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Biomedical subjects

D E Pietz

Publications and source records attributed to D E Pietz.

18 recordsLinked to original sources
PubMed1984

Application of an enzyme-linked immunosorbent assay for detection of Brucella antigens in vaginal discharge of cows.

An enzyme-linked immunosorbent assay (ELISA) was developed for detecting Brucella abortus. Carbodiimide cyanamide was used as an antigen-coupling agent and NH4Cl was used to neutralize residual active charges present on the cuvette surface. Suitable ELISA reactions were observed in vaginal swabs placed in phosphate-buffered saline solution. The ELISA described may be useful under controlled laboratory conditions for detecting B abortus-infected cows shedding bacteria in vaginal secretions.

Animals↗
PubMed1983

Detection of brucella antibodies of different immunoglobulin classes in cow milk by enzyme-linked immunosorbent assay.

Enzyme-linked immunosorbent assays were conducted on milk of cows from which Brucella abortus was isolated and that of noninfected controls. Horseradish peroxidase-labeled rabbit antibovine immunoglobulins IgG, IgG1, and IgA were used as conjugates. A heat-killed whole-cell suspension of B abortus strain 19 was used as the antigen. Differences in antibody profiles were observed in milk of cows from which B abortus was isolated and in milk of noninfected cows. Antibody profiles were similar in milk of cows infected with B abortus and that of cows from which B abortus strain 19 was isolated.

Animals↗
PubMed1980

Development of an enzyme-linked immunosorbent assay for detecting antibodies in sera of Brucella suis-infected swine.

An enzyme-linked immunosorbent assay was developed using a heat-killed Brucella suis antigen for detecting antibodies in the sera of swine from which B. suis was isolated. Optimal enzyme-linked immunosorbent assay reactions were obtained using heat-killed B. suis antigen at a concentration comparable to McFarland Standard No. 1. Statistically significant differences were observed in the enzyme-linked immunosorbent assay results of 40 animals from which B. suis was isolated and the results for 48 noninfected swine at serum dilutions of 1:25 and 1:50 (P < 0.0001). The enzyme-linked immunosorbent assay is a rapid reproducible test which can be readily automated that appears to have practical value for screening large numbers of breeding and slaughter swine for brucellosis.

Animals↗
PubMed1979

Enzyme immunoassay for detecting Brucella antibodies in cow's milk.

An enzyme immunoassay (EIA) was developed for detecting Brucella antibodies in milk of cows infected with Brucella abortus. The enzyme immunoassay using heat-killed cells of B. abortus strain 19 was of comparable sensitivity to the Brucella ring test in detecting antibodies in milk of a reference positive control cow experimentally infected with B. abortus strain 2308 and in milk of 16 naturally infected cows from which B. abortus was isolated. No detectable enzyme immunoassay reactions were present in milk of 11 noninfected controls. The enzyme immunoassay is a procedure which can be readily automated so that screening tests for brucellosis could be conducted at a reference laboratory where uniform conditions can be maintained.

Animals↗
PubMed1979

Temporal cell-mediated immune responses of cattle following experimental and natural exposure to living Brucella abortus.

A study on cell-mediated immune responses in cattle with different exposure experiences to Brucella abortus was conducted by an in vitro lymphocyte stimulation assay. The purpose of this study was to determine how soon the cell-mediated immune responses would be detected following experimental exposure to B. abortus and to study the cell-mediated immune trend following experimental and natural exposure of cattle to B. abortus. The first positive cell-mediated immune responses occurred one to two weeks after experimental inoculation with living B. abortus strain 2308. The cell-mediated immune responses in these animals appeared at least one week before the appearance of of B. abortus serum agglutinating antibodies. Animals which were naturally infected with B. abortus biotypes 1 and 2 demonstrated positive cell-mediated immune responses throughout the study.

Animals↗
PubMed1978

Whole-blood lymphocyte stimulation assay for measurement of cell-mediated immune responses in bovine brucellosis.

A study was conducted to develop an in vitro whole-blood lymphocyte stimulation assay for measurement of cell-mediated immune response in bovine brucellosis. A soluble antigen (BASA) prepared from killed cells of Brucella abortus 1119-3 was used. Cattle infected with B. abortus field strains, B. abortus 19 calfhood- and adult-vaccinated cattle, and nonexposed cattle were tested. Blood was diluted 10-fold in RPMI-1640 medium (without added serum) and cultured with BASA (at a concentration of 2.2 microgram per culture) at varying times of incubation. Results were assayed for [3H]thymidine incorporation into deoxyribonucleic acid. A 6-day period was found to be optimal for incubating blood cultures to achieve maximum specific lymphocyte stimulation. Serological tests and bacteriological isolation attempts were conducted simultaneously with lymphocyte stimulation tests, and there was a significant correlation between cell-mediated immune response and bacteriological findings. There was a significant correlation between cell-mediated immune response and the level of serum antibodies on a group basis, but there was little correlation between the two systems on individual infected animals. Among vaccinated animals there was little or no correlation between cell-mediated immune and humoral responses. The whole-blood assay was found to be simple, fast, sensitive, and reproducible.

Animals↗
PubMed1978

Kinetics of in vitro bovine lymphocyte immunostimulation with a Brucella abortus antigen.

A Brucella abortus-soluble antigen was investigated, using in vitro assay of lymphocyte immunostimulation, to determine which concentration of this antigen and which period of incubation of the lymphocyte cultures would induce maximum specific lymphocyte immunostimulation as an additional method for further study of B abortus infection in cattle. Soluble antigen was prepared from autoclaved cells of B abortus strain 1119-3. Peripheral blood lymphocytes were obtained from cattle infected with B abortus and from healthy control cattle not infected with B abortus. The lymphocytes were prepared by the Ficoll-Hypaque density gradient technique, suspended in RPMI 1640 medium (1.5 X 10(6)/ml), cultured with several dilutions of soluble antigen, and incubated. Prior to termination of incubation, cultures were labeled with 1 muCi of [3H]thymidine and, after harvesting, assayed for [3H]thymidine incorporation in DNA by a liquid scintillation spectrometer. Maximum specific immunostimulation of lymphocytes from B abortus-infected cattle was induced in this assay system with 6 days' incubation and 22 microgram of protein/ml/1.5 X 10(6) lymphocytes, using protein content to express concentration of soluble antigen in this system.

Animals↗
PubMed1978

Specific lymphocyte stimulation in cattle naturally infected with strains of Brucella abortus and cattle vaccinated with Brucella abortus strain 19.

Cell-mediated immune responses in cattle naturally infected with strains of Brucella abortus and in cattle vaccinated with B abortus strain 19 during calfhood were studied by an in vitro lymphocyte-stimulation procedure. Lymphocytes were prepared from peripheral bovine blood by the Ficoll-diatrizoate technique, suspended in RPMI-1640 medium (1.5 X 10(6) lymphocytes/ml), cultured with B abortus-soluble antigen or phytohemagglutinin, and incubated for 6 days. Sixteen hours prior to termination of incubation, cultures were labeled with 1 muCi of [3H]thymidine (3HdT) and, after harvesting, assayed for 3HdT incorporation into DNA by liquid scintillation spectrometry. Lymphocytes from cattle with bacteriologically confirmed isolation of B abortus underwent a significantly higher lymphocyte stimulation with B abortus-soluble antigen than did cattle vaccinated with B abortus strain 19 during calfhood (P less than 0.005). Standard seroagglutination tests were conducted simultaneously with lymphocyte-stimulation tests, but there was no apparent correlation between levels of humoral antibodies and the cell-mediated immune responses as measured by in vitro specific lymphocyte stimulation.

Agglutination Tests↗
PubMed1978

Cell-mediated immune responses in swine from a herd infected with Brucella suis.

Cell-mediated immune (CMI) responses in swine naturally infected with Brucella suis biotype 3, swine suckling an infected sow and Brucella-noninfected swine were studied by an in vitro lymphocyte transformation procedure. The antigen used was a soluble antigen prepared from killed cells of B suis biotype 3. Lymphocytes were prepared from peripheral swine blood by the Ficoll-Hypaque technique, suspended in RPMI-1640 medium (1.5 X 10(6) lymphocytes/ml), cultured with B suis-soluble antigen or concanconcanavalin A, and incubated for 6 days. Sixteen hours prior to termination of incubation, cultures were labeled with 1 muCi of [3H]thymidine and, after harvesting, were assayed for [3H]thymidine incorporation into DNA by liquid scintillation spectrometry. Agglutination tests were conducted on sera collected simultaneously with samples for lymphocyte-stimulation tests. The B suis-soluble antigen elicited specific stimulation in lymphocytes from infected pigs. On a group basis, there was high correlation between the amount of serum antibodies and specific lymphocyte stimulation, but on an individual animal basis, there was little correlation of the results of both systems in infected swine. There was high correlation between recovery of Brucella from the tissues of animals and the degree of CMI response. Suckling pigs from an infected sow did not develop CMI responses, as measured by our system.

Agglutination Tests↗
PubMed1976

Evaluation of five mediums for the stabilization of Brucella abortus strain 19 dessicated by lyophilization.

Lyophilized Brucella abortus Strain 19 vaccines were prepared using five recommended mediums. These five vaccines were compared with each other and with liquid vaccine relative to their suitability for (1) maintaning distinguishing cultural characteristics; (2) maintaining viability during lyophilization; (3) quality of restoration; (4) maintaining viability under various storage temperatures and (5) maintaining stable, antigenic and virulence qualities.

Brucella Vaccine↗