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PubMed2000

Overlap of the anti-cardiolipin and anti-nucleosome responses of the (NZW X BXSB)F1 mouse strain: a new pattern of cross-reactivity for lupus-related autoantibodies.

The association of anti-nuclear antigen (ANA) and anti-cardiolipin (CL) antibodies is often observed during systemic lupus erythematosus (SLE) or the primary anti-phospholipid syndrome, thereby raising the possibility of a relationship between these two autoantibody populations. To determine whether ANA and anti-CL antibodies can overlap, we derived, from a male (NZW x BXSB)F1 mouse, 14 hybridomas selected based on their capacities to react with CL and to label HEp-2 cell nuclei. Four of these anti-CL were IgG and bound to CL and phosphatidylserine in a cofactor-dependent manner and reacted strongly with nucleosomes. Variable region sequence analysis indicated that these four monoclonal antibodies (mAb) were derived from three independent B cell clones that used recurrent heavy and/or light chain immunoglobulin rearrangements, as assessed by comparison with each other and prototypic anti-CL mAb previously derived from different lupus mouse strains. These results indicate that anti-CL mAb can have overlapping cross-reactivities with nucleosomes, thereby defining a new category of SLE-related autoantibodies characterized by their capacities to recognize distinct supramolecular complexes, formed by the association of an anionic structure and a protein, that exert a strong selective pressure on autoreactive B cell clones.

Amino Acid Sequence↗

PubMed1995

Human immunodeficiency virus rather than hepatitis C virus infection is relevant to the development of an anti-cardiolipin antibody.

We have investigated whether or not a relationship exists between anti-cardiolipin antibody (aCL) positivity and human immunodeficiency virus type 1 (HIV) and/or hepatitis C virus (HCV), and we have attempted to clarify which virus has close association with the development of aCL. We found that aCL positivity in HIV-infected patients was significantly higher than in HCV-infected patients. Furthermore, HIV/HCV dual-infected patients exhibited a higher aCL positivity than patients infected by HCV alone. From these results, we conclude that HIV rather than HCV plays an important role in the development of aCL.

Antibodies, Anticardiolipin↗

PubMed2006

Regulation of cardiolipin synthase levels in Saccharomyces cerevisiae.

The Saccharomyces cerevisiae cardiolipin (CL) synthase encoded by the CRD1 gene catalyses the synthesis of CL, which is localized to the inner mitochondrial membrane and plays an important role in mitochondrial function. To investigate how CRD1 expression is regulated, a lacZ reporter gene was placed under control of the CRD1 promoter and the 5'-untranslated region of its mRNA (P(CRD1)-lacZ). P(CRD1)-lacZ expression was 2.5 times higher in early stationary phase than in logarithmic phase for glucose grown cells. Non-fermentable growth resulted in a two-fold elevation in expression relative to glucose grown cells. A shift from glycerol to glucose rapidly repressed expression, whereas a shift from glucose to glycerol had the opposite effect. The derepression of P(CRD1)-lacZ expression by non-fermentable carbon sources was dependent on mitochondrial respiration. These results support a tight coordination between translation and transcription of the CRD1 gene, since similar effects by the above factors on CRD1 mRNA levels have been reported. In glucose-grown cells, P(CRD1)-lacZ expression was repressed 70% in a pgs1delta strain (lacks phosphatidylglycerol and CL) compared with wild-type and rho- cells and elevated 2.5-fold in crd1delta cells, which have increased phosphatidylglycerol levels, suggesting a role for phosphatidylglycerol in regulating CRD1 expression. Addition of inositol to the growth medium had no effect on expression. However, expression was elevated in an ino4delta mutant but not in ino2delta cells, suggesting multiple and separate functions for the inositol-responsive INO2/INO4 gene products, which normally function as a dimer in regulating gene function.

Base Sequence↗

PubMed1993

Association of anti-cardiolipin antibodies with vascular thrombosis and neurological manifestation of Behçets disease.

We have studied 44 patients with Behçet's Disease (BD) to look for any correlation of arterial and venous thrombosis or central nervous system (CNS) manifestations with anti-cardiolipin antibodies (ACLA). Twenty patients were positive for ACLA by MELISA method. Ten patients had IgG antibody, four had IgM and six had both IgG and IgM. Of these patients, 11 had a history of vascular thrombosis and thrombophlebitis and nine had CNS manifestations. The association of ACLA with vascular thrombosis or CNS manifestation of Behçet's disease was statistically not significant.

Antibodies, Anticardiolipin↗

PubMed1993

Circulating intercellular adhesion molecules-1 and autoantibodies including anti-endothelial cell, anti-cardiolipin, and anti-neutrophil cytoplasma antibodies in patients with vasculitis.

Circulating intercellular adhesion molecule-1 (ICAM-1) and 3 types of autoantibodies were measured in 30 patients with angiographical or pathological proved vasculitis. There were 18 patients with systemic vasculitis and 12 patients with cutaneous vasculitis. The measured antibodies included anti-endothelial cell antibodies (AECA), anti-cardiolipin (ACL) antibodies of 3 isotypes and anti-neutrophil cytoplasma antibodies (ANCA). The results showed that patients with systemic vasculitis had elevated levels of ICAM-1, AECA and IgG isotype antibody as compared with none or lower in patients with cutaneous vasculitis. Levels of ICAM-1 and IgG isotype ACL antibodies also decreased significantly after disease activity subsided in patients with systemic vasculitis. Measurement of ICAM-1 and autoantibodies may be useful in evaluating the extent of involvement and in following the disease activity of patients with vasculitis.

Adolescent↗

PubMed2005

IgG and IgM isotypes of anti-cardiolipin and anti-beta2-glycoprotein i antibodies reflect different forms of recent thrombo-embolic events.

We correlated the distribution and levels of serum anti-cardiolipin (aCL) and anti-beta(2)-glycoprotein-1 antibodies (anti-beta(2)-GPI) of the IgG and IgM isotypes to the clinical spectrum of recent (<6 months) thrombo-embolic events in a cohort of 162 patients. Clinical information was obtained by questionnaires from the referring physicians. Cerebro-vascular infarction (CVI) had taken place in 82 patients, deep venous thrombosis (DVT) in 34, pulmonary embolism (PE) in 14, myocardial infarction (MI) in four, and other thromboses in 28 patients. SLE was the most commonly associated rheumatic disease and accounted for 20 (12%) patients. In 124 (77%) patients no underlying rheumatic disease was identified. Isolated IgG aCL was found in 31 of 48 patients with DVT/PE (65%), but in only 21 of 82 patients with CVI (26%); p<0.0001. IgG anti-beta(2)-GPI were detected in 23 (48%) DVT/PE patients, but in only 13 (16%) CVI patients; p<0.001. The IgG class anti-beta(2)-GPI positive patients had significantly higher levels of IgG aCL (mean 65 units) compared to IgG anti-beta(2)-GPI negative patients (mean 29 units); p<0.0001. In contrast, isolated IgM aCL was found in nine (19%) patients with DVT/PE, but in 46 (56%) CVI patients; p<0.0001. Only ten patients had IgM anti-beta(2)-GPI. The present study shows that the IgG and IgM aCL isotypes seem to define different clinical subsets of patients with thrombo-embolic events with IgG aCL being most prevalent in the group having DVT/PE, IgM aCL being found primarily among CVI patients.

Adolescent↗

PubMed1990

Biochemical characterization and regulation of cardiolipin synthase in Saccharomyces cerevisiae.

Cardiolipin (CL) synthase activity was characterized in mitochondrial extracts of the yeast Saccharomyces cerevisiae and was shown for the first time to utilize CDP-diacylglycerol as a substrate. CL synthase exhibited a pH optimum of 9.0. Maximal activity was obtained in the presence of 20 mM magnesium with a Triton X-100: phospholipid ratio of 1:1. The apparent Km values for phosphatidylglycerol and CDP-diacylglycerol were 1 mM and 36 microM, respectively. CL synthase activity was maximal at 45 degrees C and heat inactivation studies showed that the enzyme retained greater than 75% of its activity at temperatures up to 55 degrees C. To study the regulation of CL synthase, the enzyme was assayed in cells grown under conditions known to affect general phospholipid synthesis. Unlike many phospholipid biosynthetic enzymes including PGP synthase, which catalyzes the initial step in CL biosynthesis, CL synthase was not repressed in cells grown in the presence of the phospholipid precursor inositol. Detailed procedures for the enzymatic synthesis of 32P-labelled substrates are described.

Cytidine Diphosphate Diglycerides↗

PubMed1993

Crossreaction between antibodies to oxidised low-density lipoprotein and to cardiolipin in systemic lupus erythematosus.

Serum lipoproteins contain phospholipids and modified low-density lipoprotein (LDL) may thus act as a target for antiphospholipid antibodies. Raised concentrations of IgG antibodies against oxidised LDL were found in 47 of 61 (80%) patients with systemic lupus erythematosus (SLE). 46% of patients also had raised concentrations of IgG anticardiolipin antibodies. Binding of anticardiolipin antibodies to solid-phase cardiolipin was inhibited by oxidised LDL but not by native LDL in 16 of 21 sera from SLE patients. These observations suggest crossreactivity between antiphospholipid antibodies, which are closely associated with thrombosis in SLE, and antibodies to oxidised LDL, thus providing a possible link between thrombotic and atherosclerotic complications in SLE.

Adult↗

PubMed1991

Genetic regulation of cardiolipin synthase in Escherichia coli.

Recombinant DNA and genetic techniques were used to construct Escherichia coli strains SOH92 [phi(cls-lacZ+)] and SOH93 [phi(cls-'lacZ)hyb]. beta-Galactosidase (116 kDa) synthesized by strain SOH92 was primarily present in the particulate fraction. Strain SOH92 produced about 20-fold more beta-galactosidase activity than strain SOH93. Expression of clsphilacZ in both SOH92 and SOH93 was influenced by the terminal electron acceptor (increasing in the order oxygen, nitrate, fumarate) when the cells were cultured in minimal medium with glycerol as the sole carbon source. As strains SOH92 and SOH93 progressed from early to late log growth phase under aerobic conditions in LB broth, clsphilacZ expression increased about 2.5-fold. Fusion strains containing a pss-1 allele had an increased cardiolipin (CL) level, but no corresponding increase in clsphilacZ expression was observed. A cls::Tn10dTet null mutation was introduced into SOH92 and SOH93. The strains produced less CL, but no corresponding changes in clsphilacZ expression were observed. A high copy number plasmid bearing the cls gene had no effect on clsphilacZ expression. Taken together, these results indicate that cls is not subject to autogenous regulation.

Blotting, Southern↗

PubMed1993

Production of anti-cardiolipin antibody in AKR/J mice with streptozocin-induced insulitis and diabetes.

We herein report that anti-cardiolipin antibodies (ACA) were detected in AKR/J mice treated with multiple low doses of streptozocin (STZ)-induced insulitis and diabetes. Daily intraperitoneal (i.p.) injections of 40 mg/kg body wt. of STZ for five consecutive days in the AKR/J mice resulted in hyperglycemia and mononuclear cell infiltrations of islets (insulitis). ACA appeared on day 14, when hyperglycemia began to occur, at a rate of 13.3% (4/30). The rate increased to 83.3% (25/30) on day 21, when diabetes developed, and then fell to 10% (3/30) on day 28. Neither the diabetic AKR/J mice treated with a single high dose of STZ (200 mg/kg body wt.) nor the non-diabetic insulitis free Balb/c mice and B10.S(9R) mice treated with multiple low doses of STZ (40 mg/kg body wt.) produced ACA. The IgG subclass of the ACA belonged mainly to IgG2a. These findings suggest that ACA are produced in association with the development of insulitis, but not induced by either hyperglycemia or STZ.

Animals↗

PubMed2004

Identification of an Arabidopsis thaliana gene for cardiolipin synthase located in mitochondria.

Cardiolipin (CL) is an anionic phospholipid with a dimeric structure. In eukaryotes, it is primarily localized in the inner membranes of mitochondria. Although the biosynthetic pathway of CL is well known, the gene for CL synthase has not been identified in any higher organisms. In this study, the CLS gene for a CL synthase has been identified in a higher plant, Arabidopsis thaliana. We have shown that the CLS gene encodes a CL synthase by demonstrating its ability to catalyze the reaction of CL synthesis from CDP-diacylglycerol and phosphatidylglycerol, and that CLS is targeted into mitochondria. These findings demonstrate that CLS is a CL synthase located in mitochondria.

Amino Acid Sequence↗

PubMed2000

Coronary calcium and anti-cardiolipin antibody are elevated in patients with typical chest pain.

The aim of this study was to examine whether detection of coronary calcium and the autoimmune response associated with atherosclerosis, either solely or in combination, are different in patients with typical and atypical chest pain. Coronary calcium as detected by spiral computerized tomography and levels of antibodies against cardiolipin (CL), oxidized low-density lipoprotein (ox-LDL), and beta2-glycoprotein-I (beta2-GPI) were studied in patients with typical chest pain (n = 52), atypical chest pain (n = 19), or without chest pain (n = 21). Patients with typical chest pain had higher mean levels of coronary calcium (expressed as natural transformation of total coronary calcium score) compared with patients with atypical chest pain and controls (5.04 vs 3.21 and 2.75, respectively; p < 0.001). The levels of anti-CL were (mean +/- SD of optical density multiplied by 1,000): 262 +/- 140, 170 +/- 82, and 230 +/- 115 for patients with typical chest pain, atypical chest pain, and controls, respectively (p = 0.016). No significant difference was found between groups regarding anti-ox-LDL and anti-beta2-GPI autoantibody levels. In the typical chest pain group, there was a higher prevalence of high total coronary calcium scores (p = 0.03) and high anti-CL levels (p = 0.01) than in the atypical chest pain group. Eighteen of 52 patients with typical chest pain (35%) had both high calcium scores and high antibody levels, whereas none of the 19 patients (0%) who had atypical chest pain had high levels of both (p = 0.003). A combination of both coronary calcium and anti-CL was associated with higher area under the receiver operator characteristic curves than for each separately. High coronary calcium scores or high anti-CL levels are found more often in typical than in atypical chest pain patients, but a combination of high levels of both can better differentiate typical from atypical chest pain patients.

Aged↗

PubMed1998

Abnormal IgG cardiolipin antibody titers in patients with Raynaud's phenomenon and/or related disorders: prevalence and clinical significance.

BACKGROUND: The prevalence of antibodies to cardiolipin (a-CL) in patients with Raynaud's phenomenon (RP) and/or related disorders (rD) is not known. OBJECTIVE: The purpose of this study was to determine the prevalence of these antibodies. METHODS: We assayed IgG a-CL in 230 consecutive patients with RP/rD and compared the results with those in a series of 230 control blood donors. RESULTS: Sixteen percent of patients were a-CL positive versus 7.8% of the control donors (p = 0.014). The prevalence of a-CL positivity was 8.7% for primary RP, 10.5% for secondary RP, 8% for chilblains, 25% for essential acrocyanosis, 20% for connective tissue diseases, and 17% for undifferentiated connective tissue diseases. Among patients with digital necrosis, 24% were a-CL positive. CONCLUSION: The prevalence of abnormal a-CL titers is higher in patients with RP/rD than in control donors, especially in patients with a connective tissue disease.

Antibodies, Anticardiolipin↗

PubMed1995

Increased thromboembolic incidence in anti-cardiolipin-positive patients with malignancy.

This study was undertaken to determine the prevalence of anti-cardiolipin antibodies (ACLAs) in patients with malignancy and to investigate a possible association of ACLAs with thromboembolic events in such patients. The study included 216 patients with solid and non-solid malignancies and an age-matched control group of 88 healthy subjects. ACLA levels were measured and related to thromboembolic phenomena (diagnosed by imaging methods) that occurred within 12 months of the diagnosis of cancer. Forty-seven patients (approximately 22%) with cancer were ACLA positive as compared with only three subjects (approximately 3%) in the control group (P < 0.0001). The ACLA-positive cancer patients had a significantly higher rate of thromboembolic events than ACLA-negative cancer patients: 13 of 47 (28%) vs 24 of 169 (14%), respectively (P < 0.05). High titres of either IgG-ACLA or IgM-ACLA were found in 10 out of 13 ACLA-positive cancer patients with thrombotic complications, but in only 2 out of 34 cancer ACLA-positive patients without thromboembolic events (P < 0.0001). In four cancer patients in whom ACLA levels were followed ACLA decreased after successful surgery/chemotherapy treatment and remained negative and thromboembolic free for 12 months of follow-up. Patients with malignancies show an increased prevalence of ACLA. Furthermore, ACLA-positive patients, mainly those with high titres, are much more prone to thromboembolic events.

Adult↗

PubMed1998

Factors affecting cardiolipin antibody assays: modification with polyethylene glycol compound.

Anti-cardiolipin antibody (aCL) measurement is only semi-reproducible, and current assays detect irrelevant as well as clinically significant antibodies. Factors found to influence results included the source of the enzyme-linked immunoabsorbent assay (ELISA) plate, and its pretreatment with solvents; the nature of the blocking solution; and the composition of the diluent used for reagents. Fetal calf serum (FCS) in the diluent appeared to reduce nonspecific (clinically irrelevant) binding and was not simply a source of beta2-glycoprotein-1 (beta2GP1). A polyethylene glycol compound was found to be an effective blocking agent, and its use enhanced the discrimination between positive and negative samples. A high level of variation may be inherent in aCL-ELISA methodology, but the use of polyethylene glycol compound appeared to be a helpful modification.

Antibodies, Anticardiolipin↗

PubMed1998

Anti-mitochondrial M5 type antibody represents one of the serological markers for anti-phospholipid syndrome distinct from anti-cardiolipin and anti-beta2-glycoprotein I antibodies.

The aim of this study was to characterize the antigen specificity and to evaluate the diagnostic and prognostic value of anti-mitochondrial M5 type antibodies (AMA M5). Fifty-eight patients selected on the basis of their AMA M5 positivity were investigated in relationship to their clinical and serological profile. Cross-absorption studies, Western blotting and immunoprecipitation analysis were carried out for AMA M5 antigen specificity characterization. Most patients had a diagnosis of systemic lupus erythematosus (SLE) (65.5%) or of primary anti-phospholipid syndrome (PAPS) (24%); all the patients were positive for IgG or IgM anti-cardiolipin (anti-CL) antibodies and 49% of them also displayed lupus anticoagulant (LA) activity. Anti-beta2-glycoprotein I (beta2-GPI) IgG were detectable in 30/38 sera (78.9%) and IgM in 34/38 (89.4%). While anti-CL and anti-beta2-GPI IgG antibodies were significantly associated with history of thrombosis and fetal loss, AMA M5 displayed a statistical association only for thrombocytopenia and recurrent fetal loss. Absorption with human beta2-GPI both in free solution or in solid phase as well as with CL liposomes or CL/beta2-GPI liposome complexes did not affect AMA M5 fluorescence. While AMA M5 activity is absorbed by whole mitochondrial preparations, no specific reactivities against several human, bovine and rat mitochondrial proteins could be detected in Western blotting and immunoprecipitation studies. AMA M5 appear to be detectable in both primary and secondary APS, displaying a strong association with the presence of thrombocytopenia and fetal loss. Although strictly related to anti-phospholipid antibodies, AMA M5, anti-CL and anti-beta2-GPI antibodies represent distinct serological markers of the APS.

Adolescent↗

PubMed2000

Increased transforming growth factor-beta (TGF-beta)-secreting T cells and IgA anti-cardiolipin antibody levels during acute stage of childhood Henoch-Schönlein purpura.

Henoch-Schönlein purpura (HSP) is a small vessel vasculitis characterized by increased serum IgA and IgA-dominant immune complex deposition in lesions. The involvement of IgA implies a probable role for TGF-beta, a major factor in IgA production, in the pathogenesis of HSP. Among IgA antibodies, serum IgA anti-cardiolipin antibodies (aCL) have been found in many diseases, including vasculitis. In addition to the clinical presentations and laboratory parameters, we further investigated the roles of IgA aCL and TGF-beta in childhood HSP. Twenty-six Chinese children with the diagnosis of HSP were enrolled. Blood samples from these patients were collected at both acute and convalescent stages. Intracellular staining of lymphocytes was performed to enumerate type 1 (interferon-gamma-secreting), type 2 (IL-4-secreting), and type 3 (TGF-beta-secreting) helper T cells. Serum levels of TGF-beta were detected by ELISA. Serum IgA aCL of 21 of 26 patients at the acute stage, 11 of them at the convalescent stage, were measured by ELISA. The data showed that IgA aCL serum levels were significantly elevated in patients compared with healthy controls (P < 0.001), and those patients at the convalescent stage (P < 0.001). In addition, TGF-beta-secreting T cells were significantly elevated during the acute stage, and decreased at the convalescent stage. Although more studies are needed, the high prevalence of IgA aCL and increased TGF-beta-secreting T cells in children with acute HSP revealed some points which should permit a better understanding of the pathogenesis of HSP.

Acute Disease↗

PubMed2003

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

BACKGROUND: 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. MATERIALS AND METHODS: One hundred and seventy-four consecutive HCV patients, 50 HBV patients and 267 healthy were investigated for the presence of anti-CLAbs and antibodies against beta2-glycoprotein I (beta2-GPI), which is the most important cofactor of the 'pathogenic' anti-CLAbs in APLS. IgG anti-CLAbs were determined by an in-house quantitative ELISA and anti-beta2-GPIAbs using a commercial ELISA kit. RESULTS: 21.3% of the HCV and 14% of the HBV patients tested positive for IgG anti-CLAbs (P < 0.0001 compared with healthy controls). Neither age, sex, certain epidemiologic and laboratory parameters nor the clinical status and the histologic findings were associated with anti-CLAbs detection in both diseases. 2.3% of the HCV (P < 0.05 compared with healthy controls) and 2% of the HBV patients tested positive for anti-beta2-GPIAbs. Presence of anti-CLAbs was not associated with features of APLS. CONCLUSIONS: A significant proportion of the HCV and HBV patients had detectable IgG anti-CLAbs. However, the anti-CLAbs titres were relatively low, and in most cases seem to be cofactor-independent ('nonpathogenic'). The latter is further supported by the lack of their association with clinical features of APLS. Furthermore, anti-CLAbs appear to be detected irrespective of the demographic, laboratory, clinical and histologic status in both HCV and HBV. However, prospective studies of longer duration may be required in order to address whether anti-CLAbs in patients with chronic viral hepatitis are or are not of clinical importance.

Abortion, Habitual↗