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.
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.
Authors: Pearl Toy; Mark A Popovsky; Edward Abraham; Daniel R Ambruso; Leslie G Holness; Patricia M Kopko; Janice G McFarland; Avery B Nathens; Christopher C Silliman; David Stroncek Journal: Crit Care Med Date: 2005-04 Impact factor: 7.598
Authors: A Davoren; B R Curtis; I A Shulman; A F Mohrbacher; J Bux; B J Kwiatkowska; J G McFarland; R H Aster Journal: Transfusion Date: 2003-05 Impact factor: 3.157
Authors: Christopher C Silliman; Brian R Curtis; Patricia M Kopko; Samina Y Khan; Marguerite R Kelher; Randy M Schuller; Baindu Sannoh; Daniel R Ambruso Journal: Blood Date: 2006-10-12 Impact factor: 22.113
Authors: H Take; Y Tomiyama; Y Shibata; T Furubayashi; S Honda; H Mizutani; T Nishiura; T Tsubakio; Y Kurata; T Yonezawa Journal: Br J Haematol Date: 1990-11 Impact factor: 6.998