GAD-grafted IgG Molecule Induces Immune Tolerance To Treat Diabetes In NOD Mice

This study was designed to induce immune tolerance by GAD-grafted IgG, with view to evaluate the affection on treating diabetes in NOD mice.The study was carried out in two parts: 1, GAD-grafted IgG molecule by retroviral-mediated gene expression is used to induce immune tolerance to control diabetes in NOD mice. (1) Identifying female NOD mice model. (2) Identifying GAD-Ig/BSSK, Ig/BSSK, GAD-Ig/MBAE, GAD/MBAE, Ig/MBAE vectors. Retroviral are packaged. (3) That these retro viral transferred J558 cells is testified by RT-PCR, Western blot, ELISA assays. (4) That these retroviral transferred B lineage cells is by RT-PCR, Western blot, ELISA assays. A gene transferred GAD-IgG fusion expressed in B lineage cells can delay diabetes taking on and decrease incidence of diabetes in NOD mice. 2, GAD-grafted IgG molecule by AAV-mediated gene expression is used to induce immune tolerance to control diabetes in NOD mice. (1) Ig, GAD, GAD-Ig gene are inserted into pSNAV. These AAV are packaged. (2) That these AAV transferred BHK-21 cells is testified by RT-PCR, Western blot, ELISA assays. (3) After EGFP/AAV is injected into muscle, EGFP protein can be expressed. (4) Injecting these AAV into muscle in NOD mice in 7 weeks. The results show that GAD-grafted IgG can decrease the incidence of diabetes in NOD mice.Conclusion:We successfully construct GAD-grafted IgG which is used to treat diabetes in NOD mice. Elementary results show that GAD-grafted IgG can delay diabetes taking on and dicrease the incidence of diabetes in NOD mice.