Mitochondrial Dysfunction in the Liver and Antiphospholipid Antibody Production Precede Disease Onset and Respond to Rapamycin in Lupus-Prone Mice.

OBJECTIVE: Antiphospholipid antibodies (aPL) constitute a diagnostic criterion of systemic lupus erythematosus (SLE), and aPL have been functionally linked to liver disease in patients with SLE. Since the mechanistic target of rapamycin (mTOR) is a regulator of oxidative stress, a pathophysiologic process that contributes to the development of aPL, this study was undertaken in a mouse model of SLE to examine the involvement of liver mitochondria in lupus pathogenesis. METHODS: Mitochondria were isolated from lupus-prone MRL/lpr, C57BL/6.lpr, and MRL mice, age-matched autoimmunity-resistant C57BL/6 mice as negative controls, and transaldolase-deficient mice, a strain that exhibits oxidative stress in the liver. Electron transport chain (ETC) activity was assessed using measurements of oxygen consumption. ETC proteins, which are regulators of mitochondrial homeostasis, and the mTOR complexes mTORC1 and mTORC2 were examined by Western blotting. Anticardiolipin (aCL) and anti-ß2 -glycoprotein I (anti-ß2 GPI) autoantibodies were measured by enzyme-linked immunosorbent assay in mice treated with rapamycin or mice treated with a solvent control. RESULTS: Mitochondrial oxygen consumption was increased in the livers of 4-week-old, disease-free MRL/lpr mice relative to age-matched controls. Levels of the mitophagy initiator dynamin-related protein 1 (Drp1) were depleted while the activity of mTORC1 was increased in MRL/lpr mice. In turn, mTORC2 activity was decreased in MRL and MRL/lpr mice. In addition, levels of aCL and anti-ß2 GPI were elevated preceding the development of nephritis in 4-week-old MRL, C57BL/6.lpr, and MRL/lpr mice. Transaldolase-deficient mice showed increased oxygen consumption, depletion of Drp1, activation of mTORC1, and elevated expression of NADH:ubiquinone oxidoreductase core subunit S3 (NDUFS3), a pro-oxidant subunit of ETC complex I, as well as increased production of aCL and anti-ß2 GPI autoantibodies. Treatment with rapamycin selectively blocked mTORC1 activation, NDUFS3 expression, and aPL production both in transaldolase-deficient mice and in lupus-prone mice. CONCLUSION: In lupus-prone mice, mTORC1-dependent mitochondrial dysfunction contributes to the generation of aPL, suggesting that such mechanisms may represent a treatment target in patients with SLE.

Pregnancy outcome in women with antiphospholipid syndrome on low-dose aspirin and heparin: a retrospective study.

This retrospective review of hospital records analysed pregnancy outcome with 2 different treatments for women with recurrent miscarriage diagnosed with antiphospholipid syndrome in the index pregnancy. Of 64 women, 29 had received aspirin and 35 aspirin plus heparin. Pregnancy-induced hypertension, prematurity, intrauterine growth restriction and neonatal death were considered as maternal and fetal complications. There were no significant differences in antenatal and maternal complications between the groups. HOwever, there were significant differences in mean anticardiolipin IgG antibody levels. Aspirin alone or in combination with parin was equally efficacious in women with antiphospholipid syndrome and recurrent miscarriage.

Successful pregnancy after rituximab in a women with recurrent in vitro fertilisation failures and anti-phospholipid antibody positive.

We report a case of successful pregnancy after rituximab in a patient with a history of in vitro fertilisation (IVF) failures and positive anti-cardiolipin antibody (ACA). Following a course of rituximab, her ACA became negative and she successfully conceived with IVF treatment. This is the first case in literature describing the use of rituximab therapy in this clinical scenario.

[Suppressive effect of recombinant immunomodulator ronkoleukin on the blood level of antiinflammatory cytokines, anticardiolipin autoantibodies and heart failure].

AIM: To elucidate clinical efficacy and immunocorrecting properties of recombinant immunomodulator ronkoleukin in patients with postinfarction cardiac dysfunction with NYHA FC II-III CHF. MATERIAL AND METHODS: In a 6-months prospective comparative clinically controlled study we observed 33 survivors of myocardial infarction divided into 2 groups according to FC of chronic heart failure (CHF): group I (n=17) with FC II CHF with LVEF > 45% (mean age 52 +/- 2.9 years) and group II (n=16) with FC III CHF and lowered ( 40%) LVEF (mean age 53.7 +/- 3.3 years). Comparison group comprised practically healthy subjects. Clinico-laboratory and functional assessment of state of patients was carried out before initiation of therapy with ronkoleukin and in 2 - 3 days after completion of 2 courses of therapy with 3 months interval. Immunological study included determination of subpopulation content of peripheral blood lymphocytes, blood plasma immunoglobulins, antiinflammatory cytokines Il-1a, Il-1b, Il-2, Il-6, Il-8, TNFa and AB to Cl. RESULTS: It was found that ronkoleukin is an effective immunocorrector producing no adverse effects in patients with FC II-III CHF. Most pronounced effect ronkoleukin manifested in relation to humoral immunity lowering dysimmunoglobulinemia, blood levels of IgA, IgG, CIC and antibodies to cardiolipin, inhibiting excessive cytokine activation in dependence on degree of severity of CHF. CONCLUSION: Administration of ronkoleukin to patients with postinfarction dysfunction of the heart with FC II-III CHF for correction of secondary immunodeficient state in addition to basic therapy provides positive changes of hematological, immunological parameters, intracardiac hemodynamics, facilitates regression of symptoms of CHF and improves quality of life.

Aspirin reduces anticardiolipin antibodies in patients with coronary artery disease.

BACKGROUND: Anticardiolipin antibodies (aCL) have been found to be elevated in patients with coronary artery disease (CAD) and have been associated with an adverse outcome owing to their prothrombotic activity. The aim of this study was to investigate the effect of aspirin treatment on aCL levels in patients with chronic CAD. MATERIALS AND METHODS: Forty patients with chronic CAD scheduled for elective coronary artery bypass graft surgery (CABG) and 40 healthy controls participated in the study. Patients were treated with 300 mg of aspirin once daily (o.d.) for the first 12 days and placebo for the following 12 days before CABG in a double-blind, cross-over trial. Immunoglobulin (Ig) G-, IgM-, IgA-aCL and C-reactive protein (CRP) levels were measured in the controls and at the end of each treatment period in the patients with CAD. RESULTS: The IgA- and IgG-aCL levels were greater in patients with CAD than in the controls. Compared with the placebo, IgA, IgG subtypes and CRP levels were reduced after aspirin treatment (P = 0.001, P = 0.02, P = 0.04, respectively). The percentage reduction of IgA- and IgG-aCL was related to the percentage reduction of CRP after aspirin (P < 0.05). CONCLUSION: Aspirin treatment with 300 mg o.d. reduced the serum levels of IgA and IgG subtypes in patients with chronic CAD in parallel to a reduction in CRP. These findings offer an additional pathophysiological mechanism of the beneficial effects of aspirin in patients with chronic CAD.

Effects of unfractionated and low molecular weight heparin on antiphospholipid antibody binding in vitro.

OBJECTIVE: To compare the efficacy of unfractionated heparin and low molecular weight heparin in the in vitro binding of antiphospholipid antibodies obtained from the sera of patients with recurrent pregnancy loss. METHODS: Women with immunoglobulin (Ig) G antibodies to the phospholipids cardiolipin and phosphatidylserine were selected based on a positive test by a standard enzyme-linked immunosorbent assay (ELISA). The sera were reassayed for antiphospholipid antibodies in a modified ELISA using increasing doses of unfractionated heparin or low molecular weight heparin (0, 16, 32, 64, 128, and 256 IU). Sera were fractionated by unfractionated and low molecular weight heparin affinity chromatography to compare the binding avidity and antiphospholipid antibody activity. RESULTS: All sera demonstrated a dose-dependent inhibition in measured antiphospholipid antibody activity with the addition of unfractionated or low molecular weight heparin. Levels of IgG cardiolipin and IgG phosphatidylserine were significantly inhibited in the presence of 32 IU of low molecular weight heparin (P <.001 and P <.05, respectively) and in the presence of 64 IU of unfractionated heparin (P <.001 and P <.05, respectively). Antiphospholipid antibody binding activity in serum as measured in the ELISA was maximally reduced 76-89% with 256 IU of either heparin derivative. Affinity chromatography with unfractionated or low molecular weight heparin columns absorbed 72% and 66% of IgG cardiolipin activity, respectively, and 46% and 54% of IgG phosphatidylserine activity, respectively. CONCLUSION: These data suggest that low molecular weight heparin and unfractionated heparin reduce the in vitro binding of antiphospholipid antibodies on a per unit basis. Both heparins demonstrate binding activity similar to that of antiphospholipid antibodies in vitro.

Antiphospholipid antibodies in young women with and without oral contraceptive use.

The role of thrombophilia in the elevated risk of thromboembolism during oral contraceptive use has been established. We performed a cross-sectional study among young women to survey the occurrence of antiphospholipid antibodies among users and non-users of oral contraceptives. Serum levels of immunoglobulin (Ig)G, IgA and IgM isotypes of anti-beta2-glycoprotein I and anticardiolipin antibodies were measured by validated enzyme-linked immunosorbent assay methods. Combining all types of antiphospholipid antibodies, pill-users had an elevated antibody titre more than twice as frequently as non-users (odds ratio, 2.3; 95% confidence interval, 1.1-5.1). The higher frequency of elevated antibody titre was related most commonly to IgG type anti-beta2-glycoprotein I antibodies. Oral contraceptive use increases the risk of elevated antiphospholipid antibody levels among asymptomatic young women.

Effect of Salvia miltiorrhiza Bunge injection on anticardiolipin antibody production induced by beta2 glycoprotein.

AIM: To explore the therapeutic effect and the mechanism of Chinese herbs on antiphospholipid syndrome (APS) by observing the effect of Salvia miltiorrhiza Bunge injectio (SmBI) on anticardiolipin antibody (aCL) induced by beta2 glycoprotein I (beta2-GP I). METHODS: Sixty female mice randomly fell into 6 groups: group A, B, C, D was injected through abdominal cavity with different dosage of SmBI daily; after 14 d, group A, B, C, E was immunized with 150 microg of purified human beta2-GP I in complete Freund's adjuvant subcutaneously; group F as control. The titre of aCL were detected by enzyme linked immunosorbent assay; subsets of T cell were grouped by streptavidin-biotin complex technique; and the activity of IL-2 was measured by MTT chromatometry. RESULTS: (1) Compared with group E, the absorbance (A) of aCL in group A, B, and C was decreased (P < 0.05 or P < 0.01). By linear correlation, the dosage is negatively correlated with the A values of aCL in 1, 2, and 3 weeks (P < 0.01). (2) Compared with group E, TH/TS ratio was reduced in group A, B, and C (P < 0.05 or P < 0.01); there is no significant differences between group D and F (P>0.05). By linear correlation, the dosage is negatively correlated with TH/TS ratio (P < 0.01). (3) Compared with E, the activity of IL-2 in group B and C decreased significantly (P < 0.01). By linear correlation, there is negative correlation between dosage and IL-2 activity (P < 0.01). There is no significant difference between D and F (P > 0.05). (4) There is positive correlation between TH/TS ratio and IL-2 activity in different dilutions (P<0.01). CONCLUSION: The mechanism of suppressive effect of SmBI on aCL induced by beta2-GP I may be realized by resuming the elevated TH/TS ratio and IL-2 activity. The state that SmBI have no effect on normal mice indicates that SmBI has selective immunoregulative functive.

Síndrome anticuerpo antifosfolípido: cómo llegamos a su diagnóstico? / Anti-phosppholipid antibody syndrome: how do we arrive to its diagnosis?

Nuestro objetivo es presentar un razonamiento diagnóstico para la detección del síndrome anticuerpo antifosfolípido, teniendo en cuenta las variadas manifestaciones cutáneas del mismo. La sintomatología clínica, la biopsia de piel, el análisis hematológico e inmunológico incluyendo anticuerpos anticardiolipinas y el estudio de la coagulación constituyen la metodología para llegar al diagnóstico de certeza del síndrome. Las manifestaciones clínicas más relevantes son trombosis arteriales y venosas en cualquier segmento del árbol vascular y abortos recurrentes. Además puede presentar cefaleas e hipertensión arterial. Hacemos hincapié en el estudio de coagulación, sus pruebas de selección (APTT-dRVVT-TCK) y la detección de anticuerpos anticardiolipinas por enzimoinmunoanálisis

Síndrome anticuerpo antifosfolípido: cómo llegamos a su diagnóstico? / Anti-phosppholipid antibody syndrome: how do we arrive to its diagnosis?

Nuestro objetivo es presentar un razonamiento diagnóstico para la detección del síndrome anticuerpo antifosfolípido, teniendo en cuenta las variadas manifestaciones cutáneas del mismo. La sintomatología clínica, la biopsia de piel, el análisis hematológico e inmunológico incluyendo anticuerpos anticardiolipinas y el estudio de la coagulación constituyen la metodología para llegar al diagnóstico de certeza del síndrome. Las manifestaciones clínicas más relevantes son trombosis arteriales y venosas en cualquier segmento del árbol vascular y abortos recurrentes. Además puede presentar cefaleas e hipertensión arterial. Hacemos hincapié en el estudio de coagulación, sus pruebas de selección (APTT-dRVVT-TCK) y la detección de anticuerpos anticardiolipinas por enzimoinmunoanálisis (AU)

Neonatal estrogen treatment and its consequences for thymus development, serum level of autoantibodies to cardiolipin, and the delayed-type hypersensitivity response.

Eight-week-old female and male NMRI mice treated neonatally with the synthetic estrogen diethylstilbestrol (DES), estradiol-17beta, or tamoxifen displayed an enlarged thymus when compared with controls (approximately 1.5-fold). In control females, either ovariectomy or adrenalectomy increased thymus weight to the level characteristic for DES-treated females, but these endocrine ablations had no significant effect in DES females. The serum estrogen levels were similar in intact DES, ovariectomized DES, and ovariectomized female controls; serum corticosterone was similar in controls and DES females. The expression of the Thy1.2+ marker and the percentages of CD4+CD8+ DP and CD4+ and CD8+ SP cell subsets were similar in thymocyte populations from 8-wk-old controls and DES females; the CD4+ and CD8+ SP subsets were similar in splenocyte populations. The levels of serum immunoglobulin (Ig) G and IgM autoantibodies to cardiolipin showed age-dependent fluctuations but were similar in controls and DES females; however, the IgG autoantibodies in DES females were qualitatively different from those in controls with respect to sensitivity to bovine serum (a source of beta2-glycoprotein I). Contrary to females, DES-treated males had higher levels of autoantibodies than controls. The delayed-type hypersensitivity (DTH) response to oxazolone was similar in controls and DES animals at 8 wk, increased in DES females and males at 6 mo, but was reduced in DES females at 1 yr. Thus, even though adult mice with thymus enlargement after neonatal estrogen treatment do not differ from controls with respect to the expression of the Thy1.2 marker or percentages of CD4+/CD8+ DP or SP subsets in thymus and spleen, qualitative and quantitative differences occur in immune parameters (autoantibodies to cardiolipin) and a T-cell-dependent immune response (DTH).

Dietary polyunsaturated fatty acids decrease anti-dsDNA and anti-cardiolipin antibodies production in idiotype induced mouse model of systemic lupus erythematosus.

OBJECTIVE: To examine the effect of diets with different polyunsaturated fatty acid contents, including linseed oil which contains 70% omega-3 fatty acids, on autoantibody production in idiotype induced mouse model of systemic lupus erythematosus (SLE). METHODS: Five different fats were fed to mice with induced SLE and antibody titers to anti-DNA and anti-cardiolipin were determined and histological examination of kidneys were carried out. RESULTS: SLE mice fed linseed oil showed lower titers of antibodies to DNA and to cardiolipin and less severe kidney damage than mice fed other diets, including fish oil. CONCLUSION: Use of linseed oil may attenuate the severity of SLE and this diet may be recommended for other auto-immune diseases as well.

Tratamiento del factor inmunológico en el aborto recurrente y en el retardo de crecimiento intrauterino / Treatment of the immunologic factor in the recurrent abortion and the intrauterine growth retardation

Objetivo: Describir los resultados del tratamiento anti-agregante plaquetario en un grupo de pacientes con auto-anticuerpos y antecedentes de aborto recurrente (AR) y/o retardo de crecimiento intrauterino (RCIU). Material y Método: Catorce pacientes que consultaron por AR y/o RCIU, en cuyo estudio inmunológico se detectaron autoanticuerpos, constituyen el material del presente trabajo. A las pacientes que tenían como antecedentes un RCIU, una vez descartados todos los cuadros clínicos que pudieran ocasionarlo, se les efectuó determinación de autoanticuerpos. Las parejas que consultaban por AR eran sometidas a estudio para descartar un factor uterino, infeccioso, endocrinológico, metabólico, genético, inmunológico y espermático. En algunas pacientes se realizó una laparoscopia para descartar endometriosis. Todas las pacientes fueron tratadas con Acido Acetilsalicílico (AAS) 80 mg. diarios, a partir del momento de la detección de autoanticuerpos. A una sola paciente se le administró además Heparina subcutánea durante dos embarazos. Resultados: De las catorce pacientes en las que se detectaron autoanticuerpos, doce embarazaron, algunas de ellas en más de una oportunidad, sumando un total de 19 embarazos. De ellos, 5 concluyeron en aborto espontáneo y catorce concluyeron en el tercer trimestre con recién nacido vivo. De éstos, cinco presentaron un crecimiento fetal normal, mientras que los nueve restantes desarrollaron RCIU. La vía de terminación fue vaginal en cinco casos y abdominal en los nueve restantes. Conclusiones: El tratamiento del síndrome antifosfolipídico con antiagregantes plaquetarios (AAS o heparina), parecería ofrecer un porvenir reproductivo alentador a las portadoras de dicho síndrome

Tratamiento del factor inmunológico en el aborto recurrente y en el retardo de crecimiento intrauterino / Treatment of the immunologic factor in the recurrent abortion and the intrauterine growth retardation

Objetivo: Describir los resultados del tratamiento anti-agregante plaquetario en un grupo de pacientes con auto-anticuerpos y antecedentes de aborto recurrente (AR) y/o retardo de crecimiento intrauterino (RCIU). Material y Método: Catorce pacientes que consultaron por AR y/o RCIU, en cuyo estudio inmunológico se detectaron autoanticuerpos, constituyen el material del presente trabajo. A las pacientes que tenían como antecedentes un RCIU, una vez descartados todos los cuadros clínicos que pudieran ocasionarlo, se les efectuó determinación de autoanticuerpos. Las parejas que consultaban por AR eran sometidas a estudio para descartar un factor uterino, infeccioso, endocrinológico, metabólico, genético, inmunológico y espermático. En algunas pacientes se realizó una laparoscopia para descartar endometriosis. Todas las pacientes fueron tratadas con Acido Acetilsalicílico (AAS) 80 mg. diarios, a partir del momento de la detección de autoanticuerpos. A una sola paciente se le administró además Heparina subcutánea durante dos embarazos. Resultados: De las catorce pacientes en las que se detectaron autoanticuerpos, doce embarazaron, algunas de ellas en más de una oportunidad, sumando un total de 19 embarazos. De ellos, 5 concluyeron en aborto espontáneo y catorce concluyeron en el tercer trimestre con recién nacido vivo. De éstos, cinco presentaron un crecimiento fetal normal, mientras que los nueve restantes desarrollaron RCIU. La vía de terminación fue vaginal en cinco casos y abdominal en los nueve restantes. Conclusiones: El tratamiento del síndrome antifosfolipídico con antiagregantes plaquetarios (AAS o heparina), parecería ofrecer un porvenir reproductivo alentador a las portadoras de dicho síndrome (AU)

beta 2-Glycoprotein I reactivity of monoclonal anticardiolipin antibodies from patients with the antiphospholipid syndrome.

OBJECTIVE: To elucidate the specificity of anticardiolipin antibodies (aCL) from patients with the antiphospholipid syndrome (APS) to various phospholipids (PLs), DNA, and beta 2-glycoprotein I (beta 2-GPI). METHODS: Five monoclonal aCL were established from peripheral blood lymphocytes of 3 patients with the APS. The reactivity of monoclonal aCL with various PLs, with DNA, and with beta 2-GPI was examined by enzyme-linked immunosorbent assay (ELISA). RESULTS: All of the monoclonal aCL bound to anionic PLs, only in the presence of beta 2-GPI. Neither monoclonal aCL nor beta 2-GPI bound to DNA. Monoclonal aCL bound to solid-phase beta 2-GPI on polystyrene ELISA plates that had carboxyl groups on their surface, but did not react with solid-phase beta 2-GPI on ordinary polystyrene plates. A mixture of beta 2-GPI and CL inhibited the binding of monoclonal aCL to beta 2-GPI, but CL or beta 2-GPI alone did not. CONCLUSION: Monoclonal aCL may recognize a cryptic epitope, which appears as a result of beta 2-GPI binding to anionic PLs or to polystyrene with carboxyl groups.