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HNA-3a-specific antibodies recognize choline transporter-like protein-2 peptides containing arginine, but not glutamine at Position 154. Transfusion 2011 Oct;51(10):2168-74 PMID: 21517890 PMCID: PMC3592927

Pubmed ID

21517890

Abstract

BACKGROUND: Antibodies specific for the neutrophil antigen HNA-3a cause severe, sometimes fatal transfusion-related acute lung disease (TRALI) when transfused, but it has not been possible to screen blood donors for anti-HNA-3a because using neutrophils as targets was impractical and molecular properties of the antigen were unknown. Recently it was shown that HNA-3a is carried on choline transporter-like protein-2 (CTL2) and that the HNA-3a/b phenotype is closely correlated with an R154Q amino acid polymorphism in CTL2. However, it has not been shown by direct experiment that R154 is essential for the HNA-3a epitope.

STUDY DESIGN AND METHODS: Preliminary attempts to express recombinant full-length CTL2 (R154) recognized by anti-HNA-3a were unsuccessful. We therefore tested HNA-3a-specific antibodies from donors implicated in TRALI reactions for reactivity against chemically synthesized linear and cyclic CTL2 peptides containing R154 or Q154.

RESULTS: Nine of 20 HNA-3a antibodies recognized the R154, but not the Q154 version of a cyclic 36-residue CTL2 peptide (D131-K166). However, 11 others failed to distinguish between the two versions of this peptide.

CONCLUSION: The findings provide direct evidence that R154 in the context of CTL2 D131-K166 is necessary to create the HNA-3a epitope but, in the context of cyclic CTL2 peptide D131-K166, is sufficient to detect only about one-half of the HNA-3a-specific antibodies implicated in TRALI. It is likely that fragments of CTL2 longer than can be made on a large scale with an automated synthesizer will be needed to produce a target capable of detecting all examples of anti-HNA-3a in donated blood.

Author List

Curtis BR, Sullivan MJ, Holyst MT, Szabo A, Bougie DW, Aster RH

Authors

Richard H. Aster MD Professor in the Medicine department at Medical College of Wisconsin
Brian Curtis PhD Director in the Platelet & Neutrophil Immunology Laboratory department at BloodCenter of Wisconsin
Aniko Szabo PhD Associate Professor in the Institute for Health and Equity department at Medical College of Wisconsin




Scopus

2-s2.0-80054762701   16 Citations

MESH terms used to index this publication - Major topics in bold

Arginine
Blood Transfusion
Enzyme-Linked Immunosorbent Assay
Epitope Mapping
Epitopes
Glutamine
Humans
Isoantibodies
Isoantigens
Membrane Glycoproteins
Membrane Transport Proteins
jenkins-FCD Prod-299 9ef562391eceb2b8f95265c767fbba1ce5a52fd6