Faculty Collaboration Database

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

10/29/2011

Pubmed ID

22032286

Pubmed Central ID

PMC3564575

DOI

10.1111/j.1537-2995.2011.03411.x

Scopus ID

2-s2.0-84861097580 (requires institutional sign-in at Scopus site)   8 Citations

Abstract

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 RH

Author

Brian 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

Arginine
Blood Donors
Epitopes
HEK293 Cells
Humans
Isoantibodies
Isoantigens
Membrane Glycoproteins
Membrane Transport Proteins
Protein Processing, Post-Translational
Recombinant Proteins