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Journal ArticleDOI

A platelet and granulocyte membrane defect in paroxysmal nocturnal hemoglobinuria: usefulness for the detection of platelet antibodies

01 Jul 1969-Journal of Clinical Investigation (American Society for Clinical Investigation)-Vol. 48, Iss: 7, pp 1199-1210
TL;DR: It is concluded that, in PNH, platelets and granulocytes share the membrane defect characteristic of erythrocytes in this disorder, and these observations support the concept that PNH arises as the result of a somatic mutation in a primitive cell capable of differentiating into ery Throttleblast, myeloblast, and megakaryoblast lines.
Abstract: The tendency of platelets and leukocytes to lyse after their interaction with antibody and complement was studied by measuring the release of 51Cr from cells labeled with this isotope. Platelets from six patients with paroxysmal nocturnal hemoglobinuria (PNH) were 15-230 times more sensitive to antibodies and 10-32 times more sensitive to complement than normal platelets or platelets from patients with other types of thrombocytopenic or hemolytic disorders. Mixed white blood cell (WBC) preparations from patients with PNH were 3-20 times more sensitive to anti-WBC antibodies and 5-10 times more sensitive to C′ than were WBC preparations from normal subjects, but PNH lymphocytes showed normal immunologic reactivity. PNH platelets, like PNH erythrocytes, lysed more readily than normal platelets in acidified serum and in media of reduced ionic strength, but these characteristics were not demonstrable with PNH WBC's under the conditions of study. In PNH, platelets appear to comprise a single population with respect to their sensitivity to immune lysis, yet their survival time as measured with 51Cr falls within normal limits. PNH granulocytes likewise appear to consist of a single, uniformly sensitive population. It is concluded that, in PNH, platelets and granulocytes share the membrane defect characteristic of erythrocytes in this disorder. These observations support the concept that PNH arises as the result of a somatic mutation in a primitive cell capable of differentiating into erythroblast, myeloblast, and megakaryoblast lines. PNH platelets or enzymatically treated normal platelets permit the detection of some types of platelet antibodies in dilutions up to 2000-fold greater than is possible with currently available methods, a finding suggesting that the immune lysis technique will prove useful for the study of platelet immunology.

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Citations
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Journal ArticleDOI
01 Sep 1980-Blood
TL;DR: Adult autoimmune throbocytopenic purpura (ATP) is a platelet disorder that develops in certain individuals with a genetic as well as sex (female) predisposition following an environment event, which leads to the rapid clearance and destruction of opsonized platelets by the reticuloendothelial system, particularly the spleen.

436 citations

Journal ArticleDOI
TL;DR: Quantitation of surface IgG of thrombocytopenic platelets was useful in predicting response to treatment and correlated with failure to respond to prednisone therapy.
Abstract: We studied the clinical applicability of a recently developed technic that determines antiplatelet antibody directly on the platelet surface or in serum. The technic is a quantitative complement lysis-inhibition assay. Normal platelets have less than 0.4 pg of surface IgG. All patients with idiopathic thrombocytopenic purpura who were studied had greater than that value. Surface IgG was increased in inverse proportion to the platelet count. Surface levels of greater than 1.1 pg correlated with failure to respond to prednisone therapy. Incubation of normal serums with normal platelets did not increase surface IgG of such platelets, but the incubation with thrombocytopenic serums increased their surface IgG 0.5 to 100 times. The degree of increase did not predict response to treatment. However, quantitation of surface IgG of thrombocytopenic platelets was useful in predicting response to treatment.

433 citations

Journal ArticleDOI
TL;DR: It appears that the lesion leading to PNH can occur at various stages in the differentiation of hematopoietic cells.
Abstract: Decay-accelerating factor (DAF) is a 70,000 Mr protein that has been isolated from the membrane of red cells. The function of DAF is to inhibit the assembly of amplifying enzymes of the complement cascade on the cell surface, thereby protecting them from damage by autologous complement. We raised monoclonal antibodies to DAF and used them to study its distribution in cells from the peripheral blood of normal individuals and of patients with paroxysmal nocturnal hemoglobinuria (PNH), a disease characterized by the unusual susceptibility of red cells to the hemolytic activity of complement. The results of immunoradiometric assays and of fluorescence-activated cell sorter analysis showed that DAF was present not only on red cells but was widely distributed on the surface membrane of platelets, neutrophils, monocytes, and B and T lymphocytes. By Western blotting, we observed small but consistent differences in the Mr of DAF from the membranes of various cell types. Quantitative studies showed that phagocytes and B lymphocytes, which presumably enter more frequently in contact with immune complexes and other potential activators of complement, had the highest DAF levels. As previously reported by others, the red cells from PNH patients were DAF deficient. When the patients' red cells were incubated in acidified serum (Ham test), only the DAF-deficient cells were lysed. In addition, we detected defects in DAF expression on platelets and all types of leukocytes. The observed patterns of DAF deficiency in these patients were consistent with the concept that the PNH cells were of monoclonal origin. In one patient, abnormal and normal cells were found only in the erythroid, myeloid, and megakaryocytic lineages. In two other patients, the lymphocytes were also DAF deficient, suggesting that a mutation occurred in a totipotent stem cell. It appears, therefore, that the lesion leading to PNH can occur at various stages in the differentiation of hematopoietic cells.

376 citations

Journal ArticleDOI
TL;DR: In the human body, most tissues consist primarily of differentiated cells that normally show little evidence of proliferation and self-renewal as discussed by the authors, however, the gastrointestinal mucosal cel...
Abstract: (First of Three Parts) IN man, most tissues consist primarily of differentiated cells that normally show little evidence of proliferation and self-renewal. However, the gastrointestinal mucosal cel...

207 citations

References
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Journal ArticleDOI
01 Jun 1964-Blood
TL;DR: The viability of column-separated cells was shown by their non-staining with trypan blue, motility, phagocytic ability, oxygen consumption, and survival or development in tissue culture, whereas Nowell’s blast-like, dividing, phytohemagglutinin cells were produced only in cultures containing lymphocytes.

437 citations


"A platelet and granulocyte membrane..." refers methods in this paper

  • ...Granulocytes and monocytes were removed by incubating the WBCon a glass bead column essentially as described by Rabinowitz (10)....

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Journal ArticleDOI
TL;DR: It is shown that the red cells of patients with paroxysmal nocturnal hemoglobinuria (PNH) are unusually susceptible to lysis by hetero- and isoantibodies and complement, and the acidified serum test or Ham's test, is of major importance in the diagnosis of the disease.
Abstract: time that the red cells of patients with paroxysmal nocturnal hemoglobinuria (PNH) are unusually susceptible to lysis by hetero- and isoantibodies and complement. These observations have since been confirmed by many other studies using a variety of antibodies (2). The susceptibility of these cells to lysis by antibody and complement has been utilized in the detection of hemolytic antibodies in human serum (3) and in the analysis of the kinetics of hemolysis of human cells by specific antibody and complement (4). The red cells of patients with PNH are also lysed by fresh normal serum, especially when the pH is reduced to 6.5 to 7.0 (5, 6). This reaction, the acidified serum test or Ham's test, is of major importance in the diagnosis of the disease. Lysis by acidified normal serum is abolished when the serum is treated with ammonia or heated to 56° C

271 citations


"A platelet and granulocyte membrane..." refers background or methods or result in this paper

  • ...In some studies the log of y/1l - y was plotted against the log of complement or antibody concentration as done by Rosse and Dacie in analyzing the lysis curves of PNH erythrocytes (6)....

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  • ...of two populations: one highly sensitive to immune lysis and the other only slightly more sensitive than normal (6)....

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  • ...9) are comparable to observations made with PNH erythrocytes (6) and indicate that with platelets, as with red cells, there is...

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Journal ArticleDOI

250 citations


"A platelet and granulocyte membrane..." refers background or result in this paper

  • ...lets in vivo to a variety of antibodies (11, 41, 42)....

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  • ...05 ml of platelet antibody having anti-P1Al specificity (11) which was sufficiently potent to lyse any contaminating platelets but did not react with WBC....

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  • ...trast to isoantibodies reacting with specific platelet antigens which may vary widely in reactivity with platelets from different donors (11, 18)....

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  • ...to six drug-dependent platelet antibodies and two isoantibodies with PlAl specificity2 (11) is compared with that of normal platelets in Table II....

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