| Multiparameter flow cytometry for the diagnosis and monitoring of small GPI-deficient cellular populations. | |
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MedLine Citation:
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PMID: 20533383 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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BACKGROUND: Glycosyl-phosphatidylinositol (GPI)-negative blood cells are diagnostic for Paroxysmal Nocturnal Hemoglobinuria (PNH). Marrow failure states are often associated with GPI-negative cell populations. Quantification of small clonal populations of GPI-negative cells influences clinical decisions to administer immunosuppressive therapy in marrow failure states (aplastic anemia or myelodysplastic syndrome) and to monitor minimal residual disease after allogeneic blood or marrow transplantation (BMT). We studied the reliability of high-resolution flow cytometry markers operating at the limits of detection. METHODS: We performed serial quantification of the PNH clone size in 38 samples using multiparameter flow cytometry. Granulocytes, monocytes, and RBCs were gated using forward and side scatter as well as lineage-specific markers. The GPI-linked markers fluorescent aerolysin (FLAER), CD55, and CD59 were comparatively evaluated. We also evaluated CD16 on granulocytes and CD14 on monocytes. The sensitivity of detection by each marker was further defined by serial dilution experiments on a flow-sorted sample. Two patients had quantification of their GPI-negative clones before and after allogeneic BMT. RESULTS: FLAER was the most discriminant marker and allowed identification of 0.1% of GPI-negative cells despite other markers having superior signal-to-noise characteristics. CD14 and CD16 were inferior to CD55 at lower concentrations and in clinical application. CONCLUSIONS: Multiparameter flow cytometry permits quantification of small GPI-negative clones with a sensitivity limit of about 0.1%. The single most reliable marker to monitor small granulocyte or monocyte PNH clones is FLAER, especially in conditions such as myelodysplastic syndromes or BMT, when traditional GPI-linked surface marker expression can be significantly altered. © 2010 International Clinical Cytometry Society. |
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Authors:
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Minoo Battiwalla; Mehmet Hepgur; Dalin Pan; Philip L McCarthy; Manmeet S Ahluwalia; Susan H Camacho; Petr Starostik; Paul K Wallace |
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Publication Detail:
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Type: Evaluation Studies; Journal Article; Research Support, N.I.H., Extramural Date: 2010-06-07 |
Journal Detail:
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Title: Cytometry. Part B, Clinical cytometry Volume: 78 ISSN: 1552-4957 ISO Abbreviation: Cytometry B Clin Cytom Publication Date: 2010 Sep |
Date Detail:
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Created Date: 2010-11-05 Completed Date: 2011-02-28 Revised Date: 2011-09-13 |
Medline Journal Info:
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Nlm Unique ID: 101235690 Medline TA: Cytometry B Clin Cytom Country: United States |
Other Details:
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Languages: eng Pagination: 348-56 Citation Subset: IM |
Copyright Information:
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Copyright © 2010 International Clinical Cytometry Society. |
Affiliation:
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Department of Medicine, Roswell Park Cancer Institute, Buffalo, New York, USA. kunashni@yahoo.com |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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Adolescent Adult Aged Antigens, CD14 / analysis, immunology Antigens, CD55 / analysis, immunology Antigens, CD59 / analysis, immunology Bacterial Toxins / analysis Biological Markers / analysis Blood Cells / chemistry*, immunology Bone Marrow Transplantation / immunology Female Flow Cytometry / methods* Fluorescent Dyes / analysis GPI-Linked Proteins / analysis, immunology Glycosylphosphatidylinositols / analysis*, deficiency*, immunology Granulocytes / immunology Hemoglobinuria, Paroxysmal / diagnosis*, immunology Humans Male Middle Aged Monitoring, Physiologic / methods* Monocytes / immunology Pore Forming Cytotoxic Proteins / analysis Receptors, IgG / analysis, immunology Reproducibility of Results Young Adult |
| Grant Support | |
ID/Acronym/Agency:
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5 P30 CA016056-29/CA/NCI NIH HHS; P30 CA016056-329022/CA/NCI NIH HHS; P30 CA016056-339022/CA/NCI NIH HHS; P30 CA016056-33S59022/CA/NCI NIH HHS; P30 CA016056-33S69022/CA/NCI NIH HHS; S10 RR022335-01A1/RR/NCRR NIH HHS; S10 RR022335-01A10001/RR/NCRR NIH HHS |
| Chemical | |
Reg. No./Substance:
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0/Antigens, CD14; 0/Antigens, CD55; 0/Antigens, CD59; 0/Bacterial Toxins; 0/Biological Markers; 0/FCGR3B protein, human; 0/Fluorescent Dyes; 0/GPI-Linked Proteins; 0/Glycosylphosphatidylinositols; 0/Pore Forming Cytotoxic Proteins; 0/Receptors, IgG; 101754-01-2/CD59 protein, human; 53126-24-2/aerolysin |
| Comments/Corrections | |
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