Impaired Mitogen-Induced Expression of High-Affinity Interleukin 2 Receptors on Spleen Cells From NOD/Shi/Kbe Mice Free

The immune abnormalities of NOD mice, a model of human type I (insulin-dependent) diabetes, have been postulated to be T-lymphocyte dependent. We measured responsiveness to exogenous interleukin 2 (IL-2) and IL-2 production in spleen mononuclear cells from female NOD/Shi/Kbe mice after stimulating the cells with concanavalin A (ConA blasts) or phytohemagglutinin (PHA blasts). Exogenous IL-2 produced significantly lower proliferative responses in each blast from 3- and 10-wk-old NOD/Shi/Kbe mice than from control strains. IL-2 production in NOD/Shi/Kbe mice was inclined to decrease but not significantly compared with controls. Even sufficient amounts of recombinant IL-2 (rlL-2) or IL-1 (rlL-1), added with mitogens to the preculture medium, failed to provoke normal proliferative responses from NOD/Shi/Kbe mouse cells. To clarify the reason for this defect, we investigated the expression of IL-2 receptors (IL-2Rs) on mitogen-activated cells with anti- IL-2R monoclonal antibody (PC61) and radiolabeled IL-2. Cytofluorometry showed no significant difference between strains in the number of PC61⁺ ConA and PHA blasts. However, Scatchard analysis with ¹²⁵I-labeled IL-2 showed that the number of high-affinity IL-2Rs (H-IL-2Rs), the mediators of the biological activity of IL-2, was decreased in NOD/Shi/Kbe mice compared with controls, whereas the number of lowaffinity IL-2Rs (L-IL-2Rs) was not different. Separating the L3T4⁺ and Lyt-2⁺ populations of T lymphocytes by cell sorting showed both to be deficient in H-IL-2Rs. Chemical cross-linking of ¹²⁵I-IL-2 bound to H-IL-2Rs on ConA blasts generated the labeling of both IL-2R p70 and p55 proteins (proteins of 70,000 and 55,000 M^r, respectively). These proteins were quantifiably less radioactive in the NOD/Shi/Kbe mice. These studies suggest that the impaired H-IL-2R expression on mitogen-activated splenic T lymphocytes may be involved in the pathogenesis of type I diabetes in NOD/Shi/Kbe mice.