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E Wiegeshaus

Publications and source records attributed to E Wiegeshaus.

14 recordsLinked to original sources

Animal models for experimental tuberculosis.

Animal models for tuberculosis vaccine assays have evolved through a series of sequential experiments aimed at optimizing the activity of a particular vaccine product. As a result, studies that use different animal models do not agree on the potency ranking of antituberculosis vaccines, and each major test-system variable contributes to the disagreement. Disagreements among laboratories about the efficacy of vaccines are in part due to differences in test systems. A survey of potency assays of tuberculosis vaccines suggests that, based on the choice of a specific combination of variables in the test model, an investigator can show that any given vaccine product has superior potency. In view of these problems with animal models for research on tuberculosis vaccines, we recommend that attention should be focused on those animal models that replicate the key aspects of the natural history of human tuberculosis. The development of vaccines by means of new technologies requires animal models to assay the protective potency of vaccines that, ideally, inhibit tubercle bacilli at the point of infection. The development of new tuberculosis vaccines may be handled most efficiently in a joint venture that includes both laboratories involved in vaccine production and laboratories concerned with animal models.

Animals↗

An analysis of some hypotheses related to the Chingelput bacille Calmette-Guérin trial.

Investigation of several hypotheses related to the outcome of the Chingelput Bacille Calmette-Guérin (BCG) Trial suggests at least 2 factors that might explain the major scientific puzzle of a protective effect expected of 80% and a protective effect observed of 0% (i.e., equivalent protection for BCG and placebo). One factor that explains some of the low efficacy observed for BCG in this trial is the virtual saturation level of exposure to environmental mycobacteria (EM). Studies in animal models demonstrated that the protection afforded by infection with EM was equivalent to the protection that resulted from BCG vaccination. The second factor, pathogenetic pathway, explains why there was still a high case rate for tuberculosis, even though the population was fully vaccinated by EM. This hypothesis states that tuberculosis in India, as well as in most developing countries, results primarily from exogenous reinfection, a pathway against which BCG (or EM) exerts no protective effect beyond that induced by the first episode of infection with Mycobacterium tuberculosis.

BCG Vaccine↗

Virulence of Mycobacterium tuberculosis for guinea pigs: a quantitative modification of the assay developed by Mitchison.

One of the more accepted methods of assay of virulence of tubercle bacilli is one developed by Mitchison in which guinea pigs were infected by the intramuscular route with 1.0 mg of tubercle bacilli freshly harvested from Lowenstein Jensen medium and in which virulence was based on a subjective score of the extent of gross disease in the animal 6 weeks after infection. Due to the practical difficulties involved in such an assay when routinely performed, the following modifications were made: frozen stocks of the culture were prepared prior to the date of infection, the inoculum was quantitated by means of colony forming units, and virulence was based on the number of tubercle bacilli recovered from the spleen of infected animals 6 weeks after infection. A significant correlation was obtained between the findings from the Mitchison assay of virulence and the modification. The latter assay is recommended for its quantitative and objective features.

Animals↗

A guinea pig model of experimental airborne tuberculosis for evaluation of the response to chemotherapy: the effect on bacilli in the initial phase of treatment.

The purpose of this study was to develop and evaluate a guinea pig model of experimental airborne tuberculosis for its ability to assess chemotherapeutic regimens for their efficacy against virulent tubercle bacilli in vivo during the initial phase of treatment. The tissues examined included primary lung lesions and the metastatic foci in lung and spleen which result from the naturally occurring bacillaemia. The treatments examined, INH + RIF, INH + EMB, EMB + RIF, were initiated 4 weeks after infection and were continued for 8 weeks. Although minor differences were observed in the time of onset of a significant bactericidal effect or in the rate of decline in the microbial population, all three treatment combinations resulted in a significant reduction in the number of M. tuberculosis H37Rv recovered from primary lung lesions, primary lesion-free lung lobes and spleen. X-rays taken of excised inflated lung lobes showed a relationship between the degree of calcification of primary lung lesions and the number of surviving bacilli.

Animals↗

Virulence for guinea pigs of tubercle bacilli isolated from the sputum of participants in the BCG trial, Chingleput District, South India.

This study, conducted in Madras, India and in Madison, Wisconsin, USA, was concerned with the virulence of isolates of Mycobacterium tuberculosis obtained from the sputum of individuals living in the Chingleput district of south India. The following results were obtained. 1. The findings of Mitchison with respect to the predominance of low virulence for guinea pigs among isolates from persons living Madras, were confirmed on isolates from the sputum of residents of the Chingleput district. 2. A high correlation was found between the log10 number of tubercle bacilli recovered from the spleen of guinea pigs infected intramuscularly with 1.0 mg of tubercle bacilli and the root index of virulence. 3. A high correlation was found between the log10 number of tubercle bacilli recovered from the spleen of guinea pigs infected intramuscularly with 1.0 mg of tubercle bacilli and the number recovered from the spleen of guinea pigs infected by the respiratory route with 5-10 tubercle bacilli. 4. Relatively low correlations were found between RIV and the susceptibility of isolates to thiophene-2 carboxylic acid hydrazide or to hydrogen peroxide.

Animals↗

Resolution and identification of ribosomes from mycobacteria.

Ribosomes were isolated from mycobacteria (BCG) by differential centrifugation by use of a modification of Nirenberg's procedure. Individual components of the ribosomal spectrum were resolved in linear sucrose density gradients. Sedimentation constants of the individual components were determined by analytical ultracentrifugation. Ribosomes from escherichia coli labeled with (14)C-uracil were used as markers for an independent confirmation of the identity of individual peaks. The typical ribosomal spectrum of 70S units and 50S and 30S subunits was obtained in tris(hydroxymethyl)aminomethane buffer with 0.01 m Mg(CH(3)COO)(2). This corresponds to the ribosomal spectrum obtained in E. coli under comparable conditions. In 0.0001 m Mg(2+), the 70S units were dissociated to 50S and 30S particles. Pancreatic ribonuclease produced no significant changes in main ribosomal components. The isolation techniques used in this work precluded the recognition of polyribosomes.

Bacterial Proteins↗

Host-parasite relationships in experimental airborne tuberculosis. I. Preliminary studies in BCG-vaccinated and nonvaccinated animals.

Smith, D. W. (University of Wisconsin, Madison), E. Wiegeshaus, R. Navalkar, and A. A. Grover. Host-parasite relationships in experimental airborne tuberculosis. I. Preliminary studies in BCG-vaccinated and nonvaccinated animals. J. Bacteriol. 91:718-724. 1966.-Previous studies from this laboratory on immunogenicity and allergenicity of defatted mycobacterial vaccines involved subcutaneous challenge of guinea pigs and killing of the animals 6 weeks later to evaluate the amount of disease. This type of experiment has discontinued in this laboratory in favor of an airborne challenge type of experiment, with the advantages that animals can be challenged with small numbers of bacilli by a natural route, and the number of primary lesions, the rate of spread from those lesions, and the rate of bacillary multiplication can be used to evaluate protection. Experiments to determine uniformity of infection showed that a fair degree of uniformity resulted when seven guinea pigs were exposed simultaneously, and were studied 3 weeks later to determine numbers of primary lesions and bacilli in the tissues. A less satisfactory degree of uniformity was obtained when more animals were exposed at one time. BCG-vaccinated and nonvaccinated animals were studied to determine the earliest time and the optimal time for killing the animals to detect the effects of vaccination. In guinea pigs, the degree of protection assessed by lesion counts is time-dependent, but the degree of protection assessed by viable counts of bacilli in the tissues was relatively constant 3 to 12 weeks after infection. Mice vaccinated subcutaneously with BCG were not protected against infection at any interval between 2 and 19 weeks. Guinea pigs vaccinated subcutaneously with the same lot of vaccine were protected as judged by counts of viable bacilli in the tissues 3 weeks after infection.

Air↗

Mycoside G, a Specific Glycolipid in Mycobacterium marinum (Balnei).

Navalkar, R. G. (University of Wisconsin, Madison), E. Wiegeshaus, E. Kondo, H. K. Kim, and D. W. Smith. Mycoside G, a specific glycolipid in Mycobacterium marinum (Balnei). J. Bacteriol. 90:262-265. 1965.-A new specific glycolipid in extracts prepared from strains designated Mycobacterium marinum and M. balnei has been demonstrated by use of the techniques of column chromatography and infrared spectroscopy. Since there is now agreement among many workers that M. marinum and M. balnei are identical, the demonstration of the same specific glycolipid in both species is not surprising. This substance, which we have designated mycoside G, is chemically similar to mycosides A and B, and apparently differs only in the sugar moiety. In addition, the lipids extracted from these cultures contain phthiocerol dimycocerosate, a wax component found also in M. tuberculosis and M. bovis.

Journal Article↗