Abstract
CD157, encoded by BST-1, has been described as a useful flow cytometric marker for the analysis of paroxysmal nocturnal hemoglobinuria (PNH) as it is a glycosylphosphatidylinositol (GPI)-linked molecule highly expressed on normal monocytes and neutrophils. We and others observed isolated CD157 signal dropouts during intended PNH analysis. We hypothesize that these negative populations occur due to an antibody failure. To investigate the reason for this finding, we compared two different anti-CD157 antibody clones for PNH analysis. We sequenced BST-1 of CD157-negative probands that are not suffering from PNH and expressed wild type and a discovered variant form of CD157 in HEK293 cells. We compared the binding patterns of two different anti-CD157 antibody clones (SY11B5 and RF3) by flow cytometry and western blot analysis. When sequencing two CD157-negative probands we detected a common SNP (p.Arg145Gln) in exon 3 of BST-1. We found that only anti-CD157 antibody clone RF3 but not the more widely used clone SY11B5 was able to detect both, the wild type and the variant form of CD157 in flow cytometric experiments. The failure of anti-CD157 antibody clone SY11B5 to detect a common SNP can explain some CD157-negative cytometric data. This provides crucial knowledge for laboratories performing PNH analyses as such results can potentially lead to false-positive PNH interpretation. Our results confirm the importance of published PNH guidelines. © 2018 International Clinical Cytometry Society.
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Disclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.