FIBRINOGEN SYNTHESIS BUT DEFECTIVE STORAGE IN THE PLATELETS OF A PATIENT WITH GLANZMANN’S THROMBASTHENIA

1987 ◽  
Author(s):  
F Belloc ◽  
P Hourdille ◽  
M Boisseau ◽  
A T Nurden
Keyword(s):  
Polyacrylamide Gel Electrophoresis ◽  
Type I ◽  
Membrane Glycoproteins ◽  
Chase Period ◽  
Glanzmann’S Thrombasthenia ◽  
Multiple Protein ◽  
Severe Deficiency ◽  
Glanzmann's Thrombasthenia ◽  
Sds Page ◽  
History Of

Patient A.M. has a lifelong history of bleeding episodes and platelet function defects typical of type I Glanzmann's thrombasthenia. Analysis of platelet membrane glycoproteins (GP) by SDS-polyacrylamide gel electrophoresis (SDS-PAGE) or by studying the binding of monoclonal antibodies to intact platelets, confirmed the presence of only trace amounts of GP lib and GP Ilia. SDS-PAGE also revealed a severe deficiency of the alpha-granule pool of platelet fibrinogen although an immunoblot performed using a monospecific rabbit anti-fibrinogen antibody showed the presence of residual amounts of fibrinogen. This was estimated to be approximately 10 % of the normal range by an ELISA procedure. Incubation of washed platelets from A.M. with (35S) methionine for 3 h at 37°C resulted in the incorporation of radioactivity into multiple protein bands as revealed by SDS-PAGE followed by fluorography. Immunoprecipitation experiments with affinity purified anti-fibrinogen IgG bound to ultrogel confirmed the synthesis of fibrinogen. The amounts of radioactivity obtained were similar to those immunoprecipitated from control platelet extracts under the same conditions. However, unlike for control platelets, the neosynthesized fibrinogen in A.M. platelets was no longer detected after a 18 h cold chase. In contrast, neosynthesized thrombospondin of the patient was normally preserved during the same chase period. When the fate of the neosynthesized fibrinogen in A.M. platelets was studied it was found to be lost at a faster rate than in normal platelets. Immunoblotting experiments confirmed that cytoplasmic proteases may degrade fibrinogen when exposed to the protein. Our results suggest that the fibrinogen deficiency in the platelets of patients with type I thrombasthenia may be related to a storage abnormality in megakaryocytes or platelets and not from a synthesis defect.

Detection of Carriers in Glanzmann's Thrombasthenia

1983 ◽  
Vol 49 (03) ◽  
pp. 182-186
Author(s):  
G T E Zonneveld ◽  
E F van Leeuwen ◽  
A Sturk ◽  
J W ten Cate
Keyword(s):  
Bleeding Time ◽  
Periodic Acid ◽  
Type I ◽  
Clot Retraction ◽  
Periodic Acid Schiff ◽  
Glanzmann’S Thrombasthenia ◽  
Aggregation Response ◽  
Glanzmann's Thrombasthenia ◽  
Sds Page ◽  
Reduced State

SummaryQuantitative glycoprotein (GP) analysis of whole platelets or platelet membranes was performed by SDS-polyacrylamide gelelectrophoresis (SDS-PAGE) and periodic acid Schiff staining in the families of two unrelated Glanzmann’s thrombasthenia (GT) patients. Each family consisted of two symptom free parents, a symptom free daughter and a GT daughter. All symptom free members had a normal bleeding time, clot retraction and platelet aggregation response to adenosine 5’-diphosphate (ADP), collagen and adrenalin. Platelet Zw* antigen was normally expressed in these subjects. GT patiens, classified as a type I and II subject, showed reduced amounts of GP lib and of GP nia. Analysis of isolated membranes in the non-reduced state, however, showed that the amount of GP Ilia was also reduced in three of the four parents, whereas one parent (of the GT type I patient) and the two unaffected daughters had normal amounts of GP Ilia. Quantitative SDS-PAGE may therefore provide a method for the detection of asymptomatic carriers in GT type I and II.

scholarly journals Monocytes and platelets share the glycoproteins IIb and IIIa that are absent from both cells in Glanzmann's thrombasthenia type I

1983 ◽  
Vol 214 (2) ◽  
pp. 331-337 ◽  
Author(s):  
G Gogstad ◽  
Ø Hetland ◽  
N O Solum ◽  
H Prydz
Keyword(s):  
Platelet Membrane ◽  
Type I ◽  
Membrane Glycoproteins ◽  
Edta Treatment ◽  
Glanzmann’S Thrombasthenia ◽  
Glanzmann's Thrombasthenia

By means of an antiserum specific to the complex of the platelet membrane glycoproteins IIb and IIIa we demonstrate here that monocytes and purified monocyte membranes share these glycoproteins with platelets. The monocyte glycoprotein IIb-IIIa complex showed complete immunological identity with the platelet counterpart and, furthermore, dissociated after EDTA treatment exactly as did the platelet complex. In Glanzmann's thrombasthenia type I, monocytes as well as platelets lack this antigen completely.

GLANZMANN'S THROMBASTHENIA IN PREGNANCY: CLINICAL FEATURES AND PLATELET SEROLOGY

1987 ◽  
Author(s):  
M Picó ◽  
A Ribera ◽  
C Martin ◽  
J Zuazu ◽  
J Monasterio
Keyword(s):  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Antigenic Site ◽  
Type I ◽  
Glanzmann’S Thrombasthenia ◽  
Negative Results ◽  
Normal Platelet ◽  
Glanzmann's Thrombasthenia ◽  
Neonatal Thrombocytopenia ◽  
Platelet Antibody

A 24-year-old woman with a diagnosis of Glanzmann's Thrombasthenia (GT) type I (confirmed in our Hemostasis Section by sodium dodecyl sulphate-polyacrylamide gel electrophoresis) and with a history of several transfusions, was seen in our Service since the second month of her first pregnancy.In the first visit a strong anti-Rh(D) antibody was found in her serum. In 28th week an ammiocentesis was done. Seventy two hours later, she was hospitalized because of disminution of foetal movements. Then, a platelet-allo-antibody, IgG+IgM type (title IgG 1/512, IgM 1/2) was detected in her scrum by immunofluorescence test, as well as an anti-HLA antibody (by lymphocytotoxicity test) which reacted with her husband lyin-phocites.The platelet antibody wasn't EDTA or Paraformaldehide (PFA) dependent and reacted with all normal platelets of a panel of known platelet phenotypes and, also although significantly weakly, with GT type II platelets. The negative results were only observed with GT type I platelets. Therefore, it seemed to recognize an antigenic site located in GP IIb-IIIa. Besides, this antibody inhibited the normal platelet aggregation.A caesarian was advised because of foetal suffering an was performed without immediate complications. The patient was protected by our hemotherapy established support and delivered a boy with a severe hemolytic disease (Hematocrit: 6%, bilirrubine: 1,9 mgs./lOCimls., platelets: 40.000/ mm3), who died 48h. later. His platelets had a direct positive test IgG type and in his serum the platelet antibody as in his mother's was detected.We discuss the specificity of the antibody detected and also, its possible implication in the neonatal thrombocytopenia.

scholarly journals Residual amounts of glycoproteins IIb and IIIa may be present in the platelets of most patients with Glanzmann's thrombasthenia

Blood ◽  
1985 ◽  
Vol 65 (4) ◽  
pp. 1021-1024 ◽  
Author(s):  
AT Nurden ◽  
D Didry ◽  
N Kieffer ◽  
RP McEver
Keyword(s):  
Bleeding Disorder ◽  
Platelet Membrane ◽  
Type I ◽  
Membrane Glycoproteins ◽  
Glanzmann’S Thrombasthenia ◽  
Polyclonal Rabbit ◽  
Glanzmann's Thrombasthenia ◽  
Genes Encoding ◽  
Or Genes

Abstract Glanzmann's thrombasthenia is an inherited bleeding disorder characterized by abnormalities of platelet membrane glycoproteins (GP) IIb and IIIa. Most patients, usually designated as type I, have been reported to have undetectable levels of GP IIb and GP IIIa with the assay used. We have used polyclonal rabbit antibodies against GP IIb and GP IIIa in a sensitive immunoblot procedure capable of revealing trace amounts of these glycoproteins. Platelets from nine thrombasthenic patients, including seven with type I disease, were studied. GP IIIa, although decreased, was clearly detectable in platelets of eight patients and GP IIb was identified in five. Our findings suggest that residual quantities of GP IIb and GP IIIa are present in most patients with thrombasthenia and therefore that major deletions in the gene or genes encoding these proteins are uncommon.

Megakaryocytes from the Marrow of a Patient with Glanzmann’s Thrombasthenia Lacked GPII b-III a Complexes

1986 ◽  
Vol 56 (01) ◽  
pp. 066-070 ◽  
Author(s):  
P Hourdillé ◽  
P Fialon ◽  
F Belloc ◽  
M Namur ◽  
M R Boisseau ◽  
...  
Keyword(s):  
Polyacrylamide Gel Electrophoresis ◽  
Fluorescein Isothiocyanate ◽  
Type I ◽  
Igg Antibodies ◽  
Immunogold Staining ◽  
Glanzmann’S Thrombasthenia ◽  
Normal Individuals ◽  
Glanzmann's Thrombasthenia ◽  
Von Willebrand ◽  
Willebrand Factor

SummaryAlthough it is recognized that glycoprotein (GP) IIb-IIIa complexes are deficient in platelets in Glanzmann’s thrombasthenia, little is known of the origin of the defect. We have examined the megakaryocytes in a bone marrow aspirate obtained from a thrombasthenia patient during surgery. Analysis of platelet proteins by SDS-polyacrylamide gel electrophoresis confirmed the patient to be of the type I subgroup. The megakaryocytes were examined by immunofluorescence or by immunocytochemical procedures combined with electron microscopy. Antibodies used included the murine monoclonal antibody, AP-2 and the human allo-antibody, IgG L, both of which recognize determinants on GP IIb-IIIa complexes. Bound antibody was detected by anti-IgG antibodies coupled to fluorescein isothiocyanate or adsorbed on gold particles. In the immunofluorescence studies, permeabilized megakaryocytes were identified by double staining using an antibody to von Willebrand factor (vWF). Whereas mature megakaryocytes and their small precursor cells from normal individuals were strongly fluorescent with AP-2 and IgG L, most vWF positive cells from the Glanzmann’s thrombasthenia patient were negative and the remainder gave but a weak background fluorescence. Immunogold staining on the surface of marrow cells was severely reduced. Our results confirm a deficiency of GP IIb-IIIa complexes in megakaryocytes in thrombasthenia.

A Variant of Glanzmann's Thrombasthenia Characterized by Abnormal Glycoprotein IIb/IIIa Complex Formation

1989 ◽  
Vol 62 (03) ◽  
pp. 977-983 ◽  
Author(s):  
Dominique J Fournier ◽  
Arnold Kabral ◽  
Peter A Castaldi ◽  
Michael C Berndt
Keyword(s):  
Monoclonal Antibodies ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Glanzmann’S Thrombasthenia ◽  
Crossed Immunoelectrophoresis ◽  
Fibrinogen Binding ◽  
Glanzmann's Thrombasthenia ◽  
History Of ◽  
And Control ◽  
Increased Susceptibility

SummaryGlanzmann's thrombasthenia is a congenital bleeding abnormality characterized by absent platelet aggregation due to the failure of fibrinogen to bind to activated thrombasthenic platelets. In the majority of cases, this defect is caused by the absence or marked reduction of a specific fibrinogen-binding aggregation receptor, the GP IIb/IIIa complex. E.T., an 18-year-old female with a life-long history of bleeding and easy bruising, had the normal clinical features of Glanzmann's thrombasthenia. Surprisingly, sodium dodecyl sulphate-polyacrylamide gel electrophoresis of her platelets showed no apparent abnormality of the GP IIb/IIIa complex. Control platelets washed in the presence of 2 mM EDTA and control and patient platelets washed in the presence of 2 mM calcium ions showed normal reactivity with anti-GP IIb, anti-GP Ilia, and anti-GP IIb/IIIa complex specific monoclonal antibodies as evaluated by flow cytometry. In contrast, patient's platelets washed in the presence of 2 mM EDTA reacted with anti-GP IIb, anti-GP Ilia, but not with the complex-specific monoclonal antibodies. The increased susceptibility of the patient's GP IIb/IIIa complex to EDTA dissociation was confirmed by crossed immunoelectrophoresis (CIE). CIE analysis further indicated that the patient's GP IIb/IIIa complex did not bind fibrinogen. The combined results suggest that this patient has Glanzmann's thrombasthenia due to an abnormal association of the GP IIb/IIIa complex which results in the failure of the complex to bind fibrinogen.

Glanzmann’s Thrombasthenia : Surface Labelling by Various Techniques and Analysis by Gel Electrophoresis.

1979 ◽  
Author(s):  
J.L. McGregor ◽  
K.J. Clemetson ◽  
E. James ◽  
M. Dechavanne
Keyword(s):  
Sialic Acid ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Surface Membrane ◽  
Polyacrylamide Gel ◽  
Galactose Oxidase ◽  
Membrane Glycoproteins ◽  
Platelet Surface ◽  
Glanzmann’S Thrombasthenia ◽  
Glanzmann's Thrombasthenia

The surface membrane glycoproteins of glanzmann’s thrombasthenia and normal whole platelets were labelled by techniques specific for either sugar or protein moieties. The labelled platelets were solubilized and electrophoresed in reduced or unreduced state by discontinuous SDS-polyacrylamide gel electrophoresis. Galactose oxidase + NaB3H4, labelling, showed with reduced samples 4 glycoprotein bands : a high M.W. glycoprotein and GP Ia, GP Ib, GP IIIb, more intensely labelled than with control platelets but with similar M.W. After treatment with neuraminidase + galactose oxidase + NaB3H4 to remove terminal sialic acid and label penultimate galactose residues the gels showed on both unreduced and reduced samples the absence of PG IIb and IIIa and a relatively broad and intensely labelled GPIb band compared with control platelets. The use of sodium periodate + NaB3H4, to label predominantly sialic acid moieties gave essentially the same number of GP bands in both reduced and unreduced samples as in normal platelets. Lactoperoxidase iodination showed in thrombasthenic platelets both in the reduced and unreduced states the absence of GPIIb, GPIIIa and more intensely labelled GPIb and CPIIIb than with control platelets. The combination of multilabelling and discontinuous Polyacrylamide gel system provides a reliable method for investigating the platelet surface.

scholarly journals Residual amounts of glycoproteins IIb and IIIa may be present in the platelets of most patients with Glanzmann's thrombasthenia

Blood ◽  
1985 ◽  
Vol 65 (4) ◽  
pp. 1021-1024 ◽  
Author(s):  
AT Nurden ◽  
D Didry ◽  
N Kieffer ◽  
RP McEver
Keyword(s):  
Bleeding Disorder ◽  
Platelet Membrane ◽  
Type I ◽  
Membrane Glycoproteins ◽  
Glanzmann’S Thrombasthenia ◽  
Polyclonal Rabbit ◽  
Glanzmann's Thrombasthenia ◽  
Genes Encoding ◽  
Or Genes

Glanzmann's thrombasthenia is an inherited bleeding disorder characterized by abnormalities of platelet membrane glycoproteins (GP) IIb and IIIa. Most patients, usually designated as type I, have been reported to have undetectable levels of GP IIb and GP IIIa with the assay used. We have used polyclonal rabbit antibodies against GP IIb and GP IIIa in a sensitive immunoblot procedure capable of revealing trace amounts of these glycoproteins. Platelets from nine thrombasthenic patients, including seven with type I disease, were studied. GP IIIa, although decreased, was clearly detectable in platelets of eight patients and GP IIb was identified in five. Our findings suggest that residual quantities of GP IIb and GP IIIa are present in most patients with thrombasthenia and therefore that major deletions in the gene or genes encoding these proteins are uncommon.

Abnormal Processing of the Glycoprotein IIb Transcript due to a Nonsense Mutation in Exon 17 Associated with Glanzmann’s Thrombasthenia

1995 ◽  
Vol 73 (05) ◽  
pp. 756-762 ◽  
Author(s):  
Yoshiaki Tomiyama ◽  
Hirokazu Kashiwagi ◽  
Satoru Kosugi ◽  
Masamichi Shiraga ◽  
Yoshio Kanayama ◽  
...  
Keyword(s):  
Dna Analysis ◽  
Molecular Genetic ◽  
Genetic Defect ◽  
Nucleotide Sequence Analysis ◽  
Type I ◽  
Normal Amount ◽  
Immunoblot Assay ◽  
Glanzmann’S Thrombasthenia ◽  
Glanzmann's Thrombasthenia ◽  
Pcr Products

SummaryWe analyzed the molecular genetic defect responsible for type I Glanzmann’s thrombasthenia in a Japanese patient. In an immunoblot assay using polyclonal anti-GPIIb-IIIa antibodies, some GPIIIa (15% of normal amount) could be detected in the patient’s platelets, whereas GPIIb could not (<2% of normal amount). Nucleotide sequence analysis of platelet GPIIb mRNA-derived polymerase chain reaction (PCR) products revealed that patient’s GPIIb cDNA had a 75-bp deletion in the 3’ boundary of exon 17 resulting in an in-frame deletion of 25 amino acids. DNA analysis and family study revealed that the patient was a compound heterozygote of two GPIIb gene defects. One allele derived from her father was not expressed in platelets, and the other allele derived from her mother had a 9644C → T mutation which was located at the position -3 of the splice donor junction of exon 17 and resulted in a termination codon (TGA). Moreover, quantitative analysis demonstrated that the amount of the abnormal GPIIb transcript in the patient’s platelets was markedly reduced. Thus, the C → T mutation resulting in the abnormal splicing of GPIIb transcript and the reduction in its amount is responsible for Glanzmann’s thrombasthenia.

Sign in / Sign up

Export Citation Format

Share Document