Multiparameter flow cytometry for the diagnosis and monitoring of small GPI-deficient cellular populations

Autor: Susan H. Camacho, Petr Starostik, Minoo Battiwalla, Mehmet Hepgur, Paul K. Wallace, Dalin Pan, Manmeet S. Ahluwalia, Philip L. McCarthy
Rok vydání: 2009
Předmět:
Adult
Male
Pore Forming Cytotoxic Proteins
Pathology
medicine.medical_specialty
Histology
Adolescent
Glycosylphosphatidylinositols
Bacterial Toxins
Hemoglobinuria
Paroxysmal
Lipopolysaccharide Receptors
CD59 Antigens
Biology
Granulocyte
GPI-Linked Proteins
Monocytes
Article
Pathology and Forensic Medicine
Flow cytometry
Young Adult
medicine
Humans
Aplastic anemia
Bone Marrow Diseases
Aged
Bone Marrow Transplantation
Fluorescent Dyes
Monitoring
Physiologic
Blood Cells
medicine.diagnostic_test
CD55 Antigens
Myelodysplastic syndromes
Receptors
IgG
Anemia
Aplastic
Reproducibility of Results
Cell Biology
Bone Marrow Failure Disorders
Middle Aged
medicine.disease
Flow Cytometry
Minimal residual disease
medicine.anatomical_structure
Immunology
Paroxysmal nocturnal hemoglobinuria
Female
Clone (B-cell biology)
Cytometry
Biomarkers
Granulocytes
Zdroj: Cytometry. Part B, Clinical cytometry. 78(5)
ISSN: 1552-4957
Popis: 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
Databáze: OpenAIRE
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