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PubMed1988

Differential binding properties of Ga1NAc and/or Ga1 specific lectins.

Grouping of lectin binding properties, based on determinant structure rather than monosaccharide inhibition pattern, should facilitate the selection of lectins as structural probes for glycans as well as for the interpretation of the distribution and the properties of the carbohydrate chains on the cell surface. Based on the binding specificities studied with glycan by precipitin-inhibition, competitive-binding and hemagglutinin-inhibition assays, twenty Ga1 and/or Ga1NAc specific lectins have been divided into six classes according to their specificity for the disaccharide as all or part of the determinants and Ga1NAc alpha 1----Ser(Thr) of the peptide chain. The differential affinities of these lectins were characterized by quantitative precipitin assay. Abbreviation of the following six lectin determinants can also be used to classify these lectins. (1) F determinant (GalNAc alpha 1----3GalNAc, Forssman specific disaccharide). (2) A (Af) determinant (GalNAc alpha 1----3Gal, Human blood group A specific disaccharide; Af, fucosylated A, (GalNAc alpha 1----3 [LFuc alpha 1----2]Gal). (3) Tn determinant (GalNAc alpha 1----0 to Ser (Thr) of the protein core, Tn antigen). (4) T determinant (T antigen, Gal beta 1----3GalNAc alpha 1----0 to Ser (Thr) of the protein core, the mucin type sugar sequence on the human erythrocyte membrane or Gal beta 1----3GalNAc beta 1---- at the nonreducing end of ganglioside). (5) I and II determinants (human blood group type I and II carbohydrate sequences). Most of the lectins reactive to Gal beta 1----4GlcNAc (II) are also reactive to Gal beta 1----3GlcNAc (I). Lectin I (II) determinants (i.e. Gal beta 1----3 (4) GlcNAc residues) can be found at the nonreducing end of the carbohydrate chains derived from either N-glycosidic or O-glycosidic linkages. (6) B determinant (Gal alpha 1----3Gal, Human blood group B specific disaccharide). Their carbohydrate specificities are classified as following: (Table see text). The differential binding properties of lectins can be defined from comparisons of their carbohydrate specificities listed above.

Acetylgalactosamine↗
PubMed1987

Prognostic implications of circulating immune complexes and Pseudomonas aeruginosa-specific antibodies in cystic fibrosis.

We re-examined the role of circulating immune complexes (CIC) in cystic fibrosis, by their serial measurement in 16 patients who had advanced lung disease and persistent specific antibodies to Pseudomonas aeruginosa in their serum. CIC were detected by 125I C1q-BA (IgG + IgM-IC) and Raji RIA (IgG-IC), and IgA-IC by a modification of the Raji RIA method. Serum precipitins to Ps. aeruginosa were detected by crossed-immunoelectrophoresis (XIE) and antibodies to alkaline protease (AP), elastase (EL) and exotoxin-A (EA) of Ps. aeruginosa were detected by ELISA technique. CIC were positive in greater than 50% of the samples in 8 patients (group A) and in less than 25% in the other 8 (group B); follow-up averaged 22 mo in group A and 26 mo in group B. The numbers of precipitins to Pseudomonas were equally high, and levels of antibodies to AP, EL, and EA of Ps. aeruginosa were similarly positive, in the 2 groups. In group A the CIC were mostly IgG-IC by Raji RIA (76%) and IgA-IC, whereas in group B C1q-binding IC (79%) and IgA-IC were more prevalent than IgG-IC by Raji RIA. Four group A children died during follow-up, together with one group B child in whom the CIC level had suddenly risen precipitously. We postulate that a high level of CIC in association with persistent specific-antibody response to Ps. aeruginosa heralds a poor prognosis in cystic fibrosis.

Adolescent↗
PubMed1988

Stage-specific antibody response of Mastomys natalensis during the course of Dipetalonema viteae infection.

Stage-specific haemagglutinating (HA) and precipitin antibodies (PA) to infective larvae (L3), adult worms and microfilariae (mf) have been demonstrated in sera of Mastomys natalensis during the course of Dipetalonema viteae infection using indirect haemagglutination (IHA) and Ouchterlony's gel-diffusion tests. L3-specific HA antibody titre was shown to be quite low and lasted for shorter period whereas adult-specific antibody response was significantly higher and persisted for longer duration (beyond day 240 post exposure). No precipitin antibody to L3 stage could be detected, however, significant adult-specific antibody was detected which ultimately tapered off by day 210 post exposure (p.e.). In contrast, mf-specific PA which appeared at the beginning of patency, never disappeared even at the late stage of infection. Mf-specific HA antibody appeared at early incubation period (day 15 p.e.) and after exhibiting two peaks, one in the midst of prepatent period (day 30 p.e.) and the other on day 180 p.e. persisted at low level during the late stage of infection when amicrofilaraemia developed.

Animals↗
PubMed1988

[A case of semi-delayed hypersensitivity to dry sausage dust].

The authors record a case of semi-delayed hypersensitivity to sausage dust in an employee of a wholesale pork butcher and curing business. The symptoms combined cough and fever with rigors; there were no clinical or laboratory data to suggest alveolar disease. The immunoallergic "work-up" suggested a precipitin mediated disease with notably 10-14 precipitation arcs to the products of sausage sweepings. A microbiological analysis of the flora of the sausages, as well as an extensive search for precipitins and cutaneous tests have not enabled us to confirm the antigen responsible with certainty.

Alveolitis, Extrinsic Allergic↗
PubMed1988

The E. coli immunosorbent as used in serodiagnosis of Legionella infections studied by crossed immunoelectrophoresis.

In this study we investigated an immunosorbent, E. coli blocking fluid (BF), proposed for use in the Legionella Indirect Fluorescent Antibody Test (IFA). With crossed immunoelectrophoresis (CIE) of clinically relevant Legionella species, only one heat-stable antigen (no. 1) cross-reacted with the BF preparation. Patients' sera with elevated Legionella IFA titres did not react with this antigen in CIE. Out of 23 IFA positive patients' sera, six had titres lowered significantly to negative, when BF was applied as serum diluent for the titration (IFA BF negative sera). All six sera were negative in the micro agglutination test (MA). None of the IFA BF negative sera contained any Legionella precipitins in CIE, whereas nine out of the remaining 17 IFA BF positive sera unchanged by BF contained one or more precipitins. CIE results could not explain the effect of BF in Legionella IFA, and further studies are needed to sufficiently define the use of immunosorbents in diagnostic Legionella serology.

Antibodies, Bacterial↗
PubMed1977

Preparation of a specific antiserum towards the microfibrillar protein of elastic tissues.

The microfibrillar protein (MFP) of bovine foetal ligamentum nuchae was prepared and used to induce an antiserum (I) in rabbit. Antibody (I), reacted with a dispersion of MFP in Ouchterlony plates, produced one major precipitin band (A) and two minor bands (B and C). The major band (A) was used to prepare antiserum (II) in a second rabbit. Antibody (II) was monospecific for precipitin band (A) and was shown to be selectively directed against MFP of bovine and chick elastic fibres by immunofluorescent and immunoperoxidase techniques. Immunoperoxidase-labelling techniques demonstrated the regular organization of microfibrillar protein on the surface of elastic fibres and suggested the ability of MFP to assemble into a framework defining the three dimensional polymerization of elastin monomers during the formation of elastin fibres. It was observed that collagen fibrils, which were in close association with the microfibrillar protein on the surface of elastic fibres, also possessed a protein which was immunologically related to MFP.

Animals↗
PubMed1986

Modified counterimmunoelectrophoresis procedure for easy identification of antibodies to extractable nuclear antigens.

Much effort is being expended to detect and to characterize the multiplicity of antibodies directed against extractable nuclear antigens in connective tissue diseases. Hemagglutination and counterimmunoelectrophoresis are highly sensitive but lack specificity and must be followed by double diffusion. Double diffusion, however, suffers from lack of sensitivity and often from lack of clarity for interpretation of the resulting precipitin lines. We describe here a modified counterimmunoelectrophoresis procedure utilizing offsetting wells, followed by diffusion, which provides the clarity of precipitin lines for easy interpretation as well as overnight interpretation and the use of small amounts of reagents.

Antibodies, Antinuclear↗
PubMed1986

[Investigation on effectiveness and safety of fosfomycin in treatment of patients with allergy induced by antibacterial agents].

Twenty-four patients with urinary tract infections were treated with fosfomycin (FOM) to evaluate its effectiveness and safety. They all had shown allergic reactions, mainly to beta-lactams. Lymphocyte stimulation test (LST), leukocyte migration inhibition test (LMT), passive cutaneous anaphylaxis (PCA) and the precipitin reaction were performed to test whether FOM would also cause an allergic reaction. FOM was judged by the physician-in-charge to be effective in all 6 patients (100%) with acute simple cystitis and 8 (72.7%) of 11 patients with chronic complicated urinary tract infections. The drug was also effective in 15 (83.3%) of 18 patients with epididymitis, etc. Regarding the usefulness of FOM, it was judged to be useful in 21 (91.3%) of 23 patients. FOM was very useful in 8 (34.8%) of these 23. In patients tested for LST, the value was lower in cases given FOM than in cases given ampicillin (ABPC), cefazolin (CEZ) or latamoxef (LMOX); there was an especially significant (p less than 0.01) difference with ABPC. All the drugs tested were negative for the LMT, PCA and precipitin reaction tests. No subjective or objective abnormalities were attributed to the FOM treatment, and there were also no abnormal laboratory test values. FOM was evaluated to be an effective and safe antibacterial agent without in vitro or clinical allergic reactions.

Adolescent↗
PubMed1986

Persistent bronchoalveolar lymphocytosis in asymptomatic farmers.

We previously demonstrated that 14 of 23 asymptomatic dairy farmers with positive precipitins and 5 of 20 without precipitins had an alveolar lymphocytosis (greater than 22%). To verify the outcome of this lymphocytosis, we restudied, 2 or 3 yr later, 27 of these subjects. No subject had had symptoms suggestive of farmer's lung disease either prior to the initial study or between the 2 studies. All were still on their farm. Physical examination and chest roentgenograms were normal for all subjects at both studies. Pulmonary function tests showed a small but significant change in RV (101.8 +/- 20.4% to 118.7 +/- 27.6% of predicted, p less than 0.05) and FVC (98.0 +/- 12.6% to 94.3 +/-9.5%, p less than 0.05), whereas TLC, FEV1, and DLCO showed no changes. Lymphocytes from BAL were still increased in 9 of 12 subjects, whereas 3 had returned to normal; of the 15 subjects with previous normal values, 3 now had an abnormal lavage lymphocytosis. There was no correlation between lung function variations and the percentage of lymphocytes in the previous or the present BAL. We conclude that a bronchoalveolar lymphocytosis is a persistent phenomenon in a large number of asymptomatic dairy farmers, and that this finding is not related to significant disease, at least in the time span studied.

Adult↗
PubMed1975

Studies on cathepsins of rat liver lysosomes. II. Comparative studies on multiple forms of cathepsin A.

The multiple forms of cathepsin A (AI, AII, and AIII) purified from the lysosome fraction of rat liver by Sephadex G-200 and DEAE-Sephadex chromatographies were studied comparatively. Forms AI, AII and AIII were stable between pH 3.0 and 5.5, and had pH optima for CBZ-Glu-Phe at 5.6, 5.8, and 5.9, respectively. These activities were rapidly lost on heating above 60 degrees. Their isoelectric points were at 4.7, 4.8, and 4.9, and the Michaelis constants for CBZ-Glu-Phe were calculated as 10, 6.6, and 4.2 X 10(-4)M, respectively. Activity was inhibited by Ag+, Au3+, Hg2+, iodine, and p-chloromercuribenzoate (PCMB). Diisopropyl fluorophosphate (DFP), phenylmethanesulfonyl fluoride (PMSF), toluenesuffonyl fluoride (TSF), and sodium dodecyl sulfate (SDS) were inhibitory at a concentration of 10(-3)M. Soybean trypsin inhibitor, pepstatin, leupeptins, and antipain were not inhibitory, while chymostatin caused slight inhibition. No distinct difference was observed in the effects of these compounds on the multiple forms of cathepsin A despite differences in the molecular weights of these forms (100,000, 200,000, and 420,000, respectively). In immuno-diffusion analysis, cathepsin AI, AII, and AIII which had been treated with EDTA, dithiothreitol, PCMB, and a high concentration of NaCl, gave the same precipitin patterns as the untreated enzymes, but treatment with 6 M urea caused a slight alteration of the pattern. After SDS-treatment (50 mM or more), the precipitin lines of these multiple forms fused and gave a single, identical line. This suggests that the different forms of the cathepsin A are all composed of subunits which are immunologically identical or closely related, and that the subunits are mainly bound by hydrophobic forces. This conclusion is supported by results obtained by poliacrylamide gel electrophoresis in the presence of SDS.

Animals↗
PubMed1987

Concanavalin A interactions with asparagine-linked glycopeptides. Bivalency of bisected complex type oligosaccharides.

In the preceding paper (Bhattacharyya, L., Ceccarini, C., Lorenzoni, P., and Brewer, C.F. (1987) J. Biol. Chem. 262, 1288-1293), we have demonstrated that certain high mannose and bisected hybrid type glycopeptides are bivalent for concanavalin A (ConA) binding. In the present study, we have investigated the interactions of ConA with a series of synthetic nonbisected and bisected complex type oligosaccharides and related glycopeptides. The modes of binding of the carbohydrates were studied by nuclear magnetic relaxation dispersion techniques, and their affinities were determined by hemagglutination inhibition measurements. We find that certain bisected complex type oligosaccharides are capable of binding and precipitating the lectin. The corresponding nonbisected analogs, however, bind but do not precipitate the protein. The stoichiometries of the precipitin reactions were investigated by quantitative precipitation analyses. The equivalence zones (regions of maximum precipitation) of the precipitin curves indicate that the bisected complex type oligosaccharides are bivalent for lectin binding. Data for the nonbisected analogs are consistent with their being univalent. The nuclear magnetic relaxation dispersion and precipitation data indicate that nonbisected and bisected complex type carbohydrates bind with different mechanisms and conformations. The former class binds by extended site interactions with the protein involving the 2 alpha-mannose residues on the alpha(1-6) and alpha(1-3) arms of the core beta-mannose residue. The latter class binds by only 1 of these 2 mannose residues, which leaves the other mannose residue free to bind to a second ConA molecule. The role of the bisecting GlcNAc residue in affecting the binding properties of complex type carbohydrates to ConA is discussed, and the results are related to the possible structure-function properties of complex type glycopeptides on the surface of cells.

Asparagine↗
PubMed1985

Identification of common antigens in ribosome-rich extracts from Fusobacterium necrophorum.

Ribosome-rich extracts (RRE) were prepared by differential and ultracentrifugation from 25 bovine and 6 ovine isolates of Fusobacterium necrophorum (FN) including both biotypes A and B. A pooled rabbit antiserum was prepared against whole-cell and sonicated whole-cell bacterins of F necrophorum isolate FN 3080, and a 2nd pooled rabbit antiserum was prepared against a RRE of FN 3080. The RRE of the 25 bovine isolates were tested against the FN 3080 whole-cell antiserum, using Ouchterlony double-immunodiffusion procedures. One to 3 precipitin lines were observed with the 25 isolates. The individual bovine isolates were found to have lines of identity with 5 to 21 of the other 24 isolates. The 25 bovine isolates and the 6 ovine isolates were then compared, using the FN 3080 RRE antiserum. One to 3 precipitin lines were observed for the 31 isolates with the RRE antiserum, and lines of identity were observed between all 31 of the isolates. These results indicated that common antigens are present in the RRE from a wide variety of F necrophorum isolates including both A and B biotypes.

Animals↗
PubMed1985

Hyperlipoproteinemia with albumin-lipid complex: a novel type of hyperlipoproteinemia associated with insulin-resistant diabetes mellitus.

We describe a new phenotype of hyperlipoproteinemia in two members of a family with a high degree of consanguinity. Both have a history of uncontrolled diabetes mellitus without ketoacidosis, and a family history of coronary artery disease at a relatively early age. A high degree of insulin resistance was found. The abnormal lipoprotein(s) has alpha-lipoprotein mobility on cellulose acetate electrophoresis and has a relative density of less than 1.006 as determined by ultracentrifugation of serum collected after a short fast. The fraction isolated by ultracentrifugation contains about half of the serum cholesterol and triglycerides and most of the phospholipids; the major protein component is albumin. Immunoelectrophoresis showed low concentrations of beta-lipoproteins in both sera, and two abnormal precipitin bands against monospecific antiserum to antilipoprotein A; a third member of the family showed only one abnormal precipitin band against the same antibody. We tentatively propose an abnormal gene(s) as the underlying mechanism. The insulin-resistant diabetes mellitus, probably inherited separately, may aggravate the hyperlipidemia.

Adolescent↗
PubMed1977

The bovine immune response to Brucella abortus I. A water soluble antigen precipitated by sera of some naturally infected cattle.

Selected sera from cattle naturally infected with Brucella abortus precipitate water soluble antigens extracted by sonication from B. abortus. One of these antigens resembles antigen E (Baughn and Freeman) as it is excluded from Sephadex G-200 gels, migrates anodally when electrophoresed at pH 8.6, resists heating at 100 degrees C for ten minutes and appears to be susceptible to papain digestion. Precipitins specific for this antigen remained in sera from which all detectable Brucella agglutinating antibody had been removed by adsorption with live or heat killed B. abortus. The antigen has been extracted from smooth and rough strains of B abortus. Precipitins specific for this antigen have been detected in antisera produced against Brucella canis.

Agglutination Tests↗
PubMed1985

Isolation and characterization of a newly identified antigen from ragweed pollen by organic solvent extraction.

An allergenically active protein, designated as RFB2, was isolated from short ragweed pollen using an aqueous-organic solvent system followed by gel chromatography. Antigen RFB2 was found to be homogeneous based on several criteria, e.g., column chromatography, gel electrophoresis and immunodiffusion. The molecular weight of antigen RFB2 was estimated to be approximately 85,000 daltons based on the results of gel filtration on Sepharose CL-6B. Amino acid analysis revealed that antigen RFB2 contains about twice the amount of leucine and proline and about one-third the amount of cysteine and aspartic acid found in the water soluble antigens AgE, AgK and AgRa3. Antigen RFB2 showed total immunological identity with one of the AgE and the AgK precipitin bands and partial immunological identity with the AgRa3 precipitin band.

Allergens↗
PubMed1972

African swine fever: application of immunoelectroosmophoresis for the detection of antibody.

Thirty-three pigs in three groups of nineteen, ten, and four pigs were infected with three different African swine fever (ASF) virus isolates, respectively. All virus isolates were attenuated to varying degrees by passaging in cell cultures, and they retained sufficiently low virulence to produce subacute and chronic infections in pigs. Sera collected at various intervals were tested for antibody activity by the immunoelectroosmophoresis, agar gel diffusion precipitin, and complement-fixation tests using a modified Kolmer technique. Results clearly indicated that the immunoelectroosmophoresis test is a rapid (30 minute) and accurate method with extreme sensitivity and superior to the complement-fixation and agar gel diffusion precipitin tests in detecting antibody against ASF virus. Possible use of this method in detecting ASF virus infection is suggested.

African Swine Fever↗
PubMed1978

Immunochemical studies on induction of rat liver mitochondrial serine: pyruvate aminotransferase by glucagon.

The 7- to 10-fold increase in the rat liver serine:pyruvate aminotransferase activity after glucagon administration was shown to occur mainly in the mitochondrial matrix of parenchymal cells. The enzyme was purified from glucagon-treated rat liver mitochondria to apparent homogeneity as judged by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. A specific rabbit antibody was prepared against the purified enzyme. Upon Ouchterlony double diffusion analysis, the mitochondrial extracts of glucagon-treated rat liver produced a single and fused precipitin line between the purified enzyme against the antibody. The supernatant fraction of glucagon-treated rat liver and the mitochondrial extracts of normal liver were also shown to make a single and fused precipitin line with the purified enzyme, when applied in large quantities. The quantitative immunotitration demonstrated that the glucagon-induced increase in the activity of liver serine:pyruvate aminotransferase were accompanied by the parallel increase in the amount of the enzyme antigen. Isotopic leucine incorporation studies showed that the relative rate of synthesis of the enzyme was increased approximately 10-fold by glucagon administration under the conditions employed. The rate of the degradation of the aminotransferase in the normal rat liver was a relatively slow process with a half-life of approximately 30 h. Thus the accumulation of serine:pyruvate aminotransferase in rat liver mitochondria by glucagon treatment can be ascribed mainly to the rise in the rate of enzyme synthesis.

Animals↗
PubMedDate not supplied

Late asthmatic reactions to inhalation of fractions from extracts of Candida albicans and Asperguillus fumigatus.

To investigate the mechanism of late respiratory reactions after exposure to extracts of C.albicans and A.fumigatus, patients with asthma were studied with skin tests, inhalation tests and serologic tests including precipitin tests, using purified polysaccharide fraction (PS) and protein fraction (P). Thirty five inhalation tests in 30 patients separated reactors with late onset into two groups, 7 patients with precipitins, intermediate dermal and respiratory reactions, and 10 patients with delayed skin reaction and delayed dyspnea. Exposure to PS could elicit immediate and intermediate respiratory reactions but not delayed reactions, which had been observed after exposure to Prut or crude extract, suggesting that the delayed respiratory reactions observed were immunologically different from those caused by Arthustype allergy. Skin test with PS elicited immediate, intermediate, or both reactions, and those with P evoked immediate, intermediate, or delayed reactions or some combinations of these. The same relationship of purified fractions to patterns of skin reactions as well as respiratory reactions existed in guinea pigs immunized with C.albicans. Taking the results to animal experiments into consideration it would appear that some of the late responses after exposure to Prot or crude extract of C.albicans and A.fumigatus were due to allergic mechanism of the delayed type.

Adult↗