Intracellular localization of glycosyl-phosphatidylinositol-anchored CD67 and FcRIII (CD16) in affected neutrophil granulocytes of patients with paroxysmal nocturnal hemoglobinuria.

Immunoelectron microscopical studies performed in healthy human neutrophils showed the presence of glycosyl-phosphatidylinositol (GPI)-linked CD67 in granules. The use of immunogold double-labeling of CD67 and lactoferrin (LF; as marker for specific granules) or CD67 and myeloperoxidase (MPO; as marker for azurophilic granules) showed that CD67 occurred only in the specific granules. Furthermore, flow cytometry showed that CD67 has a low level of expression on the plasma membrane of these cells. In paroxsymal nocturnal hemoglobinuria (PNH)-affected neutrophils, CD67 was not detected in any intracellular compartment by immunoelectron microscopy, and flow cytometry showed no CD67 on the plasma membrane. In earlier studies, FcRIII was found on the plasma membrane, in electron-lucent vesicles, and in the Golgi complex of healthy neutrophils, and in the Golgi complex of some of the PNH-affected neutrophils. Here we have studied FcRIII in PNH-affected cells of three other patients and found, by immunoelectron microscopy, that the receptor can not be detected in these cells. However, flow cytometry showed that FcRIII was not completely absent on the plasma membrane of the affected cells, but that the level of expression on these cells was low. Thus, PNH patients can differ from one another with respect to the occurrence of affected neutrophils that have a detectable level of FcRIII in the Golgi complex. In summary, these findings show not only that the expression of the two GPI-linked proteins, CD67 and FcRIII, is markedly lower on the plasma membrane, but also that neither occurred in any of the intracellular compartments of affected neutrophils of the PNH patients examined in this study.

[1]  J. Robinson,et al.  A novel intracellular compartment with unusual secretory properties in human neutrophils , 1991, The Journal of cell biology.

[2]  T. Huizinga,et al.  Intracellular localization and de novo synthesis of FcRIII in human neutrophil granulocytes. , 1990, Blood.

[3]  I. Singer,et al.  Adhesomes: specific granules containing receptors for laminin, C3bi/fibrinogen, fibronectin, and vitronectin in human polymorphonuclear leukocytes and monocytes , 1989, The Journal of cell biology.

[4]  M. Lisanti,et al.  A glycophospholipid membrane anchor acts as an apical targeting signal in polarized epithelial cells , 1989, The Journal of cell biology.

[5]  Robert B Sim,et al.  Studies on the sensitivity to complement‐mediated lysis of erythrocytes (Inab phenotype) with a deficiency of DAF (decay accelerating factor) , 1989, British journal of haematology.

[6]  D. Brown,et al.  Mechanism of membrane anchoring affects polarized expression of two proteins in MDCK cells. , 1989, Science.

[7]  J. Lippincott-Schwartz,et al.  Pre-Golgi degradation of newly synthesized T-cell antigen receptor chains: intrinsic sensitivity and the role of subunit assembly , 1989, The Journal of cell biology.

[8]  M. G. Low.  Glycosyl‐phosphatidylinositol: a versatile anchor for cell surface proteins 1 , 1989, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[9]  M. Lisanti,et al.  Polarized apical distribution of glycosyl-phosphatidylinositol-anchored proteins in a renal epithelial cell line. , 1988, Proceedings of the National Academy of Sciences of the United States of America.

[10]  P. Weller,et al.  Release of C8 binding protein (C8bp) from the cell membrane by phosphatidylinositol-specific phospholipase C. , 1988, Blood.

[11]  J. Lippincott-Schwartz,et al.  Degradation from the endoplasmic reticulum: Disposing of newly synthesized proteins , 1988, Cell.

[12]  T. Huizinga,et al.  The Pi-linked receptor FcRIII is released on stimulation of neutrophils , 1988, Nature.

[13]  T. Springer,et al.  The major Fc receptor in blood has a phosphatidylinositol anchor and is deficient in paroxysmal nocturnal haemoglobinuria , 1988, Nature.

[14]  J. Atkinson,et al.  Normal polymorphic variations and transcription of the decay accelerating factor gene in paroxysmal nocturnal hemoglobinuria cells. , 1988, Proceedings of the National Academy of Sciences of the United States of America.

[15]  R. Doms,et al.  Regulation of protein export from the endoplasmic reticulum. , 1988, Annual review of cell biology.

[16]  M. Ferguson,et al.  Cell-surface anchoring of proteins via glycosyl-phosphatidylinositol structures. , 1988, Annual review of biochemistry.

[17]  T. Springer,et al.  Chemoattractant-regulated mobilization of a novel intracellular compartment in human neutrophils. , 1987, Science.

[18]  H. Müller-Eberhard,et al.  Deficiency of the homologous restriction factor in paroxysmal nocturnal hemoglobinuria , 1987, The Journal of experimental medicine.

[19]  K. Tokuyasu.  Application of cryoultramicrotomy to immunocytochemistry , 1986, Journal of microscopy.

[20]  M. Klokker,et al.  Non-specific binding of protein-stabilized gold sols as a source of error in immunocytochemistry. , 1986, European journal of cell biology.

[21]  A. von dem Borne,et al.  Neutrophil activation detected by monoclonal antibodies. , 1986, Journal of immunology.

[22]  J. Slot,et al.  A new method of preparing gold probes for multiple-labeling cytochemistry. , 1985, European journal of cell biology.

[23]  J. Metcalf,et al.  Evidence for distinct intracellular pools of receptors for C3b and C3bi in human neutrophils. , 1985, Journal of immunology.

[24]  U. Testa,et al.  Ultrastructural localization of lactoferrin and myeloperoxidase in human neutrophils by immunogold. , 1985, Blood.

[25]  Berger,et al.  Human neutrophils increase expression of C3bi as well as C3b receptors upon activation. , 1984, The Journal of clinical investigation.

[26]  J. Dubochet,et al.  On the preparation of cryosections for immunocytochemistry. , 1984, Journal of ultrastructure research.

[27]  K. Austen,et al.  Affected erythrocytes of patients with paroxysmal nocturnal hemoglobinuria are deficient in the complement regulatory protein, decay accelerating factor. , 1983, Proceedings of the National Academy of Sciences of the United States of America.

[28]  D. Roos,et al.  Characterization of hereditary partial myeloperoxidase deficiency. , 1982, The Journal of laboratory and clinical medicine.

[29]  J. Slot,et al.  Use of colloidal gold particles in double-labeling immunoelectron microscopy of ultrathin frozen tissue sections , 1981, The Journal of cell biology.

[30]  J. Gallin,et al.  Secretory responses of human neutrophils: exocytosis of specific (secondary) granules by human neutrophils during adherence in vitro and during exudation in vivo. , 1979, Journal of immunology.

[31]  M. Leffell,et al.  Character of azurophil and specific granules purified from human polymorphonuclear leukocytes. , 1974, Laboratory investigation; a journal of technical methods and pathology.

[32]  J. Dacie.  Paroxysmal nocturnal haemoglobinuria. , 1972, The Scientific basis of medicine annual reviews.

[33]  M. Farquhar,et al.  THE DEVELOPMENT OF NEUTROPHILIC POLYMORPHONUCLEAR LEUKOCYTES IN HUMAN BONE MARROW , 1971, The Journal of experimental medicine.

[34]  R. Horn,et al.  Ultrastructural aspects of neutrophil granulocyte development in humans. , 1970, Laboratory investigation; a journal of technical methods and pathology.

[35]  E. Weibel,et al.  PRACTICAL STEREOLOGICAL METHODS FOR MORPHOMETRIC CYTOLOGY , 1966, The Journal of cell biology.