PHARMACEUTICAL STUDY FOR THERAPY OF LUNG DISEASES USING HUMAN SOD GENE TRANSFORMED CELLS

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Purpose. The purposes of this work were to construct a secretable SOD protein using recombinant DNA technics and to evaluate the anti-inflammatory effects of SOD delivered by genetically modified skin fibroblasts and lung epithelial cells in vitro and in vivo.Methods. To secrete SOD protein into extracellular space, we constructed the plasmid with interleukin-2 signal peptide and human SOD (pRc/CMV-ILSOD). Rat skin fibroblasts and lung epithelial cells were transfected with pRc/CMV-ILSOD including secretable SOD-coding cDNA.The effects of host and transformants on oxidativestress using the xanthine/xanthine oxidase (X/XO) system were examined by two in vitro models to study the autocrine and paracrine SOD action. The anti-inflammatory effects by transplantation of host and transformants were evaluated in 3 kinds of acute inflammation models, carrageenin-induced paw edema, liquid nitrogen-induced edema and paraquat induced lung damage, in rats.Results. The transformants (ILSOD cells) se … More creted SOD protein into the extracellular space, and the extracellular SOD activity in ILSOD cells cultures was significantly increased in comparison with that in host cell cultures. ILSOD cells diminished the cytotoxic activity by X/XO,in autocrine and paracrine fashions. These protective effects of ILSOD cells against X/XO-induced cytotoxicity correlated well with the decreaase in lipid peroxidation in the damaged cells. The in vivo study showed that transplantation of ILSOD cell suspensions into the hind paw in rats inhibited carrageenin-induced paw edema for at least 7 days, and the degrff and the durability of these inhibitory effects were dependent on the number of ILSOD cells transplanted. These inhibitory effects of ILSOD cell suspensions were reduced by coadministration of hSOD antiserum. The healing of paw edema caused by carrageenin was markedly enhancec by transplantation of ILSOD cells into the edematous hind paw. And transplantation of ILSOD cell suspensions into the subcutaneous tissue in rats inhibited liquid nitrogen-induced edema. Furthermore, tansplantation of ILSOD cell suspensions into the thorax in rats inhibited liquid paraquat-induced lung damage.Conclusions. The results suggested that genetically modified skin fibroblasts and lung epithelial cells area suitable delivery system for obtaining efficient and continuous supply of SOD at the target site, and this strategy may be useful as a drug delivery system for other therapeutic proteins. Less