Immunophenotypic analysis of reticulocytes in paroxysmal nocturnal hemoglobinuria

Blood. 1995 Aug 15;86(4):1586-9.

Abstract

The hematologic disorder paroxysmal nocturnal hemoglobinuria (PNH) occurs following an acquired somatic mutation in the Piga gene within a bone marrow stem cell. The progeny of this mutated cell cannot synthesize glycosylphosphatidylinositol (GPI) anchors, with a resultant deficiency in surface expression of all GPI-linked proteins. The protean clinical manifestations of PNH presumably result from the deficiency of these GPI-linked surface proteins. To explain the observation that neutrophils are affected at a significantly higher percentage than circulating erythrocytes and to analyze the proliferative rates of erythroid production in PNH, we studied 25 patients using flow cytometry. The fluorescent dye thiazole orange was used to detect reticulocytes, and CD59 monoclonal antibody was used to identify GPI-deficient cells. In contrast to the mature circulating erythrocytes, the percentage of abnormal reticulocytes was similar to the percentage of affected neutrophils. However, the vast majority of reticulocytes was completely GPI-deficient, ie, were type III cells, even in patients with only modest numbers of circulating type III erythrocytes. In addition, greater than 5% type II reticulocytes were identified in only 3 patients, although greater than 5% type II mature erythrocytes were identified in 10 of 25 patients. The results show that the erythroid and neutrophil bone marrow precursors have an equivalent proliferative advantage in PNH. The data also have important implications for the origin of type-II erythrocytes in PNH.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Adolescent
  • Adult
  • Antigens, CD / metabolism
  • CD59 Antigens
  • Female
  • Glycosylphosphatidylinositols / metabolism
  • Hemoglobinuria, Paroxysmal / immunology*
  • Hemoglobinuria, Paroxysmal / pathology
  • Humans
  • Immunophenotyping
  • Male
  • Membrane Glycoproteins / metabolism
  • Middle Aged
  • Reticulocytes / immunology*

Substances

  • Antigens, CD
  • CD59 Antigens
  • Glycosylphosphatidylinositols
  • Membrane Glycoproteins