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2019, Number 1

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Rev Cubana Hematol Inmunol Hemoter 2019; 35 (1)

Diagnosis of paroxysmal nocturnal hemoglobinuria by flow cytometry

Díaz DG, Marsán SV, Sánchez BA

Language: Spanish
References: 30
Page: 1-17
PDF size: 596.22 Kb.


ABSTRACT

Introduction: The paroxysmal nocturnal hemoglobinuria (PNH) is a clonal, acquired disease and not malignant hematopoietic stem cell. In this condition, the anchor to the cell membrane of molecules such as the CD55 and CD59 is affected, This antigens are fundamental in the regulation of the complement-mediated lysis. By its high specificity and sensitivity multiparametric flow cytometry (MFC) is the goal standard for the diagnosis of this disease.
Objective: To establish a diagnosis of PNH by MFC algorithm.
Methods: A sample of peripheral blood of a patient with suspicion of PNH was analyzed by MFC. The cell immunophenotyping was carried out using a panel of monoclonal antibodies directed against antigens that are expressed in the cytoplasmic membrane through its the glycosylphosphatidylinositol anchor. The samples were read in a Cytometer GALLIOS, Beckman Coulter and the data obtained were analyzed with the use of the Kaluza software.
Results: We identified four clones HPN. A HPN clone of approximately 90%, was observed in granulocytes with deficiency in the expression of CD16, CD24, CD55, CD59. In the monocytes were two clones: (1) CD14-CD59- clone and (2) CD14-CD59 + clone, with size clone of 59.77% and 19.45%, respectively. A clone of 19.98% was identified in erythrocytes and determined the degree of involvement of the same.
Conclusions: The proposed analysis algorithm allows to identify cellular populations with clones PNH. In addition, these clones can be quantified in terms of size clonal and expressiveness of anchor to glycosylphosphatidylinositol antigen dependent. With the MFC is achieved with high sensitivity to determine the degree of involvement of the erythrocytes in the expression of CD59 as a direct measure of susceptibility undergoing lysis by complement.


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