Platelet Activation Via Binding of Monoclonal Antibodies to the Fcγ Receptor II

1993 ◽  
pp. 221-228
Author(s):  
J. Michael Wilkinson ◽  
Edward J. Hornby ◽  
Kalwant S. Authi
Keyword(s):  
Monoclonal Antibodies ◽  
Platelet Activation ◽  
Fcγ Receptor

Field flow fractionation for assessing neonatal Fc receptor and Fcγ receptor binding to monoclonal antibodies in solution

2011 ◽  
Vol 414 (1) ◽  
pp. 88-98 ◽  
Author(s):  
Joey Pollastrini ◽  
Thomas M. Dillon ◽  
Pavel Bondarenko ◽  
Robert Y.-T. Chou
Keyword(s):  
Monoclonal Antibodies ◽  
Receptor Binding ◽  
Fc Receptor ◽  
Field Flow Fractionation ◽  
Fcγ Receptor ◽  
Neonatal Fc Receptor ◽  
Flow Fractionation

Platelet activation studies with CD41/61 monoclonal antibodies

1996 ◽  
Vol 52 (4) ◽  
pp. 357-362 ◽  
Author(s):  
A. Mateo ◽  
J.M. Perez de la Lastra ◽  
J.J. Garrido ◽  
D. Llanes
Keyword(s):  
Monoclonal Antibodies ◽  
Platelet Activation

DETECTION OF ACTIVATED PLATELETS IN WHOLE BLOOD BY FLOW CYTOMETRY

1987 ◽  
Author(s):  
S J Shattil ◽  
J A Hoxie ◽  
M Cunningham ◽  
C S Abrahms ◽  
J O’Brien ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Monoclonal Antibodies ◽  
Platelet Activation ◽  
Whole Blood ◽  
Thrombus Formation ◽  
Membrane Surface ◽  
White Blood Cells ◽  
Color Flow ◽  
Platelet Surface ◽  
Activated Platelets

Platelets may become activated in a number of clinical disorders and participate in thrombus formation. We have developed a direct test for activated platelets in whole blood that utilizes dual-color flow cytometry and requires no washing steps. Platelets were distinguished from erythrocytes and white blood cells in the flow cytometer by labeling the platelets with biotin-AP1, an antibody specific for membrane glycoprotein lb, and analyzing the cells for phycoerythrin-streptavidin fluorescence. Membrane surface changes resulting from platelet activation were detected with three different FITC-labeled monoclonal antibodies: 1) PAC1, an antibody specific for the fibrinogen receptor on activated platelets; 2) 9F9, which binds to the D-domain of fibrinogen and detects platelet-bound fibrinogen; and 3) S12, which binds to an alpha-granule membrane protein that associates with the platelet surface during secretion. Unstimulated platelets demonstrated no PAC1, 9F9, or S12-specific fluorescence, indicating that they did not bind these antibodies. Upon stimulation with agonists, however, the platelets demonstrated a dose-dependent increase in FITC-fluorescence. The binding of 9F9 to activated platelets required fibrinogen. Low concentrations of ADP and epinephrine, which induce fibrinogen receptors but little secretion, stimulated near-maximal PAC1 or 9F9 binding but little S12 binding. On the other hand, a concentration of phorbol myristate acetate that evokes full platelet aggregation and secretion induced maximal binding of all three antibodies. When blood samples containing activated and non-activated platelets were mixed, as few as 0.8% activated platelets could be detected by this technique. There was a direct correlation between ADP-induced FITC-PAC1 binding and binding determined in a conventional 125I-PAC1 binding assay (r = 0.99; p < 0.001). These studies demonstrate that activated platelets can be reliably detected in whole blood using activation-dependent monoclonal antibodies and flow cytometry. This method may be useful to assess the degree of platelet activation and the efficacy platelet inhibitor therapy in thrombotic disorders.

Production of Monoclonal Antibodies Specific for Platelet Activation Antigens and Their Use in Evaluating Platelet Function

Hybridoma ◽  
1990 ◽  
Vol 9 (6) ◽  
pp. 631-641 ◽  
Author(s):  
MARYBETH W. CARMODY ◽  
KENNETH A. AULT ◽  
JANE G. MITCHELL ◽  
NEAL S. ROTE ◽  
AH-KAU NG
Keyword(s):  
Monoclonal Antibodies ◽  
Platelet Activation ◽  
Platelet Function ◽  
Activation Antigens

Detection of plasmapheresis-induced platelet activation using monoclonal antibodies

Transfusion ◽  
1992 ◽  
Vol 32 (6) ◽  
pp. 534-540 ◽  
Author(s):  
T Wun ◽  
T Paglieroni ◽  
K Sazama ◽  
P Holland
Keyword(s):  
Monoclonal Antibodies ◽  
Platelet Activation

Neutrophil activation by anti-β 2 glycoprotein I monoclonal antibodies via Fcγ receptor II

1995 ◽  
Vol 57 (3) ◽  
pp. 387-394 ◽  
Author(s):  
Josiane Arvieux ◽  
Marie-Christine Jacob ◽  
Bernard Roussel ◽  
Jean-Claude Bensa ◽  
Maurice G. Colomb
Keyword(s):  
Monoclonal Antibodies ◽  
Neutrophil Activation ◽  
Fcγ Receptor ◽  
Glycoprotein I

scholarly journals Platelet activation by cross-linking HLA class I molecules and Fc receptor

Blood ◽  
1992 ◽  
Vol 79 (11) ◽  
pp. 2901-2908 ◽  
Author(s):  
E Rubinstein ◽  
I Urso ◽  
C Boucheix ◽  
RC Carroll
Keyword(s):  
Monoclonal Antibodies ◽  
Platelet Activation ◽  
Hla Class I ◽  
Fc Receptor ◽  
Class I ◽  
Cross Linking ◽  
Chain Molecule ◽  
Kinase C ◽  
Hla Class I Molecules ◽  
Higher Sensitivity

Abstract The effect on platelet activation of monoclonal antibodies directed against common determinants of the HLA class I heavy chain molecule was studied. Cross-linking W6/32, an anti-HLA class I of IgG2a subclass, led to platelet activation. Two other antibodies of the same subclass did not have this effect on platelets. The lack of activity of the F(ab')2 fragments suggests that the activation signal is mediated by the platelet Fc receptor (Fc gamma RII). Indeed, except for a higher sensitivity of W6/32 to aspirin and apyrase, activations by cross- linking IV-3 (an anti-Fc gamma RII) and W6/32 are similar at the level of InsP3 formation, calcium mobilization, pH modifications, and activation of protein kinase C and myosin kinase. When HLA class I molecules and Fc gamma RII are cross-linked together, platelet activation occurs. This is not observed when a control IgG2a is substituted for W6/32 or when CD9 and Fc receptor are cross-linked together. This suggests that HLA class I molecules and Fc gamma RII synergize to activate platelets.

scholarly journals Human platelet aggregation by murine monoclonal antiplatelet antibodies is subtype-dependent

Blood ◽  
1993 ◽  
Vol 81 (7) ◽  
pp. 1792-1800 ◽  
Author(s):  
S De Reys ◽  
C Blom ◽  
B Lepoudre ◽  
PJ Declerck ◽  
M De Ley ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
Platelet Aggregation ◽  
Platelet Activation ◽  
Human Platelet ◽  
Human Platelets ◽  
Antigen Recognition ◽  
Metabolic Inhibitors ◽  
Antigen Binding ◽  
Human Platelet Aggregation ◽  
Murine Monoclonal Antibodies

Abstract Twenty murine monoclonal antibodies (MoAbs) generated against different human platelet antigens induced clumping of human platelets in plasma and buffer. Whereas one MoAb could agglutinate platelets, clumping for 19 MoAbs was blocked by metabolic inhibitors, indicating that these induce platelet activation. Fifteen MoAbs were of IgG1, two of IgG2a, and two of IgG2b subtype. F(ab')2 fragments of these did not evoke an aggregatory response, but specifically inhibited aggregations by and binding of their respective intact MoAbs to platelets. Single-platelet counting technology indicated that the MoAbs bind through their antigen- binding and Fc domains mainly to the surface of the same platelet, rather than cause interplatelet-binding. Despite these similarities, the mechanism of action was nevertheless subtype-dependent. Aggregation induced by all IgG1 antibodies could consistently be prevented by blocking the Fc gamma II-receptor, whereas aggregations induced by all IgG2 antibodies still occurred with blocked Fc-receptor, provided functional complement was present. We therefore conclude that platelet activation by MoAb-binding is initiated by antigen recognition followed by an Fc domain-dependent step, which involves the Fc gamma II-receptor for IgG1-type MoAbs and complement-binding for IgG2-type MoAbs. Thus, antibodies of different subtypes can aggregate platelets via different pathways.

scholarly journals Monoclonal antibodies to human platelet glycoprotein IIb beta that initiate distinct platelet responses

Blood ◽  
1985 ◽  
Vol 65 (5) ◽  
pp. 1112-1119 ◽  
Author(s):  
LK Jennings ◽  
DR Phillips ◽  
WS Walker
Keyword(s):  
Monoclonal Antibodies ◽  
Platelet Aggregation ◽  
Platelet Activation ◽  
Human Platelet ◽  
Platelet Glycoprotein ◽  
Membrane Glycoprotein ◽  
Spleen Cells ◽  
Glanzmann’S Thrombasthenia ◽  
Glanzmann's Thrombasthenia ◽  
Mouse Myeloma

Abstract Hybridomas secreting monoclonal antibodies (MoAbs) to human platelet membrane glycoprotein IIb (GPIIb) were prepared by fusing cells of a mouse myeloma line to spleen cells from a BALB/c mouse immunized with purified GPIIb. Six of the hybridomas secreted MoAbs that recognized epitopes on the 23,000-dalton, disulfide-linked subunit of GPIIb, GPIIb beta. All six of these MoAbs agglutinated platelets in the absence of calcium. The agglutination titers of three of the MoAbs, however, were enhanced between 2 and 6 log2 dilutions when titrated in the presence of mmol/L of calcium. The enhancement in titer was the result of MoAb- induced platelet activation followed by platelet aggregation, a reaction that could also be initiated by the monovalent Fab fragments prepared from one of the MoAbs. The MoAbs did not significantly agglutinate platelets from patients with Glanzmann's thrombasthenia, confirming biochemical evidence that there is a paucity of GPIIb beta in the membranes of these cells. Our results show that MoAbs to epitopes on GPIIb beta initiate distinct platelet responses; therefore, they should be useful for studying the ways in which regions of surface glycoproteins are involved in platelet-platelet interactions. In addition, these reagents may prove of value in diagnosing and typing patients with Glanzmann's thrombasthenia.

PLATELET ACTIVATION INDUCED BY A MONOCLONAL ANTIBODY AGAINST THE PLATELET GP Ilb/IIIa COMPLEX

1987 ◽  
Author(s):  
P W Modderman ◽  
J G Huisman ◽  
J A van Mourik ◽  
A E G Kr ◽  
v d Borne
Keyword(s):  
Monoclonal Antibody ◽  
Monoclonal Antibodies ◽  
Platelet Aggregation ◽  
Medical Research ◽  
Platelet Activation ◽  
Dense Granules ◽  
Fibrinogen Binding ◽  
Platelet Release ◽  
Induced Aggregation ◽  
Aggregation Of Platelets

A receptor for fibrinogen on the platelet GP Ila/lIIb complex is induced by ADP, thrombin and other agonists. To study functional domains on GP Ilb/IIIa, the effects of anti-GP Ilb/IIIa monoclonal antibodies (Mab’s) on platelet function were determined. One of these Mab’s, 6C9, induced platelet aggregation. The antibody binds to the intact GP Ilb/IIIa complex only, not to free GP lib or free GP Ilia. Its epitope is different from that of C17, a Mab that inhibits ADP-induced fibrinogen binding and platelet aggregation. 6C9 induces fibrinogen-mediated aggregation rather than agglutination since 6C9-induced platelet interactions were blocked by treatments that also inhibited the effects of ADP etc., without inhibiting binding of 6C9 itself. 6C9 induces binding of 125I-fibrinogen (35.000 ± 7.300 molecules/platelet, Kd = 1.3 ± 0.4 µM) to unstirred platelets. Binding of fibrinogen was 60 to 80% inhibited by apyrase, which indicates that 6C9-induced fibrinogen binding is largely mediated via ADP released from platelets. In addition, 6C9 induced aggregation of platelets in the absence of extracellular fibrinogen. Mediation of this process by platelet fibrinogen or other a-granule proteins, released upon activation by 6C9, was implicated by the finding that aggregation of washed platelets, but not of platelets to which fibrinogen was added, could be blocked by PGI2. Platelet release was also assessed directly by measuring β-thromboglobulin (α-granules) and (14C) serotonin (dense granules) in the medium of unstirred platelets incubated with 6C9. F(ab')2 fragments of 6C9 only aggregated platelets in the presence of fibrinogen and did not release (14C) serotonin. Moreover, release induced by intact 6C9 was inhibited by anti-GP Ilb/IIIa Mab C17 but not by C17 F(ab’)2, although the latter inhibited ADP-induced platelet aggregation. These data indicate that binding of antibodies to specific sites on GP Ilb/IIIa may induce Fc-dependent platelet activation.This study was supported by the Foundation for Medical Research MEDIGON (grant no. 900-526-057.

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