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#27753626   2016/10/18 Save this To Up

A unique antiphospholipid assay recognizing phospholipid mixture compared with criteria antiphospholipid immunoassays in lupus patients.

Background While essential for the classification of antiphospholipid syndrome (APS), anticardiolipin (aCL) assays lack specificity and anti-β2glycoproteinI (anti-β2GPI) assays lack sensitivity in this regard. Our aim was to perform a comparative analysis of the APhL ELISA assay (IgG/IgM) and criteria antiphospholipid (aPL) immunoassays in identifying APS-related clinical manifestations in a large group of patients with systemic lupus erythematosus (SLE). Methods Serum samples from 1178 patients from the Hopkins ( n = 543), LUMINA ( n = 588) and Jamaican SLE cohorts ( n = 47) were examined for IgG/IgM positivity in aCL (in-house), anti-β2GPI (two commercial kits) and APhL (Louisville APL) ELISA assays. Correlation of assay positivity with clinical manifestations and sensitivity, specificity, positive and negative predictive values and likelihood ratios were evaluated. A case series analysis was also performed in patients for whom there was isolated positivity in the specific aPL assays. Results The prevalence of aCL positivity was 34.9%, anti-β2GPI kit A was 22.6%, APhL was 11.5% and anti-β2GPI kit B was 7.6% in the study population. Anti-β2GPI kit B, aCL and APhL assays were correlated with venous thrombosis, while only APhL was significantly correlated with arterial thrombosis and consistently correlated with pregnancy-related morbidity. No significant correlations were noted for anti-β2GPI kit A. Sensitivity was greatest for aCL assays followed by anti-β2GPI kit A, APhL and anti-β2GPI kit B, while specificity was greatest and equal for anti-β2GPI kit B and APhL assays. Conclusions Overall, APhL antibodies, especially IgG, represent a promising biomarker for the classification of APS patients in the context of autoimmunity and in risk assessment with regards to pregnancy morbidity and thrombotic manifestations.

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#24417081   2014/01/14 Save this To Up

[Inhibitory effects of fluvastatin on activation of THP-1 cells induced by anti-beta2GPI/beta2GPI complex].

This study is to explore the interventional effects of fluvastatin on anti-beta2GPI/beta2GPI-induced activation in THP-1 mononuclear cells. In vitro, human mononuclear cells THP-1 were treated with fluvastatin, LPS and anti-beta2GPI/beta2GPI, then the TF expression on THP-1 cells was detected by real-time quantitative PCR (RT-qPCR) or TF activity was detected by kit. TNF-alpha mRNA and its protein expression were investigated by RT-PCR and ELISA kit. The expression of phospho-NF-kappaB p65 and inhibitory protein of NF-kappaB (IkappaB-alpha) were measured by Western blotting. The results suggested that the expression of TF and TNF-alpha on THP-1 cells was significantly up-regulated with treatment of anti-beta2GPI/beta2GPI complex (100 mg x L(-1)), compared with that of untreated cells (P < 0.05). Fluvastatin (50 mg x L(-1)) could decrease TF (mRNA and activity) expression and the level of TNF-alpha (mRNA and protein) in THP-1 cells with anti-beta2GPI/beta2GPI complex. The expression of TF and TNF-alpha was shown in a concentration-dependent manner. Moreover, anti-beta2GPI/beta2GPI complex could downregulate IkappaB-alpha levels and increase the levels of phospho-NF-kappaB p65. And these effects of anti-beta2GPI/beta2GPI complex could be blocked by fluvastatin. In conclusion, fluvastatin may interfere the expression and regulation of NF-kappaB signal transduction pathway, thereby inhibit the effects of anti-beta2GPI/beta2GPI on activation of THP-1 cells, by decreasing the expression of TF and TNF-alpha.

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Anti AGO2 Human, Monoclon Anti AGO2 Mouse, Monoclon Anti-AICDA(Activation-ind Anti AICDA(Activation ind Anti C Reactive Protein A Anti AGO2 Human, Monoclon Anti AGO2 Mouse, Monoclon anti HCMV IE pp65 IgG1 (m anti HCMV gB IgG1 (monocl Mouse Anti-Mouse Major Hi Mouse Anti-Nuclear Pore C anti CD4 T helper cells

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#22193216   2012/11/26 Save this To Up

Reevaluation of predictive value of ACL and anti-β2GP1 antibody for thrombosis in patients with systemic lupus erythematosus: from a perspective of a practical world.

Detection of ACL (anticardiolipin, ACL) and anti-β2GP1 (beta2 glycoprotein1, β2GP1) antibody has been widely used, and the criteria of APS (Antiphospholipid syndrome, APS) have been used for the prediction of thrombosis in patients with SLE. What is the exact predictive value of these two antibodies? Is it really necessary to apply the criteria of APS to each patient just for the purpose of prediction of thrombosis? The aim of this retrospective study is to reevaluate the predictive value of different combination of ACL and anti-β2GP1 antibody for thrombosis formation in Chinese patients with SLE. Patients fulfilling the 1997 ACR classification criteria for SLE were enrolled and retrospectively analyzed. Thrombosis was confirmed by ultrasound, cerebral MRI, computed tomography pulmonary angiogram and angiography. Both IgG and IgM isotype of ACL and anti-β2GP1 antibody were detected with ELISA kit. ROC curves and other parameters of diagnostic test for different combination of ACL and anti-β2GP1 were analyzed and compared. 175 patients were recruited and thrombosis was diagnosed in 49 patients. In patients with thrombosis, 95.9% had been treated with glucocorticoids before detection of the two antibodies, 44.9% had hypertension and 53.1% had hyperlipidemia. ACL was positive in 28 patients (16%), and anti-β2GP1 antibody was positive in 21 patients (12%). The presence of a low or higher titer of either ACL (>12 RU/ml) or anti-β2GP1 antibody (>20 RU/ml) once has the highest predictive accuracy. The sensitivity, the specificity, the Youden's index and the area under ROC curve are 61.11%, 81.11%, 0.4222 and 0.711 respectively. A transient low or higher titer of ACL or anti-β2GP1 antibody had a good predictive value for thrombosis in patients with SLE, especially in those with other traditional risk factors for thrombosis and those treated with glucocorticoids.

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#17020782   2007/04/23 Save this To Up

Antiphospholipid antibody ELISAs: survey on the performance of clinical laboratories assessed by using lyophilized affinity-purified IgG with anticardiolipin and anti-beta2-Glycoprotein I activity.

Lupus anticoagulant (LA) and anticardiolipin (aCL) antibodies are the classical tests used to diagnose the antiphospholipid syndrome (APS). Unfortunately, since these are nonspecific and standardization is lacking, the results of laboratory work-ups upon which diagnosis are made are often misleading. The performance of clinical laboratories in detecting LA using lyophilised affinity purified immunoglobulin has been previously reported. The same material was used to investigate the inter-laboratory variability of aCL and anti-beta(2)-Glycoprotein I (beta(2)-GPI) antibody measurements. Laboratories were asked to test normal plasma spiked with purified IgG or distilled water in order to obtain 3 samples positive for aCL and anti-beta(2)-GPI at different antibody concentration (A, B and C) and 3 samples of normal plasma. Thirty-five laboratories participated and interpreted their test results. All performed an ELISA for IgG aCL antibodies, while 17 also tested samples using IgG anti-beta(2)-GPI antibody ELISA. Sensitivity and specificity were calculated on the basis of the responses provided by each laboratory. Overall, 99/105 samples were correctly interpreted as positive and 97/101 as negative for the presence of IgG aCL, corresponding to a sensitivity and specificity of 94% and 96%, respectively. Likewise, 46/51 samples were correctly defined as positive and 50/51 as negative for the presence of IgG anti-beta(2)-GPI corresponding to a sensitivity and specificity of 90% and 98%, respectively. A wide variability in results pertaining to the positive samples was found for aCL-ELISA (coefficient of variation of 79%, 59%, and 53% for samples A, B, and C, respectively) as well as for abeta(2)-GPI-ELISA (coefficient of variation of 85%, 95%, and 50% for samples A, B, and C, respectively). This was confirmed when the analysis was restricted to those centres using the same commercial kit. Median antibody concentrations reported by centres for positive samples were consistent with the prolongation of coagulation tests assessing lupus anticoagulant (LA). Among these, dRVVT showed a good sensitivity and linear correlation with aCL antibody concentration. In conclusion, on the whole this survey found correct interpretation of positive and negative samples by both ELISAs. Nonetheless the high variability of reported data remains a major problem that only a consensus on the part of laboratories and manufacturers to utilize standard, uniform materials and procedures can hope to overcome.

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#16855163   2006/07/20 Save this To Up

Anti-beta-2 glycoprotein I antibodies affect Bcl-2 and Bax trophoblast expression without evidence of apoptosis.

Antiphospholipid antibodies (aPLs) reacting with beta-2 glycoprotein I (beta2GPI) have been associated with recurrent fetal loss and pregnancy complications. The aim of the study was to investigate whether aPLs with anti-beta2GPI specificity induce apoptosis of human trophoblasts in vitro. To this end, human anti-beta2GPI monoclonal IgM derived from a patient with antiphospholipid syndrome and a human irrelevant monoclonal IgM were incubated with human trophoblast cell cultures for 24, 48, and 72 h. In all the cultures we evaluated: (i) Bcl-2 and Bax mRNA and protein expression by Western blot and reverse transcription polymerase chain reaction (RT-PCR), respectively; (ii) DNA fragmentation by a commercial ELISA kit and by agarose gel electrophoresis; and (iii) the percentage of cells reactive with the monoclonal antibody (MAb) M30 by indirect immunofluorescence. The results were: Bcl-2/Bax ratio increased in untreated trophoblast cells during the time of culture, showing the highest values detectable after 72 h (2.68 and 2.28 at protein and mRNA levels, respectively). Cell incubation with anti-beta2GPI MAbs induced a significant Bcl-2/Bax ratio reduction in comparison with untreated cells (1.22 and 1.28 at protein and mRNA levels, respectively, after 72 h incubation). No significant difference was detected after cell exposure to irrelevant MAbs. However, neither DNA fragmentation nor increase in cells positive for the caspase-cleaved epitope of cytokeratin 18 cytoskeletal protein (M30) was found. In Conclusion, anti-beta2GPI antibodies react with trophoblast cells and reduce the Bcl-2/Bax ratio, but without any clear apoptotic effect.

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#15893301   2005/06/20 Save this To Up

Clinical laboratory testing for the antiphospholipid syndrome.

The first aCL test was developed in 1983 and subsequently standardized. Although in the last 6 to 7 years, new and more specific tests have become available, the aCL ELISA and the LA tests are still the first choice to be used in diagnosis of APS. While there is now doubt that the anticardiolipin test is useful in the diagnosis of APS, limitations of the assay have caused uncertainty and misinterpretation of the value of the test. Utilization of validated ELISA kits with well-tested calibrators and an "in-house standard" may enable more reproducible measurements. Reporting results semiquantitatively preserves the clinical utilize of the test without the misinterpretation of a quantitative result that may lack precision. The development of newer tests such as the beta2GPI ELISA and the APhL ELISA Kit, utilizing the phospholipid mixture, give promise to a more specific and reliable diagnosis of APS, while retaining good sensitivity. Other tests such as ELISA for prothrombin antibodies and annexin V antibodies are still under development and will require standardization and extensive evaluation. The aCL test should continue to be done and included in the Sapporo criteria. The aCL test is not as specific as the anti-beta2GPI test, but it is very sensitive and together with the LA test should capture the majority of the APS patients. IgA aCL and anti-beta2GPI positivity alone is rare but occasionally found and shown to be associated with major clinical manifestations of APS. Therefore, it is now recommended to include both tests, IgA aCL and IgG, IgM and IgA anti-beta2GPI to confirm diagnosis of APS.

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#15653437   2005/01/17 Save this To Up

Apolipoprotein H (apoH)-dependent autoantibodies and apoH protein polymorphism in selected patients showing lupus anticoagulant activity.

Apolipoprotein H (apoH) is considered to be a necessary cofactor for the binding of certain antiphospholipid antibodies to anionic phospholipids. Some apoH-dependent antiphospholipid antibodies also exert lupus anticoagulant (LA) activity, which seems to depend on antiphospholipid antibody epitope specificity. The aim of this study was to evaluate whether the presence of less frequent apoH alleles may induce structural or conformational changes in these "LA-dependent" regions that may initiate more frequent autoimmune responses in subjects. We selected patients with confirmed LA activity and none or low titers of anticardiolipin antibodies that had been sent to the laboratory for routine antiphospholipid antibody determination. Many of them had some clinical manifestation of antiphospholipid syndrome. Antibodies to apoH were determined with a commercially available anticardiolipin/apoH ELISA kit. ApoH protein polymorphism (apoH phenotype) was demonstrated by isoelectric focusing and immunoblotting. Our results showed that 47/74 (63.5%) of our selected LA-positive patients also had elevated apoH-dependent antiphospholipid antibody titers. These results point to two subgroups of patients according to the LA potency of apoH-dependent antibodies. A strong positive correlation (non-linear or linear) for apoH-dependent antibody titers and LA activity was observed in both subgroups of patients. In this study, we did not find significant differences in the distribution of apoH phenotypes among control subjects and patients with apoH-dependent/LA-positive auto- antibodies.

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#15617625   2004/12/24 Save this To Up

Anti-phospholipid and anti-DNA antibodies are not associated with the elevated release of circulatory fetal DNA in pregnancies affected by preeclampsia.

We have previously shown that the levels of circulatory fetal DNA are elevated in preeclampsia and that these increases correspond to disease severity. Several reports have indicated that increased levels of antiphospholipid (anti-PL) and anti-DNA antibodies may be associated with preeclampsia, in particular with the severe forms of the disorder. Since the release of cell-free DNA by the placenta is attributed to some form of cell death or damage and as anti-PL and anti-double-stranded DNA (dsDNA) antibodies have been proposed to lead to placental damage, we have studied the relationship between these parameters in preeclampsia.

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#12588291   2003/02/17 Save this To Up

Anti-cardiolipin antibodies in patients with chronic viral hepatitis are independent of beta2-glycoprotein I cofactor or features of antiphospholipid syndrome.

Although controversial, some authorities have implicated hepatitis C virus (HCV) as a cause of anti-phospholipid syndrome (APLS). Anti-cardiolipin antibodies (anti-CLAbs) in APLS are cofactor-dependent ('pathogenic' antibodies). We conducted a study in order to determine the prevalence of anti-CLAbs in HCV patients, and furthermore to address whether these autoantibodies are cofactor-dependent or not and whether they are associated with features of APLS. Patients with hepatitis B virus (HBV) were also evaluated in order to assess whether there are differences in the prevalence and the clinical significance of anti-CLAbs between these two major types of chronic viral hepatitis.

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#12082318   2002/06/25 Save this To Up

Beta2-glycoprotein I dependent anticardiolipin antibodies and lupus anticoagulant in patients with recurrent pregnancy loss.

The present study was aimed to define the incidence of antiphospholipid antibodies of different types lupus anticoagulant (LAC), venereal disease research laboratory test (VDRL) and Beta2-glycoprotein I dependent anticardiolipin antibodies Beta2 I aCL) in our cohort of population experiencing recurrent pregnancy loss (RPL) from Andhra Pradesh, South India.

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