Cell-growth control by monomeric antigen: the cell surface expression of lysozyme-specific Ig V-domains fused to truncated Epo receptor

• Published in: Journal of immunological methods Vol. 241; no. 1; pp. 159 - 170
• Main Authors: Ueda, Hiroshi, Kawahara, Masahiro, Aburatani, Takahide, Tsumoto, Kouhei, Todokoro, Kazuo, Suzuki, Eiji, Nishimura, Hajime, Schueler, Paula A, Winter, Greg, Mahoney, Walt C, Kumagai, Izumi, Nagamune, Teruyuki
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier B.V 31-07-2000; Elsevier
• Subjects: Antibody variable region; Biological and medical sciences; Cell Membrane - metabolism; DNA-Binding Proteins - metabolism; erythropoietin receptors; Fundamental and applied biological sciences. Psychology; Fundamental immunology; Fusion protein; Growth control; Growth Substances - genetics; Growth Substances - immunology; Growth Substances - pharmacology; Immunoglobulin Fragments - biosynthesis; Immunoglobulin Fragments - genetics; Immunoglobulin Variable Region - biosynthesis; Immunoglobulin Variable Region - genetics; Ligands; Milk Proteins; Molecular immunology; Muramidase - genetics; Muramidase - immunology; Muramidase - pharmacology; Phosphorylation; Protein engineering; Protein Engineering - methods; Receptor dimerization; Receptors, Erythropoietin - biosynthesis; Receptors, Erythropoietin - genetics; Recombinant Fusion Proteins - biosynthesis; Signal Transduction; STAT5 Transcription Factor; Techniques; Trans-Activators - metabolism; Receptor dimerization; Protein engineering; Antibody variable region; EpoR, erythropoietin receptor; Growth control; V H, antibody variable heavy chain domain; Fusion protein; V L, antibody variable light chain domain; Epo, erythropoietin; HEL, hen egg lysozyme; Immunoglobulins; Enzyme; Variable region; Method; Antigen; Glycosidases; Hydrolases; O-Glycosidases; Dimerization; Lysozyme; Biological receptor
• ISSN: 0022-1759; 1872-7905
• DOI: 10.1016/S0022-1759(00)00202-7

Previously we have shown that the V
H and V
L fragments of an anti-hen egg lysozyme (HEL) antibody HyHEL-10 are weakly associated but can be driven together by antigen. By joining these antibody variable domains to the cytoplasmic portion of the murine erythropoietin receptor, we created a chimeric growth factor receptor that could be activated by HEL. After co-transfection with two plasmids encoding the respective chimeric receptors in IL-3 dependent murine pro-B Ba/F3 cells, a portion of the cells survived under antigen dependent stimulation without IL-3. These surviving cells all showed coexpression of the two chimeric receptor chains and demonstrated HEL dose-dependent growth stimulation without IL-3. When another IL-3 dependent cell line 32D was transfected with a variant of such chimeric receptor with a linker peptide (Gly–Ser–Gly) inserted between V
H/V
L and EpoR domains, an improved growth response was attained. These observations suggest the utility of heterodimeric Fv chimeric receptors in creating cells that respond to monomeric antigen.