kaolinite and Autoimmune-Diseases

Anti-beta2-glycoprotein I (beta2-GPI) antibodies behave as classical Lupus Anticoagulants (LA), as they inhibit phospholipid-dependent coagulation reactions and their activity disappears in the presence of excess exogenous phospholipids (PLs). We have recently shown that a certain amount of PLs in the dilute Russell Viper Venom Time (dRVVT) test system is required to express LA activity of anti beta2-GPI antibodies. We have now extended this observation to two other tests, i.e., Kaolin Clotting Time (KCT) in which PLs are not added, and Tissue Thromboplastin Inhibition test (TTI) in which PLs are extremely diluted. In fact, affinity-purified antibody preparations from 5 patients with antiphospholipid syndrome did not express or only weakly expressed anticoagulant activity in both tests; the mean ratios of coagulation times obtained with purified antibodies and that of control buffer were 1.11 and 1.0 for KCT and TTI, respectively. On the contrary, the mean ratios in dRVVT were 1.31 and 1.49 at a PLs dilution of 1:8 and 1:64, respectively. Therefore, the presence of LA activity due to autoantibodies to beta2-GPI is characterized by a positive dRVVT and negative or only weakly positive KCT and TTI.

Topics: Adolescent; Adult; Antibodies, Anticardiolipin; Antiphospholipid Syndrome; Autoimmune Diseases; beta 2-Glycoprotein I; Blood Coagulation Tests; Female; Glycoproteins; Humans; Immunoglobulin G; Kaolin; Lupus Coagulation Inhibitor; Male; Phospholipids; Prothrombin Time; Sensitivity and Specificity; Thromboplastin

The dilute Russell viper venom time and kaolin clotting time (KCT) are very sensitive screening tests for lupus anticoagulant activity. However, due to the high turbidity of the kaolin reagent it is difficult to accommodate the KCT on the optical end point automation of today. We evaluated five recently reported screening tests (the silica clotting time, the Textarin/Ecarin ratio, the Taipan venom time, the factor V ratio, and a kaolin clotting time using low-turbidity kaolin) as potential alternatives to the KCT. The sensitivity and specificity of the silica clotting time compared well to KCT, detecting 10/12 KCT positive samples and showing equal sensitivity to dilution of lupus positive plasma. In addition, the silica clotting time allows for a confirmatory phospholipid correction procedure. False-positive results were seen in 2 of 15 warfarinised samples. A second assay utilising the ratio of extrinsic/intrinsic factor V assays was not affected by either warfarin or heparin. This assay also gave positive results with 3 of 23 samples previously screened as lupus negative but exhibiting anticardiolipin positivity. It was therefore concluded that a combination of the silica clotting time and dilute Russell viper venom time met the requirements of lupus sensitivity with demonstration of phospholipid dependence and optical end point compatibility. The factor V ratio is a useful second-line screen for both anticoagulated patients and anticardiolipin antibody-positive samples.

Topics: Anticoagulants; Antiphospholipid Syndrome; Autoimmune Diseases; Automation; Blood Coagulation Tests; Elapid Venoms; Endopeptidases; Evaluation Studies as Topic; Factor V; False Positive Reactions; Heparin; Humans; Kaolin; Liver Diseases; Lupus Coagulation Inhibitor; Lupus Erythematosus, Systemic; Nephelometry and Turbidimetry; Partial Thromboplastin Time; Peptide Hydrolases; Postoperative Period; Prothrombin; Prothrombin Time; Silicon Dioxide; Warfarin

Antiprothrombin and anti-beta2-glycoprotein I (beta2-GPI) antibodies belong to the family of antiphospholipid (APL) antibodies and represent the phospholipid-dependent inhibitors of coagulation. They may be distinguished by analyzing the coagulation profiles generated by the comparison of the ratios of two coagulation tests, the Kaolin Clotting Time (KCT) and the dilute Russell's Viper Venom Time (dRVVT), commonly adopted for their diagnosis. The KCT profile is caused by antiprothrombin antibodies, whereas anti-beta2-GPI antibodies are responsible for the dRVVT coagulation profile. The presence of aPL antibodies is frequently associated with acquired resistance to activated Protein C (APC-R), but limited information is available regarding the role of the different antibodies in its development. We studied the time-course of activated Factor V (FVa) generation and inactivation in the plasma of 42 patients with well-defined phospholipid-dependent inhibitors of coagulation: 24 displayed the dRVVT coagulation profile, whereas the other 18 cases showed the KCT profile. In normal pooled plasma, the peak values of FVa (mean +/- standard deviation, [SD]: 16.307 +/- 4.372 U/mL) were reached in 4 to 5 minutes and an almost complete inactivation (0.088 +/- 0.123 U/mL) was obtained within 20 minutes. At this time point, values of residual FVa exceeding 2 SD the mean of controls (0.344 U/mL) were considered abnormal. Patients belonging to the KCT coagulation profile group reached the maximal amount of FVa in plasma (22.740 +/- 7.693 U/mL, P = not significant v controls) within 4 to 5 minutes; at 20 minutes, the residual amount of FVa in plasma ranged from 0 to 1.09 U/mL (0.293 +/- 0.298; P = .027), but it was found abnormal in only six of the 18 cases. The time-course of FVa in plasma of patients belonging to the dRVVT coagulation profile group differed from that of normal controls in that the peak values (10.955 +/- 5.092 U/mL) were reached at 10 minutes and the amount of residual FVa at 20 minutes ranged from 0.320 to 14.450 U/ml (2.544 +/- 3.580 U/mL; P = .0191 v normal controls and P = . 0114 v KCT group patients). Twenty of the 24 patients belonging to the dRVVT profile group had an abnormal inactivation of FVa (chi2 = 0.001 v KCT group patients). History of venous thrombosis was experienced by 15 patients: an abnormal rate of FVa inactivation was found in 11 of them (73%) versus 15 of the 27 cases without thrombosis (56%) (x2 = 0.2556). The effect of affinity-pu

Topics: Adult; Aged; Antibodies, Antiphospholipid; Antibody Specificity; Antiphospholipid Syndrome; Autoimmune Diseases; beta 2-Glycoprotein I; Blood Coagulation; Blood Coagulation Tests; Enzyme Activation; Factor Va; Female; Glycoproteins; Humans; Immunoglobulin G; Immunoglobulin M; Kaolin; Male; Middle Aged; Protein C; Prothrombin; Prothrombin Time; Thrombophilia

The lupus anticoagulant was identified in three patients. Laboratory studies gave evidence of inhibitory activity directed against phospholipid and prekallikrein. Inhibition of prekallikrein has not been reported previously. When exposed to kaolin, all three patients' plasmas failed to develop the level of fibrinolytic activity achieved by similarly treated normal plasma. The data suggest that compromised fibrinolytic capacity may be a contributing factor in the development of thrombosis in patients with the lupus anticoagulant.

Topics: Adolescent; Adult; Autoimmune Diseases; Blood Coagulation Factors; Female; Femoral Vein; Fibrinolysis; Humans; Kallikreins; Kaolin; Lupus Coagulation Inhibitor; Lupus Erythematosus, Systemic; Phospholipids; Prekallikrein; Procainamide; Thrombophlebitis

Topics: Adjuvants, Immunologic; Aluminum; Aluminum Silicates; Animals; Antigens; Asbestos; Autoimmune Diseases; Beryllium; Brain; Encephalomyelitis, Autoimmune, Experimental; Immunization, Passive; Iron; Iron-Dextran Complex; Kaolin; Latex; Lymph Nodes; Microspheres; Nerve Tissue; Omentum; Pertussis Vaccine; Spinal Cord; Spleen; Starch