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Electrotransfer of human IL‐1Ra into skeletal muscles reduces the incidence of murine collagen‐induced arthritis
Author(s) -
Jeong JaeGyun,
Kim JongMook,
Ho SeongHyun,
Hahn Woong,
Yu Seung Shin,
Kim Sunyoung
Publication year - 2004
Publication title -
the journal of gene medicine
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.689
H-Index - 91
eISSN - 1521-2254
pISSN - 1099-498X
DOI - 10.1002/jgm.599
Subject(s) - electroporation , plasmid , in vivo , microbiology and biotechnology , complementary dna , arthritis , recombinant dna , biology , transfection , naked dna , dna , gene , immunology , genetics
Background It has previously been demonstrated that high levels of gene expression in skeletal muscles can be achieved after direct in vivo electrotransfer of naked plasmid DNA. The purpose of this study is to examine the potential of in vivo electroporation of plasmid DNA encoding human IL‐1Ra for the prevention of murine collagen‐induced arthritis (CIA). Methods DBA/1 mice were injected in gastrocnemius muscles with plasmid DNA followed by in vivo electroporation. To uncover the optimum conditions of gene transfer, various electric field strengths and different amounts of plasmid DNA were applied. Calf muscles around the injected areas were investigated with histological methods for damage to muscle tissue. The levels of human IL‐1Ra expression in the injected area and also in the serum were determined with ELISA for human IL‐1Ra. Based on these data, the effects of electrotransfer of plasmid DNA were tested using the murine CIA model. DBA/1 mice were immunized with bovine collagen type II at the base of the tail. On day 21, mice were given a booster injection with the same antigen. Mice were divided into two groups on day 26. One group of mice received plasmid containing the IL‐1Ra cDNA sequence, while control mice were given plasmid lacking the IL‐1Ra coding sequence. The incidence of arthritis was evaluated by macroscopic analysis, histological analysis, and the levels of inflammatory cytokines. Results IL‐1Ra expression increased as a function of the electrical field strength and the amount of DNA. 200 V/cm (eight pulses; 20 ms per pulse; 1 Hz) and 15 µg of plasmid DNA per mouse were found to be optimum for gene transfer. After in vivo electroporation, gene expression in both muscle and serum increased gradually, reaching a peak value on day 10. Significant levels of human IL‐1Ra expression were maintained for 20 days. Macroscopic analysis showed that the onset of CIA was significantly inhibited by direct electrotransfer of plasmid DNA encoding human IL‐1Ra. Histological analysis of knee joints showed that the incidence of arthritis in knee joints was also prevented. The levels of mouse IL‐1β and IL‐12 in paws were significantly lower in the group treated with IL‐1Ra than those in the control group. Conclusions These results demonstrate that direct electrotransfer of plasmid containing the human IL‐1Ra cDNA sequence to skeletal muscle can reduce the incidence of CIA in mice. Copyright © 2004 John Wiley & Sons, Ltd.