Developments in biologicals最新文献

The 2009 Viral Clearance Symposium (Indianapolis, IN, USA) was held to interactively discuss methods for virus removal and inactivation during biopharmaceutical manufacture. Its origin was the result of worldwide regulatory and industry recognition that challenges, gaps, and opportunities for improvement exist, which if formally addressed could benefit the field as a whole. The symposium began with presentations by the FDA (USA) and the Paul Ehrlich Institute (PEI, Germany), which highlighted viral clearance study information reported in regulatory submissions. In these two presentations, and a subsequent series of brief industry presentations covering various unit operations, it was made clear that many unit operations are quite effective in clearing viruses. This was particularly true of low pH inactivation, anion exchange chromatography, and virus filtration. Moreover, the follow-up discussions at the end of each session, and the wrap-up at the end of the symposium, aimed to synthesize the regulatory data mining knowledge base with the industry-generated data. The symposium also revealed a number of unknowns in the field which were defined and prioritized, and served as potential action items for future experimental studies.

Proteomics technology was employed to profile host responses to rabies virus (RABV) infection in order to understand how RABV infection results in neuronal dysfunction. In mice infected with wild-type (wt) RABV, the expression of proteins involved in ion homeostasis was altered. H+ ATPase and Na+/K+ ATPase were up-regulated while Ca2+ ATPase was downregulated, which resulted in reduction of intracellular Na+ and Ca2+ concentrations. Furthermore, infection with wt RABV resulted in down-regulation of SNAREs such as alpha-SNAP, TRIM9, syntaxin, and pallidin, all of which are involved in docking and fusion of synaptic vesicles to and with the presynaptic membrane. As a consequence, the accumulation of synaptic vesicles was observed in the presynapses of mice infected with wt RABV. These data demonstrate that infection with wt RABV results in the alteration of host protein expression, particularly those involved in ion homeostasis and docking and the fusion of synaptic vesicles to the presynaptic membrane, which may lead to neuronal dysfunction.

The National Reference Centre for Rabies (NRC) was created at the Pasteur Institute after the fox epizootic reached the French territory. The missions of the NRC include, among others, the surveillance of rabies cases in humans and rabies post-exposure prophylaxis (PEP) treatments. The surveillance has been effective since 1982. A Bulletin on the Epidemiology and the Prophylaxis of Rabies in Humans in France is published every year. This Bulletin is now available on the Internet for Human Health and Veterinary national and local Authorities. Since 2005, data is collected with new software, Voozanoo, directly via the Internet. Twenty cases of rabies in humans have been reported since 1970. There were no indigenously acquired cases. The number of PEP treatments peaked in 1990, when the number of cases in the wild fauna was at its acme. Following the decrease of rabies cases in the wild fauna, PEP decreased by 60%. Nevertheless, about4,000 PEP treatments are still carried out. These patients have been exposed to bats or to rabid animals illegally introduced onto the French territory, or during a stay in rabies enzootic countries, or to unobservable animals. The study of this database leads to a number of conclusions: canine variants acquired directly in canine enzootic areas, that are translocated, or acquired through iatrogenic exposure, are responsible for the majority of cases; bats appear to be an increasing source of exposure; PEP surveillance is of utmost importance to monitor and to improve the quality of case management.

Veterinary rabies vaccines produced in BHK-21/C13 permanent cell cultures have been used for a long period of time and have been proven as efficacious and safe. A candidate vaccine for human use (YU BHK Rabivak) was developed at the Pasteur Institute, Novi Sad, Serbia on the basis of the fixed rabies virus strain "L. Pasteur 2061/Vero 15 pas" using BHK 21/C13 as a cell substrate for vaccine production. To test the vaccine immunogenicity, a clinical trial was conducted involving 164 subjects between 18 and 60 years of age, immunized either with the YU BHK Rabivak vaccine candidate orwith a commercially available vaccine (Rabipur). Three groups of subjects were immunized with either vaccine by intramuscular administration in the deltoid region, following a pre-exposure regimen on days 0, 7 and 21, or the Essen or Zagreb post-exposure regimens. Rabies virus neutralizing antibodies (VNA) titres were determined by rapid fluorescent focus inhibition test (RFFIT) 21 and 30-45 days post vaccination. A protective titre of VNAs (>0.5 IU/ml) was found in all subjects vaccinated. Dynamics of the immune response showed that 96.4% of the subjects developed protective VNA titres after two doses, 99.3% after three doses and 100% after four and five doses of the candidate YU BHK Rabivak vaccine. There was a low reactogenicity without serious adverse events indicating a satisfactory safety profile in humans. Results obtained in this study indicate that BHK 21 cells offer the possibility of producing an efficacious and safe cell-culture rabies vaccine for humane use.

The immunogenic properties of ERA G 333 attenuated rabies virus strain in foxes and raccoon dogs by the oral route of administration were examined. This strain is a proposed candidate for oral rabies vaccine in Russia. As was previously demonstrated, the advantage of this virus, provided by the US Centers for Disease Control and Prevention in the framework of the Biotechnology Engagement Program, is apathogenicity for 3-week old and adult mice, as well as for other target and non-target species, even when challenged via the intracerebral route. A group of 12, 7-8 month-old foxes and eight wild captured raccoon dogs were given 2.0 ml of ERA G 333 (titre 107.5 FFU) orally on the tongue. Another three foxes and three raccoon dogs were retained as controls. Blood samples were collected before immunization and on day 60 thereafter. The antibody titres were determined by the fluorescent antibody virus neutralization (FAVN) test. Eight foxes and three raccoon dogs showed seroconversion on day 60 postvaccination. Eleven foxes and four raccoon dogs survived challenge with a virulent rabies virus strain. This study has demonstrated the possibility of using ERA G 333 for the oral vaccination of red foxes and raccoon dogs. Further experiments are needed to determine the effective dose, virus stability, and the strength and duration of immunity.

Rabies remains endemic within a number of countries in Southeast Europe including Romania, Bulgaria and Turkey. With the probable expansion of the European Union eastwards, it is likely that rabies elimination programs will be increased to reduce the burden of disease in new accession countries. A clear understanding of the epidemiology of the virus in this area of Europe is vital before such programs are introduced. With the exception of Turkey, the red fox (Vulpes vulpes) is the principal disease reservoir in Southeastern Europe. However, cases of rabies in the dog (Canis familiaris) are regularly reported. In contrast to Northern Europe, the raccoon dog (Nyctereutes procyonoides) does not appear to be a vector in the south. This study summarises the current rabies situation in Southeast Europe and demonstrates the phylogenetic relationships between the viruses in a number of the countries within the region. Rabies virus RNA was extracted from original samples and a fragment of the nucleoprotein gene amplified by reverse-transcriptase PCR. Automated sequencing was used to derive nucleoprotein gene sequences and these were used to prepare a molecular phylogeny of rabies viruses in Southeast Europe. In Bulgaria, the dog is the main vector bringing rabies into contact with humans and livestock. However, other species may also act as reservoirs for the disease, complicating the development of elimination strategies. The fox is the principal reservoir species for rabies in Romania although cases in dogs are regularly reported. Despite a gradual decline in dog rabies, urban pockets of the disease remain in many regions of Turkey. Furthermore, there is some evidence that the fox has been a significant vectorfor rabies and may be responsible for increases in rabies in cattle in the Aegean region of the country. Throughout the region there is evidence for cross-border movement of rabies by both wildlife and canine vectors.

Rabies is a serious problem in the 11 states that make up CentralAsia. More information is needed to make rabies control effective in Central Asia. It is necessary to assess the national programmes, identify the main rabies carriers, increase the number of vaccinated dogs and cats with the aim of preventing rabies cases, and to use the worldwide experience of rabies eradication by the oral vaccination of wildlife.

Different rhabdoviruses have been found in healthy bats, suggesting asymptomatic infection. The aim of this study was to focus on the epidemiology and pathogenesis of EBLV1 infection in the meridional serotine bat (Eptesicus isabellinus), as well as to search for other rhabdoviruses in this bat, which is the responsible for more than 95% of cases of human exposure to lyssaviruses in Europe. RT-PCR on oropharyngeal swabs was used together with antibody detection by the Rapid Fluorescent Focus Inhibition Test (RFFIT) to investigate EBLV1 circulation in 19 natural colonies of meridional serotine bats in Andalusia (Spain) from 1998 to 2003. The survey was based on 1,227 different captures of 1,033 individuals that were ring banded, sampled and released. Individuals that were repeatedly captured were always found in the same colony, despite the fact that some colonies were less than five km apart. Viral circulation was detected in ten colonies either by RT-PCR, serology or both. Each colony showed a different temporal pattern of viral circulation suggesting independent endemic circulation. Some positive individuals were captured healthy in following campaigns providing evidence for survival after viral infection. RNA from two apparently new Dimarhabdoviruses was also found in the pharyngeal cavity of two healthy bats.