into the Langerhans islets and delayed the onset and decreased the incidence of overt diabetes. In addition, we found that transfer of highly purified and activated CD8(+) CD122(+) PD-1(-) cells, together with diabetogenic splenocytes from NOD donors to NOD SCID recipients, accelerates the diabetes development in these mice. Together, these results demonstrate that CD8(+) CD122(+) PD-1(-) T cells from NOD mice are effector cells that are involved in the pathogenesis of autoimmune diabetes.

CD122 blockade restores immunological tolerance in autoimmune type 1 diabetes via multiple mechanisms

“…After the treatment, all mice were monitored for spontaneous development of diabetes until 40 weeks of age. The incidence of diabetes onset in ChMBC7-treated mice was significantly lower than that in the control group ( Figure 1B), consistent with previous reports (13)(14)(15)(16). Using the same treatment protocol, separated cohorts of mice were sacrificed at 10 weeks old, and the pancreata were excised and processed for histopathology analysis.…”

“…Using the NOD mouse strain, the primary animal model for T1D basic research and preclinical studies (8,17), we showed that a Fc-silent rat/mouse chimeric anti-CD122 mAb (clone ChMBC7) effectively suppressed T1D development, consistent with previous reports (13)(14)(15)(16). Using this engineered anti-CD122 mAb to modulate IL-2/IL-15Rβ signaling, we have investigated how T1D-associated autoimmunity at a prediabetic stage was regulated.…”

“…Several studies have reported that an in vivo treatment with an anti-mouse CD122 mAb (clone TM-β1, rat IgG2b) (18) prevented the development of diabetes in NOD mice (13)(14)(15)(16). However, the underlying mechanisms remain poorly defined.…”

“…It thus can be speculated that modulating CD122 signaling may impact the activities of all CD122-bearing cells in normal and diseased conditions. Indeed, a mAb against CD122 (clone TM-β1, rat IgG2b) has been reported to suppress diabetes development in nonobese diabetic (NOD) mice (13)(14)(15)(16), suggesting a therapeutic potential of CD122 blockade for human T1D patients. However, the underlying mechanisms remain largely undefined.…”

LEA29Y expression in transgenic neonatal porcine islet-like cluster promotes long-lasting xenograft survival in humanized mice without immunosuppressive therapy

“…After lysis of erythrocytes cells were washed in PBS with 10% FCS (FACS buffer), stained with fluorochrome-labeled monoclonal antibodies (mAb) to hCD45-APC, hCD3-PE, hCD4-APC (ebioscience, Germany), hCD8-PE (Exbio Praha, Czech Republic), mCD45-FITC (BioLegend, CA, USA), and analyzed by FACS analysis using an Accuri C6 flow cytometer (BD Biosciences, Germany). At the end of the study cell suspensions were prepared from the spleen and bone marrow as described previously 32, 33 . After lysis of erythrocytes 3 × 10 5 cells from bone marrow or spleen were suspended in 100 µl FACS buffer and stained for 30 min with mAb to determine the frequency of murine (mCD45-FITC) and human (hCD45-APC) cells carrying leukocyte common antigen (expressed on all hematopoietic cells) and T cells (human CD3-PE, human CD4, human CD8).…”