A CD66a-specific, activation-dependent epitope detected by recombinant human single chain fragments (scFvs) on CHO transfectants and activated granulocytes
| Autor: | Fritz Grunert, Sabine von Kleist, G.A. Nagel, Michael R. Price, John A. Thompson, Peter Jantscheff, M. J. Embleton |
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| Rok vydání: | 1996 |
| Předmět: |
Neutrophils
HL60 Immunology HL-60 Cells CHO Cells Biology Transfection Epitope law.invention Epitopes chemistry.chemical_compound Antigen Antigens CD law Cricetinae Animals Humans Immunology and Allergy Immunoglobulin Fragments Respiratory Burst chemistry.chemical_classification Chinese hamster ovary cell Cell Biology Antigens Differentiation Molecular biology Recombinant Proteins Respiratory burst N-Formylmethionine Leucyl-Phenylalanine chemistry Recombinant DNA biology.protein Tetradecanoylphorbol Acetate Antibody Glycoprotein Cell Adhesion Molecules HeLa Cells |
| Zdroj: | Journal of Leukocyte Biology. 59:891-901 |
| ISSN: | 1938-3673 0741-5400 |
| DOI: | 10.1002/jlb.59.6.891 |
| Popis: | Antibodies to CD66 recognize at least five members (CD66a–e) of the carcinoembryonic antigen (CEA) family. Recombinant human single-chain Fv fragments (scFvs) that bind specifically to CD66a (biliary glycoprotein) were obtained from a naive human scFv library. The scFvs bound to the N-domain of CD66a on Chinese hamster ovary (CHO) transfectants but did not bind to freshly isolated peripheral granulocytes or to dimethylsulfoxide-treated HL-60 cells. In contrast, scFvs bound well to granulocytes that were short-term activated with N-formyl-Met-Leu-Phe or phorbol 12-myristate 13-acetate and to human HL-60 cells that were treated with all-trans-retinoic acid to induce granulocytic differentiation. Quantification of antigenic sites showed that the activation-dependent CD66a epitopes were expressed on nearly all of the CD66a molecules on CHO-biliary glycoprotein transfectants, but they were detected only on a portion of the molecules on activated polymorphonuclear neutrophils and differentiated HL-60 cells. Binding of CD66a scFvs to their neoepitopes on prestimulated PMNs induced respiratory burst, suggesting that CD66a is capable of delivering transmembrane signals in these cells. |
| Databáze: | OpenAIRE |
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