PMID- 24308361
OWN - NLM
STAT- MEDLINE
DCOM- 20140728
LR  - 20220129
IS  - 1537-2995 (Electronic)
IS  - 0041-1132 (Linking)
VI  - 54
IP  - 6
DP  - 2014 Jun
TI  - Expression of a single-chain human leukocyte antigen-DRA/DRB3*01:01 molecule and 
      differential binding of a monoclonal antibody in the presence of specifically 
      bound human platelet antigen-1a peptide.
PG  - 1478-85
LID - 10.1111/trf.12502 [doi]
AB  - BACKGROUND: Studies show that 1 in 1200 neonates have a low platelet (PLT) count 
      due to alloimmunization against human PLT antigen (HPA)-1a (beta3 -L33). This mainly 
      occurs in HPA-1a-negative mothers who are positive for the human leukocyte 
      antigen (HLA)-DRB3*01:01 allele, but only about one-third of cases will mount an 
      effective alloimmune response. The development of specific treatment modalities 
      requires that the mechanisms driving the maternal alloimmune response against the 
      fetal PLTs be further explored. An antibody reagent that has a different binding 
      affinity to HLA-DRA/DRB3*01:01 with and without the beta3 -L33 peptide would be a 
      valuable reagent to study peptide presentation on maternal antigen-presenting 
      cells. STUDY DESIGN AND METHODS: To identify such antibodies, HLA-DRA/DRB3*01:01 
      was recombinantly expressed in Drosophila S2 cells. To delineate the epitope of 
      interesting antibodies, seven mutant HLA-DRA/DRB3*01:01 molecules were generated 
      by site-directed mutagenesis introducing naturally occurring amino acid changes 
      encoded by DRB3*02 and DRB3*03 alleles. RESULTS: The murine monoclonal antibody 
      (MoAb) DA2 showed robust binding by enzyme-linked immunosorbent assay to 
      recombinant HLA-DRA/DRB3*01:01, but binding was reduced in the presence of beta3 
      -L33 peptide. The binding affinity of DA2 to the mutant HLA-DRA/DRB3*0101 in 
      which serine at Position 60 of the beta1-chain was replaced by tyrosine was greatly 
      enhanced. Interestingly the binding of DA2 to the mutant was not reduced by the 
      presence of beta3 -L33 peptide. CONCLUSION: The results of this study generate a 
      molecular model of the interaction of the HLA-DRA/DRB3*01:01 molecule with MoAb 
      DA2. This will inform functional studies with the recombinant Class II molecules.
CI  - (c) 2013 AABB.
FAU - Bouwmans, Evelien E
AU  - Bouwmans EE
AD  - Department of Haematology, University of Cambridge, Cambridge, UK.
FAU - Smethurst, Peter A
AU  - Smethurst PA
FAU - Garner, Stephen F
AU  - Garner SF
FAU - Ouwehand, Willem H
AU  - Ouwehand WH
FAU - Morley, Sarah L
AU  - Morley SL
LA  - eng
GR  - RG/09/012/28096/BHF_/British Heart Foundation/United Kingdom
GR  - RP-PG-0310-1002/DH_/Department of Health/United Kingdom
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20131206
PL  - United States
TA  - Transfusion
JT  - Transfusion
JID - 0417360
RN  - 0 (Antibodies, Monoclonal)
RN  - 0 (Antigens, Human Platelet)
RN  - 0 (HLA Antigens)
RN  - 0 (HLA-DR alpha-Chains)
RN  - 0 (HLA-DRB3 Chains)
RN  - 0 (ITGB3 protein, human)
RN  - 0 (Integrin beta3)
SB  - IM
MH  - Antibodies, Monoclonal/*metabolism
MH  - Antigens, Human Platelet/metabolism
MH  - Binding Sites
MH  - Enzyme-Linked Immunosorbent Assay
MH  - HLA Antigens/*metabolism
MH  - HLA-DR alpha-Chains/chemistry/*metabolism
MH  - HLA-DRB3 Chains/chemistry/*metabolism
MH  - Humans
MH  - Integrin beta3
MH  - Protein Binding
MH  - Protein Structure, Secondary
EDAT- 2013/12/07 06:00
MHDA- 2014/07/30 06:00
CRDT- 2013/12/07 06:00
PHST- 2013/05/31 00:00 [received]
PHST- 2013/10/06 00:00 [revised]
PHST- 2013/10/10 00:00 [accepted]
PHST- 2013/12/07 06:00 [entrez]
PHST- 2013/12/07 06:00 [pubmed]
PHST- 2014/07/30 06:00 [medline]
AID - 10.1111/trf.12502 [doi]
PST - ppublish
SO  - Transfusion. 2014 Jun;54(6):1478-85. doi: 10.1111/trf.12502. Epub 2013 Dec 6.