THE GLOBAL MANUFACTURE OF POLIO VACCINE IN THE ENDGAME OF ERADICATION

The manufacture of medicines is increasingly globalised but biological products such as vaccines are more complex and their production raises significantly different issues to that of chemical entities. New producers in any region of the world may have difficulty in recognising all the concerns involved and can benefit from technology transfer exercises and a strong and scientifically competent regulatory authority. The issues will be discussed in the context of the Global Polio Eradication Initiative in its terminal phases and future needs for the manufacture of polio vaccine

High resolution identity testing of inactivated poliovirus vaccines

AbstractBackgroundDefinitive identification of poliovirus strains in vaccines is essential for quality control, particularly where multiple wild-type and Sabin strains are produced in the same facility. Sequence-based identification provides the ultimate in identity testing and would offer several advantages over serological methods.MethodsWe employed random RT-PCR and high throughput sequencing to recover full-length genome sequences from monovalent and trivalent poliovirus vaccine products at various stages of the manufacturing process.ResultsAll expected strains were detected in previously characterised products and the method permitted identification of strains comprising as little as 0.1% of sequence reads. Highly similar Mahoney and Sabin 1 strains were readily discriminated on the basis of specific variant positions. Analysis of a product known to contain incorrect strains demonstrated that the method correctly identified the contaminants.ConclusionRandom RT-PCR and shotgun sequencing provided high resolution identification of vaccine components. In addition to the recovery of full-length genome sequences, the method could also be easily adapted to the characterisation of minor variant frequencies and distinction of closely related products on the basis of distinguishing consensus and low frequency polymorphisms

Live attenuated vaccines: Historical successes and current challenges

AbstractLive attenuated vaccines against human viral diseases have been amongst the most successful cost effective interventions in medical history. Smallpox was declared eradicated in 1980; poliomyelitis is nearing global eradication and measles has been controlled in most parts of the world. Vaccines function well for acute diseases such as these but chronic infections such as HIV are more challenging for reasons of both likely safety and probable efficacy. The derivation of the vaccines used has in general not been purely rational except in the sense that it has involved careful clinical trials of candidates and subsequent careful follow up in clinical use; the identification of the candidates is reviewed

Human coronavirus 229E encodes a single ORF4 protein between the spike and the envelope genes

BACKGROUND: The genome of coronaviruses contains structural and non-structural genes, including several so-called accessory genes. All group 1b coronaviruses encode a single accessory protein between the spike and envelope genes, except for human coronavirus (HCoV) 229E. The prototype virus has a split gene, encoding the putative ORF4a and ORF4b proteins. To determine whether primary HCoV-229E isolates exhibit this unusual genome organization, we analyzed the ORF4a/b region of five current clinical isolates from The Netherlands and three early isolates collected at the Common Cold Unit (CCU) in Salisbury, UK. RESULTS: All Dutch isolates were identical in the ORF4a/b region at amino acid level. All CCU isolates are only 98% identical to the Dutch isolates at the nucleotide level, but more closely related to the prototype HCoV-229E (>98%). Remarkably, our analyses revealed that the laboratory adapted, prototype HCoV-229E has a 2-nucleotide deletion in the ORF4a/b region, whereas all clinical isolates carry a single ORF, 660 nt in size, encoding a single protein of 219 amino acids, which is a homologue of the ORF3 proteins encoded by HCoV-NL63 and PEDV. CONCLUSION: Thus, the genome organization of the group 1b coronaviruses HCoV-NL63, PEDV and HCoV-229E is identical. It is possible that extensive culturing of the HCoV-229E laboratory strain resulted in truncation of ORF4. This may indicate that the protein is not essential in cell culture, but the highly conserved amino acid sequence of the ORF4 protein among clinical isolates suggests that the protein plays an important role in vivo

Human Papillomavirus Antibody Reference Reagents for Use in Postvaccination Surveillance Serology

Suitably controlled serosurveillance surveys are essential for evaluating human papillomavirus (HPV) immunization programs. A panel of plasma samples from 18-year-old females was assembled, the majority of the samples being from recipients of the bivalent HPV vaccine. Antibody specificities were evaluated by three independent laboratories, and 3 pools that displayed no antibodies to any HPV type tested or intermediate or high levels of antibody to HPV16, HPV18, HPV31, and HPV45 were created. These pools will be useful as control reagents for HPV serology

Novel Parvovirus and Related Variant in Human Plasma

We report a novel parvovirus (PARV4) and related variants in pooled human plasma used in the manufacture of plasma-derived medical products. Viral DNA was detected by using highly selective polymerase chain reaction assays; 5% of pools tested positive, and amounts of DNA ranged from <500 copies/mL to >106 copies/mL plasma

A Complete Spectroscopic Survey of the Milky Way Satellite Segue 1: The Darkest Galaxy

We present the results of a comprehensive Keck/DEIMOS spectroscopic survey of the ultra-faint Milky Way satellite galaxy Segue 1. We have obtained velocity measurements for 98.2% of the stars within 67 pc (10 arcmin, or 2.3 half-light radii) of the center of Segue 1 that have colors and magnitudes consistent with membership, down to a magnitude limit of r=21.7. Based on photometric, kinematic, and metallicity information, we identify 71 stars as probable Segue 1 members, including some as far out as 87 pc. After correcting for the influence of binary stars using repeated velocity measurements, we determine a velocity dispersion of 3.7^{+1.4}_{-1.1} km/s, with a corresponding mass within the half-light radius of 5.8^{+8.2}_{-3.1} x 10^5 Msun. The stellar kinematics of Segue 1 require very high mass-to-light ratios unless the system is far from dynamical equilibrium, even if the period distribution of unresolved binary stars is skewed toward implausibly short periods. With a total luminosity less than that of a single bright red giant and a V-band mass-to-light ratio of 3400 Msun/Lsun, Segue 1 is the darkest galaxy currently known. We critically re-examine recent claims that Segue 1 is a tidally disrupting star cluster and that kinematic samples are contaminated by the Sagittarius stream. The extremely low metallicities ([Fe/H] < -3) of two Segue 1 stars and the large metallicity spread among the members demonstrate conclusively that Segue 1 is a dwarf galaxy, and we find no evidence in favor of tidal effects. We also show that contamination by the Sagittarius stream has been overestimated. Segue 1 has the highest measured dark matter density of any known galaxy and will therefore be a prime testing ground for dark matter physics and galaxy formation on small scales.Comment: 24 pages, 4 tables, 11 figures (10 in color). Submitted for publication in ApJ. V3 revised according to comments from the refere

Isolation of Vaccine-Like Poliovirus Strains in Sewage Samples From the United Kingdom.

Background: Environmental surveillance (ES) is a sensitive method for detecting human enterovirus (HEV) circulation, and it is used worldwide to support global polio eradication. We describe a novel ES approach using next-generation sequencing (NGS) to identify HEVs in sewage samples collected in London, United Kingdom, from June 2016 to May 2017. Methods: Two different methods were used to process raw sewage specimens: a 2-phase aqueous separation system and size exclusion by filtration and centrifugation. HEVs were isolated using cell cultures and analyzed using NGS. Results: Type 1 and 3 vaccine-like poliovirus (PV) strains were detected in samples collected from September 2016 through January 2017. NGS analysis allowed us to rapidly obtain whole-genome sequences of PV and non-PV HEV strains. As many as 6 virus strains from different HEV serotypes were identified in a single cell culture flask. PV isolates contained only a small number of mutations from vaccine strains commonly seen in early isolates from vaccinees. Conclusions: Our ES setup has high sensitivity for polio and non-PV HEV detection, generating nearly whole-genome sequence information. Such ES systems provide critical information to assist the polio eradication endgame and contribute to the improvement of our understanding of HEV circulation patterns in humans

Gravitationally lensed quasars and supernovae in future wide-field optical imaging surveys

Cadenced optical imaging surveys in the next decade will be capable of detecting time-varying galaxy-scale strong gravitational lenses in large numbers, increasing the size of the statistically well-defined samples of multiply-imaged quasars by two orders of magnitude, and discovering the first strongly-lensed supernovae. We carry out a detailed calculation of the likely yields of several planned surveys, using realistic distributions for the lens and source properties and taking magnification bias and image configuration detectability into account. We find that upcoming wide-field synoptic surveys should detect several thousand lensed quasars. In particular, the LSST should find 8000 lensed quasars, 3000 of which will have well-measured time delays, and also ~130 lensed supernovae, which is compared with ~15 lensed supernovae predicted to be found by the JDEM. We predict the quad fraction to be ~15% for the lensed quasars and ~30% for the lensed supernovae. Generating a mock catalogue of around 1500 well-observed double-image lenses, we compute the available precision on the Hubble constant and the dark energy equation parameters for the time delay distance experiment (assuming priors from Planck): the predicted marginalised 68% confidence intervals are \sigma(w_0)=0.15, \sigma(w_a)=0.41, and \sigma(h)=0.017. While this is encouraging in the sense that these uncertainties are only 50% larger than those predicted for a space-based type-Ia supernova sample, we show how the dark energy figure of merit degrades with decreasing knowledge of the the lens mass distribution. (Abridged)Comment: 17 pages, 10 figures, 3 tables, accepted for publication in MNRAS; mock LSST lens catalogue may be available at http://kipac-prod.stanford.edu/collab/research/lensing/mocklen