Performance Evaluation of Innovance Antithrombin, Berichrom Protein C and Innovance Free Protein S Assays Using the Automated Coagulation Analyzer Sysmex CS-5100

SummaryDeficiency of protein S has been associated with an increased risk of thrombotic disease as already shown for protein C deficiency. Deficiencies of any of these two proteins predispose to venous thrombosis but have been only rarely associated with arterial thrombosis.In this study we describe a case of severe cerebral arterial thrombosis in a 44-year old woman with protein S deficiency. The defect was characterized by moderately reduced levels of total and markedly reduced levels of free protein S. C4b-bp level was normal. Protein C, AT III and routine coagulation tests were within the normal limits.In her family two other members showed the same defect. All the affected members had venous thrombotic manifestations, two of them at a relatively young age. No other risk factors for thrombotic episodes were present in the family members. The patient reported was treated with ASA and dipyridamole and so far there were no relapses.

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Assessment of protein C antigen, free protein S, and protein C activity in pregnancy: A cross-sectional study of pregnant Nigerian women

International Journal of Medicine and Health Development ◽

10.4103/ijmh.ijmh_21_20 ◽

2021 ◽

Vol 26 (2) ◽

pp. 118

Author(s):

TheresaUkamaka Nwagha ◽

HelenChioma Okoye ◽

AngelaOgechukwu Ugwu ◽

LisaIfenyinwa Eweputanna ◽

EmmanuelOnyebuchi Ugwu

Keyword(s):

Protein C ◽

Protein S ◽

Cross Sectional Study ◽

Sectional Study ◽

Cross Sectional ◽

Nigerian Women ◽

Free Protein ◽

Protein C Activity ◽

In Pregnancy

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TOTAL AND FREE PROTEIN S IN NEPHROTIC SYNDROME

10.1055/s-0038-1644296 ◽

1987 ◽

Author(s):

T Gadelha-Parente ◽

M Gouault-Heilmann ◽

G Rostoker ◽

M Levent ◽

S Rafowicz ◽

...

Keyword(s):

Nephrotic Syndrome ◽

Protein C ◽

Protein S ◽

Albumin Level ◽

Selectivity Index ◽

Control Group ◽

Free Protein ◽

Positive Correlation ◽

Small Series ◽

At Iii

25 consecutive patients (15M, 10F ; mean age 30 years) with nephrotic syndrome (NS) of different grade were studied. Control group consisted in 18 healthy adult volunteers. Total protein S antigen (TPS:Ag) and free protein S antigen (FPS:Ag) after precipitation of C4-BP-bound protein S by PEG 3-5 % final concentration were measured by Laurell's technique. PS:Ag was also searched in concentrated urine of 9 patients by ELISA method, more sensitive than the Laurell's technique. In the same plasma samples we measured C4-BP, Protein C Ag and AT III biological activity (all reagents from D.Stago). Serum albumin level, proteinuria, proteinuria selectivity index, triglycerides, cholesterol levels were recorded. TPS:Ag was found elevated in NS (1.30±0.3 U/ml) in comparison with control group (1.09±0.32 U/ml) and the difference was statistically significant (p<0.05). The mean values of FPS:Ag observed in patients and controls were not statistically different, but if we consider 95 % confidence limits (0.99-1-35 U/ml), 16 pts had normal or elevated FPS:Ag level, whereas 9 had decreased FPS:Ag level. A positive correlation was found between TPS:Ag and FPS:Ag in control group (r=0.66 ; p< 0.001) and in patients with NS (r=0.4l, p<0.05). C4-BP was significantly (p<0.01) increased in nephrotic patients ( 1.37 ± 0.36 U/ml) in comparison with control group (1.04±0.27 U/ml). A negative correlation was found between FPS:Ag and C4-BP levels in control group (r = −0.57, P< 0.01) but not in nephrotic patients. A positive correlation was found between FPS:Ag and albumin level and between FPS:Ag and cholesterol level. No correlation was found between TPS:Ag or FPS:Ag and proteinuria, proteinuria selectivity index, AT III and protein C levels. Traces of PS were found in urine (0. to 2.5 U/day) in 9 patients tested. 2/25 pts suffered thromboembolic events : one had a very low level of FPS:Ag in addition to a decreased level of AT III. The other one had normal FPS:Ag and AT III level but a borderline Protein C level. In conclusion. An acquired FPS:Ag deficiency was observed in 9/25 pts with NS despite an increased level of TPS:Ag. In this small series of patients the acquired FPS deficiency does not seem to be related either to an urinary loss of FPS or to an increased binding to C4-BP, as suggested by some authors.

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Patients with Antineutrophil Cytoplasmic Antibodies Associated Vasculitis in Remission Are Hypercoagulable

The Journal of Rheumatology ◽

10.3899/jrheum.130200 ◽

2013 ◽

Vol 40 (12) ◽

pp. 2042-2046 ◽

Cited By ~ 34

Author(s):

Marc Hilhorst ◽

Kristien Winckers ◽

Benjamin Wilde ◽

René van Oerle ◽

Hugo ten Cate ◽

...

Keyword(s):

Protein C ◽

Tissue Factor Pathway Inhibitor ◽

Severe Renal Impairment ◽

Protein S ◽

Endothelial Activation ◽

Antineutrophil Cytoplasmic Antibodies ◽

Low Grade ◽

Healthy Controls ◽

Free Protein ◽

Tissue Factor Pathway

Objectives.The risk of venous thromboembolism (VTE) is increased in patients with antineutrophil cytoplasmic antibodies (ANCA) associated vasculitides (AAV) as compared to healthy subjects. The mechanisms underlying this increased occurrence of VTE are not completely understood. We hypothesize that AAV patients in remission are more procoagulant than healthy controls.Methods.Patients with AAV in remission and no VTE for the last 6 months were included. Patients with severe renal impairment (serum creatinine > 250 μmol/l) were excluded. Age and sex matched healthy controls were included. The endogenous thrombin potential (ETP) was determined together with hemostatic variables: fibrinogen, D-dimers, factor VIII (FVIII), tissue factor pathway inhibitor (TFPI), protein C, and free protein S.Results.Thirty-one patients were included. In 27 patients not taking anticoagulants, ETP was measured and found to be elevated: 137.1% as compared to a median of 90.0% for healthy controls (p < 0.01). Fibrinogen and D-dimer levels were not elevated in patients (median 3.5 g/l and 279 μg/l, respectively). FVIII and TFPI levels were also significantly increased in patients as compared to healthy controls (159% vs 137%; 122.5% vs 101%, respectively), whereas protein C and free protein S levels were not elevated (126.5% vs 118.6% and 124.6% vs 118.3%, respectively).Conclusion.Patients with AAV in remission are more procoagulant than healthy controls, as indicated by an increased ETP. The increased FVIII level measured in these patients suggests persistence of endothelial activation and/or dysfunction. This endothelial dysfunction may cause a continuous low-grade procoagulant state.

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Coagulation Activation and Tissue Necrosis in Meningococcal Septic Shock: Severely Reduced Protein C Levels Predict a High Mortality

Thrombosis and Haemostasis ◽

10.1055/s-0038-1651669 ◽

1995 ◽

Vol 73 (01) ◽

pp. 015-020 ◽

Cited By ~ 117

Author(s):

Karin Fijnvandraat ◽

Bert Derkx ◽

Marjolein Peters ◽

Rob Bijlmer ◽

Augueste Sturk ◽

...

Keyword(s):

Septic Shock ◽

Protein C ◽

Purpura Fulminans ◽

Clinical Signs ◽

Protein S ◽

Coagulation System ◽

Protein C Deficiency ◽

Free Protein ◽

Meningococcal Septic Shock ◽

Protein C Activity

SummaryIn 35 consecutively admitted children (mean age: 4.3 years) with a clinical diagnosis of meningococcal septic shock (MSS), activation of the coagulation and fibrinolytic pathways was evaluated directly at admittance to the paediatric intensive care unit (ICU). The association of clinical signs and haemostatic abnormalities was assessed.All patients had signs of extensive activation of the coagulation system. The 28-day mortality was 26%. Protein C activity was strongly reduced, especially in non-survivors in whom it was significantly lower than in survivors (5% versus 23%; p <0.0001). There was a strong negative correlation between protein C activity and the mean size of the skin lesions (r = -0.71, p <0.001). These results suggest that an acquired protein C deficiency in MSS is related to the pathogenesis of purpura fulminans.Furthermore, no increase in C4b-binding protein (C4BP) and no decrease in the ratio free protein S/total protein S was observed, suggesting that a deficiency of free protein S as a result of increased C4BP does not play a pathogenetic role in meningococcal septic shock.

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PROTEIN C/PROTEIN S IN THE FOETAL BLOOD. ABSENCE OF BOUND PROTEINS AND C4 BINDING PROTEIN

10.1055/s-0038-1644290 ◽

1987 ◽

Author(s):

E Melissari ◽

M F Scully ◽

C Parker ◽

K H Nicolaides ◽

V V Kakkar

Keyword(s):

Pregnant Women ◽

Protein C ◽

Binding Protein ◽

Protein S ◽

First Trimester ◽

Normal Adult ◽

Blood Protein ◽

Free Protein ◽

The Mean ◽

Foetal Blood

Protein C, free and bound protein S and C4 binding protein levels (C4bp), were measured by electroimmunoassay in 7 pregnant women aged 22-29 years at 16-18 weeks of gestation, immediately prior to termination of pregnancy for social reasons. Protein C and protein S levels were also measured in their foetuses from blood taken through the umbilical cord. In this group of pregnant women the mean levels for protein C were 104% of normal adult mean (range 80-128%), for C4bp 100% (52-150%), for free protein S 66% (43-89%). In the foetuses the mean value for protein C was 15.3% (10.5-21%) and for free protein S 36.85% (27-47%) of the normal adult mean. Bound protein S and C4bp levels were zero. Conclusions: (1) free protein S is significantly decreased (< 2SD below the normal adult mean) in women after the first trimester of gestation whereas no change is seen in protein C concentration; (2) C4bp levels are at zero in the foetus as also are the levels of bound protein S; (3) foetal blood protein S level is approximately 2.5 times higher than protein C. Since all other vitamin K-dependent factors have been observed to be in the range of 10-20% of normal at this stage of gestation, our findings may be further proof of a non hepatic (endothelial) source of plasma protein S.

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Protein C, Protein S and c4b-Binding Protein in Severe lnfection and Septic Shock

Thrombosis and Haemostasis ◽

10.1055/s-0038-1647469 ◽

1991 ◽

Vol 65 (02) ◽

pp. 126-129 ◽

Cited By ~ 98

Author(s):

J F Hesselvik ◽

J Malm ◽

B Dahlbäck ◽

M Blombäck

Keyword(s):

Septic Shock ◽

Total Protein ◽

Protein C ◽

Blood Donors ◽

Antithrombin Iii ◽

Severe Infection ◽

Protein S ◽

C Protein ◽

Normal Value ◽

Free Protein

SummaryWe measured concentrations of the natural anticoagulant protein C; its cofactor, protein S; and the carrier protein C4bbinding protein (C4BP), in 24 patients with severe infection and 13 with septic shock. Decreased antithrombin III levels were found in 16 of 24 infection patients and all shock patients; high thrombin-antithrombin (TAT) complexes were present in 16 of 24 infection and 12 of 13 shock patients. Protein C concentrations were significantly reduced compared to healthy blood donors, to 60 ± 14% (infection) and 47 ± 20% (septic shock) (mean ± 1 SD). Total protein S levels were not reduced (119 ± 36.7 and 88 ± 20.0%, normal value 96±15%). Free protein S was also normal (27 ± 9.4 and 30 ± 8.7%, normal value 29 ± 9%). The percentage free of total protein S was normal in shock patients (35 ± 8.5%), but significantly reduced in patients without shock (23 ± 5.3%). C4BP was significantly higher than normal in the latter group (135 ± 43%), but not in the shock group (118 ± 40%), possibly due to increased consumption. Thus, no deficiency of total or free protein S was found in these patients, who had evidence of activated coagulation but no clinical DIC.

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Free protein S levels are elevated in familial C4b-binding protein deficiency

Blood ◽

10.1182/blood.v76.12.2527.2527 ◽

1990 ◽

Vol 76 (12) ◽

pp. 2527-2529 ◽

Cited By ~ 8

Author(s):

PC Comp ◽

J Forristall ◽

CD West ◽

RG Trapp

Keyword(s):

Total Protein ◽

Protein C ◽

Binding Protein ◽

Activated Protein C ◽

Protein S ◽

Protein Deficiency ◽

Normal Range ◽

Free Protein ◽

Protein Levels

Abstract In plasma, 40% of the protein S is free and functions as a cofactor for the anticoagulant effects of activated protein C. The remaining 60% of protein S is complexed to C4b-binding protein and is functionally inactive. A family with hereditary C4b binding protein deficiency has been identified with C4b-binding protein levels in an affected father and daughter of 37 micrograms/mL and 23 micrograms/mL, respectively; these values are significantly below the normal range for this protein of 180 micrograms/mL +/- 44 micrograms/mL (mean +/- 2 SD). The total protein S (free + bound) is normal in these individuals (23.2 micrograms/mL and 17.8 micrograms/mL, respectively; normal 19.1 micrograms/mL +/- 6.0 micrograms/mL). The free protein S levels are markedly increased at 22.5 micrograms/mL and 17.4 micrograms/mL, respectively (normal 5.9 micrograms/mL +/- 2.4 micrograms/mL). This experiment of nature shows that total protein S levels in plasma are not affected by the absence of C4b-binding protein and that chronic elevation of free protein S is not associated with increased hemorrhagic tendencies.

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Coagulation Factors, Postmenopausal Hormone Replacement Therapy and the Risk of Venous Thrombosis: The WHI Clinical Trials of Postmenopausal Hormone Therapy.

Blood ◽

10.1182/blood.v110.11.127.127 ◽

2007 ◽

Vol 110 (11) ◽

pp. 127-127 ◽

Cited By ~ 1

Author(s):

Mary Cushman ◽

Joseph Larson ◽

Frits R. Rosendaal ◽

Lawrence S. Phillips ◽

Barbara V. Howard ◽

...

Keyword(s):

Risk Factors ◽

Relative Risk ◽

Venous Thrombosis ◽

Protein C ◽

Coagulation Factor ◽

Protein S ◽

Postmenopausal Hormone ◽

Free Protein ◽

Coagulation Marker ◽

D Dimer

Abstract Background. Postmenopausal estrogen (E) therapy, especially in combination with progestin (P) doubles the relative risk of venous thrombosis (VTE). Risk with hormones is higher with increasing age, obesity and with factor V Leiden. We studied coagulation markers as susceptibility factors for postmenopausal hormone-related VTE. Methods. The Women’s Health Initiative program included two placebo-controlled double-blind randomized trials of two E regimens, E (conjugated equine estrogens) or E+P (E + medroxyprogesterone acetate), in 16,608 postmenopausal women aged 50–79. We performed a nested case control study that measured baseline levels of coagulation markers in 215 women who developed VTE during follow up and 867 age-matched controls. The joint effects of treatment assignment to either E regimen vs placebo and prespecified abnormal levels of each coagulation factor on relative risk of VTE were estimated by logistic regression adjusting for age, race, body-mass index and type of E regimen. Results. Low levels of protein C and free protein S (<5th percentile), high D-dimer (top quartile), and high plasmin antiplasmin complex (PAP) and prothrombin fragment 1–2 (top decile) were all associated with risk of VTE with adjusted odds ratios (95% CI) of 2.0 (1.0–4.1), 2.9 (1.5–5.6), 2.8 (2.0–4.0), 2.5 (1.6–4.0) and 1.9 (1.2–3.1), respectively. Elevated factors II, VIII, IX and fibrinogen were not VTE risk factors. Compared to women with normal coagulation marker levels assigned to placebo, the joint odds of VTE with either E regimen plus an abnormal coagulation marker were more than additive compared to the separate effects of hormones and coagulation abnormalities for low protein C, low free protein S, and elevated D-dimer, PAP and F1–2. The odds ratios of VTE with the combination of an abnormal coagulation factor and assignment to hormones were (in order listed in prior sentence), 4.5 (95% CI 2.0–10.2), 6.7 (3.0–14.5), 6.1 (3.7–10), 5.8 (3.2–10.5) and 4.4 (2.4–7.7). Conclusions. We report new findings of elevated F1-2 and PAP as VTE risk factors in women in this prospective study nested in trials of E or E+P versus placebo. Protein C or S values below the 5th percentile were also clinically relevant even though they do not represent inherited deficiency. Lower protein C and free protein S, and higher D-dimer, F1-2 and PAP all identified women at increased risk of VTE with hormones. If our findings are confirmed in management studies, measurement of these factors might assist women with decision-making on safety of E or E+P.

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Circulating Microparticles, Plasma Protein C and Free Protein S Levels in Children with Sickle Cell Anemia.

Blood ◽

10.1182/blood.v112.11.1431.1431 ◽

2008 ◽

Vol 112 (11) ◽

pp. 1431-1431

Author(s):

Andrea Piccin ◽

Elva Eakins ◽

Ciaran V Murphy ◽

William G Murphy ◽

Corrina McMahon ◽

...

Keyword(s):

Steady State ◽

Platelet Activation ◽

Sickle Cell ◽

Sickle Cell Anemia ◽

Protein C ◽

Annexin V ◽

Protein S ◽

Free Protein ◽

Sub Saharan ◽

Lower Correlation

Abstract Microparticles (MP) originate from blebbing and shedding from cell membrane surfaces in physiological and pathological conditions. Increased levels are generated by a number of mechanisms including platelet activation, vascular endothelial damage, thrombin activity, C5b-9 activation, and PF4-heparin-antibody interaction. Increased circulating MP have been described in patients with sickle cell anemia (SCA). Elevated monocyte-derived MP expressing tissue factor have been reported in patients in crisis. The lack of a standardised method for MP quantification remains problematic. We measured MP (numbers and functional markers), protein C and free protein S, in a large cohort of pediatric patients to investigate the role of MP in SCA and their relation to protein C and free protein S plasma levels. One hundred and eleven children of sub-Saharan African ethnicity with SCA (hemoglobin (Hb) SS) were studied: 51 without previous history of crisis in steady state (mean age 5.3 years); 15 in crisis (9 chest crises, 6 other, mean age 5.4 years); 30 on hydroxyurea (mean age 7.2 years); 15 on transfusion therapy (mean age 6.5 years); 17 children of sub-Saharan African ethnicity of similar age (mean age 4.6 years) were used as control group (Hb AA). MP were analyzed by flow cytometry, according to Biró et al (J Thromb Haemost.2004; 2(10):1842–51), using Annexin V and antibodies against, CD61, CD42a, CD62P (P-selectin), CD235a, CD14, CD142 (tissue factor), CD201 (endothelial protein C receptor or EPCR), CD62E (E-selectin), CD36 (thrombospondin or TSP-1), CD47 (TSP-1 receptor), CD31 PECAM (platelet-endothelial cellular adhesion marker), CD144 (VE-cadherin). Protein C (chromogenic) and free protein S (latex based assay) were measured in all subjects. Correlation was measured by Pearson Rank test, and comparisons between groups were analyzed by Mann-Whitney test. Total MP AV were lower in crisis (1.26 × 106/ml; 0.56–2.44 × 106) and steady state (1.35 × 106/ml; 0.71– 3.0 × 106) compared to transfusion (4.33 × 106/ml; 1.6–9.2 × 106p<0.01). Protein C levels were significantly lower in crisis (median 0.52 IU/ml; interquartile range 0.43–0.62) compared to all other groups: HbAA (0.72 IU/ml; 0.66–0.82, p<0.001); hydroxyurea (0.67 IU/ml; 0.58–0.77, p<0.001); steady state (0.63 IU/ml; 0.54–0.70, p<0.05) and transfusion (0.60 IU/ml; 0.54–0.70, p<0.05). In addition levels were significantly reduced in steady state (0.63 IU/ml; 0.54–0.70) compared to HbAA (0.72 IU/ml; 0.66–0.80, p<0.01). Protein S levels were significantly higher in HbAA (0.85 IU/ml; 0.72–0.97) compared with crisis (0.49 IU/ml; 0.42–0.64, p<0.001), hydroxyurea (0.65 IU/ml; 0.56–0.74, p<0.01), and transfusion (0.59 IU/ml; 0.47–0.71, p<0.01). There was also a significant difference in crisis patients compared to steady state (0.49 IU/ml; 0.42–0.64 v 0.68 IU/ml; 0.58–0.79, p<0.05). There was high correlation (R>0.9, p<0.05) between total numbers of Annexin V positive MP (MP AV) and platelet MP expressing non-activation platelet markers. There was a lower correlation between MP AV and MP CD62P (R=0.73, p<0.05) (platelet activation marker), and also a lower correlation between percentage of MP expressing CD201 (%MP CD201) and %MP CD14 (R=0.627, p<0.001). %MP CD201 was higher in crisis (11.6%) compared to HbAA (3.2%, p<0.05); %MP CD144 was higher in crisis (7.6%) compared to transfusion (2.1%, p<0.05); %CD14 (0.77%) was higher in crisis compared to transfusion (0.0%, p<0.05) and steady state (0.0%, p<0.01); MP CD14 was detectable in a higher number of samples (92%) in crisis compared to the rest (40%); %MP CD235a was higher in crisis (17.9%) compared to transfusion (8.9%), hydroxurea (8.7%) and steady state (9.9%, p<0.05); %CD62E did not differ significantly across the groups and CD142 was undetectable. These studies indicate that there are significantly lower levels of protein C and free protein S in children with sickle cell crisis. In addition there are significantly lower numbers of circulating platelet MP in steady state and crisis patients; however in crisis a significantly higher percentage of MP express markers of endothelial and vascular damage, and of red cell origin. Among these are composite hybrid microparticles expressing markers of more than one cell type, probably brought about by severe vascular stress and close contact of various circulating cell types with vascular endothelium.

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