Functional Characterization of Antibodies against Neisseria gonorrhoeae Opacity Protein Loops

Background: The development of a gonorrhea vaccine is challenged by the lack of correlates of protection. The antigenically variable neisserial opacity (Opa) proteins are expressed during infection and have a semivariable (SV) and highly conserved (4L) loop that could be targeted in a vaccine. Here we compared antibodies to linear (Ab linear) and cyclic (Abcyclic) peptides that correspond to the SV and 4L loops and selected hypervariable (HV2) loops for surface-binding and protective activity in vitro and in vivo. Methods/Findings: AbSV cyclic bound a greater number of different Opa variants than AbSV linear, including variants that differed by seven amino acids. Antibodies to the 4L peptide did not bind Opa-expressing bacteria. Ab SV cyclic and Ab HV2 cyclic, but not AbSV linear or AbHV2 linear agglutinated homologous Opa variants, and AbHV2BD cyclic but not AbHV2BD linear blocked the association of OpaB variants with human endocervical cells. Only AbHV2BD linear were bactericidal against the serum resistant parent strain. Consistent with host restrictions in the complement cascade, the bactericidal activity of Ab HV2BD linear was increased 8-fold when rabbit complement was used. None of the antibodies was protective when administered vaginally to mice. Antibody duration in the vagina was short-lived, however, with,50 % of the antibodies recovered 3 hrs postadministration. Conclusions: We conclude that an SV loop-specific cyclic peptide can be used to induce antibodies that recognize a broad spectrum of antigenically distinct Opa variants and have agglutination abilities. HV 2 loop-specific cyclic peptides elicite

Rapid Publication Multiple Gonococcal Pilin Antigenic Variants Are Produced during Experimental Human Infections

Abstract Gonococcal pilin variation is thought to allow immune evasion and change the adherence properties of the pilus. We have examined the process of pilin antigenic variation in human volunteers inoculated with strain FA1090. Our data show that pilin variation occurred throughout the process of infection, that at each time sampled after inoculation multiple pilin variants were present, and that later pilin variants appear to be recombinants between previously expressed genes and the silent storage pilin copies. Thus, during infection a large repertoire of proteins are available to the population to help avoid immune responses, to provide pili with varying functions, and to transmit to a new host. (J. Clin. Invest. 1994. 93:2744-274

PAMELA Measurements of Cosmic-ray Proton and Helium Spectra

Protons and helium nuclei are the most abundant components of the cosmic radiation. Precise measurements of their fluxes are needed to understand the acceleration and subsequent propagation of cosmic rays in the Galaxy. We report precision measurements of the proton and helium spectra in the rigidity range 1 GV-1.2 TV performed by the satellite-borne experiment PAMELA. We find that the spectral shapes of these two species are different and cannot be well described by a single power law. These data challenge the current paradigm of cosmic-ray acceleration in supernova remnants followed by diffusive propagation in the Galaxy. More complex processes of acceleration and propagation of cosmic rays are required to explain the spectral structures observed in our data.Comment: 13 pages, 4 figures, link to SOM (with tables) in the references. This manuscript has been accepted for publication in Science. This version has not undergone final editing. Please refer to the complete version of record at http://www.sciencemag.org/ [www.sciencemag.org

The cosmic-ray electron flux measured by the PAMELA experiment between 1 and 625 GeV

Precision measurements of the electron component in the cosmic radiation provide important information about the origin and propagation of cosmic rays in the Galaxy. Here we present new results regarding negatively charged electrons between 1 and 625 GeV performed by the satellite-borne experiment PAMELA. This is the first time that cosmic-ray electrons have been identified above 50 GeV. The electron spectrum can be described with a single power law energy dependence with spectral index -3.18 +- 0.05 above the energy region influenced by the solar wind (> 30 GeV). No significant spectral features are observed and the data can be interpreted in terms of conventional diffusive propagation models. However, the data are also consistent with models including new cosmic-ray sources that could explain the rise in the positron fraction.Comment: 11 pages, 3 figures, accepted for publication in PR

In-flight validation of Metis Visible-light Polarimeter Coronagraph on board Solar Orbiter

Context. The Metis coronagraph is one of the remote-sensing instruments of the ESA/NASA Solar Orbiter mission. Metis is aimed at the study of the solar atmosphere and solar wind by simultaneously acquiring images of the solar corona at two different wavelengths; visible-light (VL) within a band ranging from 580 nm to 640 nm, and in the HI Ly-alpha 121.6 +/- 10 nm ultraviolet (UV) light. The visible-light channel includes a polarimeter with electro-optically modulating Liquid Crystal Variable Retarders (LCVRs) to measure the linearly polarized brightness of the K-corona to derive the electron density. Aims. In this paper, we present the first in-flight validation results of the Metis polarimetric channel together with a comparison to the on-ground calibrations. It is the validation of the first use in deep space (with hard radiation environment) of an electro-optical device: a liquid crystal-based polarimeter. Methods. We used the orientation of the K-corona's linear polarization vector during the spacecraft roll maneuvers for the in-flight calibration. Results. The first in-flight validation of the Metis coronagraph on-board Solar Orbiter shows a good agreement with the on-ground measurements. It confirms the expected visible-light channel polarimetric performance. A final comparison between the first pB obtained by Metis with the polarized brightness (pB) obtained by the space-based coronagraph LASCO and the ground-based coronagraph KCor shows the consistency of the Metis calibrated results.Comment: 8 pages, 13 figures, 3 tables, pape

PAMELA results on the cosmic-ray antiproton flux from 60 MeV to 180 GeV in kinetic energy

The satellite-borne experiment PAMELA has been used to make a new measurement of the cosmic-ray antiproton flux and the antiproton-to-proton flux ratio which extends previously published measurements down to 60 MeV and up to 180 GeV in kinetic energy. During 850 days of data acquisition approximately 1500 antiprotons were observed. The measurements are consistent with purely secondary production of antiprotons in the galaxy. More precise secondary production models are required for a complete interpretation of the results.Comment: 11 pages, 3 figures, 1 table. Accepted for publication in Physical Review Letter

Observations of the December 13 and 14, 2006, Solar Particle Events in the 80 MeV/n - 3 GeV/n range from space with PAMELA detector

We present the space spectrometer PAMELA observations of proton and helium fluxes during the December 13 and 14, 2006 solar particle events. This is the first direct measurement of the solar energetic particles in space with a single instrument in the energy range from $\sim$ 80 MeV/n up to $\sim$ 3 GeV/n. In the event of December 13 measured energy spectra of solar protons and helium were compared with results obtained by neutron monitors and other detectors. Our measurements show a spectral behaviour different from those derived from the neutron monitor network. No satisfactory analytical fitting was found for the energy spectra. During the first hours of the December 13 event solar energetic particles spectra were close to the exponential form demonstrating rather significant temporal evolution. Solar He with energy up to ~1 GeV/n was recorded on December 13. In the event of December 14 energy of solar protons reached ~600 MeV whereas maximum energy of He was below 100 MeV/n. The spectra were slightly bended in the lower energy range and preserved their form during the second event. Difference in the particle flux appearance and temporal evolution in these two events may argue for a special conditions leading to acceleration of solar particles up to relativistic energies.Comment: Accepted for publication on Astrophysical journa

Multiple gonococcal opacity proteins are expressed during experimental urethral infection in the male

The opacity (Opa) proteins of Neisseria gonorrhoeae are a family of outer membrane proteins demonstrating phase and antigenic variation. N. gonorrhoeae strain FA0190 has 11 opa loci that encode at least 8 antigenically distinct Opa proteins. To determine if expression of one Opa protein or a subset of them is favored during gonococcal infection, we inoculated Opa-negative variants of strain FA1090 intraurethrally into male volunteers. The Opa phenotype of gonococci isolated from urine and urethral swab cultures from nine infected subjects was determined. Opa proteins were expressed in a large proportion of the reisolates from the infected subjects. Gonococci cultured from urine or urethral swab samples from six of the subjects were uniformly Opa positive, with the predominant Opa variants differing among subjects. Three different Opa proteins were represented as the predominant type in at least one subject each. In three subjects, there was more heterogeneity in Opa phenotype of the reisolates, including the presence of Opa-negative variants. An increase in the proportion of isolates expressing multiple Opa proteins occurred over time in most subjects. Passage of the inoculum in vitro did not result in similar changes in Opa expression. There was no detectable difference in infectivity of an Opa-negative variant and one expressing an Opa protein (OpaF) that was highly represented in reisolates from the original nine subjects. Reisolates from three infected volunteers inoculated with the OpaF variant showed continued expression of OpaF alone or in conjunction with other Opa proteins. These results demonstrate that there is strong selection for expression of one or more Opa proteins by strain FA1090 in vivo, but that no single protein is preferentially expressed during early infection in the male urethra

Control of gdhR Expression in Neisseria gonorrhoeae via Autoregulation and a Master Repressor (MtrR) of a Drug Efflux Pump Operon

ABSTRACT The MtrCDE efflux pump of Neisseria gonorrhoeae contributes to gonococcal resistance to a number of antibiotics used previously or currently in treatment of gonorrhea, as well as to host-derived antimicrobials that participate in innate defense. Overexpression of the MtrCDE efflux pump increases gonococcal survival and fitness during experimental lower genital tract infection of female mice. Transcription of mtrCDE can be repressed by the DNA-binding protein MtrR, which also acts as a global regulator of genes involved in important metabolic, physiologic, or regulatory processes. Here, we investigated whether a gene downstream of mtrCDE , previously annotated gdhR in Neisseria meningitidis , is a target for regulation by MtrR. In meningococci, GdhR serves as a regulator of genes involved in glucose catabolism, amino acid transport, and biosynthesis, including gdhA , which encodes an l -glutamate dehydrogenase and is located next to gdhR but is transcriptionally divergent. We report here that in N. gonorrhoeae , expression of gdhR is subject to autoregulation by GdhR and direct repression by MtrR. Importantly, loss of GdhR significantly increased gonococcal fitness compared to a complemented mutant strain during experimental murine infection. Interestingly, loss of GdhR did not influence expression of gdhA , as reported for meningococci. This variance is most likely due to differences in promoter localization and utilization between gonococci and meningococci. We propose that transcriptional control of gonococcal genes through the action of MtrR and GdhR contributes to fitness of N. gonorrhoeae during infection. IMPORTANCE The pathogenic Neisseria species are strict human pathogens that can cause a sexually transmitted infection ( N. gonorrhoeae ) or meningitis or fulminant septicemia ( N. meningitidis ). Although they share considerable genetic information, little attention has been directed to comparing transcriptional regulatory systems that modulate expression of their conserved genes. We hypothesized that transcriptional regulatory differences exist between these two pathogens, and we used the gdh locus as a model to test this idea. For this purpose, we studied two conserved genes ( gdhR and gdhA ) within the locus. Despite general conservation of the gdh locus in gonococci and meningococci, differences exist in noncoding sequences that correspond to promoter elements or potential sites for interacting with DNA-binding proteins, such as GdhR and MtrR. Our results indicate that implications drawn from studying regulation of conserved genes in one pathogen are not necessarily translatable to a genetically related pathogen