scholarly journals Evaluation by quantitative acid elution and radioimmunoassay of multiple classes of immunoglobulins and serum albumin associated with platelets in idiopathic thrombocytopenic purpura

Blood ◽  
1986 ◽  
Vol 67 (4) ◽  
pp. 1126-1131 ◽  
Author(s):  
AJ Hotchkiss ◽  
CA Leissinger ◽  
ME Smith ◽  
JV Jordan ◽  
CA Kautz ◽  
...  
Keyword(s):  
Serum Albumin ◽  
Idiopathic Thrombocytopenic Purpura ◽  
Solid Phase ◽  
Similar Group ◽  
Thrombocytopenic Purpura ◽  
Large Numbers ◽  
Wide Range ◽  
Platelet Concentration ◽  
Associated Proteins ◽  
Solid Phase Radioimmunoassay

Abstract Immunoglobulins (Igs) and serum albumin were eluted from normal platelets and platelets from patients with idiopathic thrombocytopenic purpura (ITP) with a quantitative acid elution procedure followed by solid-phase radioimmunoassay (SPRIA). Acid elution was shown to release a reproducible fraction of platelet-associated Igs, and the amounts released per platelet were independent of the platelet concentration over a wide range of concentrations. This procedure is suitable for sensitive, reproducible, and specific quantitation of large numbers of samples. Washed platelets from 13 normal donors contained the following components (expressed in femtograms per platelet, mean +/- 2 SEM): IgG, 1.40 +/- 0.26; IgA, 0.72 +/- 0.36; IgM 0.078 +/- 0.036; albumin 7.7 +/- 1.5. Immunoglobulins and albumin eluted from the platelets of ten ITP patients (two in remission), expressed as femtograms per platelet, mean (range), were: IgG 104 (0.3 to 750); IgA 90 (0.9 to 715); IgM 162 (1.2 to 1,300); and albumin 34 (6.8 to 199). All platelet-associated Igs from thrombocytopenic ITP patients were found to be elevated twofold to 2,300-fold with one Ig class occasionally elevated 50-fold to 100-fold higher than the others. A similar group of ten thrombocytopenic ITP patients was found to have twofold to 26-fold elevations of platelet- associated albumin. This demonstration of increases in multiple classes of Igs as well as serum albumin associated with platelets from ITP patients suggests that some nonimmune process may be contributing to the phenomenon of increased platelet-associated proteins in ITP.

scholarly journals Evaluation by quantitative acid elution and radioimmunoassay of multiple classes of immunoglobulins and serum albumin associated with platelets in idiopathic thrombocytopenic purpura

Blood ◽  
1986 ◽  
Vol 67 (4) ◽  
pp. 1126-1131
Author(s):  
AJ Hotchkiss ◽  
CA Leissinger ◽  
ME Smith ◽  
JV Jordan ◽  
CA Kautz ◽  
...  
Keyword(s):  
Serum Albumin ◽  
Idiopathic Thrombocytopenic Purpura ◽  
Solid Phase ◽  
Similar Group ◽  
Thrombocytopenic Purpura ◽  
Large Numbers ◽  
Wide Range ◽  
Platelet Concentration ◽  
Associated Proteins ◽  
Solid Phase Radioimmunoassay

Immunoglobulins (Igs) and serum albumin were eluted from normal platelets and platelets from patients with idiopathic thrombocytopenic purpura (ITP) with a quantitative acid elution procedure followed by solid-phase radioimmunoassay (SPRIA). Acid elution was shown to release a reproducible fraction of platelet-associated Igs, and the amounts released per platelet were independent of the platelet concentration over a wide range of concentrations. This procedure is suitable for sensitive, reproducible, and specific quantitation of large numbers of samples. Washed platelets from 13 normal donors contained the following components (expressed in femtograms per platelet, mean +/- 2 SEM): IgG, 1.40 +/- 0.26; IgA, 0.72 +/- 0.36; IgM 0.078 +/- 0.036; albumin 7.7 +/- 1.5. Immunoglobulins and albumin eluted from the platelets of ten ITP patients (two in remission), expressed as femtograms per platelet, mean (range), were: IgG 104 (0.3 to 750); IgA 90 (0.9 to 715); IgM 162 (1.2 to 1,300); and albumin 34 (6.8 to 199). All platelet-associated Igs from thrombocytopenic ITP patients were found to be elevated twofold to 2,300-fold with one Ig class occasionally elevated 50-fold to 100-fold higher than the others. A similar group of ten thrombocytopenic ITP patients was found to have twofold to 26-fold elevations of platelet- associated albumin. This demonstration of increases in multiple classes of Igs as well as serum albumin associated with platelets from ITP patients suggests that some nonimmune process may be contributing to the phenomenon of increased platelet-associated proteins in ITP.

scholarly journals Enhanced Surveillance of Foodborne Mycotoxins by Immunochemical Assay

1988 ◽  
Vol 71 (6) ◽  
pp. 1075-1081 ◽  
Author(s):  
James J Pestka
Keyword(s):  
Quality Control ◽  
Solid Phase ◽  
Protein Conjugates ◽  
Sample Extract ◽  
Large Numbers ◽  
Wide Range ◽  
Enzyme Conjugate ◽  
Aflatoxin B ◽  
Sampling Procedures ◽  
Instrumental Methods

Abstract Mycotoxins are a chemically diverse group of fungal secondary metabolites with a wide range of toxic effects. Conventional thin-layer and instrumental methods of mycotoxin analysis are time-consuming and make routine safety and quality control screening of these compounds in agricultural commodities difficult. As an alternative, specific polyclonal and monoclonal antibodies have been raised against mycotoxin-protein conjugates and used in sensitive radioimmunoassays (RIAs) and enzyme-linked immunosorbent assays (ELISAs). One of the simplest ELISA approaches involves competition for a solid-phase antibody between a mycotoxin-enzyme conjugate and an unconjugated mycotoxin in the sample extract. ELISAs have been developed for aflatoxins B, and M„ zearalenone, T-2 toxin, and deoxynivalenol, which are highly specific, rapid (10 min), easily adaptable for analyzing large numbers of samples, and directly applicable to assaying methanol-water extracts of a wide range of foods. Several commercial mycotoxin ELISAs using this approach (most typically for aflatoxin B,) are currently being marketed. Since ELISAs will be used in large part by personnel with limited technical expertise, individual kits must be critically evaluated by analytical chemists for suggested sampling procedures, efficiency of extraction, crossreactivity, mycotoxin recovery, assay reproducibility, and product shelf-life prior to routine use in food safety and quality control screening

scholarly journals Therapy of chronic idiopathic thrombocytopenic purpura in adults [see comments]

Blood ◽  
1989 ◽  
Vol 74 (7) ◽  
pp. 2309-2317 ◽  
Author(s):  
P Berchtold ◽  
R McMillan
Keyword(s):  
Idiopathic Thrombocytopenic Purpura ◽  
Clinical Studies ◽  
Morbidity And Mortality ◽  
Cooperative Group ◽  
Chronic Idiopathic Thrombocytopenic Purpura ◽  
Therapeutic Approaches ◽  
Group Setting ◽  
Thrombocytopenic Purpura ◽  
Chronic Itp ◽  
Large Numbers

Abstract Chronic ITP is a common hematologic illness. Approximately three fourths of the patients respond to corticosteroids or splenectomy and need no further treatment. Patients refractory to these two therapeutic approaches are relatively resistant to present forms of treatment and are at much greater risk for morbidity and mortality. Future clinical studies evaluating therapy in this refractory group would be best performed in a cooperative group setting in which large numbers of patients could be treated in a prospective randomized manner.

Platelet-associated autoantibodies as detected by a solid-phase modified antigen capture ELISA test (MACE) are a useful prognostic factor in idiopathic thrombocytopenic purpura

Blood ◽  
2004 ◽  
Vol 103 (12) ◽  
pp. 4562-4564 ◽  
Author(s):  
Fabrizio Fabris ◽  
Raffaella Scandellari ◽  
Elisabetta Ruzzon ◽  
Maria Luigia Randi ◽  
Guido Luzzatto ◽  
...  
Keyword(s):  
Idiopathic Thrombocytopenic Purpura ◽  
Clinical Course ◽  
Solid Phase ◽  
Elisa Test ◽  
Patient Admission ◽  
Enzyme Linked Immunosorbent Assay ◽  
Thrombocytopenic Purpura ◽  
Antigen Capture ◽  
Clinical Worsening ◽  
Platelet Autoantibodies

Abstract There were 50 consecutive idiopathic thrombocytopenic purpura (ITP) adult patients (platelet count < 100 × 109/L) grouped according to positivity or negativity of a solid-phase modified antigen capture enzyme-linked immunosorbent assay (ELISA) test (MACE) against glycoprotein IIb/IIIa (GPIIb/IIIa), Ib/IX, and IIa/IIIa. Observation started on the day of MACE assay and lasted at least 6 months. Clinical worsening was defined as the need for starting or modifying therapy because of thrombocytopenia lower than 20 × 109/L or patient admission due to bleeding symptoms. MACE-positive patients had a higher probability of clinical worsening than MACE-negatives (P < .004). The proportion of patients worsening was 18 (72%) of 25 among MACE-positives and 8 (32%) of 25 among MACE-negatives. The median time to clinical worsening was 2.1 months for MACE-positive patients and 27.7 months for MACE-negatives. The assay of specific platelet autoantibodies may be a useful prognostic tool for the clinical course of ITP. (Blood. 2004;103:4562-4564)

scholarly journals Heavy-chain subclass of round antiplatelet IgG in autoimmune thrombocytopenic purpura

Blood ◽  
1980 ◽  
Vol 56 (1) ◽  
pp. 84-87 ◽  
Author(s):  
K Hymes ◽  
PH Schur ◽  
S Karpatkin
Keyword(s):  
Heavy Chain ◽  
Solid Phase ◽  
Protein A ◽  
Normal Serum ◽  
Staphylococcal Protein A ◽  
Antiplatelet Antibody ◽  
Thrombocytopenic Purpura ◽  
Autoimmune Thrombocytopenic Purpura ◽  
Solid Phase Radioimmunoassay ◽  
Monospecific Antisera

Abstract The gamma heavy-chain subclass of bound antiplatelet antibody was examined in six patients with autoimmune thrombocytopenic purpura (ATP) by a solid-phase radioimmunoassay. Monospecific antisera for gamma G1, gamma G2, gamma G3, and gamma G4 subclasses were employed in a “sandwich” technique, utilizing the binding of 126I-staphylococcal protein A. We have previously reported that serum antiplatelet antibody was restricted to be gamma G3 subclass in ATP. In contrast, all 4 IgG subclasses were found bound to platelets of ATP patients in the same distribution as that present in normal serum. It is suggested that the differences noted between serum antiplatelet IgG and platelet-bound IgG may represent different mechanisms of platelet injury.

scholarly journals Changes in splenic microcirculatory pathways in chronic idiopathic thrombocytopenic purpura

Blood ◽  
1991 ◽  
Vol 78 (6) ◽  
pp. 1485-1489 ◽  
Author(s):  
EE Schmidt ◽  
IC MacDonald ◽  
AC Groom
Keyword(s):  
Idiopathic Thrombocytopenic Purpura ◽  
Organ Transplant ◽  
Lymphoid Hyperplasia ◽  
White Pulp ◽  
Chronic Idiopathic Thrombocytopenic Purpura ◽  
Thrombocytopenic Purpura ◽  
Marginal Sinus ◽  
Antiplatelet Antibodies ◽  
Large Numbers ◽  
Flow Through

Abstract The spleen plays a central role in the pathogenesis of chronic idiopathic thrombocytopenic purpura (ITP); it produces massive quantities of antiplatelet antibodies, leading to accelerated phagocytosis of platelets. Lymphoid hyperplasia typically occurs in the spleen, characterized by large numbers of lymphatic nodules with active germinal centers. Whether changes in splenic microcirculatory pathways also occur is not known. We have studied this question by scanning electron microscopy of corrosion casts, comparing spleens removed from patients with ITP with normal spleens obtained from organ transplant donors. The casts demonstrate two major changes in microcirculatory pathways in ITP. Firstly, a striking proliferation of arterioles and capillaries is found in the white pulp and marginal zone (MZ), seen as extensive vascularization in 92.3% of lymphatic nodules (n = 191) versus 0.6% (n = 224) in normal spleens. Secondly, the marginal sinus, a series of flattened, anastomosing vascular spaces between the white pulp and MZ, is absent in 89.4% of lymphatic nodules versus 4.9% in normal spleens. The cause of these microcirculatory changes, which may not be exclusive to ITP, is presently unknown. Absence of the marginal sinus may affect distribution of blood flow through the MZ such that platelets spend increased amounts of time in the proximity of macrophages. In the presence of antiplatelet antibodies found in ITP spleens, this delayed transit would lead to greatly increased platelet destruction.

scholarly journals Heavy-chain subclass of round antiplatelet IgG in autoimmune thrombocytopenic purpura

Blood ◽  
1980 ◽  
Vol 56 (1) ◽  
pp. 84-87
Author(s):  
K Hymes ◽  
PH Schur ◽  
S Karpatkin
Keyword(s):  
Heavy Chain ◽  
Solid Phase ◽  
Protein A ◽  
Normal Serum ◽  
Staphylococcal Protein A ◽  
Antiplatelet Antibody ◽  
Thrombocytopenic Purpura ◽  
Autoimmune Thrombocytopenic Purpura ◽  
Solid Phase Radioimmunoassay ◽  
Monospecific Antisera

The gamma heavy-chain subclass of bound antiplatelet antibody was examined in six patients with autoimmune thrombocytopenic purpura (ATP) by a solid-phase radioimmunoassay. Monospecific antisera for gamma G1, gamma G2, gamma G3, and gamma G4 subclasses were employed in a “sandwich” technique, utilizing the binding of 126I-staphylococcal protein A. We have previously reported that serum antiplatelet antibody was restricted to be gamma G3 subclass in ATP. In contrast, all 4 IgG subclasses were found bound to platelets of ATP patients in the same distribution as that present in normal serum. It is suggested that the differences noted between serum antiplatelet IgG and platelet-bound IgG may represent different mechanisms of platelet injury.

scholarly journals Prevalence and clinical significance of elevated antiphospholipid antibodies in patients with idiopathic thrombocytopenic purpura

Blood ◽  
1994 ◽  
Vol 84 (12) ◽  
pp. 4203-4208 ◽  
Author(s):  
R Stasi ◽  
E Stipa ◽  
M Masi ◽  
F Oliva ◽  
A Sciarra ◽  
...  
Keyword(s):  
Platelet Count ◽  
Clinical Features ◽  
Idiopathic Thrombocytopenic Purpura ◽  
Solid Phase ◽  
Oral Prednisone ◽  
Complete Response ◽  
Anticardiolipin Antibodies ◽  
Common Finding ◽  
Thrombocytopenic Purpura ◽  
Significant Difference

Antibodies against phospholipid antigens (APA) have been demonstrated in idiopathic thrombocytopenic purpura (ITP), but their clinical and pathogenetic significance has remained elusive. In this study we analyzed the prevalence and clinical features of ITP patients with elevated APA. In addition, we prospectively evaluated APA levels after treatment with corticosteroids and compared them with platelet- associated immunoglobulin (PAIgG) titers. We studied 149 patients with newly diagnosed ITP. Of these, 78 had a platelet count less than 50 x 10(9)/L and received an initial treatment with oral prednisone (PDN). In 71 asymptomatic cases with platelet counts between 50 x 10(9)/L and 120 x 10(9)/L, no therapy was scheduled. However, in five of them, the platelet count fell below 50 x 10(9)/L after more than 12 months; these patients were treated with PDN. Tests for APA included the measurement of anticardiolipin antibodies (ACA) with a solid-phase immunoassay and the detection of the lupus-like anticoagulant (LA) activity with coagulation tests that included kaolin-clotting time, dilute Russel's Viper venom time, activated partial thromboplastin time (aPTT), and dilute aPTT. Controls consisted of 174 apparently healthy subjects. Either LA or elevated ACA was seen in 69 patients (46.3%) at diagnosis. LA and ACA were both elevated in 24 cases (16.1% of the overall patient population and 34.8% of patients with high APA concentrations). No correlation was found between LA ratio values and ACA-IgG or -IgM titers, or between ACA-IgG and ACA-IgM levels. The presence of these antibodies was not associated with sex, age, platelet count, or the severity of hemorrhages. PAIgG was detected in 106 of 127 cases (83%). Again, no relationship was observed with clinical parameters or with APA levels. However, all cases with elevated APA also had increased PAIgG. With regard to the clinical course, we were not able to detect any significant difference between patients with normal and elevated APA. An initial complete response to prednisone treatment was observed in 43 of 83 cases (51.8%), with 13 (15.7%) achieving a prolonged complete remission. APA levels were not significantly modified after PDN therapy and on relapse. We conclude that APA positivity is a common finding in patients with ITP and does not select a category with different clinical features. APA levels are not influenced by immunosuppressive therapy with steroids and are not related to the activity of the disease. Therefore, we do not support a role for APA in the pathogenesis of ITP.

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