PMID- 15009806
OWN - NLM
STAT- MEDLINE
DCOM- 20041104
LR  - 20190827
IS  - 0001-2815 (Print)
IS  - 0001-2815 (Linking)
VI  - 63
IP  - 4
DP  - 2004 Apr
TI  - Molecular studies of anti-HLA-A2 using light-chain shuffling: a structural model 
      for HLA antibody binding.
PG  - 345-54
AB  - Human leukocyte antigen (HLA) A2 is one of the most immunodominant HLA antigens. 
      Through a process of light-chain variable domain (VL) shuffling, we analyzed the 
      VL domains' role in anti-HLA-A2/A28-binding site diversity. This was achieved by 
      combining a VH3-30-encoded HLA-A2/A28-specific heavy-chain variable domain with 
      10(4) non-immune VL domains. Twelve HLA-A2/A28-specific antibodies were 
      subsequently identified. VL gene analysis demonstrated an absence of Vlambda 
      domains and that all have VkappaI-encoded light chains. The affinities correlated 
      with the VkappaI gene present, with the seven highest affinity antibodies using 
      Vkappa domains encoded by the O18 gene segment. A 300-fold difference in affinity 
      was observed between the 12 antibodies, and homology modeling demonstrated a 
      correlation between electrostatic surface potential of the antigen-binding site 
      and affinity for HLA. Overlap between the T-cell receptor-binding site and that 
      of the antibodies was indicated by inhibition of cytotoxic T-lymphocyte killing 
      of peptide-pulsed target cells. A model of antibody binding to HLA-A2 suggested 
      contact with both alpha helices of the HLA molecule, such that the 
      antigen-binding site spans the peptide-binding groove. These data increase the 
      understanding of antibody recognition of HLA and may facilitate the production of 
      clonotypic antibodies with peptide-specific binding.
FAU - Watkins, N A
AU  - Watkins NA
AD  - Department of Hematology, Division of Transfusion Medicine, University of 
      Cambridge, Long Road, Cambridge CB2 2PT, UK. naw23@cam.ac.uk
FAU - Dafforn, T R
AU  - Dafforn TR
FAU - Kuijpers, M
AU  - Kuijpers M
FAU - Brown, C
AU  - Brown C
FAU - Javid, B
AU  - Javid B
FAU - Lehner, P J
AU  - Lehner PJ
FAU - Navarrete, C
AU  - Navarrete C
FAU - Ouwehand, W H
AU  - Ouwehand WH
LA  - eng
PT  - Journal Article
PL  - England
TA  - Tissue Antigens
JT  - Tissue antigens
JID - 0331072
RN  - 0 (Antibodies, Monoclonal)
RN  - 0 (Epitopes)
RN  - 0 (HLA-A2 Antigen)
RN  - 0 (Immunoglobulin Fragments)
SB  - IM
MH  - Antibodies, Monoclonal/chemistry/genetics/*immunology
MH  - Antibody Affinity/genetics
MH  - *Antibody Specificity
MH  - Binding Sites, Antibody
MH  - Cloning, Molecular
MH  - Cytotoxicity Tests, Immunologic
MH  - Epitopes/*immunology
MH  - Gene Library
MH  - HLA-A2 Antigen/*immunology
MH  - Humans
MH  - Immunoglobulin Fragments/chemistry/genetics/*immunology
MH  - Protein Structure, Tertiary
MH  - Structural Homology, Protein
MH  - T-Lymphocytes, Cytotoxic/*immunology
EDAT- 2004/03/11 05:00
MHDA- 2004/11/05 09:00
CRDT- 2004/03/11 05:00
PHST- 2004/03/11 05:00 [pubmed]
PHST- 2004/11/05 09:00 [medline]
PHST- 2004/03/11 05:00 [entrez]
AID - 194 [pii]
AID - 10.1111/j.0001-2815.2004.00194.x [doi]
PST - ppublish
SO  - Tissue Antigens. 2004 Apr;63(4):345-54. doi: 10.1111/j.0001-2815.2004.00194.x.