Intramuscular injection of interleukin-10 plasmid DNA prevented autoimmune diabetes in mice.
OBJECTIVE
To investigate the effect of plasmid coding interleukin-10 (IL-10) DNA on the development of autoimmune diabetes induced by multiple low doses of streptozotocin (STZ) in mice.


METHODS
Injection of STZ (40 mg/kg, i.p.) was given daily for five consecutive days. pcDNA3-IL-10 plasmid (IL-10-treated group) or pcDNA3-null plasmid (pcDNA3-null-treated group) (100 microg DNA once a day) were injected into skeletal muscles of mice on d 1 and d 14. Blood glucose concentration was measured. After mice were killed on d 28, serum IFN-gamma level was measured by ELISA, and pancreatic IL-1beta and TNF-alpha mRNA expression was detected by semi-quantitative reverse-transcription PCR (RT-PCR). The number of CD4+ and CD8+ lymphocytes from spleen was detected using FACS. In addition, pancreatic histology was measured for determination of insulitis grades.


RESULTS
Treatment with pcDNA3-IL-10 resulted in the retention and expression of the vector in skeletal muscle, associated with a considerable elevation in the plasma level of IL-10, which was not observed in pcDNA3-null-treated mice. In IL-10-treated diabetic mice induced by STZ, delay-type hypersensitivity responses were suppressed and the glucose level was greatly lower on d 14, 21, and 28 than pcDNA3-null-treated group (P<0.05 or P<0.01). On d 21 and 28 the incidence of diabetes was 33.3% and 40.0%, respectively, which was markedly lower than that of pcDNA3-null-treated group (P<0.05). In IL-10-treated mice pancreatic IL-1beta and TNF-alpha mRNA expression was depressed, and serum IFN-gamma concentration and the number of spleen CD4+ or CD8+ lymphocytes were decreased on d 28. The insulitis grades of IL-10-treated mice were lower than that of pcDNA3-null-treated group (P<0.01).


CONCLUSIONS
Systemic administration of IL-10 plasmid DNA can alleviate insulitis of experimental autoimmune diabetes in mice and reduce incidence of diabetes.
Zhang
Zhen-Lin
ZL
Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200031, China.
Shen
Shui-Xian
SX
Lin
Bo
B
Yu
Lu-Yang
LY
Zhu
Li-Hua
LH
Wang
Wei-Ping
WP
Luo
Fei-Hong
FH
Guo
Li-He
LH
D000818:Animals; D004247:DNA / Q000627:therapeutic use; D003921:Diabetes Mellitus, Experimental / Q000139:chemically induced / Q000517:prevention & control; D003922:Diabetes Mellitus, Type 1 / Q000139:chemically induced / Q000517:prevention & control*; D015316:Genetic Therapy; D007273:Injections, Intramuscular; D007375:Interleukin-1 / Q000378:metabolism; D016753:Interleukin-10 / Q000235:genetics / Q000378:metabolism / Q000627:therapeutic use*; D051379:Mice; D008810:Mice, Inbred C57BL; D010179:Pancreas / Q000378:metabolism / Q000473:pathology; D010957:Plasmids / Q000235:genetics; D012333:RNA, Messenger / Q000378:metabolism; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism
D007375:Interleukin-1; D012333:RNA, Messenger; D014409:Tumor Necrosis Factor-alpha; D016753:Interleukin-10; D004247:DNA