Cell-growth control by monomeric antigen: the cell surface expression of lysozyme-specific Ig V-domains fused to truncated Epo receptor.
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.
Department of Chemistry and Biotechnology, Graduate School of Engineering, The University of Tokyo, Hongo, Bunkyo-ku, 113-8656, Tokyo, Japan. hueda@bio.t.u-tokyo.ac.jp
H
H
Ueda
M
M
Kawahara
T
T
Aburatani
K
K
Tsumoto
K
K
Todokoro
E
E
Suzuki
H
H
Nishimura
P A
PA
Schueler
G
G
Winter
W C
WC
Mahoney
I
I
Kumagai
T
T
Nagamune
D002462:Cell Membrane / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000378:metabolism; D006133:Growth Substances / Q000235:genetics / Q000276:immunology / Q000494:pharmacology*; D007128:Immunoglobulin Fragments / Q000096:biosynthesis / Q000235:genetics; D007135:Immunoglobulin Variable Region / Q000096:biosynthesis* / Q000235:genetics; D008024:Ligands; D008894:Milk Proteins; D009113:Muramidase / Q000235:genetics / Q000276:immunology* / Q000494:pharmacology*; D010766:Phosphorylation; D015202:Protein Engineering / Q000379:methods*; D017467:Receptors, Erythropoietin / Q000096:biosynthesis* / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D050799:STAT5 Transcription Factor; D015398:Signal Transduction; D015534:Trans-Activators / Q000378:metabolism
D004268:DNA-Binding Proteins; D006133:Growth Substances; D007128:Immunoglobulin Fragments; D007135:Immunoglobulin Variable Region; D008024:Ligands; D008894:Milk Proteins; D017467:Receptors, Erythropoietin; D011993:Recombinant Fusion Proteins; D050799:STAT5 Transcription Factor; D015534:Trans-Activators; C055966:immunoglobulin Fv; D009113:Muramidase