AET-treated platelets: their usefulness for platelet antibody detection and an examination of their altered sensitivity to immune lysis

2-Aminoethylisothiouronium bromide (AET) increases the sensitivity of blood cells to complement-mediated immune lysis. We compared the sensitivities of untreated or AET-treated platelets to immune lysis induced by different types of platelet antibody in the 51Cr platelet lysis test. AET platelets were 8–16 times more sensitive to autoantibody and alloantibody, but 8–16 times less sensitive to drug- dependent antibody. AET-platelets bound similar amounts of alloantibody but less drug-dependent antibody, and they lysed at higher complement dilutions than did untreated platelets. AET-platelets detected 10 of 25 autoantibodies, 9 of 9 alloantibodies, and 5 of 8 drug-dependent antibodies. Untreated platelets detected 1 of 25, 6 of 9, and 7 of 8 of these respective platelet antibodies. The use of AET-platelets in the 51Cr platelet lysis test increases its sensitivity for detecting non- drug-dependent platelet antibodies. AET-platelets resemble paroxysmal nocturnal hemoglobinuria (PNH) platelets in their enhanced sensitivity to complement-mediated lysis. They differ from PNH platelets in their insensitivity to immune lysis induced by drug-dependent antibodies and, in this respect, are similar to Bernard-Soulier syndrome platelets.

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AET-treated platelets: their usefulness for platelet antibody detection and an examination of their altered sensitivity to immune lysis

Blood ◽

10.1182/blood.v54.5.1101.1101 ◽

1979 ◽

Vol 54 (5) ◽

pp. 1101-1108 ◽

Cited By ~ 5

Author(s):

PL Cimo ◽

SA Gerber

Keyword(s):

Blood Cells ◽

Paroxysmal Nocturnal Hemoglobinuria ◽

Antibody Detection ◽

Enhanced Sensitivity ◽

Platelet Antibodies ◽

Different Types ◽

Drug Dependent ◽

Altered Sensitivity ◽

Bernard Soulier Syndrome ◽

Platelet Antibody

Abstract 2-Aminoethylisothiouronium bromide (AET) increases the sensitivity of blood cells to complement-mediated immune lysis. We compared the sensitivities of untreated or AET-treated platelets to immune lysis induced by different types of platelet antibody in the 51Cr platelet lysis test. AET platelets were 8–16 times more sensitive to autoantibody and alloantibody, but 8–16 times less sensitive to drug- dependent antibody. AET-platelets bound similar amounts of alloantibody but less drug-dependent antibody, and they lysed at higher complement dilutions than did untreated platelets. AET-platelets detected 10 of 25 autoantibodies, 9 of 9 alloantibodies, and 5 of 8 drug-dependent antibodies. Untreated platelets detected 1 of 25, 6 of 9, and 7 of 8 of these respective platelet antibodies. The use of AET-platelets in the 51Cr platelet lysis test increases its sensitivity for detecting non- drug-dependent platelet antibodies. AET-platelets resemble paroxysmal nocturnal hemoglobinuria (PNH) platelets in their enhanced sensitivity to complement-mediated lysis. They differ from PNH platelets in their insensitivity to immune lysis induced by drug-dependent antibodies and, in this respect, are similar to Bernard-Soulier syndrome platelets.

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Evaluation of influence of IL-6 C-572G gene polymorphism and clinical factors on positive platelet antibody test

Journal of Advanced Pediatrics and Child Health ◽

10.29328/journal.japch.1001023 ◽

2021 ◽

Vol 4 (1) ◽

pp. 006-012

Author(s):

Lin Jeong-Shi ◽

Lee Li-Hsuan ◽

Liu Hsueng-Mei ◽

Chen Ying-Ju ◽

Chiou Tzeon-Jye

Keyword(s):

Gene Polymorphism ◽

Solid Phase ◽

Antibody Test ◽

Antibody Detection ◽

Nucleotide Polymorphisms ◽

Clinical Factors ◽

Hematologic Diseases ◽

Platelet Antibodies ◽

Positive Reactivity ◽

Platelet Antibody

Background: Interleukin-6 (IL-6) promotes antibody production. The objective of this study was to investigate whether IL-6 C-572G single nucleotide polymorphisms (SNP) and clinical factors are associated with positive platelet antibody test. Materials and methods: Thirty platelet recipients with platelet antibodies (responders) and 20 platelet recipients without platelet antibodies (non-responders) were randomly selected. The -572 C>G (rs 1800796) SNPs in the promoter region of IL-6 gene were genotyped by polymerase chain reaction (PCR)-restriction fragment length polymorphism (RFLP) method. Solid phase red cell adherence assay (SPRCA) was used for platelet antibody detection. Results: Age, sex, percentage patients with benign diseases, and percentage of patients with homozygotes for the C allele at position -572 of the IL-6 gene were similar between responders and non-responders. Although the amounts of platelets pheresis transfused to patients with hematologic diseases were higher than those of non-hematologic diseases (47.2 ± 54.2 vs. 17.4 ± 13.8 units, p = 0.019), detection rate of platelet antibodies was lower in patients with hematologic diseases than that in patients with non-hematologic diseases (42.3% vs. 79.2%, p = 0.01). Conclusion: There was no association between IL-6 C-572G gene polymorphism and positive reactivity in solid phase platelet antibody detection method in platelet recipients.

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Drug-Induced Thrombocytopenia

Archives of Pathology & Laboratory Medicine ◽

10.5858/133.2.309 ◽

2009 ◽

Vol 133 (2) ◽

pp. 309-314

Author(s):

Barton Kenney ◽

Gary Stack

Keyword(s):

At Risk ◽

19Th Century ◽

Clinical Management ◽

Drug Induced ◽

Medication Withdrawal ◽

Platelet Antibodies ◽

Patients At Risk ◽

Drug Dependent ◽

The 19Th Century ◽

Prompt Diagnosis

Abstract Drug-induced thrombocytopenia was first described in the 19th century, yet our understanding of its pathogenesis continues to evolve. The list of drugs implicated in drug-induced thrombocytopenia is extensive and growing. Many, if not most, of these medications induce thrombocytopenia by immune mechanisms. Because the degree of thrombocytopenia can put patients at risk for serious bleeding, a prompt diagnosis is key to clinical management. The laboratory approach to diagnosing drug-induced thrombocytopenia is 2-pronged. First, nondrug causes of thrombocytopenia must be ruled out. Second, testing for drug-dependent platelet antibodies, available at specialized reference laboratories, often can identify the offending medication, although usually not in time for initial clinical management. Once a medication is suspected of causing thrombocytopenia, it must be discontinued promptly, and the patient should be monitored closely. Thrombocytopenia generally resolves quickly after offending medication withdrawal, and the prognosis of drug-induced thrombocytopenia is then excellent.

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ENDOTHELIAL FUNCTION AND MORPHOFUNCTIONAL STATE OF BLOOD CELLS IN THE BASINS OF THE ISCHEMIC LIMBS IN THE DIFFERENT TYPES OF SURGICAL TREATMENT OF ATHEROSCLEROSIS OBLITERANS

Clinical Practice ◽

10.17816/clinpract2141-46 ◽

2011 ◽

Vol 2 (1) ◽

pp. 41-46

Author(s):

I I Katelnitsky ◽

S A Pleskachev ◽

M A Burikov ◽

A S Matsionis ◽

P E Povilaytite

Keyword(s):

Surgical Treatment ◽

Red Blood Cells ◽

Blood Cells ◽

Lower Limbs ◽

Lumbar Sympathectomy ◽

Treatment Implementation ◽

Different Types ◽

Red Blood Cell Membranes ◽

Severity Of The Disease ◽

Atherosclerosis Obliterans

The aim of the investigation was to study the morphological condition of blood cells in the basins of the ischemic limbs and their dynamics as a result of various types of surgical treatment. Implementation of combined surgical treatment has a more normalizing effect in comparison with isolated reconstructive surgical treatment. The use of lumbar sympathectomy in patients with occlusive lesions of arteries of lower limbs and varying degrees of ischemia reduces endothelial dysfunction and normalizes a number of parameters describing the morphology and functioning of red blood cells and platelets. There was detected the deformation of red blood cells indicating that the restoration of the plasticity of red blood cell membranes significally reduced the degree of agglutination of red blood cells. According to obtained data the influence of sympathectomy depends on the severity of the disease in general the highest efficiency is observed at the II and III degree of ischemia.

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QUININE CAN INHIBIT OR ENHANCE PRODUCTION OF MURINE ANTI-HUMAN PLATELET ANTIBODIES

10.1055/s-0038-1644577 ◽

1987 ◽

Author(s):

D J Christie ◽

S S Lennon ◽

L L Wischnack

Keyword(s):

Human Platelet ◽

Serum Levels ◽

Human Platelets ◽

Mouse Serum ◽

Enzyme Linked Immunosorbent Assay ◽

Antibody Activity ◽

Low Doses ◽

Platelet Activity ◽

Platelet Antibodies ◽

Drug Dependent

Quinine (Qn) can provoke potent antibodies capable of destroying platelets and inducing life-threatening immunologic thrombocytopenia (DITP). A question critical to understanding the mechanism of DITP is why do platelets from all normal individuals express Qn-induced antigens while only relatively few individuals make the destructive drug-dependent antibodies? This problem was investigated in a murine animal model. BALB/c mice (6-12wk) were injected intraperitoneally, every other week, with saline or lOOOpg of Qn (this dose of drug gave serum levels comparable to human therapeutic serum levels). Two wk after the second injection, mice were immunized with a single dose of 108 human platelets and either 0 or 1000μg of Qn. One wk later, mouse serum was screened in the presence and absence of drug for anti-platelet activity by a complement-dependent 51Cr release assay. Results are shown in the table and represent data from at least 2 groups of five mice each.Antibody titers were more than 10-fold lower in mice receiving 1000 |lg Qn than in mice injected with platelets alone. In contrast, mice repeatedly immunized (3-6 injections) with platelets and low doses of Qn (20μg) developed enhanced antibody activity (drug-dependent and nondrug-dependent) that consistently titered two-to four-fold higher than mice injected with platelets alone. Results were confirmed by an enzyme-linked immunosorbent assay which showed that the murine antibodies were IgG. These findings demonstrate that by varying the dose, Qn can either inhibit or enhance production of anti-human platelet antibodies in mice. This suggests the possibility that most individuals fail to respond to Qn because therapeutic doses of the drug inhibit antibody formation; yet in individuals capable of responding, even low doses of Qn (as found in tonic water) can enhance production of antibodies that provoke DITP.

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Characterization of the complement sensitivity of calcium loaded human erythrocytes

Blood ◽

10.1182/blood.v78.11.3056.3056 ◽

1991 ◽

Vol 78 (11) ◽

pp. 3056-3065 ◽

Cited By ~ 17

Author(s):

ST Test ◽

P Butikofer ◽

MC Yee ◽

FA Kuypers ◽

B Lubin

Keyword(s):

Red Blood Cells ◽

Blood Cells ◽

Paroxysmal Nocturnal Hemoglobinuria ◽

Recent Observation ◽

Membrane Vesicles ◽

Intracellular Potassium ◽

Calcium Loading ◽

Increased Sensitivity ◽

Membrane Vesiculation

Abstract A deficiency of membrane proteins having a glycosylphosphatidylinositol (GPI) anchor is characteristic of the erythrocytes of paroxysmal nocturnal hemoglobinuria (PNH) and is currently believed to be the basis for the enhanced susceptibility to lysis by activated complement observed in these cells. Our recent observation that GPI-anchored proteins are preferentially lost into membrane vesicles shed from normal erythrocytes after calcium loading led us to examine the hypothesis that the remnant erythrocytes might also have increased sensitivity to complement-mediated hemolysis. Indeed, red blood cells treated in such a manner became more sensitive to lysis by antibody and complement or to lysis initiated by activated cobra venom factor complexes (CoFBb). As a consequence of membrane vesiculation, the erythrocytes lost up to approximately 50% of their immunoreactive decay- accelerating factor and 25% to 30% of their immunoreactive membrane inhibitor of reactive lysis (MIRL). Closer examination of the defect responsible for the marked increase in sensitivity to CoFBb-initiated hemolysis seen in calcium-loaded erythrocytes showed that a complex combination of factors produced the defect. These included a decrease in both functional and immunoreactive MIRL and depletion of intracellular potassium and adenosine triphosphate (ATP). These results suggest the possibility that loss of DAF and MIRL via membrane vesiculation, as well as decreases in intracellular potassium and/or ATP, might contribute to the phenotype of PNH erythrocytes. Further, normal or pathologic red blood cells might develop a PNH-like defect after membrane vesiculation if sufficient decreases in potassium and ATP also occurred.

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Hostility in Drug Dependent Individuals: Its Relation to Specific Drugs, and Oral or Intravenous Use

The British Journal of Psychiatry ◽

10.1192/bjp.128.2.188 ◽

1976 ◽

Vol 128 (2) ◽

pp. 188-193 ◽

Cited By ~ 26

Author(s):

Michael R. Gossop ◽

Alec Roy

Keyword(s):

Patient Group ◽

Patient Selection ◽

Drug Dependence ◽

Drug Effects ◽

Several Variables ◽

Different Types ◽

Drug Dependent ◽

Selection Factors ◽

Personal Functioning

SummaryAlthough a number of studies have suggested that hostility and drug dependence may be related, there are few systematic studies of this issue. Using the Hostility and Direction of Hostility Questionnaire, the present study compares several drug-dependent groups of patients. Results showed that the intravenous in-patient group was more hostile on several variables than their out-patient counterparts, and also more hostile than an oral in-patient group. Barbiturate abusers were found to have high levels of hostility; amphetamine abusers were the least hostile group, and narcotic dependent patients were intermediate between the two. Correlations between scales of the HDHQ, were all positive, and most were both high and statistically significant, suggesting that hostility represents a relatively generalized trait in drug-dependent subjects. The results are discussed both in terms of patient selection factors—the differential pressures on different types of patients, and in terms of direct drug effects. It is suggested that the hostility of drug-dependent patients may represent an important problem of personal functioning and may require special attention in treatment programmes.

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STUDIES ON THE PHYSIOLOGY OF THE WHITE BLOOD CELL

Blood ◽

10.1182/blood.v2.3.235.235 ◽

1947 ◽

Vol 2 (3) ◽

pp. 235-243 ◽

Cited By ~ 22

Author(s):

RICHARD WAGNER

Keyword(s):

Blood Cell ◽

White Blood Cell ◽

Blood Cells ◽

Glycogen Content ◽

Striated Muscle ◽

White Blood Cells ◽

Glycogen Storage ◽

Different Types ◽

Blast Cells ◽

Reducing Substances

Abstract The technic of determining glycogen in isolated white blood cells was applied to the study of the different types of leukemia and of polycythemia, in order to obtain information on the physiology of the white blood cell. From this study it is concluded that the granulated leukocyte is the only carrier of glycogen in whole blood. The "reducing substances" in lymphocytes and blast cells are not considered as true glycogen. The glycogen content of wet white blood cells in the rabbit amounts to about 1 per cent. In the human being a range of from 0.17 to 0.67 per cent was calculated. In disease higher percentages occur, in polycythemia up to 1.64 per cent and in glycogen storage disease up to 3.05 per cent. The glycogen concentration of normal white blood cells is within the same range as that of the striated muscle.

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Different distribution of decay-accelerating factor on hematopoietic progenitors from normal individuals and patients with paroxysmal nocturnal hemoglobinuria

Blood ◽

10.1182/blood.v72.2.507.507 ◽

1988 ◽

Vol 72 (2) ◽

pp. 507-511 ◽

Cited By ~ 1

Author(s):

A Kanamaru ◽

K Okuda ◽

E Ueda ◽

T Kitani ◽

T Kinoshita ◽

...

Keyword(s):

Monoclonal Antibodies ◽

Blood Cells ◽

Paroxysmal Nocturnal Hemoglobinuria ◽

Cell Surfaces ◽

Hematopoietic Progenitors ◽

Cell Sorter ◽

Decay Accelerating Factor ◽

Density Distributions ◽

Complement Dependent Cytotoxicity ◽

Normal Individuals

Abstract Deficiency of decay-accelerating factor (DAF) occurs in blood cells in paroxysmal nocturnal hemoglobinuria (PNH), characterized by an unusual susceptibility to hemolysis by complement activation. This study examined DAF expression on hematopoietic progenitors from normal individuals and PNH patients using a fluorescence-activated cell sorter (FACS) with monoclonal antibodies to DAF. Nonphagocytic mononuclear marrow cells expressing different density distributions of DAF were sorted into DAF-, DAF+/-, DAF+, and DAF++ fractions. The cells from each fraction were cultured in methylcellulose and assayed for CFU-E, BFU-E, CFU-GM, and CFU-Mix. The percentages of distribution of DAF- negative normal progenitors increased in the order of CFU-E, CFU-GM, BFU-E, and CFU-Mix, whereas those of DAF-positive cells inversely decreased in this order. These results indicate that DAF expression may accompany differentiation from CFU-Mix to CFU-E. On the other hand, most progenitors in PNH patients had little, if any, expression of DAF on their cell surfaces. These findings were supported by another approach using a complement-dependent cytotoxicity method with the anti- DAF monoclonal antibodies. Abnormal expression of DAF was found on the progenitors in the bone marrow as well as on mature cells circulating in the blood in PNH.

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The Significance of Anti-Platelet Antibodies Associated with Antibiotic Use, A Descriptive Analysis.

Blood ◽

10.1182/blood.v114.22.4470.4470 ◽

2009 ◽

Vol 114 (22) ◽

pp. 4470-4470

Author(s):

Ruta M Shah ◽

Zbigniew M Szczepiorkowski ◽

Miriam K Leach, MS, MT ◽

Richard A Zuckerman

Keyword(s):

Research Funding ◽

Immune Thrombocytopenia ◽

Solid Phase ◽

Descriptive Analysis ◽

Antibiotic Use ◽

Bleeding Complications ◽

Clear Understanding ◽

Platelet Antibodies ◽

Stop Date ◽

Drug Dependent

Abstract Abstract 4470 Introduction Antibiotics (Abx) have been implicated in immune thrombocytopenia via drug dependent platelet antibodies (DDPA). Our institution has had a DDPA assay available since 1994 which is used to guide clinical decisions. We performed a retrospective review to determine the significance of DDPA. Methods We reviewed the medical records of patients (pts) who tested positive for abx DDPA between 1994 and 2006 and performed a descriptive analysis. Detection of DDPAs was performed using a previously described, modified solid phase red cell adherence assay that detects hapten or immune complex reactions. Results A total of 71 pts were included in this analysis. Multiple classes of abx were tested. Platelet nadir was <50 in 70%, between 50 and 100 in 26% and over 100 in 4% of pts. Pts had between 1 and 4 abx tested: 37% had 1, 37% had 2, 15% had 3, and 11% had 4 tested; 65% of pts had one abx positive and 35% had >1 abx positive for DDPA. Of those with >1 abx tested (n=45), 14 (31%) had all tested abx positive. 53% of abx testing took place on or after the abx stop date and 32% of abx tested were administered for <=3 days. Only 29 of 38 pts receiving heparin were tested for heparin-associated antibodies, and 14 had positive results. 49 pts had other non-abx drugs tested, 19 with positive DDPA. Thus, 35% had alternative non-abx testing positive for DDPA. 31% of pts had bleeding complications and 35% of pts died during hospitalization. Excluding pts who died and those with non-abx positive DDPA left 25 pts, median platelet counts were: abx start=142; nadir=22, abx discontinuation=46, hospital discharge=192. Conclusions Antibiotic DDPA testing is usually performed in ill pts with multiple medical complications and comorbidities. Abx were stopped for concern of DDPA in a number of pts for whom typical immune thrombocytopenia was not present or alternative explanations could be found. Though some pts exhibited improvement in platelet count after abx were stopped, a clear understanding of which pts may benefit from DDPA testing could not be determined based on the retrospective nature of this study and the complexity of pts histories. Further research is necessary to clarify the clinical applicability of DPPA testing. Disclosures: Szczepiorkowski: Cersus Corporation: Research Funding; CaridianBCT: Research Funding; BASF: Research Funding; Terumo Corporation: Research Funding; Fenwel, Inc - Scientific Advisory Board: Research Funding.

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