In vivo γ-tocopherol supplementation decreases systemic oxidative stress and cytokine responses of human monocytes in normal and asthmatic subjects

We have recently reported that gamma tocopherol (γT) reduces allergen and zymosan-induced inflammation using rodent models. As an initial step in extending these observations to humans, we conducted an open-label, Phase I dosing study of two doses (one or two capsules/daily for one week) of a gamma tocopherol rich preparation containing 623mg of γ tocopherol, 61.1mg of d-α-tocopherol, 11.1 mg of d-β-tocopherol (11.1mg), and 231 mg of d-σ-tocopherol per capsule. Endpoints for this study include serum levels of 5-nitro-gamma tocopherol, as a marker of oxidative stress, and changes in serum gamma, alpha and delta tocopherol and γ-2′-carboxyethyl-6-hydroxychroman (CEHC) six and 24 hours after the first dose and after 1 week of treatment. To assess biological activity of this treatment, we obtained peripheral blood mononuclear cells at baseline and after 1 week of treatment with 2 capsules of a gamma tocopherol rich preparation/day, and examined the inflammatory cytokine response of these cells in culture to ex-vivo endotoxin/LPS (0.01 ng/ml) challenge. We also monitored a number of safety endpoints to examine how well this preparation is tolerated in 8 normal volunteers (4 allergic and 4 non-allergic) and 8 allergic asthmatics. We further obtained human monocytes from a subset of these volunteers and treated them ex vivo with γT, αT,γ-CEHC and α-CEHC and assessed their actions on LPS induced degradation of IkBα, and JNK signaling and ROS generation. As detailed herein, this open label study demonstrates that gamma tocopherol enriched supplementation decreased systemic oxidative stress, increased serum levels of gamma tocopherol, and inhibited monocyte responses to LPS without any adverse health effects. Further,in vitro treatment of human monocytes with γ-CEHC and α-CEHC inhibits ROS generation and LPS-induced degradation of IκB and JNK activation

Plasma Microparticle Tissue Factor Activity in Patients With Antiphospholipid Antibodies With and Without Clinical Complications

Antiphospholipid syndrome (APS) is defined by the association of autoantibodies to certain phospholipid-binding proteins with arterial or venous thrombosis (‘AT’ or ‘VT’, respectively), and/or pregnancy-related morbidity (PM). Antiphospholipid antibodies (aPLA) promote activation of several cell types including monocytes, resulting in procoagulant tissue factor (TF) expression that may contribute to the vascular complications. Since TF synthesis by monocytes is frequently accompanied by release of TF-bearing microparticles, we hypothesized that plasma microparticle TF activity (MP-TF) may be elevated in APS patients and contribute to thrombosis and/or PM. Platelet-poor plasma specimens were obtained from 30 patients with definite APS and 72 patients with asymptomatic aPLA from the Antiphospholipid Syndrome Collaborative Registry (APSCORE). MP-TF was measured by an in-house factor Xa generation assay. The two groups were well matched for gender, age, ethnicity, proportions with underlying SLE, and aPLA profiles. MP-TF (median and (IQR)) in asymptomatic aPLA subjects was 0.09 pg/mL (0.05–0.14) compared to 0.13 pg/mL (0.10–0.17) in APS (p<0.001). No differences in MP-TF levels were observed between APS subjects with PM, thrombosis, or PM + thrombosis. Similarly, among subjects with either APS or asymptomatic aPLA, MP-TF did not differ in the presence or absence of underlying SLE. Prospective studies will be required to determine if plasma MP-TF activity is causally related to thrombotic or gestational complications in APS

Delayed Apoptotic Cell Clearance and Lupus-like Autoimmunity in Mice Lacking the c-mer Membrane Tyrosine Kinase

Mice lacking the membrane tyrosine kinase c-mer have been shown to have altered macro-phage cytokine production and defective phagocytosis of apoptotic cells despite normal phagocytosis of other particles. We show here that c-mer–deficient mice have impaired clearance of infused apoptotic cells and that they develop progressive lupus-like autoimmunity, with antibodies to chromatin, DNA, and IgG. The autoimmunity appears to be driven by endogenous antigens, with little polyclonal B cell activation. These mice should be an excellent model for studying the role of apoptotic debris as an immunogenic stimulus for systemic autoimmunity

Plasma Microparticle Tissue Factor Activity in Patients With Antiphospholipid Antibodies With and Without Clinical Complications

Antiphospholipid syndrome (APS) is defined by the association of autoantibodies to certain phospholipidbinding proteins with arterial or venous thrombosis ('AT' or 'VT', respectively), and/or pregnancy-related morbidity (PM). Antiphospholipid antibodies (aPLA) promote activation of several cell types including monocytes, resulting in procoagulant tissue factor (TF) expression that may contribute to the vascular complications. Since TF synthesis by monocytes is frequently accompanied by release of TF-bearing microparticles, we hypothesized that plasma microparticle TF activity (MP-TF) may be elevated in APS patients and contribute to thrombosis and/or PM. Platelet-poor plasma specimens were obtained from 30 patients with definite APS and 72 patients with asymptomatic aPLA from the Antiphospholipid Syndrome Collaborative Registry (APSCORE). MP-TF was measured by an in-house factor Xa generation assay. The two groups were well matched for gender, age, ethnicity, proportions with underlying SLE, and aPLA profiles. MP-TF (median and (IQR)) in asymptomatic aPLA subjects was 0.09 pg/mL (0.05-0.14) compared to 0.13 pg/mL (0.10-0.17) in APS (p <0.001). No differences in MP-TF levels were observed between APS subjects with PM, thrombosis, or PM + thrombosis. Similarly, among subjects with either APS or asymptomatic aPLA, MP-TF did not differ in the presence or absence of underlying SLE. Prospective studies will be required to determine if plasma MP-TF activity is causally related to thrombotic or gestational complications in APS. (C) 2013 Elsevier Ltd. All rights reserve

Rapid Publication Lupus Anticoagulant Activity of Autoimmune Antiphospholipid Antibodies Is Dependent upon fl2-Glycoprotein I

Abstract It has been reported that antiphospholipid autoantibodies do not recognize phospholipid alone, but rather the plasma protein j2-glycoprotein I (,B2GPI), or a ,B2GPI-phospholipid complex. In vitro #2GPI binds to anionic phospholipids and inhibits the prothrombinase activity of procoagulant membranes. In light of the fact that lupus anticoagulants, a type of antiphospholipid antibody, have similar anticoagulant properties, the relationship of ,B2GPI to lupus anticoagulant activity was investigated. IgG from patients with autoimmune diseases or syphilis were tested for anticardiolipin reactivity and lupus anticoagulant activity in the presence and absence of #2GPI. As expected, anticardiolipin reactivity associated with autoimmune disease was ,B2GPI dependent. In contrast, IgG from a patient with syphilis recognized cardiolipin alone and binding was inhibited by ,B2GPI. Autoimmune antiphospholipid antibodies prolonged the dilute Russell viper venom time of normal plasma, but had no effect onf2GPI-depleted plasma. Antiphospholipid antibodies associated with syphilis had no anticoagulant effect. RP-1, an anti-,B2GPI mAb, had anticoagulant effects similar to those ofautoimmune antiphospholipid antibodies. These data demonstrate that antiphospholipid autoantibodies exert lupus anticoagulant activity via an interaction with ,82GPI. These antibodies and RP-1 appear to amplify the anticoagulant effect of ,B2GPI itself. (J. Clin. Invest. 1992. 90:1100-110

Plasma Microparticle Tissue Factor Activity in Patients With Antiphospholipid Antibodies With and Without Clinical Complications

Antiphospholipid syndrome (APS) is defined by the association of autoantibodies to certain phospholipidbinding proteins with arterial or venous thrombosis ('AT' or 'VT', respectively), and/or pregnancy-related morbidity (PM). Antiphospholipid antibodies (aPLA) promote activation of several cell types including monocytes, resulting in procoagulant tissue factor (TF) expression that may contribute to the vascular complications. Since TF synthesis by monocytes is frequently accompanied by release of TF-bearing microparticles, we hypothesized that plasma microparticle TF activity (MP-TF) may be elevated in APS patients and contribute to thrombosis and/or PM. Platelet-poor plasma specimens were obtained from 30 patients with definite APS and 72 patients with asymptomatic aPLA from the Antiphospholipid Syndrome Collaborative Registry (APSCORE). MP-TF was measured by an in-house factor Xa generation assay. The two groups were well matched for gender, age, ethnicity, proportions with underlying SLE, and aPLA profiles. MP-TF (median and (IQR)) in asymptomatic aPLA subjects was 0.09 pg/mL (0.05-0.14) compared to 0.13 pg/mL (0.10-0.17) in APS (p <0.001). No differences in MP-TF levels were observed between APS subjects with PM, thrombosis, or PM + thrombosis. Similarly, among subjects with either APS or asymptomatic aPLA, MP-TF did not differ in the presence or absence of underlying SLE. Prospective studies will be required to determine if plasma MP-TF activity is causally related to thrombotic or gestational complications in APS. (C) 2013 Elsevier Ltd. All rights reserve

Antiphospholipid Syndrome: Clinical Characteristics of Patients and Affected Family Members From Multiplex Families

Abstract Abstract 2972 Poster Board II-950 Background: Antiphospholipid syndrome (APS) is characterized by venous and/or arterial thromboembolic events, recurrent fetal loss, and persistently elevated antiphospholipid antibody (aPL) levels. Familial clustering of individuals with elevated aPL levels occurs, and up to 37% of patients with APS have one or more relatives with at least one clinical feature of APS. Individuals with elevated aPL levels or APS are also frequently identified in families with other autoimmune (AI) disorders, such as lupus or rheumatoid arthritis. Individuals with different autoimmune disorders appear to share common susceptibility loci, suggesting that a common set of susceptibility genes may contribute to clinically distinct autoimmune disorders. To investigate the heritability of APS, we are enrolling patients with APS who have one or more family members affected by APS (multiplex APS) or by other, non-APS, autoimmune disorders (multiplex AI). This study summarizes clinical characteristics of probands and family members enrolled to date in these two groups. Methods: Probands meeting clinical and laboratory criteria for APS (Miyakis, et al. J.Thromb.Haemost, 2006;4: 295-306) who had at least one clinically affected relative positive for either APS or another autoimmune disorder (e.g., lupus, rheumatoid arthritis) were recruited and enrolled into the study. A detailed personal and family history was obtained and relevant family members were also approached to participate in the study. Blood specimens were collected for genetic, serologic, and coagulation testing. Results: Review of more than 200 potential participants identified 13 probands with multiplex APS families and 49 with multiplex AI families. Probands from both groups more frequently had primary rather than secondary APS, and thromboembolic events were the most common clinical manifestation. Catastrophic APS was reported in 1 multiplex APS proband and 4 multiplex AI probands. Proband characteristics are summarized in the Table. In the multiplex APS families, 1 to 3 family members had APS, and the most common clinical manifestation was thromboembolism. In addition, 8 multiplex APS families also had one or more family members who were affected with other autoimmune disorders, most commonly lupus and rheumatoid arthritis. In the multiplex AI families, 1 to 8 family members were affected by a variety of autoimmune disorders, including lupus, Hashimoto's disease, Sjögren's syndrome, rheumatoid arthritis, myasthenia gravis, type I diabetes mellitus, and other diseases. Affected family members most commonly included siblings and/or parents of the probands. Conclusions: In the participants enrolled to date, probands with APS who belonged to multiplex APS or multiplex AI families most commonly had primary APS, and thromboembolic complications were the most common clinical manifestation. Families that were multiplex AI were more common than families that included more than one family member affected with APS, and families that were multiplex APS frequently included members that had other autoimmune disorders. Disclosures: No relevant conflicts of interest to declare