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At least 1,531 records · Page 85Linked to original sources
PubMed1998

Argininosuccinate lyase: a new autoantigen in liver disease.

Anti-liver cytosol 1 autoantibody (LC1) characterizes a severe form of autoimmune hepatitis (AIH), staining the cytoplasm of periportal hepatocytes and targeting an unidentified 60-kD liver cytosolic antigen. To identify its target, we used high-titre anti-LCI+ sera from two patients with AIH to screen 18 cytoplasm enzymes with periportal location by double immunodiffusion (DDI). Both sera gave a broad precipitin line against human liver cytosol, suggesting that they may recognize two distinct antigens, a possibility confirmed by the appearance of two precipitin lines when DDI conditions were optimized (0.8% agarose and 3% polyethylene glycol (PEG)). Experiments by DDI and Western blot (WB) identified a liver cytosolic autoantigen of 50 kD, different from LC1, giving a line of identity with argininosuccinate lyase (ASL). Reactivity to ASL was then investigated by DDI and WB in 57 patients with AIH, 17 with primary biliary cirrhosis (PBC), 15 with chronic hepatitis B virus (HBV) infection, 13 with alphal-antitrypsin deficiency, 17 with Wilson's disease, 18 with extrahepatic autoimmune disorders, and in 48 healthy controls. Anti-ASL was found in 16% of AIH and 23% of PBC patients by DDI and in 14% of AIH, 23% of PBC and 20% of HBV patients by WB. No argininosuccinate was present in the urine of four anti-ASL+ patients tested, excluding an inhibition of enzymatic activity by anti-ASL. The addition of anti-ASL+ serum to human fibroblast cultures induced a significant increase in ASL activity. ASL is a new autoantigen in liver disease and its clinical relevance warrants further investigation.

Argininosuccinate Lyase↗
PubMed2002

Invasive aspergillosis in cancer.

Ten confirmed cases of invasive aspergillosis (IA) in cancer patients were analysed retrospectively. Eight were pulmonary, one was sinonasal and one was cutaneous. The majority of patients had haematological malignancies (7), the remaining three were cases of solid tumours. Fever was present in all 10 cases. Cough and lung signs were present in all eight cases of invasive pulmonary aspergillosis. Haemoptysis was encountered in three of nine cases of pulmonary and sinonasal aspergillosis. Mortality was low (2%). While corticosteroids, antibiotics and anticancer chemotherapy/radiotherapy were factors predisposing the patients to IA, neutropenia was perhaps responsible for their mortality. Seven of the patients had other associated pathogens isolated in culture in addition to Aspergillus spp. Aspergillus fumigatus was the predominant species, followed by A. flavus, A. glaucus, A. nidulans and A. niger. Direct microscopic examination (in six of seven cases) and culture (six of seven cases) correlated well with radiographic and clinical findings in cases with lung involvement. Serology for anti-Aspergillus antibodies performed by gel diffusion precipitin test was positive in one case of sinonasal aspergillosis, wherein only one precipitin band was observed. Correlation of clinical symptoms, consistent radiographic findings and microbiological work-up (the latter including a triad of direct microscopy, culture and serology) are required to arrive at a diagnosis of IA, especially where histology cannot form the mainstay of diagnosis.

Adolescent↗
PubMed2003

Antinuclear antibody testing in a regional immunopathology laboratory.

A systematic review has been undertaken of antinuclear antibody testing over a 6-year period in a regional immunotherapy laboratory servicing a population of 400 000. Twenty-eight per cent of the 20 205 antinuclear antibody tests performed on a hyperexpressing Ro transfected cellular substrate were positive (titre >/= 1 : 80) with the most common immunofluorescent patterns being homogeneous (39%), speckled (20%), mixed (17%), nucleolar (8%), Ro (7%) and centromere (4%). Ro antibody as detected by immunofluorescence was strongly concordant with anti-Ro detected by counter immunoelectrophoresis precipitation; of 261 anti-Ro counter immunoelectrophoresis precipitation positive patients surveyed, only 15 were missed and 20 masked (with homogenous pattern) by immunofluorescence. Ro antibodies were found in patients with a variety of immune disorders, particularly connective tissue disorders, whilst a clinical survey of the anticentromere sera revealed that 67% were derived from patients with limited scleroderma. Extractable nuclear antibodies and their characterization was performed on 10 939 occasions with 12.9% being positive with anti-Ro constituting 30.2%, anti-Ro/La 25.7%, unidentified precipitin line 17.8%, anti-ribo nuclear protein 12.5%, respectively, with anti-Scl70, anti-Jo-1 and anti-Sm and various combinations making up the remainder. Unidentified precipitin lines were particular prominent in patients with connective tissue disorders. DNA quantification was performed on 12 068 occasions with 11% giving elevated values, the majority from patients with systemic lupus erythematosus. Of these positive sera 44% also demonstrated one or more extractable nuclear antibodies and 25% anticardiolipin antibodies. Regular participation in a Quality Assurance Program revealed accurate and consistent performance of antinuclear antibody testing. In conclusion antinuclear antibody detection and characterization for systemic immune disorders can provide the clinician with useful diagnostic and prognostic information; it is important that the laboratory results are relevant, timely, accurate and precise. Systematic reviews as demonstrated in this report, can provide such evidence.

Antibodies, Antinuclear↗
PubMed2001

Three-year follow-up study of allergy in workers in a mushroom factory.

Exposure to mushroom spores may cause many respiratory allergic diseases, however, there has been no serial study in a mushroom factory to address this problem. The aim of this study was to investigate the serial changes in respiratory allergy and the incidence of hypersensitivity pneumonitis (HP) in mushroom workers. A 3-year follow-up study, beginning in June 1996, was conducted in a newly operating mushroom factory in which one kind of mushroom is produced: Hypsizigus marmoreus (Bunashimeji). Allergic symptoms, chest roentgenogram, serum precipitins to the spores and soluble adhesion molecules in sera were evaluated once a year in 60 workers and 20 controls. Three out of the 60 subjects were diagnosed as having HP caused by inhalation ofthe mushroom spore and they were therefore excluded from this study, and the 57 non-HP subjects were evaluated. In this study 24 workers quit because of intolerable cough, runny nose, wheezing, sputum, fever elevation and/or shortness of breath at their place of work. During each year of this study as many as 70-80% of employees suffered some ofthe above symptoms, cough being the most frequent, and positive rate of serum precipitins to the spore revealed 30% in 1996, 93% in 1997 and 94% in 1998. From the June 1996 examination until the following May, serum soluble intercellular adhesion molecule-1 levels of the 15 workers who quit during that period were significantly higher than those in the 42 workers still employed in 1997 (P < 0.05). Workers in Bunashimeji mushroom factories might be at critical risk of developing respiratory allergy. In our 3-year study over 90% workers were sensitized to the spore, 40% quit because of the symptoms and 5% developed HP. It was suggested that workers should be counselled about the risk of mushroom allergy and precautionary measures should be taken to prevent its occurrence.

Adult↗
PubMed1981

Heterogeneity of human prekallikrein deficiency (Fletcher trait): evidence that five of 18 cases are positive for cross-reacting material.

We studied the plasma of 18 patients with a functional deficiency of plasma prekallikrein (Fletcher trait). Samples from 13 subjects contained less than 1 per cent of normal levels of prekallikrein antigen recognized by a specific antiserum (cross-reacting-material negative [CRM-]). In the five other subjects, however, nonfunctional material immunologically indistinguishable from normal prekallikrein was detected by radioimmunoassays at concentrations of 13 to 30 per cent (CRM+ variant). None of the plasma samples contained detectable circulating anticoagulants against prekallikrein. On immunodiffusion against antiserum to kallikrein, each of the five CRM+ samples formed a single precipitin line of complete identity with normal plasma or purified prekallikrein. On immunoelectrophoresis, the precipitin line had the same mobility as that for normal prekallikrein. These studies demonstrate the molecular heterogeneity of human prekallikrein deficiency and show that persons with the CRM+ variant have a nonfunctional form of prekallikrein in their plasma.

Blood Coagulation Disorders↗
PubMed1976

A fetal protein in chromatin of Novikoff hepatoma and Walker 256 carcinosarcoma tumors that is absent from normal and regenerating rat liver.

Antibodies to chromatin proteins of Novikoff hepatoma cells formed precipitin bands in the double-diffusion immunoprecipitation assay with chromatin proteins of Novikoff hepatoma, Walker 256 carcinosarcoma, and 18-day fetal rat liver. The antigen used for preparation of antiserum was the chromatin proteins initially extracted with 3 M NaCl-7 M urea and soluble after dialysis to 0.14 M NaCl-0.35 M urea. The chromatin proteins used for analytical studies were extracted with 0.6 M NaCl containing 0.01 M Tris-HCl (pH 8) and 100 muM phenylmethylsulfonyl fluoride. Corresponding chromatin proteins of normal and 18-hr regenerating rat liver, heart, and kidney did not form precipitin bands. The antigen was purified from the chrmatin of Novikoff hepatoma cells by exclusion chromatography on Sephadex G-150 and preparative nondenaturing polyacrylamide gel electrophoresis. Its migration on denaturing sodium dodecyl sulfate-polyacrylamide gels corresponded to a molecular weight of 26,000. Amino acid analysis showed that the ratio of acidic to basic amino acids was 1.4 to 1.0. Evidence for its homogeneity included its migration as a single protein spot on two-dimensional polyacrylamide gel electrophoresis and its single lysine amino-terminal amino acid. This protein is a glycoprotein, as shown by the presence of 15 moles of galactosamine per mole of antigen. These studies demonstrate the presence of a fetal glycoprotein in the chromatin of two tumors that may have an important role in determining their gene products.

Animals↗
PubMed1978

Examination of the immunoglobulin classes involved in the serological response of pregnant sheep to Aspergillus fumigatus.

Pregnant sheep inoculated with Aspergillus fumigatus conidia developed agglutinating and precipitating antibodies for mycelial antigens. The agglutinins were initially exclusively of the IgM class, but were later supplemented by IgG antibodies, although IgM production was usually sustained throughout the serological response. Precipitins active in the immunodiffusion test were of the IgG class. They developed later in the immune response than agglutinins and declined more rapidly. The precipitins and IgG agglutinins were more closely associated with recent active infection than IgM agglutinins.

Animals↗
PubMed1979

Antibodies to Candida species after operations on the large intestine: observations on the association with oral and faecal yeast colonization.

The occurrence of agglutinating and precipitating antibodies to Candida species was determined in 50 patients undergoing operations on the large intestine. The number of patients with oral or faecal yeast colonization and the amount of faecal colonization increased during the period after operation. No patient developed overt fungal infection but 15 patients developed precipitins to mannan antigens and 9 patients developed percipitins to cytoplasmic antigens during this period. The association between the formation of precipitins and the extent of faecal colonization is discussed.

Adult↗
PubMed1985

The development and evaluation of an ELISA for the detection of antibodies to caprine arthritis-encephalitis virus in goat sera.

An enzyme-linked immunosorbent assay (ELISA) was developed for the detection of antibodies to caprine arthritis encephalitis virus (CAEV) in goat sera. The system was evaluated using some 1500 sera from flocks of known clinical history. From this data the interpretation limits of the system were determined. The ELISA system was compared with a gel precipitin test using 5800 sera. Of the positive sera, ELISA detected 97.3% and AGPT 61%. Further evaluation was made using 60 sera of known CAEV reactivity from the USA, and results agreed 100%. Indications are that antibody to the envelope glycoprotein gp135 is being detected. The ELISA system is more sensitive than the precipitin test and is presently being used in a CAEV flock accreditation scheme.

Journal Article↗
PubMed1990

Comparison of antigens from agents of actinomycetoma by immunodiffusion and electrophoresis procedures.

Antigenic extracts were prepared from culture filtrates of the principal agents known to cause actinomycetoma, namely Actinomadura madurae, Actinomadura pelletieri, Nocardia asteroides, Nocardia brasiliensis, Nocardia otitidis-caviarum, and Streptomyces somaliensis. These antigenic preparations were compared by immunodiffusion (ID), counterimmunoelectrophoresis (CIE), line immunoelectrophoresis (LIE) and rocket line immunoelectrophoresis (RLIE), with rabbit antisera prepared against each of the extracts. Cross-reactivity between antigenic extracts from the different actinomycetes, measured by determining the number of precipitin lines in homologous and heterologous systems, was common. Reactions were always stronger and more precipitin lines were present when antigenic extracts were tested against homologous antisera. Similarities between A. madurae and A. pelletieri antigens were of a low order and cross-reactivity did not exceed 33%. A. pelletieri resembled N. asteroides more closely than A. madurae, with 44% of detectable antigenic components cross-reacting. The three species of Nocardia had common antigenic epitopes, but the overall degree of similarity was of a low order (between 12 and 27% by LIE and RLIE). Antigenic extracts of S. somaliensis had few components in common with the other species tested and only one of the 34 lines present in the RLIE system for N. asteroides showed any reaction of identity with an antigenic component in the S. somaliensis extract. Single cross-reacting lines were also present in the CIE and LIE systems.

Actinomycetales Infections↗
PubMed2003

Respiratory diseases in French cork workers.

A cross-sectional study on suberosis was conducted in the Champagne-Ardenne County, France, to determine the prevalence of respiratory symptoms, the level of pulmonary function, and the presence of precipitins against Penicillium frequentans. Thirteen of the 33 workers exposed to cork dust had respiratory symptoms excluding hypersensitivity pneumonitis. The respiratory symptoms were not correlated with tobacco habits or duration of exposure. The levels of pulmonary function were not significantly impaired. No precipitin arc against Penicillium frequentans was found in the sera of exposed workers. The varied symptomatology of suberosis may point to several different diseases, each with its own determining factor. In the present study, exposure to weak humidity and low level of cork dust were related to asthma and chronic bronchitis only, excluding hypersensitivity pneumonitis.

Adult↗
PubMed1989

Investigation of organic aerosols generated during sugar beet slicing.

A description is given of a hygiene investigation made to determine workers' exposure to airborne microorganisms and antigen in the slicing area of a sugar beet refinery following complaints by the workers of work-related respiratory symptoms. Dust concentrations in the slicing area were not high (up to 3.5 mg/m3), but there were many airborne microorganisms (200,000 colony forming units/m3), mostly gram-positive bacteria. Of 15 workers, 7 had precipitins to a sugar beet extract. Of these 7, there were 5 who also had precipitins to an extract of dust collected on a 'Staplex' filter. There was a line of identity between the beet and dust extracts, indicating the dust contained appreciable quantities of beet antigen. As a result of these findings, local exhaust ventilation was installed before the next slicing season. Further investigations were made near the beginning and end of this second season. The results showed lower dust concentrations and markedly fewer numbers of airborne bacteria. Although total numbers of bacteria fell, numbers of gram-negative bacteria increased and, at the end of the season, comprised 20% of the total bacterial numbers. Specific immunoglobulin G (IgG) concentration in the workers' serums fell between the two slicing seasons and then remained constant through the next slicing season. This reflected the environmental measurements that showed lower dust concentrations after the installation of exhaust ventilation equipment.

Aerosols↗
PubMed1959

The occurrence of polyglycerophosphate as an antigenic component of various gram-positive bacterial species.

A bacterial substance has been described which gives a precipitin reaction with certain antisera to Group A streptococci. The precipitating antigen is present in various Gram-positive bacteria, including most hemolytic streptococci, staphylococci, and aerobic sporulating bacilli. It is not present in any of the Gram-negative species examined or in pneumococci, clostridia, or corynebacteria. Analysis of purified preparations obtained from Group A streptococci indicates that the antigen is a simple polymer of glycerophosphate. The identification has been confirmed by immunochemical studies, including precipitin tests and specific inhibition with synthetic polyglycerophosphates. In addition, the infrared spectra of bacterial and synthetic polyglycerophosphate are shown to be closely similar. Immunochemical analysis suggests that the amount of polyglycerophosphate present in Group A streptococci and staphylococci is approximately 1 per cent of the dry weight of the cells. The cellular localization and function of the polyglycerophosphate have not been established.

Antigens↗
PubMed1962

Physical properties of antibody to bovine serum albumin as demonstrated by hemagglutination.

Rabbit antibovine serum albumin antisera were fractionated by zone electrophoresis on starch, zone ultracentrifugation in sucrose density gradients, and diethylaminoethyl-cellulose chromatography, and were assayed by the passive HA technique. Antisera had at least two antibodies: one associated with a fraction characterized as a gamma-2 globulin with a sedimentation rate of 7.3S and low anionic binding (fraction I), and one associated with a fraction characterized as a gamma-1 globulin (or beta-globulin) with a sedimentation rate of 20.4S and high anionic binding (fraction IV). The HA titers of fractions I and IV of early sera were approximately equal. On prolonged antigenic stimulation fraction IV agglutinin decreased in concentration and the relative concentration of fraction I agglutinin increased. Chromatographic analysis of hyper-immune sera yielded 2 additional HA fractions with intermediate anionic binding. These results indicated that rabbits synthesize anti-BSA antibodies similar to rabbit anti-cellular antibodies. Rabbit anti-BSA precipitin migrated mainly as a gamma-2 globulin and the agglutinin migrated with gamma- and beta-globulins. The evidence suggested that the HA method might measure antibody not measured by the quantitative precipitin technique.

Animals↗
PubMed1963

Studies on the dimensions of the rabbit antibenzylpenicilloyl antibody-combing sites.

Rabbit antisera prepared against conjugates of the benzylpenicilloyl (BPO) bifunctional haptenic group were analyzed to determine whether the antibodies are adapted to only a portion of the large BPO molecule, or to the entire molecule, and whether specificity extends to the lysine side chain and adjoining structures of the immunizing carrier protein. No antibodies adapted to the phenylacetylamine portion of the BPO group could be detected in a pooled rabbit anti-BPO serum globulin fraction by PCA and quantitative precipitin analysis using several phenylacetylamine-protein conjugates as antigens. No antibodies adapted only to the thiazolidine carboxylic acid portion of the BPO molecule were detected in the anti-BPO globulin fraction using quantitative precipitin and hapten inhibition methods. At least the bulk of the anti-BPO antibodies was found to be adapted to the entire BPO haptenic group. By quantitative hapten inhibition of precipitation of the anti-BPO globulin fraction, the anti-BPO antibodies were found to show specificity for a 6 carbon amide side chain corresponding to the lysine side chain through which BPO groups are bound predominantly to protein. The contribution of this 6 carbon chain to antibody-hapten binding was small; (-DeltaF degrees ) was calculated to be 460 calories per mole (average). Rabbit anti-BPO antibodies prepared against BPO-rabbit serum albumin conjugates showed specificity also toward structures of the immunizing carrier protein, and possibly toward secondary or tertiary structural configurations. Penicilloyl conjugates of rabbit serum albumin precipitated from 3 individual rabbit antisera more anti-BPO antibodies than did penicilloyl conjugates of heterologous carriers (poly-L-lysine, human serum albumin, and human gamma-globulin). Anti-BPO antibodies demonstrated heterogeneity with regard to closeness of fit to the haptenic group, or with regard to the dimensions of the combining sites, or both. It was concluded that at least a large part of anti-BPO antibodies are specifically adapted to a large antigenic unit comprised of the entire BPO group, the lysine side chain, and structural configurations of the immunizing carrier protein.

Animals↗
PubMed1964

THE IDENTIFICATION AND ISOLATION OF TWO MOUSE-TOXIC PROTEIN COMPONENTS IN EXTRACTS FROM PASTEURELLA PESTIS.

The toxin activity of Pasteurella pestis cells, strain "Tjiwidej," was found to be associated with two proteins. Using a disc electrophoresis technique in conjunction with mouse lethality, two toxic proteins were isolated exhibiting intraperitoneal LD(50)'s of less than 1.0 to 1.5 microg protein. Each produced a single characteristic precipitin band on agar gel diffusion plates. The slower migrating toxin in gel diffusion or disc electrophoresis was designated as toxin A. It was shown to be sensitive to deoxycholate and digitonin, did not accumulate in 5-fluorotryptophan treated cells, and was associated with the membrane fraction of the cell. The faster migrating toxin B, apparently is resistant to surface-active agents, and is not affected by treatment of cells with 5-fluorotryptophan. Toxin B is associated with the soluble or cytoplasmic fraction of the cell. This evidence suggested that each toxin represented a distinctly different molecular species. The possibility is discussed that toxin B is synonymous with the murine toxin previously isolated by paper curtain electrophoresis which revealed only one antigen band in the Oudin precipitin reaction.

Animals↗
PubMed1968

Immunochemical studies on the cross-reactivity between streptococcal and staphylococcal mucopeptide.

Particulate mucopeptides of Group A-variant streptococci and Staphylococcus aureus, solubilized by ultrasonic treatment, give a precipitin reaction with the sera of rabbits immunized with Group A-variant streptococci. gamma-G globulin antibodies have been recovered from these sera which react with the mucopeptides but not with the Group A-variant carbohydrate. The immunochemical basis for the cross-reactivity between the streptococcal and staphylococcal mucopeptides was investigated in detail. Three chemically different fractions have been isolated from enzymatic digests of staphylococcal mucopeptide and were employed as haptenic inhibitors of the precipitin reaction. A fraction consisting of the peptide moiety of mucopeptide was the strongest inhibitor, whereas the hexosamine-rich fraction was less effective. The third fraction, rich in glycine, was least effective. It is suggested that the immunologic cross-reactivity between streptococcal and staphylococcal mucopeptide is due to the fact that these two substances contain chemically similar tetrapeptides. The hexosamine polymer which is identical for both mucopeptides may also contribute to their cross-reactivity.

Amino Acids↗
PubMed1978

Demonstration and quantitation of activation of the first component of complement in human serum.

Activation of the first component of human complement (C1) in human sera can be readily detected in double immunodiffusion studies with anti-C1q, anti- C1r, and anti-C1s as it produces a characteristic pattern quite different from that of precursor C1. Native macromolecular C1 gives a continuous line of precipitation with antisera to C1q, C1r, and C1s in double diffusion studies. After activation of C1 by incubation of serum with complement activators, three major changes occurred in the Ouchterlony pattern. First, spurring of the C1s precipitin line over that of macromolecular C1, indicating release of C1s from C1, was observed with low doses of activator. Release of C1s was quantitated by single radial diffusion and shown to be complete with the highest activator dose examined. Second, C1q was released with larger activator doses as shown also by spurring of the precipitin line due to this component over the remaining macromolecular C1. Third, and most surprising, C1r antigenicity was progressively lost as the activator dose was increased and no C1r line remained with the highest dose of activator tested. This was not true with C1s as there was no change in the total C1s concentration in serum incubated with various activator doses. These observations provide two approaches to the quantitation of C1 activation in human serum. First, C1r and C1s can be quantitated by single radial diffusion. A decrease in the C1r:C1s ratio correlates with activation. Second, C1s released by the activation can be quantitated by single radial diffusion if the agarose contains high concentrations of anti-C1q to confine C1, also containing C1s, to the area near the application well, and lesser concentrations of anti-C1s to permit free C1s to produce a measurable ring. The extent of release of C1s also correlates with activation. These immunochemical techniques to quantitate C1 activation directly inserum do not require specialized reagents. It is hoped that they will be useful in screening pathological sera and in monitoring the status of the complement system in patients.

Complement C1↗