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.
Ueda
H
H
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
Kawahara
M
M
Aburatani
T
T
Tsumoto
K
K
Todokoro
K
K
Suzuki
E
E
Nishimura
H
H
Schueler
P A
PA
Winter
G
G
Mahoney
W C
WC
Kumagai
I
I
Nagamune
T
T
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