scholarly journals Role of kidney in the catabolic clearance of human platelet antiheparin proteins from rat circulation

Blood ◽  
1981 ◽  
Vol 57 (2) ◽  
pp. 233-238
Author(s):  
CP Bastl ◽  
J Musial ◽  
M Kloczewiak ◽  
J Guzzo ◽  
I Berman ◽  
...  
Keyword(s):  
Renal Excretion ◽  
Kidney Tissue ◽  
Rat Plasma ◽  
Renal Tissue ◽  
Human Platelets ◽  
Platelet Factor 4 ◽  
Metabolic Clearance ◽  
Platelet Factor ◽  
Functioning Kidney

Stimulated platelets release at least two antiheparin proteins: platelet factor 4 (PF4) and low affinity platelet factor 4 (LA-PF4) from which beta-thromboglobulin (beta TG) is derived. We have found previously marked elevation of LA-PF4/beta TG antigen in platelet poor plasma of patients with chronic renal failure, whereas levels of PF4 remained normal. Therefore, we examined the role of the kidneys in the metabolic clearance of LA-PF4/beta TG and PF4. The supernates of aggregates of thrombin-stimulated human platelets were injected into sham operated control rats, nephrectomized rats, and into rats with acute ureteral ligation. The disappearance of human LA-PF4/beta TG antigen and PF4 in rat plasma determined by specific radioimmunoassays followed biphasic exponential curves. The half-lives (t1/2) for the fast and slow components of LA-PF4 in control rats were 6.4 and 68.4 min. Nephrectomy significantly increased these times to 9.7 and 144 min, while ureteral ligation resulted in no significant change. Comparison of the level of LA-PF4/beta TG antigen and of creatinine in aorta and in renal vein showed 25%-30% extraction of these compounds by the kidney. Less than 0.1% of the total LA-PF4 antigen injected was recovered in the urine of control rats. In contrast to these results, the clearance of PF4 was not affected by nephrectomy. In conclusion: (1) functional renal tissue is necessary for normal clearance of LA- PF4/beta TG, but renal excretion does not play a major role in its elimination suggesting that the protein is catabolized by the kidney; and (2) catabolic clearance of PF4 does not depend on functioning kidney tissue.

scholarly journals Role of kidney in the catabolic clearance of human platelet antiheparin proteins from rat circulation

Blood ◽  
1981 ◽  
Vol 57 (2) ◽  
pp. 233-238 ◽  
Author(s):  
CP Bastl ◽  
J Musial ◽  
M Kloczewiak ◽  
J Guzzo ◽  
I Berman ◽  
...  
Keyword(s):  
Renal Excretion ◽  
Kidney Tissue ◽  
Rat Plasma ◽  
Renal Tissue ◽  
Human Platelets ◽  
Platelet Factor 4 ◽  
Metabolic Clearance ◽  
Platelet Factor ◽  
Functioning Kidney

Abstract Stimulated platelets release at least two antiheparin proteins: platelet factor 4 (PF4) and low affinity platelet factor 4 (LA-PF4) from which beta-thromboglobulin (beta TG) is derived. We have found previously marked elevation of LA-PF4/beta TG antigen in platelet poor plasma of patients with chronic renal failure, whereas levels of PF4 remained normal. Therefore, we examined the role of the kidneys in the metabolic clearance of LA-PF4/beta TG and PF4. The supernates of aggregates of thrombin-stimulated human platelets were injected into sham operated control rats, nephrectomized rats, and into rats with acute ureteral ligation. The disappearance of human LA-PF4/beta TG antigen and PF4 in rat plasma determined by specific radioimmunoassays followed biphasic exponential curves. The half-lives (t1/2) for the fast and slow components of LA-PF4 in control rats were 6.4 and 68.4 min. Nephrectomy significantly increased these times to 9.7 and 144 min, while ureteral ligation resulted in no significant change. Comparison of the level of LA-PF4/beta TG antigen and of creatinine in aorta and in renal vein showed 25%-30% extraction of these compounds by the kidney. Less than 0.1% of the total LA-PF4 antigen injected was recovered in the urine of control rats. In contrast to these results, the clearance of PF4 was not affected by nephrectomy. In conclusion: (1) functional renal tissue is necessary for normal clearance of LA- PF4/beta TG, but renal excretion does not play a major role in its elimination suggesting that the protein is catabolized by the kidney; and (2) catabolic clearance of PF4 does not depend on functioning kidney tissue.

Release Of Platelet Factor-4 In Vivo After Heparin Injection

1981 ◽  
Author(s):  
A Koneti Rao ◽  
John C Holt ◽  
Pranee James ◽  
Stefan Niewiarowski
Keyword(s):  
Molecular Weight ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Human Platelets ◽  
Platelet Factor 4 ◽  
Plasma Samples ◽  
Platelet Factor ◽  
Immunoreactive Material ◽  
Elution Pattern

A single bolus of heparin administered to 8 normal volunteers resulted in a significant increase in levels in platelet poor plasma (PPP) of platelet factor-4 (PF4) but not low-affinity platelet factor-4/β-thromboglobulin (LA-PF4/βTG). However, the presence of heparin interfered with the binding of 125I-PF4 to antibody in radioimmunoassay (RIA). This effect was overcome by increasing the concentration of NaCl from 0.15 to 0.5 M in the buffer used for RIA. In order to establish that the increased amount of immunoreactive material present in PPP was indeed PF4, the protein was isolated from postheparin plasma. A bolus of 5000 units of porcine lung heparin (Upjohn) was administered intravenously to 2 volunteers and plasma samples obtained before and 5 minutes after the injection. The levels of PF4 in PPP rose from 18 and 10 ng/ml before to 185 and 454 ng/ml at 5 minutes after injection in the two volunteers, respectively. The 5 minute samples were adsorbed to heparin agarose columns and PF4 levels decreased to 16 and 10 ng/ml respectively. The immunoreactive material was eluted with 1.2 M NaCl from the heparin agarose columns, showing typical elution pattern for PF4. This material was applied to SDS-polyacrylamide gel electrophoresis in parallel with purified PF4 obtained from human platelets. RIA carried out on eluates from gel slices revealed a species of the same molecular weight as standard PF4. Thus, heparin injection results in appearance in the circulation of a material identical to PF4. LA-PF4/βTG and PF4 are located in same granules and released in parallel during platelet stimulation. Further, LA-PF4 is cleared from plasma 4 times slower than PF4. Therefore, the elevation of PF4alone suggests release from sites other than platelets.

scholarly journals Platelet Activation in Heparin-Induced Thrombocytopenia is Followed by Platelet Death via Complex Apoptotic and Non-Apoptotic Pathways

2020 ◽  
Vol 21 (7) ◽  
pp. 2556
Author(s):  
Elmira R. Mordakhanova ◽  
Tatiana A. Nevzorova ◽  
Gulnaz E. Synbulatova ◽  
Lubica Rauova ◽  
John W. Weisel ◽  
...  
Keyword(s):  
Platelet Activation ◽  
Immune Complexes ◽  
Gel Filtration ◽  
Human Platelets ◽  
Platelet Factor 4 ◽  
Platelet Factor ◽  
Heparin Induced Thrombocytopenia ◽  
Mitochondrial Membrane Depolarization ◽  
Low Platelet Count ◽  
Apoptotic Pathways

Heparin-induced thrombocytopenia (HIT) is an adverse drug reaction characterized by thrombocytopenia and a high risk for venous or arterial thrombosis. HIT is caused by antibodies that recognize complexes of platelet factor 4 and heparin. The pathogenic mechanisms of this condition are not fully understood. In this study, we used flow cytometry, fluorimetry, and Western blot analysis to study the direct effects of pathogenic immune complexes containing platelet factor 4 on human platelets isolated by gel-filtration. HIT-like pathogenic immune complexes initially caused pronounced activation of platelets detected by an increased expression of phosphatidylserine and P-selectin. This activation was mediated either directly through the FcγRIIA receptors or indirectly via protease-activated receptor 1 (PAR1) receptors due to thrombin generated on or near the surface of activated platelets. The immune activation was later followed by the biochemical signs of cell death, such as mitochondrial membrane depolarization, up-regulation of Bax, down-regulation of Bcl-XL, and moderate activation of procaspase 3 and increased calpain activity. The results show that platelet activation under the action of HIT-like immune complexes is accompanied by their death through complex apoptotic and calpain-dependent non-apoptotic pathways that may underlie the low platelet count in HIT.

Molten globule monomer to condensed dimer: role of disulfide bonds in platelet factor-4 folding and subunit association

Biochemistry ◽  
1992 ◽  
Vol 31 (48) ◽  
pp. 12255-12265 ◽  
Author(s):  
Kevin H. Mayo ◽  
Sharon Barker ◽  
Michael J. Kuranda ◽  
Anthony J. Hunt ◽  
Jill A. Myers ◽  
...  
Keyword(s):  
Disulfide Bonds ◽  
Molten Globule ◽  
Platelet Factor 4 ◽  
Platelet Factor

scholarly journals In vivo release and turnover of secreted platelet antiheparin proteins in rhesus monkey (Macaca mulatta)

Blood ◽  
1980 ◽  
Vol 56 (4) ◽  
pp. 596-607
Author(s):  
J Musial ◽  
S Niewiarowski ◽  
LH Jr Edmunds ◽  
VP Jr Addonizio ◽  
KC Nicolaou ◽  
...  
Keyword(s):  
Rhesus Monkey ◽  
Slow Component ◽  
Inhibitory Effect ◽  
Human Platelets ◽  
Platelet Factor 4 ◽  
Platelet Factor ◽  
In Vivo Release ◽  
Specific Proteins ◽  
Monkey Plasma

Human and rhesus monkey platelets secrete at least two antiheparin proteins: platelet factor 4 (PF4) and low affinity platelet factor 4 (LA-PF4). Neither of these proteins showed species-related antigenic differences. As determined by radioimmunoassay, the levels of PF4 and LA-PF4 antigen per 10(9) monkey platelets amounted to 10.7 and 20.3 microgram, respectively. One milliliter of monkey plasma prepared from blood collected into an anticoagulant composed of EDTA, prostaglandin E1, and theophylline solution contained 22.4 ng LA-PF4 and 8.0 ng PF4. Concentrations of these two platelet-specific proteins in monkeys closely resembled levels found in human platelets and plasma. Infusion of prostacyclin (PGI2) (100 or 300 ng/kg/min) into monkeys for 15 min resulted in a significant decrease of plasma levels of LA-PF4 antigen and of PF4 by 40%--60% (p < 0.0001). This decrease was related to the inhibitory effect of PGI2 on the secretion of platelets stimulated by a catheter or by venipuncture. Longer infusion of PGI2 did not produce further significant change. The supernate obtained after aggregation of human platelets stimulated by thrombin was injected into monkeys receiving PGI2 infusion. The disappearance of LA-PF4 antigen in monkey plasma followed a biphasic exponential curve with half-lives for the fast and slow components of 8.4 and 63 min. PF4 disappeared faster but followed the same pattern (half-lives for the fast and slow component of 2.1 and 70 min). Analysis of the experimental data suggests that the low levels of secreted platelet proteins in monkey plasma are related to their minimal in vivo release and to their rapid clearance.

scholarly journals The role of adiponectin and its receptor in patients with idiopathic membranous nephropathy complicated with hyperuricemia

2021 ◽  
pp. 31-31
Author(s):  
Tong Liu ◽  
Mengdi Xia ◽  
Yongji Zhang ◽  
Yibin Wang ◽  
Yun Zhou
Keyword(s):  
Membranous Nephropathy ◽  
Kidney Tissue ◽  
Renal Tissue ◽  
Idiopathic Membranous Nephropathy ◽  
Elisa Method ◽  
Adiponectin Receptor 1 ◽  
Ckd Stage ◽  
The Relationship ◽  
Nlrp3 Expression

Introduction/Objective. This study aimed to assess the changes of adiponectin (APN), IL-1?, adiponectin receptor 1 (Adipo R1), and NLRP3 expression of patients with idiopathic membranous nephropathy (IMN) complicated with hyperuricemia (HUA) and analyze the relationship between the APN pathway and the NLRP3 pathway. Methods. Forty-eight patients with IMN+HUA group, 49 patients with IMN group, 30 healthy controls, and 24 samples of healthy renal tissue were evaluated. APN and IL-1? of each group were detected by the ELISA method. AdipoR1 and NLRP3 in kidney tissue were detected by immunohistochemistry. The clinical data of each group were collected, and the relationship between APN, IL-1?, AdipoR1, NLRP3, and other indexes was analyzed. Results. (1) The concentration of UA, APN, IL-1?, and NLRP3 in the IMN+HUA group are significantly higher than those in the IMN group, but the AdipoR1 was lower. (2) With the severity of CKD stage, APN, IL-1?, and NLRP3 gradually increased in IMN+HUA group, but AdipoR1 gradually decreased. However, the above indicators did not change significantly in the IMN stages. Conclusion. The AdipoR1-AMPK and NLRP3-caspase-1-IL-1? signaling pathway may play an essential role in IMN+HUA patients. The intervention of these two pathways may make a great significance to the occurrence and progression on IMN+HUA patients.

Human Platelets Contain mRNA Transcripts for Platelet Factor 4 and Actin

1989 ◽  
Vol 62 (04) ◽  
pp. 1100-1102 ◽  
Author(s):  
Jane Sottile ◽  
Deane F Mosher ◽  
Jan Fullenweider ◽  
James N George
Keyword(s):  
Cell Lines ◽  
Human Cell ◽  
Human Lymphocytes ◽  
Human Platelets ◽  
Northern Blotting ◽  
Platelet Factor 4 ◽  
Human Cell Lines ◽  
Platelet Factor ◽  
Mrna Transcripts ◽  
Number Of Cells

SummaryRNAs from a number of cells, including platelets, were analyzed by Northern blotting for the presence of transcripts to four platelet proteins - actin, thrombospondin, fibronectin, and platelet factor 4. RNA from platelets contains considerable amounts of mRNA for platelet factor 4, easily detectable mRNA for actin, and traces of mRNA for thrombospondin. mRNA for platelet factor 4 was not detected in human lymphocytes or in any of 5 human cell lines.

scholarly journals Immunoelectron microscopic localization of platelet factor 4 and fibrinogen in the granules of human platelets.

1983 ◽  
Vol 31 (7) ◽  
pp. 905-910 ◽  
Author(s):  
T D Pham ◽  
K L Kaplan ◽  
V P Butler
Keyword(s):  
Human Platelets ◽  
Platelet Factor 4 ◽  
Water Soluble ◽  
Antigenic Determinants ◽  
Storage Site ◽  
Human Fibrinogen ◽  
Platelet Factor ◽  
Thin Sections ◽  
Fab Fragments ◽  
Platelet Proteins

To determine the storage site of platelet fibrinogen and of platelet factor 4 (PF4) in human platelets by immunoelectron microscopic techniques, washed human platelets were briefly exposed to Karnovsky's fixative and embedded in water-soluble Durcupan. Thin sections of platelets were exposed to Fab fragments of rabbit anti-human fibrinogen or of goat anti-human PF4, followed by a peroxidase conjugate of Fab fragments of antibodies to rabbit immunoglobulin (Ig) G or to goat IgG. The technique enabled preservation of the antigenic determinants of the platelet proteins, accessibility of Fab fragments to the platelet proteins, and maintenance of the ultrastructural integrity of the platelets. Using this approach, it was directly demonstrated that platelet fibrinogen and PF4 are stored in the alpha-granules of human platelets.

scholarly journals The Role of Platelet Factor 4 in Local and Remote Tissue Damage in a Mouse Model of Mesenteric Ischemia/Reperfusion Injury

PLoS ONE ◽  
2012 ◽  
Vol 7 (7) ◽  
pp. e39934 ◽  
Author(s):  
Peter H. Lapchak ◽  
Antonis Ioannou ◽  
Poonam Rani ◽  
Linda A. Lieberman ◽  
Kazuhisa Yoshiya ◽  
...  
Keyword(s):  
Mouse Model ◽  
Reperfusion Injury ◽  
Tissue Damage ◽  
Mesenteric Ischemia ◽  
Ischemia Reperfusion Injury ◽  
Ischemia Reperfusion ◽  
Platelet Factor 4 ◽  
Platelet Factor

scholarly journals Platelet factor 4 and the platelet secreted proteoglycan: immunologic characterization by crossed immunoelectrophoresis

Blood ◽  
1990 ◽  
Vol 75 (4) ◽  
pp. 902-910 ◽  
Author(s):  
SP Levine ◽  
LK Knieriem ◽  
MA Rager
Keyword(s):  
Chondroitin Sulfate ◽  
Core Protein ◽  
Human Platelets ◽  
Polyclonal Antiserum ◽  
Platelet Factor 4 ◽  
Platelet Factor ◽  
Affinity Ligand ◽  
Crossed Immunoelectrophoresis ◽  
Polyclonal Antisera ◽  
The Relationship

Abstract Platelet factor 4 (PF4) is a hydrophobic, alpha-granule protein with potent antiheparin activity. It also binds to a chondroitin sulfate- containing proteoglycan (PG) isolated from platelets. In order to evaluate further the relationship between PF4 and the chondroitin sulfate-containing proteoglycan in resting platelets, the PF4-binding proteoglycan from human platelets has been purified using purified PF4 as an affinity ligand and used to prepare polyclonal antiserum. Two antisera have been characterized: one reacts primarily with chondroitin sulfate (CS), the other reacts with the protein core of the platelet proteoglycan after chondroitinase AC digestion. PF4 and PG core protein antigen are present in separate, dissimilar precipitin arcs when triton- solubilized platelets are analyzed by crossed immunoelectrophoresis using polyclonal antisera to purified PF4 and PG. PF4 was demonstrated in a complex with a separate chondroitin sulfate antigen by crossed immunoelectrophoresis (CIE) experiments in which either anti-PF4 or anti-CS antisera was incorporated in the intermediate gel. Both the PF4- chondroitin sulfate complex and the proteoglycan are secreted from platelets when fresh, washed human platelets are stimulated by human alpha-thrombin. This second antigen may represent the PG after posttranslational modification of a precursor form of the proteoglycan.

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