Search PubMed⌕ Search

Biomedical subjects

N J Calvanico

Publications and source records attributed to N J Calvanico.

At least 37 records · Page 2Linked to original sources
PubMed1981

Characterization of Micropolyspora faeni antigens.

Culture filtrate antigens of Micropolyspora faeni grown in a synthetic medium in a stirred fermentor were characterized. The culture filtrate antigens were fractionated by preparative isoelectric focusing with a pH gradient of 3.5 to 5.5. The fractions were pooled according to their reaction with rabbit anti-M. faeni sera. A pool containing two major antigens which were resolved by analytical isoelectric focusing and polyacrylamide disc gel electrophoresis was obtained. One antigen was stainable with Coomassie blue and periodic acid-Schiff stain and was determined to have a mass of 51,000 daltons. The other antigen was stainable only with Coomassie blue and was determined to have a mass of 29,000 daltons. When used at 1 mg/ml, this pool reacted with the sera from all patients with farmer's lung disease by immunodiffusion but failed to react with control sera.

Antigens, Bacterial↗
PubMed1981

Antigens of aspergillus fumigatus. 1. Purification of a cytoplasmic antigen reactive with sera of patients with aspergillus-related disease.

An antigen has been purified from the mycelial cell sap of Aspergillus fumigatus (strain 507). The same component appears to be present in the extracellular phase (culture filtrate) in a partially degraded form. The cell sap protein has a structure composed of four polypeptides of 45,000 daltons linked through disulphide bonds. The isoelectric point (5.2-5.6) and carbohydrate content (12.5% neutral hexose) indicate that this protein is an acidic glycoprotein. It shows reactions with 75% of sera from patients with aspergilloma and allergic bronchopulmonary aspergillosis and is not reactive with sera from normal individuals of patients with other fungal diseases. It also appears to be a component of other A. fumigatus strains.

Antigens, Fungal↗
PubMed1981

The effect of IgG and IgG fragments on the absorption of inhaled antigens across the air-blood barrier of isolated perfused rabbit lungs.

The absorption of inhaled soluble protein antigens across the alveolocapillary membrane can be inhibited by passive immunization in isolated rabbit lungs. The present study was carried out to determine the immunoglobulin class and structural features (Fc-receptor binding, divalent antigen binding) required to effect the inhibition. Isolated rabbit lungs from unimmunized rabbits were perfused with autologous blood to which whole antiserum, IgG or IgG fragments specific for either ovalbumin (OA) or human serum albumin (HSA) was added. The lungs were insufflated with an aerosol containing 125I-OA and 131I-HSA and blood samples were analysed for 125I and 131I in trichloracetic acid (TCA)-precipitable and TCA-soluble forms for 4 hr after insufflation. Whole antiserum and the IgG immunoglobulin fraction of the whole antiserum were equally effective in inhibiting the antigen absorption, indicating that the IGG antibody is sufficient for the effect. The F(ab')2 and Fab' fragments of the IgG molecule were as effective as native IgG, indicating that the antigen-binding site is the only structural requirement and that Fc-receptor and divalent antigen binding are not required.

Absorption↗
PubMed1980

Passive gold agglutination. An alternative to passive hemagglutination.

A method is described for coating colloidal gold with protein for use in passive agglutination. Coating of the gold is quick, reproducible and the sterile filtered product is stable over a long period of storage at 4 degrees C. Colloidal gold coated with each of the 4 subclasses of human IgG was tested with rabbit, monkey and goat antisera and found to be compatible with each. The visual properties of coated gold particles resemble blood cells and appear to be equally sensitive.

Adsorption↗
PubMed1980

A new lymphocyte surface protein present in normal urine. I. Isolation and physicochemical properties.

A lymphocyte surface glycoprotein designated urinary acidic antigen (UA) has been isolated from normal urine by a combination of preparative isoelectric focusing and ammonium sulfate precipitation. It has an m.w. of 14,000 to 17,500 daltons, and is approximately 60% carbohydrate and 40% amino acid in content. The protein exhibited the following physical properties: S20,omega = 1.24; v = 0.693 ml/g; E1%1 cm, 278 nm = 2.08; and pI-2.5. It appears to be unrelated to beta 2-microglobulin, protein HC, urinary proteose, microglobulin, or any previously described normal urine or human serum protein.

Amino Acids↗
PubMed1980

A new lymphocyte surface protein present in normal urine. II. Cellular distribution and biologic properties.

A protein component present in normal human urine has been found on the surface of epidermal cells and lymphocytes. This protein, called urinary acidic antigen (UA), can not be detected in concentrated fractions of normal human serum by double immunodiffusion, suggesting that it is quickly cleared from the circulation. It is readily detected, however, in sera of patients with renal failure. Although it can be eliminated from the cell surface by repeated washings with PBS, it was shown to cap with anti-UA-specific antiserum. Anti-UA suppresses PWM-induced proliferation, but not the lymphocyte response to PHA, Con A, or allogeneic cells. Thus UA appears to have a specific relationship to the pokeweed response. Whether it is a structural component of the PWM receptor is uncertain.

Animals↗
PubMed1980

Immunoglobulin levels in bronchoalveolar lavage fluid from pigeon breeders.

Analysis of serum and BAL fluid immunoglobulin levels in individuals with PBD and in asymptomatic but similarly exposed pigeon breeders was carried out by immunofluorometric assays. The results indicate that the group with PBD have significantly higher levels of IgG and IgA in their BAL fluids but that IgM levels were not significantly different in the two groups. These differences were not reflected in the serum immunoglobulin levels of the two groups. The elevated BAL fluid IgG levels in individuals with PBD is associated with an increase in IgG4 subclass levels as determined PHA inhibition. These studies suggest a role for this subclass in the pathogenesis of the disease.

Adult↗
PubMed1979

An animal model of hypersensitivity pneumonitis in rabbits. Development of chronic pulmonary inflammation and cell-mediated hypersensitivity after repeated aerosol challenge.

Chronic pulmonary inflammation was produced in immunized rabbits by repeated aerosol exposure to soluble antigen. The pulmonary inflammatory response was correlated with the development of cell-mediated hypersensitivity in the lung as evaluated by migration-inhibition studies using bronchoalveolar cells. Such inflammation could be produced with either pigeon dropping extract, an etiologic agent of hypersensitivity pneumonitis, or with human gamma globulin. Development of the inflammatory response was immunospecific and could not be transferred to normal recipients with large quantities of immune serum. Collectively, these data suggest that the development of pulmonary inflammation was due to a cell-mediated immunologic reaction in the lung.

Aerosols↗
PubMed1979

IgA and IgG antibody activities of serum and bronchoalveolar fluid from symptomatic and asymptomatic pigeon breeders.

Serum IgA and IgG antibody activities against pigeon serum were measured in 16 symptomatic pigeon breeders, 20 asymptomatic pigeon breeders, and 3 normal subjects by radioimmunoassay. The IgA and IgG antibody activities against pigeon antigen of the group of patients with disease was significantly greater than those of patients in the asymptomatic and the control group. The overlap of results for the symptomatic and asymptomatic breeders limits the diagnostic value of these individual IgA or IgG antibody determinations. Bronchoalveolar fluid and serum samples from a smaller group of pigeon breeders who underwent lung lavage were available for studies of antibody activity against pigeon serum. Ten asymptomatic and 6 symptomatic breeders were available for study. Both IgG and IgA antibody activities were detected by radioimmunoassay in serum samples and bronchoalveolar fluid. The IgA antibody activity determined by the radioimmunoassay was higher in the respiratory secretions.

Alveolitis, Extrinsic Allergic↗
PubMed1979

Bullous pemphigoid antigen. II. Isolation from the urine of a patient.

This report concerns the isolation of bullous pemphigoid antigen from the nondialyzable urinary components of a patient with the disease. The isolation was accomplished by ion exchange chromatography and gel filtration. Pemphigoid antigen was found to be a basic glycoprotein that on SDS gel electrophoresis showed two major bands, one in the 18,000 m. w. region and the second with a m. w. of 74,000. Between these two bands, two additional bands appeared; one of 35,000 daltons and the other of 68,000 daltons. The 18,000 m. w. band was eluted from the gel and rerun on SDS gels. These gels showed the 18,000 m.w. band and also the appearance of the 35,000 and 74,000 m.w. bands. This finding indicates that urinary pemphigoid antigen may exist both as a single monomeric form and in polymeric aggregates.

Animals↗
PubMed1978

Studies on extracellular proteases of Streptococcus sanguis. Purification and characterization of a human IgA1 specific protease.

Extracellular caseinolytic activity was found in the culture fluid of Streptococcus sanguis ATCC 10556 grown in a dialyzed culture medium. This activity was due to multiple proteases that differed in their elution from hydroxyapatite, sensitivity to enzyme inhibitors, specificity and optimum pH. IgA protease, which splits human immunoglobulin A1 into intact Fc and Fab could be effectively separated from these relatively non-specific proteases and purified to apparent homogeneity in 20% yield by a five-step procedure. Although the bulk of the dextran sucrase activity was separated from the IgA protease, a small amount of sucrase activity remained with the final IgA protease preparation. In polyacrylamide gel electrophoresis at pH 9.5 both activities were located in the single protein band detected in this preparation. A quantitative method for the assay of IgA protease was developed, based on radial immunodiffusion to quantitate the Fab produced. This was used to follow the specific activity and yield during purification, and to characterize some of the catalytic properties of the enzyme. At an enzyme/substrate ratio of 1: 400 (w/w) the protease could effect 50% proteolysis of IgA in overnight incubation at 37 degrees C. The optimum activity was at pH 8.0, and 50% inhibition was achieved at 4 . 10(-4) M o-phenanthroline or 8 . 10(-4) M ethylene diamine tetraacetate. Concentrations of diisopropyl phosphofluoridate, phenylmethyl-sulfonyl fluoride, iodoacetate and p-chloromercuribenzoate up to 10(-2) M were without effect on the IgA protease activity. Full reactivation of the chelator inhibited enzyme could be achieved by the addition of Mg2+, Mn2+ or Ca2+.

Edetic Acid↗
PubMed1978

Phylogenetic studies with pemphigus and pemphigoid antibodies.

Indirect immunofluorescence (IF) studies were performed on skin from a variety of vertebrate specimens and IgG fractions from pemphigoid and pemphigus sera. Pemphigoid antigen was present in fish, amphibian, reptilian, avian and mammalian skin, whereas pemphigus antigen was observed in avian and mammalian skin only.

Animals↗
PubMed1977

Separation of epidermis from dermis with sodium thiocyanate.

After human skin is treated with 2 N sodium thiocyanate, epidermis is easily separated from dermis. The level of cleavage occurs at the lamina lucida of the basement membrane zone. Bullous pemphigoid antigen remains attached to the epidermis.

Histological Techniques↗
PubMed1977

Bullous pemphigoid antigen: isolation from normal human skin.

A protein moiety from epidermal PBS-soluble products was isolated by gel filtration (Bio-Gel A-1.5m) and ion exchange chromatography (DEAE-cellulose). This protein (A-1-Epid) was not retarded by DEAE-cellulose in Tris-HCl buffer, 15mM, pH 8.1. By IEP against an antiserum to epidermal antigens, it showed a single cathodal arc. On disc electrophoresis, at low pH (4.3) a single band was apparent. On SDS gels this protein demonstrated two bands, one with a molecular weight of 20,000, and the second with a molecular weight of 9,200. This purified antigen was able to block the staining of the basement membrane zone produced by bullous pemphigoid antibodies on monkey esophagus and normal human skin with the use of indirect immunofluorescence. This study also demonstrates that bullous pemphigoid antigen (A-1-Epid) and a second epidermal protein (A-2-Epid) are present in the PBS-soluble products of human esophageal mucosa, saliva, and urine. These antigens appear to be unrelated with the blood group substances or secretor status of the donors.

Animals↗
PubMed1977

PBS-soluble substances with blood group activity from human epidermis and dermis.

Phosphate-buffered saline (PBS) soluble extracts of human epidermis and dermis from secretor donors contain products with blood group antigenic activity. The predominant blood group in a sample was the same as the red cell phenotype of the donor. H substance activity was present in all PBS-soluble products (from A, B and O donors). By gel filtration chromatography on a Bio-Gel A-1.5 m column; the epidermal and dermal PBS-soluble blood group substances fractionate in the void volume.

Blood Group Antigens↗
PubMed1977

Activation of the classical complement pathway by Fc fragment of human IgA.

Intact Fcalpha fragments from five human IgA myeloma proteins were produced by two different methods, high temperature trypsinolysis and IgA protease digestion. Three of the Fcalpha fragments activated the classical pathway as determined by the titers of the individual complement components. Two other fragments did not fix complement by either pathway.

Complement System Proteins↗
PubMed1976

Bovine secretory component. Isolation, molecular size and shape, composition, and NH2-terminal amino acid sequence.

Bovine free secretory component was purified from whey by salt precipitation, gel filtration, DEAE-cellulose and phosphocellulose chromatography, and immunoadsorption. It was obtained in immunologically pure form and in 56% yield. The Stokes radius of pure free secretory component was found to be 4.3 nm by gel filtration, and an (see article) of 4.1 S was determined by the ultracentrifuge. The molecular weight was 79,000 by sodium dodecyl sulfate gel electrophoresis and by sedimentation dquilibrium in the ultracentrifuge, using a v of 0.73 determined by ultracentrifugation in D2O and H2O. A minimal axial ratio of approximately 5 was calculated. Amino acid analysis of bovine free secretory component showed remarkable similarity to that of human, dog, and rabbit but carbohydrate analysis showed significant differences. In contrast to the human, bovine free secretory compoennt has 2 methionine residues/mol. The NH2-terminal sequence was found to be Lys-Ser-Pro-Ile-PPHE-Gly-Pro-Glu-Glu-Val-Asp-Ser-Val. This sequence is identical with that the human and dog. However, the poor immunological cross-reactivity between the dog, human, and bovine proteins suggests that significant structural differences will be found in other regions of the molecule.

Amino Acid Sequence↗