Human Interleukin 10 Gene Delivery Prevents Autoimmune Diabetes In Nonobese Diabetic Mice

1. Cloning of Human Interleukin 10 Gene and its Expression in CHO Cells Objective: to get human Interleukin 10(hIL10) full-length cDNA sequence and to express functional hIL10 molecule in CHO cells. Methods: cDNA fragment encoding hIL10 gene was amplified from human peripheral normal white blood cells by RT-PCR and confirmed by DNA sequencing. The ds-DNA was inserted into pcDNA3 vector, this recombinant expressing vector was used to transfect CHO cells. The hIL10 molecules expressed were detected by ELISA and the activity of hIL10 was determined with MTT. Results: The cDNA encoding hIL10 was cloned by RT-PCR and inserted into T-easy vector and sequence analysis verified that the fragment cloned was hIL10 cDNA. hIL10 protein was expressed in the medium of CHO cells and it blocked the lymphocyte transformation.Conclusion: these results lay the foundation for further investigate the role of hIL10 gene delivery in autoimmunity,transplantation immunity and inflammatory disease.Key words hIL10 ;Clone ; CHO ; Gene expression2. IL10 Gene Delivery Prevents Autoimmune Diabetes in Nonobese Diabetic MiceObjective: to study the effectiveness of hIL10 gene delivery on type 1 diabetes in nonobese diabetic(NOD) mice. Methods: 4-week-old NOD mice,were treated with hIL10 expression plasmid or Ringer's medium without hIL10 expression plasmid by rapid tail vein injection. Serum hIL10 was detected by ELISA; blood glucose was measured weekly from 10 weeks of age. At the age of 25 weeks,the mice in two groups that were not diabetic were killed and pancreatic histopathology was studied. Results: Serum hIL10 was detectable for 14 days after injection in the mice injected with hIL10 expression plasmid.The incidence of diabetes in the hIL10 group was 25.00% which was obviously less than 71.43% in the control group(P<0.05), and the degree of insulitis in the hIL10 group was also less than that in the control group. Conclusion: Administration of hIL10 gene can partially prevent NOD mice from developing type 1 diabetes.