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Selected Publication:
SHR Neuro Cancer Cardio Lipid Metab Microb
Marget, M; Bohlsen, D; Davarnia, P; Yoo-Ott, K; Kabelitz, D; Skerra, A; Steinmann, J.
A HLA-Cw6 specific single-chain antibody fragment (scFv) recognizing a natural killer cell receptor epitope.
Mol Immunol. 2005; 42(5): 643-649.
Doi: 10.1016/j.molimm.2004.09.013
Web of Science
PubMed
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- Co-authors Med Uni Graz
- Bohlsen Dennis
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- Abstract:
- Major histocompatibility complex (MHC) class I molecules induce inhibitory signals on natural killer (NK) cells via killer cell immunoglobulin-like receptors (KIR). We recently reported a human single-chain antibody (scFv#1), which recognizes an epitope on HLA-Cw6 (genotype: *0602). Flow cytometry showed scFv#1 binding to HLA-Cw6 (strong) and also to HLA-Cw2, 4, 5 (very weak) but not to HLA-Cw1, 3, 7, 8. The presumptive epitope of the antibody fragment, which includes residues Asn77 and Lys80 was verified by introducing point mutations into HLA-Cw6 encoding cDNAs. Asn77 --> Ser77 (N77S) and Lys80- -> Asn80 (K80N) mutants of Cw6 lost scFv#1 binding capacity whereas an additional mutation at aa position 90 (Asp-->Ala, D90A) did not influence scFv#1 binding characteristics. Since residues 77 and 80 of HLA-C are directly involved in KIR/MHC interaction, we expected the induction of target cell lysis upon addition of scFv#1 when bringing NK and HLA-Cw6 positive cells together. To prove this interference, we performed Cr-release assays, using Cw*0602 and mock-transfected K562 erythroleukemia cells as targets and freshly prepared peripheral blood NK cells as effector cells. scFv#1 appeared to influence KIR on ligand binding and restored lysis at low effector to target (E/T) ratios. Pan HLA class I antibody W6/32 did not show such effects. Taken together scFv#1 binding patterns with mutagenized HLA-Cw6 and Cr-release assays are strong evidence that the scFv#1 epitope on HLA-Cw6 is at or close to the binding site of CD158a.
- Find related publications in this database (using NLM MeSH Indexing)
- Amino Acid Sequence -
- Amino Acid Substitution -
- Base Sequence -
- Binding Sites - genetics
- Cell Line - genetics
- Cytotoxicity, Immunologic - genetics
- DNA, Complementary - genetics
- Epitopes - genetics
- HLA-C Antigens - chemistry
- Humans - chemistry
- Immunoglobulin Variable Region - immunology
- Killer Cells, Natural - immunology
- Kinetics - immunology
- Models, Molecular - immunology
- Molecular Sequence Data - immunology
- Mutagenesis, Site-Directed - immunology
- Point Mutation - immunology
- Protein Conformation - immunology
- Receptors, Immunologic - immunology
- Receptors, KIR - immunology
- Receptors, KIR2DL1 - immunology
- Recombinant Proteins - chemistry
- Transfection - chemistry
- Find related publications in this database (Keywords)
- immunology
- lymphocytes
- killer cells
- receptor
- immunoglobulin fragments