Full-length recombinant choline transporter-like protein 2 containing arginine 154 reconstitutes the epitope recognized by HNA-3a antibodies. Transfusion 2012 May;52(5):1112-6
Date
10/29/2011Pubmed ID
22032286Pubmed Central ID
PMC3564575DOI
10.1111/j.1537-2995.2011.03411.xScopus ID
2-s2.0-84861097580 (requires institutional sign-in at Scopus site) 8 CitationsAbstract
BACKGROUND: Recent reports have shown that the HNA-3a leukocyte antigen, a target for antibodies that cause severe transfusion-related acute lung injury, correlates with an arginine 154 (rather than glutamine) polymorphism in choline transporter-like protein 2 (CTL2) but did not show directly that R154 determines HNA-3a. CTL2 peptides containing R154 are recognized by only half of HNA-3a antibodies studied to date. Constructs that react with all HNA-3a antibodies are needed to fully define the HNA-3a epitope.
STUDY DESIGN AND METHODS: HEK293 cells were transfected with cDNA encoding full-length CTL2 linked to green fluorescent protein (GFP). Transfectants were selected for GFP expression and tested with antibodies specific for HNA-3a and -3b.
RESULTS: Each of 20 HNA-3a antibodies reacted preferentially with HEK293 cells expressing the R154 CTL2 construct. An HNA-3b antibody reacted only with CTL2 (Q154).
CONCLUSIONS: These findings provide direct evidence that R154 in the context of full-length CTL2 is both necessary and sufficient to create the HNA-3a epitope but suggest that posttranslational modifications of the protein, for example, S-S bonds or addition of glycans, are necessary for recognition of HNA-3a by many antibodies. This could complicate development of an assay for large-scale screening of blood donors to detect anti-HNA-3a.
Author List
Kanack AJ, Peterson JA, Sullivan MJ, Bougie DW, Curtis BR, Aster RHAuthors
Richard H. Aster MD Professor in the Medicine department at Medical College of WisconsinBrian Curtis PhD Director in the Platelet & Neutrophil Immunology Laboratory department at BloodCenter of Wisconsin
MESH terms used to index this publication - Major topics in bold
ArginineBlood Donors
Epitopes
HEK293 Cells
Humans
Isoantibodies
Isoantigens
Membrane Glycoproteins
Membrane Transport Proteins
Protein Processing, Post-Translational
Recombinant Proteins
