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


METHODS
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.


RESULTS
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.


CONCLUSIONS
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.
Matsumoto
Hisanobu
H
First Department of Biochemistry, Fukuoka University School of Medicine, Japan.
Liao
Shaoxi
S
Arakawa
Fumiko
F
Ueno
Aruto
A
Abe
Hironori
H
Awasthi
Aradhana
A
Kuroki
Motomu
M
Kuroki
Masahide
M
D000818:Animals; D000951:Antigens, Neoplasm / Q000235:genetics* / Q000276:immunology*; D000970:Antineoplastic Agents / Q000633:toxicity; D017252:CD3 Complex / Q000235:genetics / Q000276:immunology; D015815:Cell Adhesion Molecules / Q000235:genetics* / Q000276:immunology*; D002455:Cell Division / Q000187:drug effects; D017931:DNA Primers; D000071858:Epithelial Cell Adhesion Molecule; D018390:Gene Targeting / Q000379:methods; D005822:Genetic Vectors; D006801:Humans; D007128:Immunoglobulin Fragments / Q000235:genetics; D007135:Immunoglobulin Variable Region / Q000235:genetics; D007376:Interleukin-2 / Q000235:genetics / Q000633:toxicity*; D015979:Killer Cells, Lymphokine-Activated / Q000276:immunology; D007700:Kinetics; D051379:Mice; D016513:Mice, SCID; D010843:Pichia / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000633:toxicity; D013274:Stomach Neoplasms / Q000188:drug therapy / Q000473:pathology*; D014162:Transfection; D014183:Transplantation, Heterologous; D014407:Tumor Cells, Cultured
D000951:Antigens, Neoplasm; D000970:Antineoplastic Agents; D017252:CD3 Complex; D015815:Cell Adhesion Molecules; D017931:DNA Primers; D000071858:Epithelial Cell Adhesion Molecule; D007128:Immunoglobulin Fragments; D007135:Immunoglobulin Variable Region; D007376:Interleukin-2; D011993:Recombinant Fusion Proteins