Transfectomas expressing both secreted and membrane-bound forms of chimeric IgE with anti-viral specificity.
Because of the lack of a cell line expressing on surface and secreting human IgE of known Ag specificity, the construction of a transfectoma line possessing such properties would be useful for studying the roles of surface IgE and the effects of anti-IgE antibodies on IgE-producing B cells. Toward this goal, the human genomic DNA segment encompassing the two exons encoding the membrane anchor peptide of epsilon-chain and their flanking regions was sequenced. Hybrid epsilon and kappa genomic DNA comprising the C regions of human epsilon- and kappa-chains and the H and L chain V regions of the murine mAb BAT123, which reacts with the gp120 envelope protein of HIV-1, were constructed. Mammalian expression vectors containing these fusion genes were used to transfect murine myeloma Sp2/0 cells, and transfectants stably expressing on surface and secreting into culture medium chimeric IgE were obtained. The chimeric IgE showed identical Ag-binding properties as the murine mAb BAT123. Acting in concert with the specific peptide Ag polyvalently coupled to a protein carrier, the chimeric antibody could induce histamine release from human blood basophils. These results demonstrate the potential utility of the transfectoma cells and the chimeric IgE in studying the roles of membrane-bound IgE and effects of anti-IgE antibodies on IgE-producing B cells.
Sun
L K
LK
Tanox Biosystems, Inc., Houston, TX 77025.
Liou
R S
RS
Sun
N C
NC
Gossett
L A
LA
Sun
C
C
Davis
F M
FM
MacGlashan
D W
DW
Chang
T W
TW
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001491:Basophils / Q000276:immunology; D001681:Biological Assay; D015152:Blotting, Northern; D015153:Blotting, Western; D004797:Enzyme-Linked Immunosorbent Assay; D005455:Fluorescent Antibody Technique; D005803:Genes, Immunoglobulin; D015483:HIV Antibodies / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D006636:Histamine Release; D006801:Humans; D006825:Hybridomas / Q000378:metabolism*; D007073:Immunoglobulin E / Q000235:genetics / Q000276:immunology*; D007139:Immunoglobulin epsilon-Chains / Q000235:genetics; D008562:Membrane Glycoproteins / Q000235:genetics / Q000276:immunology; D051379:Mice; D008969:Molecular Sequence Data; D012333:RNA, Messenger / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000276:immunology; D015183:Restriction Mapping; D014162:Transfection
D015483:HIV Antibodies; D007139:Immunoglobulin epsilon-Chains; D008562:Membrane Glycoproteins; D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins; D007073:Immunoglobulin E