Targeting of interleukin-2 to human MK-1-expressing carcinoma by fusion with a single-chain Fv of anti-MK-1 antibody

• Published in: Anticancer research Vol. 22; no. 4; p. 2001
• Main Authors: Matsumoto, Hisanobu, Liao, Shaoxi, Arakawa, Fumiko, Ueno, Aruto, Abe, Hironori, Awasthi, Aradhana, Kuroki, Motomu, Kuroki, Masahide
• Format: Journal Article
• Language: English
• Published: Greece 01.07.2002
• Subjects: Animals; Antigens, Neoplasm - genetics; Antigens, Neoplasm - immunology; Antineoplastic Agents - toxicity; CD3 Complex - genetics; CD3 Complex - immunology; Cell Adhesion Molecules - genetics; Cell Adhesion Molecules - immunology; Cell Division - drug effects; DNA Primers; Epithelial Cell Adhesion Molecule; Gene Targeting - methods; Genetic Vectors; Humans; Immunoglobulin Fragments - genetics; Immunoglobulin Variable Region - genetics; Interleukin-2 - genetics; Interleukin-2 - toxicity; Killer Cells, Lymphokine-Activated - immunology; Kinetics; Mice; Mice, SCID; Pichia - genetics; Recombinant Fusion Proteins - toxicity; Stomach Neoplasms - drug therapy; Stomach Neoplasms - pathology; Transfection; Transplantation, Heterologous; Tumor Cells, Cultured
• ISSN: 0250-7005

Targeting of cytokines into the tumor sites using antibody-cytokine fission proteins represents a novel approach in cancer immunotherapy. We previously reported a novel monoclonal antibody, FU-MK-1, which recognizes a glycoprotein antigen (termed MK-1 antigen) that is overexpressed on the surface of a majority of carcinomas. To target IL-2 and cytotoxicity of effector cells to MK-1-expressing tumor cells, we genetically fused recombinant human interleukin-2 (rhIL-2) to a single chain variable fragment (scFv) antibody derived from FU-MK-1. The resulting fission protein, designated FUscFv/IL-2 was expressed in Pichia pastoris, purified by Ni-affinity chromatography, and characterized for the MK-1-binding specificity and the IL-2 biological activity. The FUscFv/IL-2 fusion protein effectively introduced a specific cytotoxicity of lymphokine-activated killer cells to the tumor cells and consequently suppressed the tumor growth in a SCID mouse xenograft model. This approach may be used for in vivo administration to localize IL-2 to tumor tissues, enhancing the immune response to human MK-1-expressing tumors while reducing systemic side-effects.