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PubMed1987

Antibody to cardiolipin, lupus anticoagulant, and fetal death.

We compared the concordance and predictive powers of activated partial thromboplastin time (APTT) and of IgG and IgM antibody to cardiolipin (aCL), for predicting fetal death in 50 pregnant women with systemic lupus erythematosus (SLE) and/or lupus anticoagulant. Overall concordance of any abnormal determination of aCL during pregnancy with any abnormal determination of APTT was 76% (0.05 less than p less than 0.10). Fetal death occurred in 6/12 (50%) of patients with high APTT compared to 5/20 (16%) of patients with low APTT; fetal death occurred in 10/13 (77%) of patients with abnormal aCL and in 2/37 (5%) of patients with normal aCL. Sensitivity for predicting fetal death was .55 for APTT and .85 for aCL; specificity was .81 for APTT and .92 for aCL. Abnormalities of APTT and aCL are sufficiently frequently discordant to prevent equation of the 2 assays. ACL is the better assay for predicting fetal death.

Antibodies↗
PubMed1988

[Clinical value of determining the levels of lupus anticoagulant and anti-cardiolipin antibodies].

The authors present some literature and their own data on the antiphospholipid syndrome (venous and arterial thromboses; obstetric pathology: spontaneous abortions and intrauterine death of a fetus; CNS involvement: from headaches and disorientation to psychic disorders and cerebral infarctions; thrombocytopenia, early aseptic osteonecrosis and pulmonary hypertension). Altogether 58 patients with lupus erythematosus and 20 patients with a complicated obstetric history were examined. Lupoid anticoagulant and antibodies to cardiolipin were revealed in 27 patients. The determination of antiphospholipid antibodies was shown to be of practical value from the view-point of the knowledge of humoral immunity disorders in systemic lupus erythematosus, more detailed clinico-laboratory characterization of patients, the diagnosis and prediction of disorders of the blood coagulation system, and the development of obstetric pathology.

Adolescent↗
PubMed1988

Anti-cardiolipin antibodies in ischaemic heart disease.

IgG and IgM anti-cardiolipin antibodies (ACA) were assayed by an ELISA technique in 86 patients with ischaemic heart disease (IHD) and compared to 124 healthy controls and to 62 patients with rheumatoid arthritis (RA) and 20 with tuberculosis (TB). IgG ACA levels in IHD, RA and TB were comparable and significantly higher than in controls (P less than 0.0001). IgM ACA was significantly higher in IHD and RA than controls (P less than 0.0001) but not TB (P = 0.045). The number of IHD patients with raised ACA (IgG and/or IgM) was significantly greater than in RA or TB. (chi 2 = 30.77, P less than 0.0001). ACA were raised in 80.2% IHD patients on one or more occasions during a 1-11 day (mean 4.7) hospital admission. There was no difference in either ACA levels or in the frequency of ACA elevation in patients with stable or unstable angina pectoris or myocardial infarction. We conclude that there is a strong association between IHD and ACA with potentially important therapeutic implications.

Adult↗
PubMed1988

[Early modification of the fatty acid composition of cardiolipins and other phospholipids in rat liver mitochondria during dietary deficiency of essential fatty acids followed by repletion].

Weaned rats (21-day old, 44 +/- 2 g) were distributed into 3 groups. The first group was raised on a laboratory diet for 7 or 20 days (control group). The second was fed a diet containing 0.07% fat for 1, 2, 3, 7 or 66 days. The third one was fed the low-fat diet for 7 days and then switched to a laboratory chow diet for 1, 2, 5 or 9 days. Cardiolipin (CL) on the one hand and other mitochondrial phospholipids taken as a whole (PLm) on the other hand were prepared from liver mitochondria and their fatty acids analysed. Polyunsaturated fatty acids in PLm (18:2 (n-6), 20:4 (n-6), 22:6 (n-3) acids) decreased abruptly during the first 3 days of fat deficiency and then remained rather stable till day 7. CL behaved in a quite different way. 18:2 (n-6) acid, the major polyunsaturated fatty acid of CL, decreased continuously between day 1 and day 7 from 79% to 33%. A value of 19.6% was reached on day 66. When deficient rats were transferred to an equilibrated diet, the fatty acid profile of PLm was rapidly restored. Major effects were already achieved during the first 24 h and a fatty acid composition identical to that of control rats was reached within 2 days. A considerably longer period (about 9 days) was necessary for CL to reach a level analogous to that of control rats. The diminution of linoleic acid in CL was brought about by increases in the levels of monoenoic acids: palmitoleic (16:1 (n-7)), oleic (18:1 (n-9)) and cis-vaccenic (18:1 (n-7)) acids which accounted for 18.0%, 17.7% and 17.9% respectively on day 7. Increases in the (n-7) monoenes remained comparatively low in PLm, which showed a sharp rise in their oleic acid content (from 3.9% to 12.7% in 3 days). The proportion of cis-vaccenic acid relative to total octadecenoic acids was decreased from 75% to 50% in CL. A similar trend, but of smaller magnitude, was also noticed in PLm.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed1988

Cardiolipin antibodies and null alleles of C4 in black Americans with systemic lupus erythematosus.

Twelve of 44 black American patients with systemic lupus erythematosus (SLE) (27%) studied during periods of disease activity had increased levels of IgG antibodies against cardiolipin (IgG aCL). IgG aCL occurred almost exclusively in patients who had a partial genetic deficiency of C4A or C4B. Eleven of 29 patients (38%) with a C4A or C4B deficiency allele had IgG aCL, compared with 1/15 patients (7%) who did not have C4A or C4B deficiency allele (p = 0.04). During periods when SLE was less active clinically, IgG aCL levels returned to normal in 10/12 patients. Active SLE, rather than null alleles, appeared to be associated with low C4 levels in patients with IgG aCL.

Adult↗
PubMed1987

Anti-cardiolipin antibodies in neurological disorders: cross-reaction with anti-single stranded DNA activity.

Antiphospholipid (PL) antibodies have been detected in sera from patients with chronic neurological diseases associated with disorders of immunity. In an isotype specific radioimmunoassay for anti-cardiolipin (CL) antibodies, we found IgM anti-CL (greater than 2 s.d. above mean of controls) in 17/25 (68%) patients with myasthenia gravis (MG), 8/25 (32%) with the Lambert-Eaton myasthenic syndrome (LEMS), 5/17 (29%) with multiple sclerosis and 3/11 (27%) cases of migraine. IgG anti-CL was only found in low titres in sera from 10 patients with MG and three with LEMS. Significant anti-CL activity could not be detected in sera from nine patients with acute Guillain-Barré Syndrome (GBS), 12 chronic cases of epilepsy, 8/9 with oat cell carcinoma and 9/10 with acute stroke. Further tests on 39 sera with the highest anti-CL activity, from all of the above disease groups, showed a significant correlation between IgM anti-CL and IgM anti-ss DNA activities. In a series of competitive inhibition assays six sera from patients with MG were shown to have a proportion of both specific and cross-reactive IgM anti-CL and IgM anti-ss DNA antibodies. Anti-phospholipid antibodies occur in certain neurological diseases, at lower titres than seen in SLE, yet their cross-reactive binding to ss DNA suggests similar antibacterial origins as have been proposed for lupus auto-antibodies. In the absence of overt infection they might reflect a breakdown of tolerance for non-organ specific membrane antigens in diseases with predominantly organ specific membrane bound putative autoimmunogens.

Antibodies, Antinuclear↗
PubMed1987

High prevalence of anti-cardiolipin and other autoantibodies in a healthy elderly population.

Serum samples from 64 apparently healthy individuals (32 men and 32 women, mean age 81.0 years) were examined for the prevalence of several autoantibodies, including rheumatoid factor (RF), antinuclear antibodies (ANA), antibodies to extracted cellular antigens Ro (SSA), La (SSB), Sm, U1nRNP and Scl-70. IgG and IgM isotype-specific ELISA methods were applied for the detection of antibodies to ssDNA (anti-ssDNA), to dsDNA (anti-dsDNA) and to cardiolipin (anti-CL). The sera of this elderly population were found to contain a plethora of autoantibodies; RF was detected in 14.1%, ANA in 31.3% and anti-Ro (SSA) in 1.6% of the individuals. Precipitating antibodies to La (SSB), Sm, U1nRNP and Scl-70 were absent, while 15.6% of the sera displayed precipitating antibodies to a common undefined human spleen antigen. ELISA methods revealed anti-ssDNA in 17.2% of the individuals, anti-dsDNA in 14.1% and anti-CL in an extremely high incidence (51.6%). Notably, the above autoantibodies were exclusively of IgG isotype. Tests of 261 sera from healthy non-elderly individuals disclosed only anti-CL (IgG and IgM isotypes) in 2.3% of them. The levels of IgA and IgG immunoglobulins were increased in 23.4% and 29.7% of the elderly subjects, respectively. IgM was elevated in 3.1%, but it was also found decreased in 9.4%. This study documents the high incidence of autoantibodies in the healthy elderly, including for the first time, anti-CL antibodies. Furthermore, the relative impairment in IgM autoantibody production observed, possibly indicates the involution of the senescent immune system.

Adult↗
PubMed1978

[Immunohistochemical study of cardiolipin and phosphatidylinositol in mouse liver].

Localization of two phospholipid haptens--cardiolipin and phosphatidylinositol--in mouse liver sections was studied by the indirect method of fluorescent antibodies. Two types of liver sections--paraffin and cryostat, and two type of fixation--in acetone, and in the acetone, buffer, and formalin mixture--were used. Antiphospholipid sera stain specifically the plasma membrane of hepatocytes and predominantly the membrane region overlooking the blood capillary. A possibility of detecting the specific phospholipid haptens depends on the method of obtaining the sections and their fixation. Two types of immunization give two types of antiphospholipid sera which differ by the stability, by the possibility of monospecific antibodies isolation from them on lipid immunosorbents, and by the types of liver section staining.

Animals↗
PubMed1987

Evaluation of the anti-cardiolipin antibody test: report of an international workshop held 4 April 1986.

Thirty laboratories from institutions in Britain, France, Italy, The Netherlands, New Zealand, Sweden and the USA participated in a workshop to evaluate the anti-cardiolipin (aCL) test. Participants were asked to measure IgG and IgM aCL in seven samples on each of three separate days. The seven samples were prepared so that IgG and IgM aCL concentrations were known before distribution. Twenty-three of 30 laboratories measuring IgG aCL had significant regression slopes (P less than 0.001) when optical absorbance readings or counts per minute were compared with IgG aCL concentration. Twenty-four of 28 laboratories measuring IgM aCL had significant regression slopes (P less than 0.001). Coefficient of determination (R2) ranged from 81.1% to 98.7% for laboratories with valid IgG aCL assays and from 48.0% to 96.7% for valid IgM aCL assays. Valid assays had in common the use of 10% fetal calf or 10% adult bovine serum in PBS. Assays that were not valid had in common the use of PBS, PBS-Tween, or 0.3% gelatin as diluents. All laboratories with valid assays defined samples with high and moderate aCL levels as positive but there was no consensus about low positive samples. This study shows that properly performed ELISA or SRIA assays can be used to provide an accurate, reproducible, and quantitative measure of IgG and IgM aCL concentration in serum samples.

Autoantibodies↗
PubMed1985

Antibody-independent activation of C1, the first component of complement, by cardiolipin.

Lipid vesicles containing phospholipids known to be present in substantial amounts in mitochondrial membranes were tested for their capacity to activate C1. Among them, only cardiolipin (CL) was highly efficient in C1 activation; no such effect was observed with phosphatidylcholine, phosphatidylethanolamine, or phosphatidylinositol. CL was shown to bind specifically C1q, because only unlabeled C1q competed with 125I-C1q for binding to CL. The requirement for C1q was confirmed by the finding that only fully reconstituted macromolecular C1, containing C1q, was activated by CL. The specificity of CL-induced activation of C1 was also demonstrated by introducing adriamycin, an agent known to interact with CL. Whereas adriamycin did not decrease C1 activation induced by immune complexes, it abrogated C1 activation by CL. The latter was shown to be a strong nonimmune activator of C1, because C1-INH did not inhibit CL-induced activation. When the concentration of CL in vesicles was decreased in the presence of phosphatidylcholine, C1 activation was detected only above a critical level of 35 mol% CL, compatible with a minimal density or clustering of CL molecules in the plane of the membrane. Moreover, C1 activation by CL was modulated by the addition of cholesterol. The threshold of CL required for C1 activation was lowered by the incorporation of more than 35 mol% cholesterol into the vesicles. These results show that CL incorporated into liposomes can be a potent nonimmune activator of C1. The negatively charged phosphate groups in CL are likely candidates for Clq-binding.

Cardiolipins↗
PubMed1985

Measurement of anti-cardiolipin antibodies by an enzyme-linked immunosorbent assay (ELISA): standardization and quantitation of results.

We describe the development of a simple and highly sensitive double antibody sandwich enzyme-linked immunosorbent assay (ELISA) for measuring IgG and IgM anticardiolipin antibodies (ACA). Microtitre plates were coated with cardiolipin at a concentration of 45 micrograms/ml by evaporation under nitrogen. Non-specific binding of diluted sera was eliminated by blocking of plates with 10% fetal calf serum in phosphate buffered saline (PBS/FCS) for 2 h. Then sera (100 microliters) at a dilution of 1:100 were incubated in the wells for 1 h. Affinity purified goat anti-human IgG or IgM (100 microliters) at a concentration of 1 microgram/ml was subsequently added and allowed to incubate for 1 h; detection of ACA was achieved using an alkaline phosphatase conjugated rabbit anti-goat IgG reagent by reading the colorimetric yield at 405 nm after incubation with substrate. Reference serum pools were established to study reproducibility of the assay throughout its sensitivity range, and Standard curves were established. The quantitative normal range was 0-9.0 Anticardiolipin ELISA Units (AEU) for IgG and 0-8.0 (AEU) for IgM-ACA. A strong correlation was found between the ELISA and radioimmunoassay methods for measuring ACA of both IgG and IgM classes. Results from 65 patients with systemic lupus erythematosus (SLE) and 45 patients with seropositive rheumatoid arthritis are also reported. The advantages of the ELISA method for quantitative determination of ACA levels, should make it a useful and reliable method for clinical and experimental monitoring of patients with SLE and associated autoimmune disorders.

Arthritis, Rheumatoid↗
PubMed1994

V gene analysis of anti-cardiolipin antibodies from (NZW x BXSB) F1 mice.

In (NZW x BXSB) F1 (W/B F1) male mice, systemic lupus-like disease, thrombocytopenia and coronary vascular disease with myocardial infarction occur, due to the presence of platelet-associated antibodies, anti-platelet antibodies and anti-cardiolipin antibodies (aCL). We developed monoclonal aCL and analysed the specificity of aCL. In the W/B F1 mice, there are aCL with pathogenic properties, which have an IgG isotype and reveal a cofactor-dependent binding to CL, binding activity to platelets, and lupus anti-coagulant (LA) activity. Here, we analysed the usage of VH and V kappa genes of six aCL, including two pathogenic aCL, from W/B F1 mice, in an attempt to address the question of whether or not aCL with pathogenic properties use restricted Ig V genes. Sequence analysis of VH and V kappa genes of aCL showed that the pathogenic aCL had VHJ558 and V kappa 21 or V kappa 23 genes, whereas the other aCL without pathogenic features used mainly the 7183 VH family and the random V kappa gene group. However, two pathogenic aCL showed a 86.6% homology with the IgV region, each other, indicating that they were not closely related clones. Thus, these findings suggest the possibility that usage of Ig VH genes in pathogenic aCL is not random, but that there may exist a few epitopes of antigen recognized by the pathogenic aCL.

Amino Acid Sequence↗
PubMed1994

Induction of tissue factor-like activity in monocytes by anti-cardiolipin antibodies.

Anti-cardiolipin Abs (ACLA) are present in the sera of patients with antiphospholipid syndrome (APLS) and are associated with high incidence of thromboembolic phenomena, fetal loss, thrombocytopenia, and prolongation of the phospholipid-dependent coagulation assays (lupus anticoagulant). Recently, it has been shown that APLS can be induced experimentally by using ACLA. However, the pathophysiology of thrombus formation in this syndrome is unknown. Monocytes generate a potent procoagulant activity (PCA) after stimulation with various substances. Increased PCA has been found in monocytes from patients with diseases that are associated with high incidence of thromboembolic phenomena. In the present study, we report that the monoclonal ACLA that were shown previously by us to induce APLS stimulate mononuclear cells to generate a potent PCA. The PCA resembled tissue factor (TF) in that it accelerated clotting through the extrinsic coagulation pathway, was abolished by phospholipase C, and was inhibited by anti-TF mAbs. The induction of TF-like activity by ACLA in monocytes was dose- and time-dependent. It was induced in monocytes and monocytic cell lines, but not in lymphoid or myeloid cells, and did not require T lymphocytes for expression. The generation of PCA was dependent on protein synthesis inasmuch as it was prevented by adding puromycin to the system and was not affected by cytarabine. The TF-like activity that is induced by ACLA in monocytes may activate coagulation and thereby play a major role in the pathogenesis of thrombus formation in APLS.

Adult↗
PubMed1993

A somatic cell mutant defective in phosphatidylglycerophosphate synthase, with impaired phosphatidylglycerol and cardiolipin biosynthesis.

Phosphatidylglycerophosphate (PGP) synthase catalyzes a reaction involved in the synthesis of phosphatidylglycerol (PG), which serves as a metabolic precursor for cardiolipin (CL), found primarily in the mitochondrial membranes of eukaryotic cells. We isolated a Chinese hamster ovary cell mutant (designated PGS-S) with a specific lesion in PGP synthase by using an in situ enzymatic assay for the enzyme. This mutant was obtained by introducing a second mutation into mutant PGS-P that had been generated by first-step mutagenesis. The PGP synthase activities in cell extracts of mutant PGS-S grown at 33 and 40 degrees C were 14 and 1% of those in the wild type cells, respectively; in addition, PGP synthase in cell extracts of mutant PGS-S exhibited higher sensitivity to heat than that of the wild type. Mutant PGS-S also showed a temperature-dependent defect in the synthesis of PG and CL in vivo, together with temperature sensitivity for cell growth. A temperature-resistant revertant of mutant PGS-S simultaneously restored PGP synthase activity and the ability to synthesize PG and CL in vivo to nearly the same levels as those of mutant PGS-P. These results constitute genetic evidence that PGP synthase is responsible for PG synthesis and is essential for cell growth.

Animals↗
PubMed1996

Anticardiolipin antibodies from syphilis and systemic lupus erythematosus induce leakage in cardiolipin vesicles.

Anticardiolipin antibodies from sera of patients with systemic lupus erythematosus or syphilis induced leakage of entrapped carboxyfluorescein (CF) from cardiolipin (CL)/phosphatidylcholine(PC) vesicles prepared by sonication of equimolar mixtures of CL:PC. The sera dilution used here was 1:7500. IgG (5-20 micrograms/ml) from the same sera, not containing beta 2GPI, also produced a concentration-dependent leak. Vesicle leakage was inhibited by salt and was not detected with vesicles prepared exclusively with phosphatidylcholine. The demonstration of antibody-induced vesicle leakage offers a convenient system to investigate the mechanism of antibody-lipid binding as well as a potential diagnostic tool.

Antibodies, Anticardiolipin↗
PubMed1996

Monoclonal antibodies from NZW x BXSB F1 mice to beta2 glycoprotein I and cardiolipin. Species specificity and charge-dependent binding.

NZW x BXSB F1 mice develop a systemic autoimmune syndrome with various lupus-like manifestations. Male animals develop a degenerative coronary disease with myocardial infarction, resulting in death before 6 mo of age. The presence in these mice of anti-phospholipid Abs reacting with beta2-glycoprotein I may contribute to the pathogenesis of the cardiovascular lesions. beta2-glycoprotein I, a plasma protein implicated in various aspects of the coagulation pathway, is also the target of autoantibodies in humans with the anti-phospholipid syndrome. We obtained several mAbs from NZW x BXSB F1 mice that were selected for binding to cardiolipin. Two mAbs are specific for beta2-glycoprotein I and display a species-dependent pattern with preferential reactivity to mouse beta2-glycoprotein I. The other mAbs display charge-mediated interactions with anionic phospholipids in the absence of beta2-glycoprotein I. The analysis of the V region sequences of the mAbs suggests that cationic residues in the H chain complementarity-determining region 3 are important for their phospholipid reactivity. The structural features of the V(H)-D-J(H) junctions of these mAbs further support the view that an increased frequency of unusual V(D)J rearrangements directly contributes to the development of murine autoimmunity.

Amino Acid Sequence↗
PubMed1976

Circular DNA and cardiolipin in hydrogenosomes, microbody-like organelles of trichomonads.

Circular molecules of DNA approximately 3 mum in length were revealed by electron microscopy in deproteinized extracts prepared from purified hydrogenosomal fraction of a protozoan Tritrichomonae foetus. This fraction contained also cardiolipin amounting to approximately 14.4% of its total phospholipids, as detected by thin-layer chromatography and quantitative phosphorus measurement. These characteristics extend a number of biochemical properties of hydrogenosome shared also by mitochondria and by prokaryotic cells.

Cardiolipins↗