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PubMed1983

Ganglioside-basic protein interaction: protection of gangliosides against neuraminidase action.

The ability of acidic phospho- and sphingolipids to interact with basic proteins was studied by double diffusion analysis. The phospholipids, tri- and diphosphoinositide, and the sphingolipid, sulfatide, interacted with myelin basic protein as evidenced by precipitin line formation. Of the sialoglycosphingolipids (gangliosides) tested, only the myelin-specific monosialoganglioside, GM4, formed a precipitin line with myelin basic protein. In addition, myelin basic protein retarded the activity of Clostridium perfringens neuraminidase against GM4 and the disialoganglioside, GD1b. Examination of purified rat brain myelin suggested the presence of a neuraminidase activity intrinsic to myelin. This finding, in concert with ganglioside-myelin basic protein complexes which selectively protect against neuraminidase, may provide a physiological explanation for the simplified ganglioside pattern found in myelin.

Animals↗
PubMed2004

Antibody response to Pseudomonas aeruginosa in cystic fibrosis patients: a marker of therapeutic success?--A 30-year cohort study of survival in Danish CF patients after onset of chronic P. aeruginosa lung infection.

We studied the effects of increasingly intensive treatment regimens on anti-pseudomonal antibody response and survival in five successive cohorts of a total of 157 Danish cystic fibrosis patients after they had acquired chronic P. aeruginosa lung infection. The time periods were 1971-1975 (N = 21), 1976-1980 (N = 64), 1981-1986 (N = 27), 1987-1993 (N = 26), and 1994-2000 (N = 19). During this 30-year period, we introduced elective 2-week courses of chemotherapy every third month in all chronically infected patients, early aggressive treatment with inhalation of colistin and oral ciprofloxacin for 3 months whenever P. aeruginosa was cultured in sputum from noncolonized patients, and inhalation of recombinant human dornase alfa. There was a significant correlation between the calendar year when chronic P. aeruginosa infection was acquired and the subsequent increase in the level of precipitins (P < 0.00001). The median number of precipitins increased by 5 per year in the oldest calendar year cohort, and 1 per year in the youngest. The median age of onset of chronic P. aeruginosa increased from 9.3 years from 1981-1986 to 13.8 years from 1987-2000. Survival after acquisition of chronic P. aeruginosa lung infection improved with time (P = 0.008). Our study shows that CF patients who are treated intensively have lower antibody responses and longer survival after acquisition of chronic P. aeruginosa lung infection.

Anti-Bacterial Agents↗
PubMed1995

Native and asialo-Tamm-Horsfall glycoproteins as important ligands for the detection of GalNAc beta 1-->and Gal beta 1-->4GlcNAc active lectins.

The binding properties of human Tamm-Horsfall Sd(a+) urinary glycoprotein(THGP) and asialo-THGP with various applied lectins was investigated by quantitative precipitin and precipitin inhibition assays. Both glycoproteins completely precipitated Abrus precatorius agglutinin(APA). They also reacted well with Wistaria floribunda (WFA), Glycine max (soybean, SBA), and Ricinus communis agglutinins and precipitated over 78% of the lectin nitrogen added, but reacted poorly or weakly with all alpha-anomeric GalNAc specific lectins, such as Helix pomatia (HPA), Phaseolus lunatus (lima bean, LBL), and Maclura pomifera (MPL) lectins. The glycoprotein-lectin interaction was inhibited by GalNAc beta 1-->, Gal beta 1-->4GlcNAc, or by both. The findings suggest that Sd (a+) THGP and asialo-THGP are among the best water-soluble glycoprotein ligands for GalNAc beta 1-->and Gal beta 1-->4GlcNAc active lectins.

Asialoglycoproteins↗
PubMed1995

Affinity of Bandeiraea (Griffonia) simplicifolia lectin-I, isolectin B4 for Gal alpha 1-->4 Gal ligand.

The affinity of Bandeiraea (Griffonia) simplicifolia lectin-I isolectin B4 (BSI-B4) for the isomer of human blood group B active disaccharide (B, Gal alpha 1-->3Gal), the Gal alpha 1-->4Gal galabiose ligand, was studied by quantitative precipitin (QPA) and precipitin-inhibition assays. When human blood group B, P1 and H active glycoproteins were tested by OPA. BSI-B4 reacted strongly with both the B active glycoprotein purified from human ovarian cyst fluid and a P1 active glycoprotein isolated from sheep hydatid fluid and precipitated over 86% of the lectin nitrogen added. The P1 active glycoprotein-BSI-B4 interaction was inhibited by both Gal alpha 1-->3Gal alpha 1-->methyl and Gal alpha 1-->4Gal disaccharide indicating that BSI-B4 is not only reacting with Gal alpha 1-->3Gal disaccharide, but also recognizing Gal alpha 1-->4Gal. The galabiose sequence is frequently found in the carbohydrate chains of many glycosphingolipids located at the mammalian cell membranes such as intestinal and red blood cell membranes, for E. coli ligand binding and toxin attachment.

ABO Blood-Group System↗
PubMed1985

Immunological and biochemical properties of metallothioneins of golden hamster, mouse and rat.

Golden hamster, mouse and rat hepatic cadmium metallothioneins (MT) were purified by Sephadex G-75 gel filtration, DEAE-Sephadex A-25 chromatography and activated Thiol-Sepharose 4B affinity chromatography. Metallothioneins were separated by DEAE-Sephadex A-25 chromatography into two forms: MT-1 and MT-2. In mouse and golden hamster liver, MT-1 was the major form. The purified proteins were homogeneous as judged by polyacrylamide gel electrophoresis in the presence and absence of sodium dodecyl sulfate. In non-denaturing polyacrylamide gel electrophoresis, migration of mouse, rat and golden hamster hepatic metallothioneins were found to be different. Antibodies to mouse hepatic MT-1 was raised in rabbits. The antiserum cross reacted with mouse and hamster MT-1 and MT-2 giving a single precipitin band. Mouse, rat and hamster hepatic MTs are immunologically identical but electrophoretically different. The kidney and pancreatic MTs of rat and golden hamster were purified by Sephadex G-75 gel filtration. They were immunologically distinct. Pancreas MT formed a line of partial identity with hepatic MTs. Kidney MTs form two precipitin band one identical with the pancreatic form and another of complete identity with the hepatic MTs. This indicates the presence of tissue specific MTs.

Animals↗
PubMed1984

Differential binding characteristics and applications of DGal beta 1----3DGalNAc specific lectins.

The binding properties of Arachis hypogaea (PNA), Bauhinia purpurea alba (BPL), Maclura pomifera ( MPL ) and Sophora japonica (SJL) lectins were studied by quantitative precipitin and precipitin inhibition assays, demonstrating them to be most specific for DGal beta 1---- 3DGalNAc residues. Additionally, each lectin had its own binding characteristic such as different binding activities to DGal beta 1---- 4DGlcNAc or DGal beta 1---- 3DGlcNAc beta 1----linked oligosaccharides, and/or DGalNAc alpha 1----linked to the Ser or Thr of the protein moiety. These differential binding characteristics can be used for investigating fine differences of the carbohydrate structure of the glycoconjugates, especially those having DGal beta 1---- 3DGalNAc residues as terminal non-reducing ends.

Acetylgalactosamine↗
PubMed1987

Structural and immunochemical characterization of the O-haptens of Brucella abortus lipopolysaccharides from strains 19 and 2308.

The O-haptens of the major fraction (f5A) of B. abortus (Strains 2308 and 19) membrane bound smooth lipopolysaccharide (sLPS) were prepared by hydrolysis of f5A native sLPS in 1% acetic acid at 100 degrees C for 2 h. After hydrolysis, O-haptens were separated from Lipid A-protein complex by centrifugation, and from small fragments by ultrafiltration of molecular weight cut-off (MWCO) 1.0 X 10(3). These carbohydrate haptens were identified by precipitin-inhibition assay and further fractionated by both membrane filtration and dialysis. The size distributions of carbohydrate haptens of endotoxins (f5A) ranged from oligosaccharides up to polysacchandes of 1.0 X 10(4) MWCO. Three major fractions of MWCO 8.0-10.0 X 10(3), 3.5-5.0 X 10(3) and less than 1.0 X 10(3) from both strains 2308 and 19 contained more than 85% of the total immunoactive materials. These fractions of haptens were subjected to composition, proton and 13C NMR analysis and were found to be a homopolymer of alpha 1----2 linked, 4,5-dideoxy-4-formamido-D-mannose (N-formylperosamine), which is identical to O-haptens of B. abortus strain 119.3 and Yersinia enterocolitica serotype 0:9 and similar to Vibrio cholerae 569B (INABA). Fractions of these haptens exhibited similar inhibitory reactivities in a precipitin-inhibition assay as expressed as mumoles of monosaccharide of anhydro-N-formyl perosamine. They were about 480 times as active as Me alpha----DMan or DMan.

Brucella abortus↗
PubMed1986

Spinal arachnoiditis due to aspergillus meningitis in a previously healthy patient.

A 30-year-old, previously healthy, non-addicted man presented with a chronic spinal meningitis complicated by arachnoiditis and spinal cord compression. Biopsy showed a chronic granulomatous leptomeningitis, in which some cells contained branching septate organisms that were immunostained with an antiserum to Aspergillus fumigatus. Precipitins to A. fumigatus were detected in cerebrospinal fluid (CSF), but not in blood, and aspergillus infection was apparently restricted to the leptomeninges. Clinically successful treatment led to the disappearance of CSF precipitins and oligoclonal bands.

Adult↗
PubMed1981

Allergy to fish in fish meal factory workers.

Fifty-one fish meal factory workers in four Polish harbour cities were investigated. They had been employed in the fish meal factory for 1--33 years and 10.7 years on average. All but seven of these were smokers. The investigation consisted of respiratory symptoms questionnaire, chest X-ray, physical examination, respiratory function assessment, intradermal skin tests to ten fish genera, total serum IgE and serum precipitins to fish antigen assessment. Chronic bronchitis was diagnosed in 29 investigated patients. In 12 (23%) of the patients, according to their case history or to skin tests, allergy to fish was suspected, but was confirmed by elevated serum IgE levels in only two of these. In no cases were serum precipitins against fish antigen found. In six patients, either obstructive or restrictive changes were diagnosed according to conventional spirometry, but by MEF50% VC in 17 subjects (33%) slight airways obstruction was diagnosed and can be regarded as the result of the habit of smoking. According to the performed investigation, only two workers presented a full clinical and immunological picture of allergy to fish. From the practical point of view, this type of professional activity is less dangerous than might be theoretically expected.

Adult↗
PubMed1980

Immunological characterization of fibrous protein isolated from human stratum corneum.

Specific antibody was produced against the purified fibrous protein isolated from human stratum corneum. Two precipitin lines appeared when the antiserum was tested against fibrous protein of human stratum corneum (FP/SC). When the section of human plantar skin was incubated with antihuman FP/SC serum for 30 min at 37 degrees C for indirect immunofluorescent studies, only a poor fluorescence could be detected in the living layers. The fluorescence in these layers increased on the section that was incubated with the fluorescence in these layers increased on the section that was incubated with the antiserum for 1 h at 37 degrees C. However, no fluorescence could be observed in the stratum corneum under these conditions. On the other hand, irregular fluorescence appeared in the stratum corneum after more prolonged incubation (2 h) with antiserum at 37 degrees C. The fluorescence also appeared in the stratum corneum when the section was incubated with antiserum for 30 min following a short pretreatment with urea-buffer. Identical precipitin lines were observed on double diffusion of anti-human FP/SC with prekeratin and 0.85% NaCl-Tris acetate buffer (pH 7.5) soluble proteins of human living layers.

Antibodies↗
PubMed1984

Micropolyspora faeni antigens of three commercial extracts.

The protein composition of three commercial extracts of Micropolyspora faeni, produced in U.S.A., England and Italy has been evaluated by agarose gel electrophoresis. By crossed immunoelectrophoresis, 'tandem'-crossed immunoelectrophoresis and by a modification of this last technique, the antigenic composition and the common antigens of the extracts have been investigated. Hyperimmune rabbit serum and a pool of five human sera with precipitins to Micropolyspora faeni have been used as source of antibody. Different quantity and quality of protein content was observed in the available batches. Different antigenic composition was also observed, not directly related to the different proteins contained therein; three antigens were definitely common to all extracts and two of them represented the major antigens of each extract. Despite the total protein content, the major and common antigens were found in similar concentrations in all three products examined. Therefore, the discrepancies observed in the precipitin reactions using the three commercial Micropolyspora faeni extracts are due to differences in the minor antigen composition of the extracts.

Animals↗
PubMed1989

Immunodiffusion test for diagnosing human pythiosis.

An immunodiffusion test was developed for diagnosing subcutaneous and systemic pythiosis in humans. When culture filtrate antigen (CFA) from P. insidiosum was reacted against patient and rabbit antisera, 1-5 precipitin bands occurred both in patient and rabbit antisera, and a line of identity also occurred between patient and rabbit sera. When control P. insidiosum CFA was reacted with 30 apparently normal persons, 20 Thalassemia patients, 2 candidosis and 5 aspergillosis patients, no precipitin bands were found. P. insidiosum CFA also tested with rabbit antibodies to B. dermatitidis, C. immitis, H. capsulatum, P. brasiliensis, C. albicans, M. furfur and A. fumigatus revealed no cross reactions. This test is practical, sensitive and specific.

Antibodies, Fungal↗
PubMed1977

Counterimmunoelectrophoresis and opportunistic fungal infections.

Sera from 35 apparently normal humans, 37 compromised human patients, 30 hedgehogs and 30 sheep, were examined for precipitating antibodies to four opportunistic fungi - Absidia corymbifera, Aspergillus fumigatus, Candida albicans and Rhizopus arrhizus-using counterimmunoelectrophoresis (CIE). Precipitins to A. fumigatus were almost exclusively confined to specimens obtained from the compromised human group (51% of those examined) while Candida precipitating antibodies were detected in the sera of both normal (26%) and compromised (49%) humans and in 10% of the hedgehog specimens. Serum precipitins against the two phycomycetes included in the investigations were rare. Because of the complexity of most fungal antigen extracts, it appears the two phycomycetes included in the investigations were rare. Beause of the complexity of most fungal antigen extracts, it appears essential that sera be tested against a number of different antigen concentrations if CIE is to be used with confidence in fungal serology.

Antibodies, Fungal↗
PubMed1985

Enzymatic activity of atypical Oriental types of aldehyde dehydrogenases.

Catalytic activity of the atypical Oriental-type aldehyde dehydrogenase-2 (ALDH2) was considered to be null or severely diminished. Recently it was suggested that the atypical ALDH2(2) retained about 30% of the specific activity of the usual ALDH2(1). We reexamined the problem by two-dimensional crossed immunoelectrophoresis. The usual Caucasian livers exhibited two distinctive precipitin peaks, one corresponding to the cytosolic ALDH1 and the other corresponding to the usual mitochondrial ALDH2(1), in both protein stain and enzyme activity stain. In contrast, the atypical Oriental livers exhibited two precipitin peaks in protein stain, but only one peak, corresponding to ALDH1, in enzyme activity stain. These results support the original notion that the atypical ALDH2(2) is enzymatically inactive or far less active than the usual enzyme, refuting the idea of the atypical ALDH2(2) with substantial enzyme activity.

Aldehyde Dehydrogenase↗
PubMed1981

Serological diagnosis of pulmonary aspergillosis.

Methods for the preparation of antigens from clinically isolated cultures of Aspergillus were standardized. Sera from 25 suspected cases of pulmonary aspergillosis were tested against antigens prepared by us, from 4 strains of A. fumigatus and one strain of A. flavus, using the Ouchterlony double diffusion and immunoelectrophoretic techniques. Of the 25 sera, 18 reacted positively with antigens of A. fumigatus, one with A. flavus and 2 with both these species. Antigens of two non-pathogenic Aspergilli included in the study failed to react with any of the sera. Our antigen preparations gave more numerous as well as sharper precipitin lines than the commercial Bencard antigens which were used for comparison. Moreover, mycelial antigens from 48 to 96 h old cultures revealed precipitin lines comparable to that of the routine, 4 week old culture filtrate antigens, thus suggesting that the incubation period for obtaining antigens could be cut down considerably.

Antigens, Fungal↗
PubMed1993

Henoch-Schoenlein purpura due to streptokinase.

The syndrome of Henoch-Schoenlein purpura developed in a 74-year-old woman after receiving streptokinase as thrombolytic therapy for an acute myocardial infarction. Renal biopsy revealed mesangial hypercellularity with deposits of IgA. Skin biopsy also revealed IgA deposition. Immunological studies showed evidence of sensitization to streptokinase. Elevated IgG, IgA, IgM, and IgE antistreptokinase antibodies were detected in the acute serum. Positive immediate skin reactivity to streptokinase was also present. Serum precipitins to streptokinase disappeared when IgA was removed from the serum. Positive staining with biotinylated streptokinase was seen in the skin in the same pattern of distribution as IgA. These findings strongly support the role of streptokinase and IgA in the pathogenesis of Henoch-Schoenlein purpura in this patient. A control group of streptococcal-infected patients showed no immune response to streptokinase. Another control group of streptokinase-treated patients, who had no untoward reaction, had elevated immunoglobulin classes and precipitins to streptokinase. However, the precipitating antibody was IgG and streptokinase skin tests were negative.

Aged↗
PubMed1989

Characterization of the antigens that induce the production of antibodies in mice immediately after infection with Angiostrongylus cantonensis.

Following the infection of mice (a non-permissive host) with Angiostrongylus cantonensis, the occurrence of parasite-specific antibodies in the serum was monitored during the first 5 weeks after infection by the Ouchterlony method and by an immunoblotting technique, with special reference to the antigens recognized by host IgG antibodies. Only the serum from infected mice 7 days postinfection formed a single, distinct precipitin line when tested on Ouchterlony plates against extracts of digestive organs, reproductive organs, and body wall from adult female worms. In each case the precipitin line fused totally with that formed between the serum from infected mice and one of the other organ extracts. When the serum from infected mice was used as a blotting antibody under both reducing and nonreducing conditions, specific bands with molecular weights in the range from 60,000 to greater than 200,000 daltons were developed with each organ extract, although molecular weights varied according to the organ from which the antigenic extract was prepared. Further analyses on Ouchterlony plates and by immunoelectrophoresis and comparison with the reactivity of serum from permissive rat hosts infected 35 days postinfection with the same parasite revealed that an additional reactivity, distinct from that in rats, predominantly occurred in the serum from infected mice.

Angiostrongylus↗
PubMed1980

Anti-catechol-O-methyltransferase: demonstration of specificity and immunological cross-reactivity with the enzyme from rat liver, kidney, brain, and choroid plexuses.

An antiserum to rat liver catechol-O-methyltransferase (COMT) was utilized in the immunological characterization of COMT from rat kidney, brain, and choroid plexuses, in addition to rat liver. The presence of anti-COMT activity was confirmed by the direct inhibition of the activity of the enzyme from rat liver by small quantities of the antiserum and by the inhibition of the activity of the enzyme from rat brain. The specificity of the antiserum was demonstrated both by immunoelectrophoresis of rat liver COMT, and by a partial purification of rat liver COMT in which changes in COMT specific activity were correlated with the appearance of a precipitin line in double-immunodiffusion experiments. The antigenic similarity of the enzyme derived from rat liver, kidney, brain, and choroid plexuses was demonstrated by the formation of a precipitin line of identity when preparations from these four tissues were diffused against the antiserum.

Animals↗