Many studies have shown that cytokines play a major role in the pathogenesis and cure of malignant tumors such as melanoma. Systemic administration of certain cytokines in tumor patients have shown to augment tumor specific immune responses and regression of tumors in some patients. Tumors transfected with genes specific for cytokines have been shown to induce tumor specific immunity suggesting that tumor cells expressing cytokines will be of potential therapeutic value as tumor vaccines. We propose to test the efficacy of a method that could be used as an alternative for introduction of cytokine molecules onto tumors, where gene transfection is not feasible or desirable. The proposed method uses purified molecules and can be completed in a matter of hours with appropriate reagents. Under this proposal specifically we will: 1. construct, express and purify glycosyl-phosphatidylinositol (GPI)- anchored IL-2; 2. determine whether the melanoma cells reconstituted with purified GPI- IL-2 can induce tumor specific immunity. GPI-anchored cytokines will be an alternative to the existing molecules since it will be different from the parent molecule in terms of its ability to spontaneously incorporate into hydrophobic/amphiphilic surfaces such as cell membranes. The knowledge obtained from this seed grant proposal will be of immense value in preparing immunogenic tumor cells for treatment of skin cancers such as melanoma.
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