Reducing the Activity and Secretion of Microbial Antioxidants Enhances the Immunogenicity of BCG

Background

In early clinical studies, the live tuberculosis vaccine Mycobacterium bovis BCG exhibited 80% protective efficacy against pulmonary tuberculosis (TB). Although BCG still exhibits reliable protection against TB meningitis and miliary TB in early childhood it has become less reliable in protecting against pulmonary TB. During decades of in vitro cultivation BCG not only lost some genes due to deletions of regions of the chromosome but also underwent gene duplication and other mutations resulting in increased antioxidant production.

Methodology/Principal Findings

To determine whether microbial antioxidants influence vaccine immunogenicity, we eliminated duplicated alleles encoding the oxidative stress sigma factor SigH in BCG Tice and reduced the activity and secretion of iron co-factored superoxide dismutase. We then used assays of gene expression and flow cytometry with intracellular cytokine staining to compare BCG-specific immune responses in mice after vaccination with BCG Tice or the modified BCG vaccine. Compared to BCG, the modified vaccine induced greater IL-12p40, RANTES, and IL-21 mRNA in the spleens of mice at three days post-immunization, more cytokine-producing CD8+ lymphocytes at the peak of the primary immune response, and more IL-2-producing CD4+ lymphocytes during the memory phase. The modified vaccine also induced stronger secondary CD4+ lymphocyte responses and greater clearance of challenge bacilli.

Conclusions/Significance

We conclude that antioxidants produced by BCG suppress host immune responses. These findings challenge the hypothesis that the failure of extensively cultivated BCG vaccines to prevent pulmonary tuberculosis is due to over-attenuation and suggest instead a new model in which BCG evolved to produce more immunity-suppressing antioxidants. By targeting these antioxidants it may be possible to restore BCG''s ability to protect against pulmonary TB.     

Ion Effects on the H+-Translocating Adenosine Triphosphatase and Pyrophosphatase Associated with the Tonoplast of Chora corallina

H⁺-translocating ATPase and pyrophosphatase (PPase) associatedwith the tonoplast of Chara corallina were isolated with theaid of a perfusion technique, and the effects of ions on theiractivities were studied. All the alkali metal cations testedstimulated the ATPase and ATPdependent H⁺ pumping activitiesonly by 10 to 40%. Anions, on the other hand, strongly affectedthe activities. Potassium salts of Cl^- and Br^- stimulated them,while F^- and NO₃^- inhibited them. By contrast, the H⁺-translocatingPPase was insensitive to anions but sensitive to cations. Theorder of cation stimulation was Rb⁺=K⁺>Cs⁺>Na⁺=Li⁺>choline⁺.NO₃^- (50 mil), thought to be a specific inhibitor of the tonoplast-typeH⁺-ATPase, inhibited the ATPdependent H⁺ pumping almost completelybut the ATPase activity by only about 50%. Na⁺ inhibited thePP₁-dependent H⁺ pumping (I_5O=5OmM) in the presence of 50 mMKCl but not the ATP-dependent one. The PPase was more sensitiveto F^- (I₅₀=400µM) than the ATPase. Both the H⁺-ATPaseand the H⁺-PPase required Mg²⁺ for their activities, althoughan excess was inhibitory to both. The different sensitivitiesof the PP₁-dependent and the ATP-dependent H⁺- pumping enzymesto ions correspond to the tonoplast enzymes of higher plantsand may be used as "markers" to distinguish between these enzymesin characean cells (Received October 2, 1987; Accepted May 18, 1988)     

The Position and Length of the Steroid-Dependent Hypersensitive Region in the Mouse Mammary Tumor Virus Long Terminal Repeat Are Invariant despite Multiple Nucleosome B Frames

Stimulation of the mouse mammary tumor virus with steroids results in the generation of a DNase I-hypersensitive region (HSR) spanning the hormone responsive element (HRE) in the long terminal repeat. Restriction enzymes were used to characterize the accessibility of various sites within the HSR of mouse mammary tumor virus long terminal repeat-reporter constructions in four different cell lines. The glucocorticoid-dependent HSR was found to span minimally 187 bases, a stretch of DNA longer than that associated with histones in the core particle. Although the 5′-most receptor binding site within the HRE is downstream of −190, hypersensitive sites were found further upstream to at least −295. The relationship in the accessibility between pairs of sites in the vicinity of the HSR was further examined in one cell line by a two-enzyme restriction access assay. In the uninduced state, the accessibilities at these sites were found to be independent of each other. In contrast, when stimulated with hormone, the accessibilities at these sites were observed to become linked. That is, once a distinct promoter was activated, all of the sites within the HSR of that molecule became accessible. The HSR formed along an invariant stretch of DNA sequence despite the multiplicity of nucleosome frames in the nucleosome B region, where the HRE is located. The results indicate that the macroscopic length of the HSR does not arise from core length-remodeling events in molecules containing Nuc-B in alternative positions.     

Two common nonsynonymous paraoxonase 1 (<Emphasis Type="Italic">PON1</Emphasis>) gene polymorphisms and brain astrocytoma and meningioma

Background  

Human serum paraoxonase 1 (PON1) plays a major role in the metabolism of several organophosphorus compounds. The enzyme is encoded by the polymorphic gene PON1, located on chromosome 7q21.3. Aiming to identify genetic variations related to the risk of developing brain tumors, we investigated the putative association between common nonsynonymous PON1 polymorphisms and the risk of developing astrocytoma and meningioma.     

Effect of auxins and ectomycorrhizal elicitors on wall-bound proteins and enzymes of spruce [Picea abies (L.) Karst.] cells

Summary Elicitors of the ectomycorrhizal fungus Hebeloma crustuliniforme and auxins (IAA, NAA and 2,4-D) were tested for their effects on apoplastic proteins and enzymes of suspension cultured cells of Picea abies (L.) Karst. The ectomycorrhizal elicitor increased the amount of some ionically wall-bound proteins (36, 28, 24, 21 kDa) and decreased the amount of others (61, 22 kDa). The elicitor triggered an H₂O₂ burst and enhanced the peroxidase (EC 1.11.1.7) activity of the Picea cells by increasing one of the two wall-bound peroxidase isoforms. Auxins significantly suppressed the elicitor induction of peroxidase but did not influence the elicitor-triggered H₂O₂ burst. The elicitors and auxin did not change the amount and the pattern of wall-bound invertase isoforms (EC 3.2.1.26) of spruce cells. However, auxin reduced the uptake of glucose by spruce cells and increased the acidification of the cell culture medium. Since Hebeloma lacks apoplastic invertase as well as a sucrose uptake system, utilization of plant-derived sucrose depends on the apoplastic plant invertase activity. Although the host invertase is constitutive, the fungus might be able to increase this invertase activity within a mycorrhiza by lowering the pH of the interface towards the pH optimum of the enzyme via the action of auxin. This fungus-released hormone could increase the H⁺ extrusion of plant cells by activation of the plant membrane H⁺-ATPases. Additionally, an auxin-dependent suppression of glucose uptake by cortical root cells could improve the glucose supply for the fungus. Furthermore, the fungal auxin might suppress the elicitor induced formation of defense enzymes, such as peroxidase.     

Interannual variability in dissolved inorganic nutrients in northern San Francisco Bay estuary

Nearly two decades of seasonal dissolved inorganic nutrient-salinity distributions in northern San Francisco Bay estuary (1960–1980) illustrate interannual variations in effects of river flow (a nutrient source) and phytoplankton productivity (a nutrient sink). During winter, nutrient sources dominate the nutrient-salinity distribution patterns (nutrients are at or exceed conservative mixing concentrations). During summer, however, the sources and sinks are in close competition. In summers of wet years, the effects of increased river flow often dominate the nutrient distributions (nutrients are at or less than conservative mixing concentrations), whereas in summers of dry years, phytoplankton productivity dominates (the very dry years 1976–1977 were an exception for reasons not yet clearly known). Such source/sink effects also vary with chemical species. During summer the control of phytoplankton on nutrient distributions is apparently strongest for ammonium, less so for nitrate and silica, and is the least for phosphate. Furthermore, the strength of the silica sink (diatom productivity) is at a maximum at intermediate river flows. This relation, which is in agreement with other studies based on phytoplankton abundance and enumeration, is significant to the extent that diatoms are an important food source for herbivores.The balance or lack of balance between nutrient sources and sinks varies from one estuary to another just as it can from one year to another within the same estuary. At one extreme, in some estuaries river flow dominates the estuarine dissolved inorganic nutrient distributions throughout most of the year. At the other extreme, phytoplankton productivity dominates. In northern San Francisco Bay, for example, the phytoplankton nutrient sink is not as strong as in less turbid estuaries. In this estuary, however, river effects, which produce or are associated with near-conservative nutrient distributions, are strong even at flows less than mean-annual flow. Thus, northern San Francisco Bay appears to be an estuary in between the two extremes and is shifted closer to one extreme or the other depending on interannual variations in river flow.     

Contrasting trait responses to latitudinal climate variation in two lineages of an invasive grass

Plants are expected to respond to global environmental change through shifts in functional traits and in their ranges. These shifts could alter productivity and interactions among species or genetic lineages, ultimately leading to changes in distributions and abundance. In particular, cosmopolitan species are predicted to increase growth with decreasing latitude due to differences in climate and temperature. The pattern of changes in growth may vary among genotypes within species, leading to different responses with latitude. To evaluate whether climate can affect geographically distinct genotypes of cosmopolitan invasive species differently, we evaluated the trait responses of two lineages of the common reed, Phragmites australis, to variation in environmental conditions spanning North America’s Atlantic coast. Using three reciprocal transplant common gardens, we tested for the effects of garden location and plant lineage on traits related to biomass production, flowering frequency, leaf morphology, and leaf-level physiology. We found that aboveground biomass, stem density, and flowering frequency responded non-linearly to increasing latitude in one or both lineages. These results suggest that measures of plant traits over narrow latitudinal ranges may not accurately reflect organismal-level responses to global change at broad spatial scales. Given the responses to latitude that we observed in P. australis, we propose that feedbacks between growth and reproductive rate will influence range shifts in these two lineages. Such range shifts could lead to genetic admixtures, subsequently yielding more productive, locally-adapted genotypes.     

Versuche zur elektronenmikroskopischen Darstellbarkeit saurer Mucopolysaccharide mit kolloidalen Eisenlösungen

Zusammenfassung Der Einfluß einiger Aldehyde und Carnoyscher Flüssigkeit auf das Eindringvermögen von Lösungen verschiedener Eisenverbindungen wurde untersucht. Bei allen Fixationsmedien und Eisenlösungen reagieren nur die Randpartien der Gewebeproben. Offensichtlich werden Gewebekomponenten durch Reaktion mit dem Eisen für dieses impermeabel.

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Experiments for electron-microscopical demonstration of acid mucosubstances by colloidal iron methods

Summary The effect of some aldehydes and of Carnoy's fluid on the penetration of solutions of different iron compounds has been investigated. Independent of the fixation medium and the iron compound used only the peripheral zone of tissue samples do react. This seems to be due to the reaction of iron with tissue components making them impermeable to iron.

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Mit Unterstützung durch die Deutsche Forschungsgemeinschaft.