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Biomedical subjects

B Pau

Publications and source records attributed to B Pau.

At least 91 records · Page 5Linked to original sources
PubMed1991

Autoimmunity to human thyroglobulin. Respective epitopic specificity patterns of anti-human thyroglobulin autoantibodies in patients with Sjögren's syndrome and patients with Hashimoto's thyroiditis.

We evaluated the epitopic specificity pattern of anti-human thyroglobulin (anti-hTg) autoantibodies from patients with primary Sjögren's syndrome (SS). All of the primary SS sera tested contained both IgG and IgM anti-hTg autoantibodies recognizing at least 1 region on hTg; in 65% of the cases, 3 or more regions were recognized. A strong recognition of region II, as is seen in Hashimoto's thyroiditis, was associated with thyroid disorder in primary SS. These results emphasize the importance of region II in autoimmune thyroid disease.

Adult↗
PubMed1991

Evidence for a restricted idiotypic and epitopic specificity of anti-thyroglobulin autoantibodies in patients with autoimmune thyroiditis.

Anti-thyroglobulin (TG) autoantibodies from patients with autoimmune thyroiditis express a cross-reactive alpha idiotype (Id) termed T44 which is not expressed by IgG from normal individuals. The present study demonstrates that the expression of the T44 Id is strongly associated with the recognition by anti-TG autoantibodies of a specific epitopic cluster on human TG. The epitopic reactivity of anti-TG autoantibodies was determined using a competitive inhibition assay with a panel of 15 monoclonal antibodies that define six antigenic clusters on TG. All T44+ autoantibodies from patients recognized cluster II, whereas no anti-TG IgG from healthy individuals reacted with this region. Affinity columns of Sepharose-bound intravenous therapeutic immunoglobulins which contain anti-T44 activity, retained both T44 Id-expressing antibodies and a subset of region II-specific anti-TG autoantibodies from patients with Hashimoto's disease. Restricted idiotypic and epitopic specificity may demarcate disease-associated from natural anti-TG autoantibodies, suggesting that qualitative rather than quantitative criteria should be used to identify pathological autoantibodies.

Antibody Specificity↗
PubMed1991

Imaging of myocardial infarction in dogs and humans using monoclonal antibodies specific for human myosin heavy chains.

The use of three different monoclonal antibodies specific for human ventricular myosin heavy chains in the visualization of the location and extent of necrosis in dogs with experimental acute myocardial infarction and in humans is described. Using a classic immunohistochemical method or ex vivo analysis of heart slices in dogs with acute myocardial infarction subjected to intravenous injection of unlabeled antimyosin antibodies or antimyosin antibodies labeled with indium-111, it was observed that all antibody fragments specifically reached the targeted necrotic zone less than 2 h after antibody injection and remained bound for up to 24 h. In a limited but significant number of cases (5 of the 12 humans and 11 of 43 dogs), it was possible to image the necrotic zone in vivo as early as 2 to 4 h after antibody injection. In other cases, individual blood clearance variations retarded or even prevented in vivo necrosis detection. Higher antimyosin fixation values were obtained in the necrotic zones in dogs with a rapid blood clearance relative to that of the other dogs. It is concluded that antimyosin antibodies always reached necrotic areas within 2 h. If blood clearance was rapid, in vivo imaging of the necrotic area was possible 2 to 6 h after necrosis, even in humans. In some cases, however, uncontrolled individual variations in the timing required for sufficient blood clearance hampered this rapid in vivo detection of myocardial necrosis.

Aged↗
PubMed1991

Polyacrylic microspheres as a solid phase for microparticle enhanced nephelometric immunoassay (NEPHELIA (R)) of transferrin.

Polyfunctional hydrophilic microspheres (MS) can be produced by copolymerisation with gamma-irradiation of acrylic monomers. Transferrin (TRF) can be covalently bound to these MS by reaction between aldehyde groups of the MS and primary amino groups of the protein. MS-TRF conjugates thus obtained are agglutinated by specific antiserum and this agglutination is inhibited by free TRF. Agglutination and inhibition are quantified by measurement of the light scattered by MS-TRF conjugate clusters with a specially designed nephelometer, a process designated as microparticle enhanced nephelometric immunoassay (NEPHELIA (R)) for TRF. Recovery, correlation and reproducibility studies, simultaneously performed in three different laboratories, show that this TRF immunoassay is accurate for a large concentration range. NEPHELIA (R) may appear as an alternative method for a large variety of molecules.

Humans↗
PubMed1991

Highly specific radioimmunoassay for digoxin using a monoclonal antibody selected for lack of interference by digoxin-like immunoreactive substances in cord blood sera.

Four digoxin radioimmunoassays (RIA) were evaluated using four antidigoxin monoclonal antibodies (MAb) with different binding specificities for digoxin metabolites and other glycosides. These RIA have been used to measure the apparent digoxin concentrations in the sera of patients treated with digoxin, in cord blood sera and in rat intestine aqueous extracts. Two MAb strongly recognized digoxin-like immunoreactive substances (DLIS) in cord blood sera and in rat intestine aqueous extract. In contrast, one MAb (2C2), which recognizes active digoxin metabolites and does not cross-react with inactive metabolites nor with digotoxin, showed no reactivity with DLIS in the cord blood sera tested. This MAb was used in a new digoxin RIA that was rapid, sensitive, reproducible, and specific for digoxin and its active metabolites.

Antibodies, Monoclonal↗
PubMed1991

Immunoradiometric assay of myosin heavy chain fragments in plasma for investigation of myocardial infarction.

Estimation of the extent and location of infarct is important for the prognosis and hence therapeutic strategy in patients with acute myocardial infarction (AMI). Because cardiac myosin is the major structural protein of the myocardium, and may thus reflect the extent of injured tissue, we established a new sensitive immunoradiometric assay, using a pair of monoclonal antibodies (Mabs) that specifically bind the myosin heavy chain fragments liberated from the myocyte into plasma after a heart attack. A first Mab is linked to a magnetic solid phase. A second Mab, radiolabeled with 125I, is used to detect myosin trapped on the solid phase by the first Mab during a 3-h incubation. This assay can detect 10 micro-units of myosin per liter and is highly reproducible.

Antibodies, Monoclonal↗
PubMed1990

Antigen-capture ELISA for viral haemorrhagic septicaemia virus serotype I.

An antigen-capture ELISA for viral haemorrhagic septicaemia virus serotype I (VHSV I) was developed. The assay employs two monoclonal antibodies (mAb) directed against distinct epitopes of the viral envelope glycoprotein (Gp). The antigen bound by the capture mAb (A17) was detected by addition of a second mAb (L7) conjugated to horseradish peroxidase, followed by addition of the enzyme substrate. The technique is highly sensitive, enabling detection of the virus at a protein concentration as low as 1 ng/ml of total proteins (1.5 fmol of envelope Gp per ml) in purified preparations. VHSV I was also detected in culture supernatants (5 x 10(5) PFU/ml) and in extracts of kidney and spleen of rainbow trout infected experimentally (5 x 10(5) PFU/ml). The assay was highly specific: infectious haematopoietic necrosis virus, infectious pancreatic necrosis virus, spring viraemia of carp virus, pike fry rhabdovirus, eel rhabdovirus and perch rhabdovirus could not be detected by the antigen-capture ELISA for VHSV I.

Antibodies, Monoclonal↗
PubMed1990

An anti-Cryptococcus neoformans monoclonal antibody directed against galactoxylomannan.

Six monoclonal antibodies (mAb) were produced by immunizing mice with a Cryptococcus neoformans serotype A spheroplast lysate (CNSL). Two mAb were of the IgG1 isotype; the others were IgM. The results obtained with one IgM mAb (CN6) are reported herein. This mAb recognized the four serotypes of C. neoformans and no cross-reactions were observed with extracts from Cryptococcus melibiosum, Candida albicans, Saccharomyces cerevisiae, Torulopsis glabrata or Trichosporon beigelii. Fractionation of the CNSL by gel filtration revealed that mAb CN6 recognized high molecular weight substances as well as a range of smaller molecules. Indirect ELISA inhibition studies showed that this mAb recognized substances in a cryptococcal culture filtrate. Inhibition studies and agglutination tests using latex beads sensitized with purified CN6 showed that CN6 strongly reacted with the C. neoformans serotype A cell envelope galactoxylomannan-mannoprotein complex (GAlXM-MP) and only weakly with the glucuronoxylomannan (GXM) component. These tests also showed that purified galactoxylomannan (GalXM) from C. neoformans serotype A was more reactive than purified mannoprotein (MP). An anti-GalXM mAb might be a useful tool for monitoring the clinical course of cryptococcal infections.

Animals↗
PubMed1990

Novel anti-digoxin monoclonal antibodies with different binding specificities for digoxin metabolites and other glycosides.

Spleen cells of BALB/c mice immunized with a digoxin-bovine serum albumin conjugate were fused with P3-X63-Ag8.653 mouse myeloma cells. Seven monoclonal antibodies (MAb) selected by indirect ELISA were produced, purified and characterized. All the MAb were IgG1 isotypes. The apparent equilibrium association constants (Ka) of four of the MAb, determined by Scatchard analysis of the RIA data, ranged from 1 x 10(9) M-1 to 5.9 x 10(9) M-1. The estimated Ka values of the three other MAb were found to be between 4.8 x 10(7) M-1 and 5.9 x 10(8) M-1. Using digoxin and eighteen structurally-related compounds, the seven MAb could be divided into five groups based on their binding specificities assessed by an inhibition immunoenzymatic test. The MAb in Groups I and II, in particular, showed very different specificity profiles: the two MAb in Group I had low cross-reactivity with cardioinactive digoxin metabolites, whereas the high affinity MAb in Group II recognized all the digoxin metabolites tested. The MAb in Group I might be useful in a digoxin immunoassay and the Group II MAb in therapeutic reversal of digoxin intoxication.

Animals↗
PubMed1989

Association between high concentrations of Mr 52,000 cathepsin D and poor prognosis in primary human breast cancer.

The Mr 52,000 cathepsin D is the precursor of a lysosomal protease secreted in excess by breast cancer cells. This protease can degrade extracellular matrices and proteoglycans and is induced by estrogens in estrogen receptor-positive breast cancer cell lines. In a 4- to 6-yr retrospective cohort study, the concentration of the total cathepsin D (precursor plus intermediate and mature chains) was assayed in cytosols of primary tumors from 242 pre/perimenopausal and 154 postmenopausal breast cancer patients in a solid-phase immunoassay using two specific monoclonal antibodies. Patients were initially divided into groups with low, intermediate, or high concentrations of cathepsin D corresponding to the quartiles of the overall distribution. Using these groupings, the level of Mr 52,000 cathepsin D was not significantly associated with the recognized prognostic factors of age, lymph node involvement, tumor size, and/or grade of anaplasia. A significant association was found between cathepsin D concentrations and estrogen receptor status only among pre/perimenopausal patients. Receptor-positive tumors (greater than or equal to 10 fmol of estrogen receptor/mg of cytosol protein) had a significantly greater proportion of patients with high Mr 52,000 cathepsin D concentrations. Patients with high Mr 52,000 cathepsin D concentrations (greater than 78 pmol/mg for pre/perimenopausal and greater than 24 for postmenopausal patients) have shorter recurrence-free survival (P = 0.06 for pre/peri- and P = 0.039 for postmenopausal patients) and have a trend toward shorter overall survival (P = 0.30 and P = 0.089 for pre/peri- and postmenopausal groups, respectively). In multivariate analysis, Mr 52,000 cathepsin D status was found to be an independent prognostic factor for recurrence-free survival of about the same import as lymph node status for both menopausal groups. This first retrospective study demonstrates that the level of Mr 52,000 cathepsin D in cytosol of primary breast cancer biopsies is an independent prognostic factor in predicting relapses in both pre/peri- and postmenopausal patients.

Adult↗
PubMed1989

Significance of the recognition of certain antigenic regions on the human thyroglobulin molecule by natural autoantibodies from healthy subjects.

We have evaluated the epitope specificity of natural antihuman thyroglobulin (hTg) autoantibodies (aAb) in the plasma of healthy individuals. By an indirect ELISA technique, we selected 56 plasma samples with high anti-hTg antibody activity and used the IgG fraction isolated from these plasma to study the antigenic domains on the hTg molecule recognized by the natural anti-hTg aAb. A panel of 15 mAb, coupled to alkaline phosphatase and recognizing six regions (I to VI) on the hTg molecule, served to identify the domains recognized by the natural anti-hTg aAb using a competitive ELISA procedure. A total of 26 of the IgG fractions was found to interact with at least one of the regions defined by our battery of mAb. Region V was recognized by the majority of the IgG fractions. Interestingly, region II was rarely recognized by the same IgG fraction that reacted with region V. Inasmuch as we have previously shown that region II is mainly recognized by aAb in the serum of subjects with various thyroid disorders, we propose that recognition of region V reflects the normal physiologic state of the immune system with respect to the hTg molecule.

Adolescent↗
PubMed1989

Two-site immunoenzymometric assay for the 52-kDa cathepsin D in cytosols of breast cancer tissues.

We developed a solid-phase two-site immunoenzymometric assay (IEMA) of the estrogen-induced 52-kDa cathepsin D (EC 3.4.23.5) and its processed forms (48-kDa and 34-kDa proteins) in cytosols of breast cancer tissues, using two monoclonal antibodies directed against two different epitopes of these antigens. The first antibody is bound to a polystyrene microtiter well; the second is labeled with alkaline phosphatase. The assay involves a simultaneous incubation of the antigen with both antibodies, because we observed signal loss during sequential incubations. Alkaline phosphatase was chosen because other enzymes (peroxidase, beta-galactosidase) were inhibited by cytosol extraction buffers. The measurable range of 52-kDa-related proteins is from 0.3 to 6 nmol/L with precision (CVs) within and between runs of 3.9% and 15.8%, respectively. The sensitivity, accuracy, and rapid turnaround time of the two-site IEMA should facilitate the clinical evaluation of this new marker in oncology.

Alkaline Phosphatase↗
PubMed1989

Novel immunoradiometric assay of thyroglobulin in serum with use of monoclonal antibodies selected for lack of cross-reactivity with autoantibodies.

A multisite immunoradiometric assay for measurement of serum thyroglobulin (Tg), designated Magnogel-IRMA-Tg, has been developed, involving magnetic microbeads (Magnogel). This assay is based on the use of five anti-Tg monoclonal antibodies (MAbs) directed against three antigenic regions on the Tg molecule that are not recognized by anti-Tg autoantibodies (aAbs). Four of these MAbs, directed against two antigenic domains, were coupled to the magnetic beads and were used to trap the serum antigen. Another MAb, directed against the third region, was iodinated and served as the labeled second antibody. The Magnogel-IRMA-Tg technique is reproducible, rapid, and sensitive (lower detection limit, 3 micrograms/L). The assay reliably measures serum Tg in the presence of anti-Tg aAbs.

Antibodies, Monoclonal↗
PubMed1989

A two-site immunoenzymometric assay of 52-kDa pro-cathepsin D, and its use in human breast diseases.

After isolating monoclonal antibodies specific for the 52-kDa precursor of cathepsin D (cath-D), which is secreted in excess in both hormone-dependent and hormone-independent breast cancer, we developed a two-step double-determinant immunoenzymometric assay that is specific for this pro-enzyme. The assay combines the use of a monoclonal antibody specific for the precursor and bound to microtiter plates, and a second antibody directed against a smaller processed form of the mature enzyme, coupled to alkaline phosphatase. The specificity of the assay relies on separate and sequential additions of the antigen and the conjugated second antibody. It allows rapid measurement of the analyte in plasma and cytosols of normal and neoplastic mammary tissues, with a detection limit of 5 fmol and a maximal interassay coefficient of variation of 9%. This assay is particularly useful for tissue cytosol samples where the pro-enzyme form co-exists with large quantities of the mature processed forms of the enzyme. Comparative assays of 52-kDa pro-cath-D and total cath-D in cytosols of breast cancers and benign mastopathies indicate that the present assay better discriminates between benign and cancerous mammary tumors.

Antibodies, Monoclonal↗
PubMed1988

Electroimmunoassay of human serum lipoproteins containing Apo A-I by monoclonal antibodies.

Three monoclonal antibodies to human serum apolipoprotein (Apo) A-I (4A12, 4B11 and 2G11) were produced by Sanofi. They were directed to three distinct epitopes of the Apo A-I molecule, present on the surface of the lipoprotein particles. They were able to precipitate individually some lipoprotein units from the serum. Only the mixture of the three monoclonal antibodies could allow a precipitation of all Apo A-I containing particles. An electroimmunoassay using this oligoclonal mixture was assessed to standardize the Apo A-I measurement in serum. Results agreed well with those obtained by electroimmunoassays using polyclonal antisera. Moreover no pretreatment of serum samples with dissociating agents or detergents was required. Therefore, this specific, rapid (7-8 h), precise (within- and between-assay precisions were 4.25 and 3.1%, respectively) immunoassay is routinely available for apolipoprotein A-I measurement in serum.

Antibodies, Monoclonal↗
PubMed1987

Monoclonal antibodies to human apolipoprotein A-I: characterization and application as structural probes for apolipoprotein A-I and high density lipoprotein.

Three mouse monoclonal antibodies (Mabs) to human apo A-I were produced using apolipoprotein A-I or HDL3 as immunogens. These monoclonal antibodies, 2G11, 4A12 and 4B11, were characterized for their reactivity with isolated apolipoprotein A-I and HDL in solution. The immunoblotting patterns of the HDL3 two-dimensional electrophoresis show that these three monoclonal antibodies reacted with all the polymorphic forms of apolipoprotein A-I. Cotitration experiments indicated that they correspond to three distinct epitopes. In order to locate these three antigenic determinants on the isolated apolipoprotein A-I, the reactivity of the three monoclonal antibodies has been studied on CNBr-cleaved apolipoprotein A-I. The monoclonal antibodies 2G11 and 4A12 addressed to the amino (CNBr 1) and carboxy (CNBr 4) terminal segments, respectively. In comparison with the monoclonal antibodies characterized by Weech et al. ((1985) Biochim. Biophys. Acta 835, 390-401), monoclonal antibody 4A12 is the only one described in the literature which is specific of the carboxy terminal segment of apolipoprotein A-I. Monoclonal antibody 4B11 does not react with any CNBr fragment, its binding is temperature dependent, it could be directed to a conformational epitope. Relative differences were demonstrated in the expression of the three epitopes in HDL subfractions isolated by density gradient ultracentrifugation. According to Curtiss and Edgington ((1985) J. Biol. Chem. 260, 2982-2993) our results indicate the existence of an immunochemical heterogeneity in the organization of apolipoprotein A-I at the surface of HDL particles as well as in the soluble form of apolipoprotein A-I.

Antibodies, Monoclonal↗