Changes to the Natural Killer Cell Repertoire after Therapeutic Hepatitis B DNA Vaccination

Background

Improvements to the outcome of adaptive immune responses could be achieved by inducing specific natural killer (NK) cell subsets which can cooperate with dendritic cells to select efficient T cell responses. We previously reported the induction or reactivation of T cell responses in chronic hepatitis B patients vaccinated with a DNA encoding hepatitis B envelope proteins during a phase I clinical trial.

Methodology/Principal Findings

In this study, we examined changes in the peripheral NK cell populations occurring during this vaccine trial using flow cytometry analysis. Despite a constant number of NK cells in the periphery, a significant increase in the CD56^bright population was observed after each vaccination and during the follow up. Among the 13 different NK cell markers studied by flow cytometry analysis, the expression of CD244 and NKG2D increased significantly in the CD56^bright NK population. The ex vivo CD107a expression by CD56^bright NK cells progressively increased in the vaccinated patients to reach levels that were significantly higher compared to chronically HBV-infected controls. Furthermore, modifications to the percentage of the CD56^bright NK cell population were correlated with HBV-specific T cell responses detected by the ELISPOT assay.

Conclusions/Significance

These changes in the CD56^bright population may suggest a NK helper effect on T cell adaptive responses. Activation of the innate and adaptive arms of the immune system by DNA immunization may be of particular importance to the efficacy of therapeutic interventions in a context of chronic infections.

Trial Registration

ClinicalTrials.gov {"type":"clinical-trial","attrs":{"text":"NCT00988767","term_id":"NCT00988767"}}NCT00988767     

Structure and ultrastructure of the silk glands and spinneret of Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae)

This study provides comprehensive documentation of silk production in the pest moth Helicoverpa armigera from gland secretion to extrusion of silk thread. The structure of the silk glands, accessory structures and extrusion apparatus are reported. The general schema of the paired silk glands follows that found for Lepidoptera. Morphology of the duct, silk press, muscle attachments and spigot are presented as a three-dimensional reconstruction and the cuticular crescent-shaped profile of the silk press is demonstrated in both open and closed forms with attendant muscle blocks, allowing advances in our knowledge of how the silk press functions to regulate the extrusion of silk. Growth of the spigot across instars is documented showing a distinctive developmental pattern for this extrusion device. Its shape and structure are related to use and load-bearing activity.     

Mutations in tap uncouple RNA export activity from translocation through the nuclear pore complex

Interactions between transport receptors and phenylalanine-glycine (FG) repeats on nucleoporins drive the translocation of receptor-cargo complexes through nuclear pores. Tap, a transport receptor that mediates nuclear export of cellular mRNAs, contains a UBA-like and NTF2-like folds that can associate directly with FG repeats. In addition, two nuclear export sequences (NESs) within the NTF2-like region can also interact with nucleoporins. The Tap-RNA complex was shown to bind to three nucleoporins, Nup98, p62, and RanBP2, and these interactions were enhanced by Nxt1. Mutations in the Tap-UBA region abolished interactions with all three nucleoporins, whereas the effect of point mutations within the NTF2-like domain of Tap known to disrupt Nxt1 binding or nucleoporin binding were nucleoporin dependent. A mutation in any of these Tap domains was sufficient to reduce RNA export but was not sufficient to disrupt Tap interaction with the NPC in vivo or its nucleocytoplasmic shuttling. However, shuttling activity was reduced or abolished by combined mutations within the UBA and either the Nxt1-binding domain or NESs. These data suggest that Tap requires both the UBA- and NTF2-like domains to mediate the export of RNA cargo, but can move through the pores independently of these domains when free of RNA cargo.     

Campylobacter jejuni Contains Two Fur Homologs: Characterization of Iron-Responsive Regulation of Peroxide Stress Defense Genes by the PerR Repressor

Expression of the peroxide stress genes alkyl hydroperoxide reductase (ahpC) and catalase (katA) of the microaerophile Campylobacter jejuni is repressed by iron. Whereas iron repression in gram-negative bacteria is usually carried out by the Fur protein, previous work showed that this is not the case in C. jejuni, as these genes are still iron repressed in a C. jejuni fur mutant. An open reading frame encoding a Fur homolog (designated PerR for "peroxide stress regulator") was identified in the genome sequence of C. jejuni. The perR gene was disrupted by a kanamycin resistance cassette in C. jejuni wild-type and fur mutant strains. Subsequent characterization of the C. jejuni perR mutants showed derepressed expression of both AhpC and KatA at a much higher level than that obtained by iron limitation, suggesting that expression of these genes is controlled by other regulatory factors in addition to the iron level. Other iron-regulated proteins were not affected by the perR mutation. The fur perR double mutant showed derepressed expression of known iron-repressed genes. Further phenotypic analysis of the perR mutant, fur mutant, and the fur perR double mutant showed that the perR mutation made C. jejuni hyperresistant to peroxide stress caused by hydrogen peroxide and cumene hydroperoxide, a finding consistent with the high levels of KatA and AhpC expression, and showed that these enzymes were functional. Quantitative analysis of KatA expression showed that both the perR mutation and the fur mutation had profound effects on catalase activity, suggesting additional non-iron-dependent regulation of KatA and, by inference, AhpC. The PerR protein is a functional but nonhomologous substitution for the OxyR protein, which regulates peroxide stress genes in other gram-negative bacteria. Regulation of peroxide stress genes by a Fur homolog has recently been described for the gram-positive bacterium Bacillus subtilis. C. jejuni is the first gram-negative bacterium where non-OxyR regulation of peroxide stress genes has been described and characterized.     

Electrophoretic Mobility of Amylase in Drosophilids Indicates Adaptation to Ecological Diversity

Understanding the significance of electrophoretic variation is of interest for both ecological and evolutionary genetics. Although there has been a very active neutralist–selectionist debate about the patterns of electrophoretic variation in natural populations, it is only recently that charged amino acids have been shown to be important in enzyme adaptation. In this study we carried out a broad electrophoretic survey of amylase variation in 150 species of Drosophilids. The distribution of amylase electromorphs was found to be correlated with the geographical origin of the flies. Generally the faster migrating variants are found in warmer temperatures. There is also a correlation with the feeding habits of the species, in particular, fungus feeders consistently showed a deviating pattern of electrophoretic mobility. These correlations between ecological diversity and electrophoretic patterns indicate that at least some of the changes in charged amino acids are adaptive, and result from selection to cope with specific environments.     

Novel mechanism for high-altitude adaptation in hemoglobin of the Andean frog Telmatobius peruvianus

In contrast to birds and mammals, no information appears to be available on the molecular adaptations for O(2) transport in high-altitude ectothermic vertebrates. We investigated Hb of the aquatic Andean frog Telmatobius peruvianus from 3,800-m altitude as regards isoform differentiation, sensitivity to allosteric cofactors, and primary structures of the alpha- and beta-chains, and we carried out comparative O(2)-binding measurements on Hb of lowland Xenopus laevis. The three T. peruvianus isoHbs show similar functional properties. The high O(2) affinity of the major component results from an almost complete obliteration of chloride sensitivity, which correlates with two alpha-chain modifications: blockage of the NH(2)-terminal residues and replacement by nonpolar Ala of polar residues Ser and Thr found at position alpha131(H14) in human and X. leavis Hbs, respectively. The data indicate adaptive significance of alpha-chain chloride-binding sites in amphibians, in contrast to human Hb where chloride appears mainly to bind in the cavity between the beta-chains. The findings are discussed in relation to other strategies for high-altitude adaptations in amphibians.     

Mutational spectrum of the iduronate-2-sulfatase (IDS) gene in 36 unrelated Russian MPS II patients

We present a mutational analysis of the iduronate-2-sulfatase (IDS) gene of 36 Russian patients with Hunter syndrome. Among 29 mutant alleles, there were 19 missense mutations, 1 nonsense mutation, 6 mutations affecting splice sites, and 3 major structural alterations resulting in deletions. Of the 25 different mutations, 15 are novel and unique. Most of the missense mutations result in intermediate or severe phenotypes. Received: 1 June 1998 / Accepted: 27 July 1998     

Effect of octanoic acid on ethylene-mediated flower induction in Dutch iris

Treatment of Dutch iris (Iris × hollandica Hoog. cv. Sapphire Beauty) bulbs with ethylene prior to precooling stimulated flowering in bulbs of various sizes. In large sized bulbs exposure to ethylene followed by precooling resulted in 100% flowering over a five months period after planting. Flowering in control bulbs which were not treated with ethylene prior to precooling was limited to 67% during the same five months period. In medium sized bulbs flowering in the ethylene treatment was 90% compared to 75% in the control. However, the biggest stimulation of flowering by ethylene was found in small sized bulbs (from 16 to 56%). Application of octanoic acid for a short time period prior to exposure to ethylene stimulated flowering in all bulb sizes. After five months the final percentage flowering in large and medium sized bulbs of the octanoic acid plus ethylene treatment did not differ from that of the ethylene only treatment. However, the initial rate of flowering was higher in the former treatment. In small bulbs the percentage flowering was much higher in the octanoic acid plus ethylene treatment than in the ethylene only treatment. The results of this study indicate that, just as in certain flowers, fruit and seeds, treatment with octanoic acid stimulates ethylene sensitivity in Dutch iris bulbs. The sensitivity of untreated bulbs to ethylene was highest in large bulbs and lowest in small bulbs. This correlated well with the endogenous octanoic acid content of the bulbs. Octanoic acid levels were highest in large bulbs and lowest in small Bulbs. It appears that the endogenous levels of octanoic acid in the bulbs is determined prior to the onset of dormancy.     

Isolation and purification of cell wall-associated arginine carboxypeptidase fromStreptococcus mitis ATCC 15909

An arginine carboxypeptidase was isolated from the cell walls ofStreptococcus mitis ATCC 15909 by mutanolysin extraction of the walls. The enzyme was purified 32-fold by gel filtration on Sephacryl S-300, affinity chromatography on Arginine-Sepharose 4B and by rechromatography on Sephacryl S-300. The molecular mass of the enzyme was calculated to 122 kDa by gel filtration. The enzyme released arginine from the carboxy terminal of hippurylarginine and, at a low rate, lysine from furylacryloylalanyl-lysine and hippuryl-lysine. The carboxypeptidase seemed firmly bound to the cell wall because SDS treatment of the walls did not release measurable amounts of activity.