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PubMed1991

Clinical utility and specificity of anticardiolipin antibodies.

Established solid phase assays for anticardiolipin antibodies (aCL) are often characterized by high levels of nonspecific binding. As a result, only very high levels of aCL have been reported to be associated with a variety of clinical conditions including systemic lupus erythematosus (SLE), recurrent intravascular thrombosis and unexplained recurrent fetal loss. We have developed an ELISA replacing direct evaporation of soluble cardiolipin with cardiolipin micelles in physiological saline as the antigen binding step in the assay. Levels of IgG aCL were detected in various sera at dilutions of 1/100 to 1/3200, showing improved assay sensitivity. Assay specificity was determined using double stranded DNA and ovalbumin as irrelevant binding antigens and no crossreactivity was found. The controversial use of Tween 20 in the assay was investigated and results showed it decreases nonspecific binding without interfering in antibody detection. This assay has enabled us to identify differences in the prevalence and level of aCL antibodies in sera from healthy nonpregnant controls (0/25 positive), healthy pregnant controls (5/47 positive for IgG and 8/47 positive for IgM) and from women with unexplained recurrent fetal loss (16/62 and 14/62 positive, respectively). We support the observation that aCL are not normally distributed, and therefore nonparametric methods of statistical analysis are necessary to determine population prevalence. We confirm that aCL IgM are a relatively nonspecific finding, and extreme caution must be used in basing any clinical decisions on the presence of this antibody alone.

Adolescent↗
PubMed1976

[Changes of the phospholipid bilayer structure under the adsorption of ferricytochrome c on its surface].

Changes in the mobility of phospholipid molecules in liposomes membranes under adsorption ferricytochrome c on its surface were studied by means of NMR and EPR spectroscopy. It is found that the interaction of cytochrome molecules with vesicles causes the broadening of 1H-NMR signals of hydrophobic as well as polar groups in cardiolipin and phosphatidylcholine in the presence of lauric or phosphatidic acid. This broadening of 1H-NMR signals in hydrophobic groups may be caused by decrease in the rate of lateral diffusion of phospholipid molecules. The changes in the correlation time of hydrophobic spin-proub in liposomes containing phosphatydiloholine and cardiolipin with the increase of ferricytochrome c concentration were also observed. These changes suggest that the formation of protein-phospholipid clusters results in the impair of the regular structure of phospholipid bilayer.

Adsorption↗
PubMed1991

[Heterogeneity of anticardiolipin antibody].

The antibody to cardiolipin(ACA) was tested in patients with systemic rheumatic disease. The frequency of IgG ACA was 46/100(46.0%) in systemic lupus erythematosus(SLE). In other rheumatic disease, this was less than 20%. Significant correlation between the presence of IgG ACA and thrombosis and/or thrombocytopenia was found in patients with SLE. Eight sera containing high titered IgG ACA from lupus patients were selected for further inhibition study. Inhibitors were consisted of cardiolipin(CL), phosphatidyl(p-)serine, p-inositol, p-glycerol, p-ethanolamine, p-choline, ds-DNA, ss-DNA, fresh platelets(PLT)and fresh red blood cells(RBC). All sera were markedly inhibited by negatively charged phospholipids. In 4 sera(group B), there was moderate inhibition by ss-DNA, ds-DNA, PLT and RBC. In another 4 sera(group A), mild but significant inhibition was obtained by PLT alone. The number of platelet in group A was less than that in group B. There were some differences in inhibitory activity, suggesting heterogeneity of antibody to CL. It may be possible to speculate that heterogeneity of IgG ACA cause various combination of clinical features such as thrombocytopenia and thrombosis.

Abortion, Spontaneous↗
PubMed1976

Alterations in lipid constituents during growth of Mycobacterium smegmatis CDC 46 and Mycobacterium phlei ATCC 354.

Phospholipids of Mycobacterium phlei ATCC 354 and Mycobacterium smegmatis CDC 46 consist of cardiolipin, phosphatidyl ethanolamine, tri-acylated dimannophosphoinositide, tetra-acylated dimannophosphoinositide and tetra-acylated pentamannosphosphoinositide. A comparative study of lipid patterns of M. phlei ATCC 354 and of M. smegmatis CDC 46 in relation to age of culture revealed higher total lipid level and increased activity of malate-vitamin K reductase, a phospholipid requiring enzyme, during the early logarithmic growth phase of the former. No appreciable change occurred in the latter. The high total lipid content coincides with an increase in phospholipid, brought about apparently by the increase in malate-vitamin K reductase. Changes in cardiolipin and phosphatidyl ethanolamine appeared to be unique to M. phlei ATCC 354. However, in both bacterial species, a decrease in glyceride and a progressive increase in tuberculostearic acid with a concomitant decrease in oleic acid, occurred with ageing.

Cardiolipins↗
PubMedDate not supplied

[Influence of phospholipids of different natures on the structure of ferricytochrome c].

Influence of the two types of lipid molecules (neutral and acidic phospholipids) on the structure of ferricytochrome c in lipid-protein complexes was studied with the use of circular dichroism and absorption spectroscopy methods. It was found that interaction of ferricytochrome c with acidic phospholipids (cardiolipin and phosphatidylinositols) brings about some changes in the protein conformation, while interaction with neutral phospholipid (phosphatidylcholine plus 10% lauric acid) does not. Some difference in the mode of interaction of different acidic phospholipids with ferricytochrome c was also observed. As evidenced by visual light absorption spectra, cardiolipin induced the disruption of hem--methionine 80 bond in the protein molecule, while phosphatidylinositols do not cause such an effect.

Cardiolipins↗
PubMed1990

Analysis of factors affecting human hybridoma production.

Human hybridomas were produced by fusion of the GM 4672 cell line with lymphocytes from the peripheral blood, spleen, and bone marrow of patients with systemic lupus erythematosus. Lymphocytes were also obtained from normal human fetal liver of 12 weeks' gestation. The influence of anatomical source, fusion ratio, pre-stimulation with pokeweed mitogen, HLA match, and disease activity at the time of fusion was studied. Supernatants were screened for immunoglobulin secretion and binding to DNA, cardiolipin, poly(ADP-ribose), and histones by enzyme-linked immunoassay. A total of 28 fusions from 14 donors and 6 fusions with fetal lymphocytes were performed. The spleen was found to be the most efficient source of lymphocytes, with a fusion ratio of 1:1 resulting in a maximum yield of 27 clones/10(7) lymphocytes fused. HLA matching was a factor influencing the outcome with HLA A2 matching being the most important. Pre-stimulation with pokeweed mitogen did not improve the fusion frequency, and fusions using lymphocytes from patients with active disease were only marginally more successful. Over 95% of clones secreted immunoglobulin; autoreactivity was found against DNA, histones, cardiolipin, and poly(ADP-ribose). All hybrids with autoreactivity secreted IgM.

Antibodies, Monoclonal↗
PubMed1990

[Study of oxidative damage on mitochondrial membrane of myocardium deficient in selenium].

Keshan disease (KD) is an endemic cardiomyopathy with membrane damage of the myocardial mitochondria. This paper reports the results of feeding guinea pigs with a forage obtained from KD endemic area. The selenium (Se) content and glutathione peroxidase activity in blood, tissues, mitochondria, microsome and supernatant of myocardium and liver cells of the experimental group were decreased. Additionally, the activity of glutathione-S-transferase was also decreased to some extent, instead of being increased in normal cases, and the content of lipid peroxide and hydroperoxide was increased. At the same time, cardiolipin in mitochondria, and the activity of cytochrome C oxidase (CCO, an enzyme binding with mitochondrial membrane) were also reduced accompanied with oxidative damage and disorder of function of the membrane. Anyhow, when exogenous cardiolipin was added in the purified endemic CCO solution, it was capable of restoring CCO activity significantly. The significance of Se deficiency and unbalanced diet in pathogenesis of Keshan disease is discussed.

Animals↗
PubMed1990

[Sneddon syndrome: cardiac pathology and anti-phospholipid antibodies].

Besides cerebrovascular pathology and livedo, cardiac affections are introduced in clinical manifestations of Sneddon's syndrome. They involve coronary heart disease (up to development of MI), murmur (usually mitral), occasional cardiac arrhythmia. Patients with coronary disease and cardiac murmur show antibodies to cardiolipin significantly more frequently. The role of antibodies to cardiolipin in the genesis of cardiac alterations in Sneddon's syndrome is open to discussion.

Adolescent↗
PubMed1988

Reaction patterns of monoclonal antibodies to DNA.

Starting with spleen cells from MRL/lpr, NZB/W, and graft-vs-host-diseased mice, we prepared a total of 57 hybridomas that produce antibodies to DNA. Using various approaches, we studied the avidity of these monoclonals in relation to their behavior in four anti-DNA assays. From the results obtained, we postulate that on the basis of anti-DNA avidity the anti-DNA ELISA, the polyethylene glycol assay, the indirect immunofluorescence test on Crithidia luciliae, and the Farr assay (in this order) detect a decreasing amount of anti-dsDNA, the Farr assay being strictly selective for high avidity anti-dsDNA. mAb selected by the anti-DNA ELISA generally were of a low avidity toward DNA. Using cardiolipin and dextran sulfate, a polyanion that bears a resemblance in charge to DNA, we studied the cross-reactivity of the monoclonals. A total of 6 of the 57 monoclonals were found to cross-react with cardiolipin, and 26 with dextran sulfate. We observed an inverse relationship between anti-DNA avidity and cross-reactivity: the lower the avidity of the antibody, the more cross-reactive it is. Based on these findings, we postulate that it is at least questionable whether low avidity, cross-reactive (monoclonal) anti-DNA is representative for the anti-DNA found in patients with SLE.

Animals↗
PubMed1989

[Antiphospholipid antibodies: clinical significance in systemic lupus erythematosus].

A radioimmunoassay using cardiolipin as antigen and labelled SPA, anti-human IgG, anti-human IgM, anti-human IgA as second antibodies in detecting anti-cardiolipin antibody with the sera from 308 patients and 70 normal controls. Among them, 126 patients were of SLE, 27 systemic sclerosis, 40 rheumatoid arthritis, 40 Sjögren syndrome, 26 other connective tissue diseases, 7 syphilis and 32 with obstetric complications. The positive rate of anticardiolipin antibody were 42.9% (SLE), 29.7% (PSS), 20% (RA), 15% (SS), 26.9% (CTD), 85.7% (syphilis), 3.1% (obstetric complication), 0% (NC). In SLE the anticardiolipin antibody were well correlated with thrombocytopenia, cerebral lupus, thrombosis of vein and spontaneous recurrent abortion. Lupus anticoagulant (APTT) was found in 21.3% of SLE and biological false positive of VDRL test in 4.8%. Both of them correlated with the anticardiolipin antibody detected by the radioimmunoassay. The authors concluded that antiphospholipid antibodies is a group of commonly seen antibodies, which may play a rule in the pathogenesis of SLE. Further study is progressing.

Adolescent↗
PubMed1989

Serum monoclonal antibodies derived from patients with multiple myeloma react with mycobacterial phosphoinositides and nuclear antigens.

The sera of 46 patients with multiple myeloma were examined for the presence of anti-tuberculosis (TB) glycolipids antibodies. In positive sera samples, anti-polynucleotides, anti-histones, anti-cardiolipin, anti-Sm and anti-RNP activities were sought. Antibodies against three different mycobacterial glycolipids were detected in 10 of the 46 sera. Three (P429, P557, P5) showed high titres of antibodies against all three different TB glycolipids. Of the 10 reactive samples, two antibodies were purified (P429, P557). P557 reacted with various polynucleotides and other antigens tested (Sm, RNP, cardiolipin, histones). One immunoglobulin (P207) which showed high activity against Sm and RNP had no activity against TB glycolipids and was employed as a control. The cross-reactivity between mycobacterial glycolipids and the nuclear antigens was further established by bi-directional competition assays with P429 and P557. Our study shows a high incidence (22%) of anti-TB glycolipids antibodies in sera of patients with monoclonal gammopathies, some of which show anti-DNA and other anti-nuclear antigens activities. This is additional evidence for the mycobacterial-nuclear antigen cross-reactivity which may suggest a possible role of infection (e.g., tuberculosis) in autoimmune diseases.

Antibodies, Antinuclear↗
PubMed1989

Anticardiolipin syndrome in plastic surgery of the breast.

A 38-year-old female patient had bilateral silicone-made prosthesis implantation performed 17 years ago. Three years after surgery, she had a series of spontaneous miscarriages, later she developed recurrent venous and arterial thromboses. Laboratory tests detected antibodies against cardiolipin and the diagnosis of cardiolipin syndrome was established. The authors discuss the possible association between surgery and the patient's current disease.

Abortion, Habitual↗
PubMed1986

Adrenic acid content in rat adrenal mitochondrial phosphatidylethanolamine and its relation to ACTH-mediated stimulation of cholesterol side chain cleavage reaction.

We have isolated various phospholipids from adrenal mitochondria of adrenocorticotropic hormone (ACTH)-treated (stimulated) and cycloheximide/ACTH-treated (unstimulated) rats. When the effects of these phospholipids were examined on the formation of pregnenolone from endogenous cholesterol by adrenal mitochondria of unstimulated rats, phosphatidylethanolamine and phosphatidylserine from stimulated mitochondria were effective in enhancing the cleavage reaction in unstimulated mitochondria, whereas these phospholipids from unstimulated mitochondria were all ineffective. Cardiolipins from both stimulated and unstimulated mitochondria were effective. When the compositional changes in fatty acid moiety of phospholipids were examined, a significant increase in C22:4 (adrenic) acid was observed only for phosphatidylethanolamine under the influence of ACTH. A linear relationship between the contents of C22:4 acid in various phospholipids and respective steroidogenic activities was obtained (r = 0.880), suggesting an important role of this fatty acid moiety. The separation of active phosphatidylethanolamine by high performance liquid chromatography revealed that a fraction containing 25% C22:4 acid was most effective in the activation. Based on these results, it is most likely that 1-stearoyl-2-adrenoyl phosphatidylethanolamine is an active species. C22:4 acid was liberated together with C20:4 acid from adrenal triglycerides by the action of ACTH but the liberation was insensitive to cycloheximide inhibition. Finally, cardiolipin which enhances the transfer of cholesterol to cytochrome P-450scc may not be a physiological mediator of ACTH action.

Adrenal Glands↗
PubMed1977

In vitro cell response of Treponema pallidum-infected rabbits. I. Lymphocyte transformation.

Lymphocyte response to phytohaemagglutinin (PHA), Treponema pallidum, cultivable Reiter strain and cardiolipin reagent were examined using peripheral lymphocytes of T. pallidum-infected rabbits and control animals injected intratesticularly with saline extract of normal rabbit testes mixed with T. pallidum-free saline extract of infected testes. The response to PHA was significantly suppressed only in the infected animals, reaching the lowest level at 30 days after infection. A substantial increase of lymphocyte stimulation in the presence of cardiolipin reagent at 20 days after infection correlated well with the appearance of the maximum titre of Wassermann antibodies. Significant (P less than 0.02) increase in spontaneous response of cells incubated for 5 days made the response to T. pallidum and the Reiter strain difficult to interpret. An explanation for the spontaneous response is offered.

Animals↗
PubMedDate not supplied

[Passive hemagglutination test in the serodiagnosis of syphilis].

Passive haemagglutination test with pathogenic Treponema pallidum antigen in the serodiagnosis of syphilis, carried out on 752 sera (518 syphilitic patients and 234 subjects without evidence of syphilis) showed a greater general sensitivity as compared the cardiolipin tests and complement fixation test with Reiter protein antigen, and lower, to immunofluorescence (FTA--ABS). Its specificity is relatively lower than that of the cardiolopin and treponemal tests. Associated with the cardiolipin test it may be preferably used for the detection of syphilis, and moreover, passive haemagglutination may confirm the diagnosis in case of weak positive results with the FTA--ABS test. The latter is, however, essential for problem cases.

Antigens, Bacterial↗
PubMed1985

Crossreactivity of antiphospholipid antibodies.

Using a solid phase radioimmunoassay to detect anticardiolipin antibodies, the crossreactivity of these antibodies with phospholipids and DNA was tested in 10 patients with autoimmune disorders and 1 patient with secondary syphilis. The crossreactivity of anti-ds DNA and anti-ss DNA antibodies with cardiolipin was tested in 10 additional patients with SLE. In the 10 sera tested with high anticardiolipin antibody levels, there was marked inhibition of anticardiolipin antibody activity by negatively charged phospholipids. The zwitterionic phospholipids inhibited anticardiolipin antibody activity in some sera, and in instances where this occurred, inhibition was less than that obtained for the negatively charged phospholipids. There was minimal inhibition of anticardiolipin antibody by ss DNA in only 2 of 10 sera tested and no inhibition by ds DNA in any sera tested. Conversely, cardiolipin inhibited anti-ss DNA antibody activity in 2 of 10 sera, and anti-ds DNA antibody activity in 2 of 10 sera tested. There was no correlation between anticardiolipin antibody levels and either anti-ds DNA or anti-ss DNA antibody levels. These results suggest that antiphospholipid and anti-DNA antibodies are largely separate groups of antibodies. On the basis of these studies, we suggest that there may be various groups of antiphospholipid antibodies in patients with SLE and other autoimmune disorders.

Autoantibodies↗
PubMed1974

Skeletal muscle lipids. II. Changes in phospholipid composition in man from fetal to middle age.

Phospholipid compositions were determined in samples of gastrocnemius muscles of individuals from fetal stage to the age of 55 yr. The lipids were separated by thin-layer chromatography. In order to enable characterization of the individual phospholipids, a lipid extract was prepared from pooled samples of gastrocnemius muscles from adult males and separated by ion-exchange chromatography on TEAE-cellulose. The individual phosphoglycerides were purified by thin-layer chromatography and then characterized according to their content of fatty acids, aldehydes, phosphorus, and the identity of the bases. The relative amounts of the major phospholipids, choline and ethanolamine phosphoglycerides, changed little with age, and in adult males they constituted 47% and 24%, respectively. Cardiolipin increased from 3% in the fetal stage to 9% by the end of the first year of life. Sphingomyelin and serine phosphoglyceride decreased with increasing age, while inositol phosphoglyceride increased. In adult males, cardiolipin constituted 10% of total lipid phosphorus, inositol phosphoglyceride constituted 9%, serine phosphoglyceride 3%, and sphingomyelin 7%.

Adolescent↗
PubMed1976

Ganglioside biosynthesis. Characterization of uridine diphosphate galactose: GM2 galactosyltransferase in golgi apparatus from rat liver.

An enzyme that transfers galactose from UDP-Gal to ganglioside GM2 (Tay-Sachs ganglioside) was concentrated 50 times in Golgi apparatus from rat liver relative to total homogenates. This enzyme required detergents or phospholipids as dispersing agents. Of the numerous detergents tested, sodium taurocholate and Triton CF-54 were most effective in stimulating the reaction. Cardiolipin alone was more effective than any of the detergents tested in stimulating enzyme activity. The pH optimum for the reaction varied with the nature of the dispersing agent. With sodium taurocholate, Triton CF-54 and cardiolipin, the pH optima were 6.2, 5.9, and 5.6, respectively. The enzyme had a nearly absolute requirement for Mn2+, with maximum activity being attained at a concentration of 15 mM Mn2+. Other divalent or trivalent cations were either less effective than Mn2+ or inhibited the transferase reaction. The Km values calculated for UDP-Gal and GM2 were 1.1 X 10(-4) M and 9.9 X 10(-5) M, respectively. The enzyme could not be dissociated from Golgi apparatus fractions by treatment with ultrasound, indicating that it is tightly associated with the membrane and not part of the luminal contents. The newly synthesized GM2, the product of the reaction, was incorporated into or became tightly associated with the membranes of the Golgi apparatus.

Animals↗