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Microbiology and Immunology 1978it was shown in our previous paper that mice primed with chemically modified bacterial alpha-amylase (BaA), which was neither cross-reactive with anti-BaA antibody nor... (Comparative Study)
Comparative Study
it was shown in our previous paper that mice primed with chemically modified bacterial alpha-amylase (BaA), which was neither cross-reactive with anti-BaA antibody nor able to induce a humoral anti-BaA response, developed enhanced responses to a subsequent challenge with native BaA and that the magnitude of the immunological memory was closely related to the priming dose of modified BaA. This paper describes the experimental conditions for induction of delayed hypersensitivity (DH) by modified BaA in relation to the development of immunological memory for antibody response to native BaA. Mice primed with either an intraperitoneal (i.p.) or subcutaneous (s.c.) injection of modified BaA in complete Freunds adjuvant (CFA) developed enhanced anti-BaA as the immunogen and modified BaA as the eliciting antigen, the relationship of anti-BaA responses to a subsequent challenge with BaA. In contrast, when mice were immunized with an s.c. injection of the modified BaA only, a significant level of DH to native BaA could be induced, as measured by the footpad reaction (FPR). The highest degree of DH was observed in mice given 50 micrograms of modified BaA. DH was detectable within 5 days and persisted for 25 days after immunization. In the reciprocal combination of native BaA as the immunogen and modified BaA as the eliciting antigen, the relationship of anti-BaA responses to DH was examined. The primary anti-BaA responses induced by an i.p. injection of large doses of BaA was markedly higher than those induced by an s.c. injection, while DH was exhibited only in mice given s.c. injection of BaA in CFA. With respect to DH to native BaA induced by the modified BaA, it was shown that C3H/He mice were high and C57BL/6 mice were low responders.
Topics: Amylases; Animals; Antibodies, Bacterial; Antigens, Bacterial; Arsanilic Acid; Bacillus subtilis; Female; Hypersensitivity, Delayed; Immunologic Memory; Methylation; Mice; alpha-Amylases
PubMed: 99645
DOI: 10.1111/j.1348-0421.1978.tb00365.x -
Environmental Health Perspectives Aug 1977Humans are exposed to a number of toxic elements in the environment; however, most experiments with laboratory animals investigate only one toxic element. To determine...
Humans are exposed to a number of toxic elements in the environment; however, most experiments with laboratory animals investigate only one toxic element. To determine if concomitant exposure to lead (Pb), cadmium (Cd), and/or arsenic (As) modified the changes produced by any one metal in various parameters of toxicity, 168 male, Sprague-Dawley, young adult rats were fed nutritionally adequate diets to which had been added 0 or 200 ppm Pb as Pb acetate, or 50 ppm Cd as Cd chloride, or 50 ppm As as sodium arsenate or arsanilic acid in a factorial design for a period of 10 weeks. At these concentrations, Cd and As reduced weight gain even when differences in food intake were taken into account; administration of both Cd and As depressed weight gain more than did either metal alone. Pb did not adversely affect food consumption or weight gain. Increased numbers of red blood cells (RBCs) were observed following administration of Pb, Cd, or As; usually more cells were observed when two or three metals were administered, compared to individual metals. Despite increasing numbers of circulating RBCs, hemoglobin and hematocrit were reduced, especially with the Pb-Cd combination and the Cd-arsanilic acid combination. Specific effects of Pb on heme synthesis were observed, including increased urinary excretion of delta-aminolevulinic acid; this increase was reduced by the presence of dietary cadmium. Analyses of blood showed values for the laboratory rat within normal ranges for blood urea nitrogen, creatinine, cholesterol, calcium, albumin, total protein, and bilirubin. Uric acid was increased by Pb, with little modification by dietary Cd or As content. Serum glutamate-oxalate transaminase activity was reduced by As. Serum alkaline phosphatase was greatly reduced by either As or Cd but not Pb. Combinations of As and Cd did not further reduce the activity of this enzyme. Kidney weight and kidney weight/body weight ratios were increased by Pb alone, with no effects of Cd or As alone or as interactions. Liver weight/body weight ratios were reduced in animals fed 50 ppm dietary Cd. Kidney histology shows predominantly Pb effects, namely, intranuclear inclusion bodies and cloudy swelling. Ultrastructural evaluation of kidneys from Pb-treated animals disclosed nuclear inclusion bodies of the usual morphology and mitochondrial swelling. Concurrent administration of Cd greatly minimized Pb effects on the kidney under conditions of this experiment. Liver histology suggests an increased rate of cell turnover with either As compound, but few specific changes.
Topics: Administration, Oral; Animals; Arsenic; Blood Cell Count; Body Weight; Cadmium; Drug Combinations; Drug Interactions; Food; Inclusion Bodies; Kidney Tubules, Proximal; Lead; Male; Organ Size; Rats
PubMed: 198203
DOI: 10.1289/ehp.7719165 -
The Biochemical Journal Nov 1975A crude preparation of alkaline phosphatase (EC 3.1.3.1) from calf intestinal mucosa was purified by affinity chromatography on Sepharose-bound derivatives of arsanilic...
A crude preparation of alkaline phosphatase (EC 3.1.3.1) from calf intestinal mucosa was purified by affinity chromatography on Sepharose-bound derivatives of arsanilic acid, which was found to be a competitive inhibitor of the enzyme. Three biospecific adsorbents were prepared for the chromatography, and the best results were obtained with a tyraminyl-Sepharose derivative coupled with the diazonium salt derived from 4-(p-aminophenylazo)phenylarsonic acid. Alkaline phosphatase was the only enzyme retained by the affinity column in the absence of Pi. The enzyme eluted by phosphate buffer had a specific activity of about 1200 units per mg of protein at pH 10.0, with 5.5mM-p-nitrophenyl phosphate as the substrate.
Topics: Adsorption; Alkaline Phosphatase; Animals; Arsenic; Cattle; Chromatography, Affinity; Chromatography, Ion Exchange; Electrophoresis, Disc; Intestinal Mucosa; Kinetics; Proteins; Sepharose
PubMed: 1218082
DOI: 10.1042/bj1510291 -
Proceedings of the National Academy of... Oct 1972Modification of carboxypeptidase A(gamma) crystals (Anson) with diazotized arsanilic acid specifically labels tyrosine 248; at pH 8.2 the modified enzyme gives yellow...
Modification of carboxypeptidase A(gamma) crystals (Anson) with diazotized arsanilic acid specifically labels tyrosine 248; at pH 8.2 the modified enzyme gives yellow crystals, but a red solution. It has been suggested that arsanilazotyrosine 248 forms a complex with the Zn cofactor accounting for the red color in solution, but that a complex is not formed in the crystal. However, the crystal structure of carboxypeptidase A(gamma) is unknown. We show here that crystals of carboxypeptidase A(alpha), whose crystal structure has been determined, are red both in solution and in the crystalline state (at pH 8.2) after modification with diazotized arsanilic acid. These new data are of importance in relating the structure in the crystalline state to the catalytic mechanisms, as based on the x-ray diffraction evidence. The activity of carboxypeptidase A in the crystal and in solution has a ratio of only 1/3 for the alpha form, in contrast to the ratio of 1/300 for the gamma form, with carbobenzoxyglycyl-L-phenylalanine as a substrate.A pH-jump experiment monitored by stopped-flow kinetics in a split-beam apparatus has revealed a single exponential rate when a solution of arsanilazotyrosine 248 carboxypeptidase A(alpha) at pH 6.7 (yellow) is increased to pH 8.5 (red). The rate constants obtained in this experiment are 6.1 sec(-1) at 3.0 mg/ml and 7.2 sec(-1) at 1.6 mg/ml concentration of enzyme.
Topics: Animals; Arsenicals; Azo Compounds; Carboxypeptidases; Cattle; Chemical Phenomena; Chemistry; Crystallography; Hydrogen-Ion Concentration; Kinetics; Models, Structural; Pancreas; Protein Binding; Protein Conformation; Spectrophotometry, Ultraviolet; Tyrosine; Zinc; p-Azobenzenearsonate
PubMed: 4507609
DOI: 10.1073/pnas.69.10.2850 -
Immunology Mar 1972The serum antibody response of goldfish () to protein immunogen (bovine serum albumin, BSA) and haptenic determinants was studied. The goldfish antibody response was...
The serum antibody response of goldfish () to protein immunogen (bovine serum albumin, BSA) and haptenic determinants was studied. The goldfish antibody response was compared with rabbit antibody response to similar antigens. The goldfish received BSA only or BSA with one of three haptens coupled to it. Arsanilic acid, sulphanilic acid and para-amino-benzoic acid were used as haptens. Precipitation and passive haemagglutination were used to demonstrate the presence of antibody. Only one size of antibody could be demonstrated to all antigens (13.2S). Two populations of antibody could be demonstrated by the use of electrophoresis, and both populations of antibody reacted with BSA in fish receiving only BSA. In fish receiving BSA-hapten, the BSA antibody was found in the slower migrating population, while the haptenic antibody was found in the faster migrating population. The fish antibodies to all immunogens were poor precipitins, but gave titres similar to rabbit antibody when tested by passive haemagglutination. The specificity of fish protein and haptenic antibody is similar to rabbit antibody.
Topics: Aminobenzoates; Animals; Antibody Formation; Antibody Specificity; Antigens; Arsenicals; Cattle; Cyprinidae; Electrophoresis; Haptens; Hemagglutination Tests; Immunodiffusion; Precipitin Tests; Serum Albumin, Bovine; Sulfonic Acids; Ultracentrifugation
PubMed: 4623761
DOI: No ID Found -
Poultry Science Jan 1972
Topics: Animal Feed; Animals; Arsenic Poisoning; Body Weight; Calcium; Diet; Female; Lethal Dose 50; Male; Phosphorus; Poultry Diseases; Sodium; Tibia; Turkeys
PubMed: 4643540
DOI: 10.3382/ps.0510111 -
Proceedings of the National Academy of... Oct 1971Coupling of carboxypeptidase A crystals with diazotized arsanilic acid specifically labels tyrosine 248, an active-site residue of the enzyme. Many azophenols are yellow...
Coupling of carboxypeptidase A crystals with diazotized arsanilic acid specifically labels tyrosine 248, an active-site residue of the enzyme. Many azophenols are yellow and their zinc complexes are red; the "yellow" absorption spectrum of zinc arsanilazocarboxypeptidase crystals is characteristic of the arsanilazotyrosyl group, not of the zinc complex. This is consistent with the interpretation of x-ray data on native crystals of carboxypeptidase A, indicating that tyrosine 248 and the zinc atom are too far apart to form a complex. However, the enzyme in solution is red, denoting the formation of a complex between zinc and arsanilazotyrosine 248. The most likely interpretation of the data is that the orientation of arsanilazotyrosine 248 in solution and in the crystal is different. If the unlabeled tyrosine 248 of native carboxypeptidase undergoes similar changes, these data may bear upon the low activity of the enzyme in the crystalline state and on the catalytic mechanism of the enzyme based on the crystal structure. The opportunities for analogous spectrochemical studies of other, similar systems are pointed out.
Topics: Arsenicals; Azo Compounds; Binding Sites; Carboxypeptidases; Circular Dichroism; Crystallization; Diazonium Compounds; Oxidative Phosphorylation; Protein Conformation; Solutions; Tyrosine; Zinc; p-Azobenzenearsonate
PubMed: 5289887
DOI: 10.1073/pnas.68.10.2532 -
Infection and Immunity Nov 1970The immunogenicity, in rabbits, of homologous encephalitogenic brain basic protein (BP) can be enhanced by conjugating BP with three to five diazotized arsanilic or...
The immunogenicity, in rabbits, of homologous encephalitogenic brain basic protein (BP) can be enhanced by conjugating BP with three to five diazotized arsanilic or sulfanilic acid groups per molecule.
PubMed: 16557896
DOI: 10.1128/iai.2.5.676-678.1970 -
Immunology Jun 1970The cells synthesizing antibodies in response to immunization with (3-diazo, 5-succinylaminobenzoyl) p-aminophenylarsenic acid—bovine gamma globulin (3A, 5S-BGG) and...
The immune response to (3-amino, 5-succinylaminobenzoyl)-p-aminophenylarsenic acid. Immunofluorescent studies of cells making anti-arsanilic acid antibody or anti-m-aminosuccinanilic acid antibody.
The cells synthesizing antibodies in response to immunization with (3-diazo, 5-succinylaminobenzoyl) p-aminophenylarsenic acid—bovine gamma globulin (3A, 5S-BGG) and related antigens have been studied using fluorescent anti-arsanilic acid (anti-Ars), anti-m-aminosuccinanilic acid (anti-Suc) and anti-BGG antibodies, all fluorescent reagents having been prepared from specific precipitates. In the primary response, 1.6 per cent of all cells staining with anti-hapten antibodies after treatment of the smears with a mixture of Ars-rabbit gamma globulin (Ars-RGG) and Suc-RGG stain with both (1.9 per cent of cells staining with anti-Ars; 9.5 per cent of cells staining with anti-Suc). In the secondary response, 12.8 per cent of all cells staining with antihapten antibody stain with both (17.1 per cent of cells staining with anti-Ars; 50.6 per cent of cells staining with anti-Suc). No differences are noted in the percentages of cells staining with both anti-Ars and anti-Suc in animals given a mixture of Ars-BGG and Suc-BGG compared to animals given 3A, 5S-BGG or a doubly conjugated Ars-Suc-BGG (AS-BGG). Comparable numbers of doubly staining cells are present in controls given only Ars-BGG or only Suc-BGG. This is interpreted as indicating that most and probably all of the doubly staining cells noted in the experimental groups are related to an antigenic cross reaction rather than to the synthesis of two antibodies by a single cell. Evidence that this is a cross reaction of intracellular antibody is presented. No cells staining with both antihapten and anticarrier protein (BGG) antibodies have been observed.
Topics: Animals; Antibody Formation; Antigens; Arsenicals; Azo Compounds; Cross Reactions; Fluorescent Antibody Technique; Haptens; Immune Sera; Immunization; Lymph Nodes; Male; Plasma Cells; Rabbits; gamma-Globulins
PubMed: 4193666
DOI: No ID Found -
The Journal of Experimental Medicine Mar 1970A number of azobenzenearsonate (ABA) conjugates have been prepared and tested for ability to react with antibody, to sensitize for hapten-specific delayed...
A number of azobenzenearsonate (ABA) conjugates have been prepared and tested for ability to react with antibody, to sensitize for hapten-specific delayed hypersensitivity and to induce hapten-specific unresponsiveness. All conjugates tested by in vitro or in vivo methods show a capacity to react with preformed antibody. Conjugates of L-amino acid polymers are immunogenic and induce tolerance. Conjugates of D-amino acid polymers or conjugates with high density of ABA groups are nonimmunogenic and fail to induce tolerance. Since paired D- and L-polymer conjugates react comparably with preformed antibody but differ markedly in tolerance induction, it is argued that combination with an antibody-like receptor molecule on the surface of an immune-competent cell is not a sufficient condition for tolerance. The lack of effectiveness of sterically crowded conjugates as well as D-polymer conjugates argues for a preliminary biologic "processing" of antigen necessary for induction of immunity or tolerance. Such a processing event might well involve enzymatic attack on the antigen.
Topics: Animals; Antibody Formation; Arsenicals; Guinea Pigs; Hypersensitivity, Delayed; Immune Tolerance
PubMed: 5413329
DOI: 10.1084/jem.131.3.571