Trial confirms Ebola vaccine candidate safe, equally immunogenic in Africa

"This is the first study to show comparable safety and immune response of an experimental Ebola vaccine in an African population," says lead author Dr Julie Ledgerwood. "This is particularly encouraging because those at greatest risk of Ebola live primarily in Africa, and diminished vaccine protection in African populations has been seen for other diseases." Credit: © nito / Fotolia

Two experimental DNA vaccines to prevent Ebola virus and the closely related Marburg virus are safe, and generated a similar immune response in healthy Ugandan adults as reported in healthy US adults earlier this year. The findings, from the first trial of filovirus vaccines in Africa, are published in The Lancet.

"This is the first study to show comparable safety and immune response of an experimental Ebola vaccine in an African population," says lead author Dr Julie Ledgerwood from the National Institutes of Allergy and Infectious Diseases (NIAID) at the National Institutes of Health, USA. "This is particularly encouraging because those at greatest risk of Ebola live primarily in Africa, and diminished vaccine protection in African populations has been seen for other diseases."

Scientists from the NIAID developed the DNA vaccines that code for Ebola virus proteins from the Zaire and Sudan strains and the Marburg virus protein. The vaccines contain the construction plans for the proteins on the outer surface of the virus. Immune responses against these proteins have shown to be highly protective in non-human primate models.

In this phase 1 trial, the Makerere University Walter Reed Program enrolled 108 healthy adults aged between 18 and 50 from Kampala, Uganda between November, 2009 and April, 2010. Each volunteer was randomly assigned to receive an intramuscular injection of either the Ebola vaccine (30 volunteers), Marburg vaccine (30), both vaccines (30), or placebo (18) at the start of the study, and again 4 weeks and 8 weeks later.

The vaccines given separately and together were safe and stimulated an immune response in the form of neutralising antibodies and T-cells against the virus proteins. Four weeks after the third injection, just over half of the volunteers (57%; 17 of 30) had an antibody response to the Ebola Zaire protein as did 14 of 30 participants who received both the Ebola and Marburg vaccines. 

However, the antibodies were not long-lasting and returned to undetectable levels within 11 months of vaccination.

Both DNA vaccines were well tolerated in Ugandan adults with similar numbers of local and systemic reactions reported in all groups. Only one serious adverse event (neutropenia; low white blood cell count) was reported in a Marburg vaccine only recipient, but was not thought to be vaccine related.

According to Dr Ledgerwood, "These findings have already formed the basis of a more potent vaccine, delivered using a harmless chimpanzee cold virus, which is undergoing trials in the USA, UK, Mali, and Uganda in response to the ongoing Ebola virus outbreak."

Writing in a linked Comment, Dr Saranya Sridhar from the Jenner Institute at the University of Oxford in the UK says, "[This] study deserves to be the focal point around which the broader question of vaccine development, particularly for Africa, must be addressed. With the uncharitable benefit of hindsight in view of the evolving 2014 Ebola outbreak, we must ask ourselves whether a filovirus vaccine should have been in more advanced clinical development. The international response to the present Ebola outbreak is an exemplar of the speed and purpose with which clinical vaccine development can progress and has set the benchmark against which future vaccine development must be judged. This study is the first step on the aspirational road towards the deployment of filovirus vaccines in Africa and must serve to shake the metaphorical cobwebs that can stall our advance towards this destination."

Source: The Lancet

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For tiger populations, a new threat

This is an Amur tiger photographed by camera trap. Credit: WCS Russia Program

Along with the pressures of habitat loss, poaching and depletion of prey species, a new threat to tiger populations in the wild has surfaced in the form of disease, specifically, canine distemper virus (CDV). According to a new study from the Wildlife Conservation Society (WCS) and its partners, CDV has the potential to be a significant driver in pushing the animals toward extinction.

While CDV has recently been shown to lead to the deaths of individual tigers, its long-term impacts on tiger populations had never before been studied.

The authors evaluated these impacts on the Amur tiger population in Russia's Sikhote-Alin Biosphere Zapovednik (SABZ), where tiger numbers declined from 38 individuals to 9 in the years 2007 to 2012. In 2009 and 2010, six adult tigers died or disappeared from the reserve, and CDV was confirmed in two dead tigers -- leading scientists to believe that CDV likely played a role in the overall decline of the population. Joint investigations of CDV have been an ongoing focus of WCS and Russian scientists at Sikhote-Alin Zapovednik and veterinarians at the regional Primorye Agricultural College since its first appearance in tigers in 2003.

A key finding of this study: Modeling shows that smaller populations of tigers were found to be more vulnerable to extinction by CDV. Populations consisting of 25 individuals were 1.65 times more likely to decline in the next 50 years when CDV was present. The results are profoundly disturbing for global wild tigers given that in most sites where wild tigers persist they are limited to populations of less than 25 adult breeding individuals.

The scientists used computer modeling to simulate the effects of CDV infection on isolated tiger populations of various sizes and through a series of transmission scenarios. These included tiger-to-tiger transmission and transmission through predation on CDV-infected domestic dogs and/or infected wild carnivores (such as foxes, raccoon dogs and badgers). High and low-risk scenarios for the model were created based on variation in the prevalence of CDV and the tigers' contact with sources of exposure.

Results showed that CDV infection increased the 50-year extinction probability of tigers in SABZ as much as 55.8 percent compared to CDV-free populations of equivalent size.

"Although we knew that individual tigers had died from CDV in the wild, we wanted to understand the risk the virus presents to whole populations," said WCS veterinarian Martin Gilbert. "Tigers are elusive, however, and studying the long-term impact of risk factors is very challenging. Our model, based on tiger ecology data collected over 20 years in SABZ, explored the different ways that tigers might be exposed to the virus and how these impact the extinction risk to tiger populations over the long term."

WCS Russia Program Director Dale Miquelle said, "Tigers face an array of threats throughout their range, from poaching to competition with humans for space and for food. 

Consequently, many tiger populations have become smaller and more fragmented, making them much more susceptible to diseases such as CDV. While we must continue to focus on the primary threats of poaching and habitat destruction, we now must also be prepared to deal with the appearance of such diseases in the future."

Priorities for future research, according to the authors, include identifying the domestic and wild carnivore species that contribute to the CDV reservoir, and those that are the most likely sources of infection for tigers. Tigers are too rare to sustain the virus in the long term, so CDV must rely on more abundant carnivore species to persist in the environment. 

Understanding the structure of the CDV reservoir will be a critical first step in identifying measures that might prevent or control future outbreaks. In addition, since we now know that small tiger populations are at greater risk to diseases such as CDV than larger populations, conservation strategies focusing on connectedness between populations become all the more important. "Estimating the potential impact of canine distemper virus on the Amur tiger population (Panthera tigris altaica) in Russia," appears in the current online edition of PloSONE. Authors include: Martin Gilbert of WCS and Boyd Orr Centre for Population and Ecosystem Health at the University of Glasgow; Dale G. Miquelle of WCS; John M. Goodrich of Panthera; Richard Reeve, Sarah Cleaveland and Louise Matthews of Boyd Orr Centre for Population and Ecosystem Health at the University of Glasgow; and Damien Joly of WCS and Metabiota.

This study was made possible through generous support from Morris Animal Foundation, Zoo Boise Conservation Fund, AZA Tiger Species Survival Plan Tiger Conservation Campaign, and the Biotechnology and Biological Sciences Research Council.

"Morris Animal Foundation is thankful to Dr. Miquelle and his team for helping protect the Amur Tigers," said Diane Brown, DVM, PhD, DACVP and Chief Scientific Officer for Morris Animal Foundation. "Our Foundation values their hard work and dedication to this study and we look forward to many more partnerships with the Wildlife Conservation Society."

Source: Wildlife Conservation Society

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Experimental Ebola vaccine appears safe, prompts immune response

A 39-year-old woman, the first participant enrolled in VRC 207, receives a dose of the investigational NIAID/GSK Ebola vaccine at the NIH Clinical Center in Bethesda, Md. on September 2. Credit: NIAID

An experimental vaccine to prevent Ebola virus disease was well-tolerated and produced immune system responses in all 20 healthy adults who received it in a Phase 1 clinical trial conducted by researchers from the National Institutes of Health. The candidate vaccine, which was co-developed by the NIH's National Institute of Allergy and Infectious Diseases (NIAID) and GlaxoSmithKline (GSK), was tested at the NIH Clinical Center in Bethesda, Maryland. The interim results are reported online in advance of print in the New England Journal of Medicine.

"The unprecedented scale of the current Ebola outbreak in West Africa has intensified efforts to develop safe and effective vaccines, which may play a role in bringing this epidemic to an end and undoubtedly will be critically important in preventing future large outbreaks," said NIAID Director Anthony S. Fauci, M.D. "Based on these positive results from the first human trial of this candidate vaccine, we are continuing our accelerated plan for larger trials to determine if the vaccine is efficacious in preventing Ebola infection."

The candidate NIAID/GSK Ebola vaccine was developed collaboratively by scientists at the NIAID Vaccine Research Center (VRC) and at Okairos, a biotechnology company acquired by GSK. It contains segments of Ebola virus genetic material from two virus species, Sudan and Zaire. The Ebola virus genetic material is delivered by a carrier virus (chimpanzee-derived adenovirus 3 or cAd 3) that causes a common cold in chimpanzees but causes no illness in humans. The candidate vaccine does not contain Ebola virus and cannot cause Ebola virus disease.

The trial enrolled volunteers between the ages of 18 and 50. Ten volunteers received an intramuscular injection of vaccine at a lower dose and 10 received the same vaccine at a higher dose. At two weeks and four weeks following vaccination, the researchers tested the volunteers' blood to determine if anti-Ebola antibodies were generated. All 20 volunteers developed such antibodies within four weeks of receiving the vaccine. Antibody levels were higher in those who received the higher dose vaccine.

The investigators also analyzed the research participants' blood to learn whether the vaccine prompted production of immune system cells called T cells. A recent study by VRC scientist Nancy J. Sullivan, Ph.D., and colleagues showed that non-human primates inoculated with the candidate NIAID/GSK vaccine developed both antibody and T-cell responses, and that these were sufficient to protect vaccinated animals from disease when they were later exposed to high levels of Ebola virus.

The experimental NIAID/GSK vaccine did induce a T-cell response in many of the volunteers, including production of CD8 T cells, which may be an important part of immune protection against Ebola viruses. Four weeks after vaccination, CD8 T cells were detected in two volunteers who had received the lower dose vaccine and in seven of those who had received the higher dose.

"We know from previous studies in non-human primates that CD8 T cells played a crucial role in protecting animals that had been vaccinated with this NIAID/GSK vaccine and then exposed to otherwise lethal amounts of Ebola virus," said Julie E. Ledgerwood, D.O., a VRC researcher and the trial's principal investigator. "The size and quality of the CD8 T cell response we saw in this trial are similar to that observed in non-human primates vaccinated with the candidate vaccine."

There were no serious adverse effects observed in any of the volunteers, although two people who received the higher dose vaccine did develop a briefly lasting fever within a day of vaccination.

Source: NIH/National Institute of Allergy and Infectious Diseases

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A clear, molecular view of how human color vision evolved

Mountain Gorilla - Bwindi Uganda. “Gorillas and chimpanzees have human color vision,” Yokoyama says. “Or perhaps we should say that humans have gorilla and chimpanzee vision.” Credit: © Alexander / Fotolia

Many genetic mutations in visual pigments, spread over millions of years, were required for humans to evolve from a primitive mammal with a dim, shadowy view of the world into a greater ape able to see all the colors in a rainbow.

Now, after more than two decades of painstaking research, scientists have finished a detailed and complete picture of the evolution of human color vision. PLOS Genetics published the final pieces of this picture: The process for how humans switched from ultraviolet (UV) vision to violet vision, or the ability to see blue light.

"We have now traced all of the evolutionary pathways, going back 90 million years, that led to human color vision," says lead author Shozo Yokoyama, a biologist at Emory University. 

"We've clarified these molecular pathways at the chemical level, the genetic level and the functional level."

Co-authors of the PLOS Genetics paper include Emory biologists Jinyi Xing, Yang Liu and Davide Faggionato; Syracuse University biologist William Starmer; and Ahmet Altun, a chemist and former post-doc at Emory who is now at Fatih University in Istanbul, Turkey.

Yokoyama and various collaborators over the years have teased out secrets of the adaptive evolution of vision in humans and other vertebrates by studying ancestral molecules. The lengthy process involves first estimating and synthesizing ancestral proteins and pigments of a species, then conducting experiments on them. The technique combines microbiology with theoretical computation, biophysics, quantum chemistry and genetic engineering.

Five classes of opsin genes encode visual pigments for dim-light and color vision. Bits and pieces of the opsin genes change and vision adapts as the environment of a species changes.

Around 90 million years ago, our primitive mammalian ancestors were nocturnal and had UV-sensitive and red-sensitive color, giving them a bi-chromatic view of the world. By around 30 million years ago, our ancestors had evolved four classes of opsin genes, giving them the ability to see the full-color spectrum of visible light, except for UV.

"Gorillas and chimpanzees have human color vision," Yokoyama says. "Or perhaps we should say that humans have gorilla and chimpanzee vision."

For the PLOS Genetics paper, the researchers focused on the seven genetic mutations involved in losing UV vision and achieving the current function of a blue-sensitive pigment. 

They traced this progression from 90-to-30 million years ago.

The researchers identified 5,040 possible pathways for the amino acid changes required to bring about the genetic changes. "We did experiments for every one of these 5,040 possibilities," Yokoyama says. "We found that of the seven genetic changes required, each of them individually has no effect. It is only when several of the changes combine in a particular order that the evolutionary pathway can be completed."

In other words, just as an animal's external environment drives natural selection, so do changes in the animal's molecular environment.

In previous research, Yokoyama showed how the scabbardfish, which today spends much of its life at depths of 25 to 100 meters, needed just one genetic mutation to switch from UV to blue-light vision. Human ancestors, however, needed seven changes and these changes were spread over millions of years. "The evolution for our ancestors' vision was very slow, compared to this fish, probably because their environment changed much more slowly," 
Yokoyama says.

About 80 percent of the 5,040 pathways the researchers traced stopped in the middle, because a protein became non-functional. Chemist Ahmet Altun solved the mystery of why the protein got knocked out. It needs water to function, and if one mutation occurs before the other, it blocks the two water channels extending through the vision pigment's membrane.

"The remaining 20 percent of the pathways remained possible pathways, but our ancestors used only one," Yokoyama says. "We identified that path."

In 1990, Yokoyama identified the three specific amino acid changes that led to human ancestors developing a green-sensitive pigment. In 2008, he led an effort to construct the most extensive evolutionary tree for dim-light vision, including animals from eels to humans. At key branches of the tree, Yokoyama's lab engineered ancestral gene functions, in order to connect changes in the living environment to the molecular changes.

The PLOS Genetics paper completes the project for the evolution of human color vision. "We have no more ambiguities, down to the level of the expression of amino acids, for the mechanisms involved in this evolutionary pathway," Yokoyama says.

Source: Emory Health Sciences

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Flu virus key machine: First complete view of structure revealed

The complete structure allows researchers to understand how the polymerase uses host cell RNA (red) to kick-start the production of viral messenger RNA. Credit: © EMBL/P.Riedinger

Scientists looking to understand -- and potentially thwart -- the influenza virus now have a much more encompassing view, thanks to the first complete structure of one of the flu virus' key machines. The structure, obtained by scientists at EMBL Grenoble, allows researchers to finally understand how the machine works as a whole, and could prove instrumental in designing new drugs to treat serious flu infections and combat flu pandemics.

If you planned to sabotage a factory, a recon trip through the premises would probably be much more useful than just peeping in at the windows. Scientists looking to understand -- and potentially thwart -- the influenza virus have now gone from a similar window-based view to the full factory tour, thanks to the first complete structure of one of the flu virus' key machines. The structure, obtained by scientists at the European Molecular Biology Laboratory (EMBL) in Grenoble, France, allows researchers to finally understand how the machine works as a whole. Published in two papers in Nature, the work could prove instrumental in designing new drugs to treat serious flu infections and combat flu pandemics.

The machine in question, the influenza virus polymerase, carries out two vital tasks for the virus. It makes copies of the virus' genetic material -- the viral RNA -- to package into new viruses that can infect other cells; and it reads out the instructions in that genetic material to make viral messenger RNA, which directs the infected cell to produce the proteins the virus needs. Scientists -- including Cusack and collaborators -- had been able to determine the structure of several parts of the polymerase in the past. But how those parts came together to function as a whole, and how viral RNA being fed in to the polymerase could be treated in two different ways remained a mystery.

"The flu polymerase was discovered 40 years ago, so there are hundreds of papers out there trying to fathom how it works. But only now that we have the complete structure can we really begin to understand it," says Stephen Cusack, head of EMBL Grenoble, who led the work.

Using X-ray crystallography, performed at the European Synchrotron Radiation Facility (ESRF) in Grenoble, Cusack and colleagues were able to determine the atomic structure of the whole polymerase from two strains of influenza: influenza B, one of the strains that cause seasonal flu in humans, but which evolves slowly and therefore isn't considered a pandemic threat; and the strain of influenza A -- the fast-evolving strain that affects humans, birds and other animals and can cause pandemics -- that infects bats.

"The high-intensity X-ray beamlines at the ESRF, equipped with state-of-the-art Dectris detectors, were crucial for getting high quality crystallographic data from the weakly diffracting and radiation sensitive crystals of the large polymerase complex," says Cusack. "We couldn't have got the data at such a good resolution without them."

The structures reveal how the polymerase specifically recognises and binds to the viral RNA, rather than just any available RNA, and how that binding activates the machine. They also show that the three component proteins that make up the polymerase are very intertwined, which explains why it has been very difficult to piece together how this machine works based on structures of individual parts.

Although the structures of both viruses' polymerases were very similar, the scientists found one key difference, which showed that one part of the machine can swivel around to a large degree. That ability to swivel explains exactly how the polymerase uses host cell RNA to kick-start the production of viral proteins. The swivelling component takes the necessary piece of host cell RNA and directs it into a slot leading to the machine's heart, where it triggers the production of viral messenger RNA.

Now that they know exactly where each atom fits in this key viral machine, researchers aiming to design drugs to stop influenza in its tracks have a much wider range of potential targets at their disposal -- like would-be saboteurs who gain access to the whole production plant instead of just sneaking looks through the windows. And because this is such a fundamental piece of the viral machinery, not only are the versions in the different influenza strains very similar to each other, but they also hold many similarities to their counterparts in related viruses such as lassa, hanta, rabies or ebola.

The EMBL scientists aim to explore the new insights this structure provides for drug design, as well as continuing to try to determine the structure of the human version of influenza A, because although the bat version is close enough that it already provides remarkable insights, ultimately fine-tuning drugs for treating people would benefit from/require knowledge of the version of the virus that infects humans. And, since this viral machine has to be flexible and change shape to carry out its different tasks, Cusack and colleagues also want to get further snapshots of the polymerase in different states.

"This doesn't mean we now have all the answers," says Cusack, "In fact, we have as many new questions as answers, but at least now we have a solid basis on which to probe further."

Source: European Molecular Biology Laboratory (EMBL)

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Flu at the zoo and other disasters: Experts help animal exhibitors prepare for the worst

After experiencing power outages during a 2007 ice storm in Springfield, Mo., Dickerson Park Zoo officials improved their backup power and heating systems to keep animals -- like Henry, pictured here -- safe and warm. Credit: Dickerson Park Zoo

Here are three disaster scenarios for zoo or aquarium managers: One, a wildfire lunges towards your facility, threatening your staff and hundreds of zoo animals. Two, hurricane floodwaters pour into your basement, where thousands of exotic fish and marine mammals live in giant tanks. Three, local poultry farmers report avian influenza (bird flu) in their chickens, a primary source of protein for your big cats.

What do you do?

These are among the many potential disasters the managers of zoos and aquariums ponder in their emergency preparedness drills and plans. But these stories are not just worst-case scenarios: The events described above actually happened, and the aftermath -- often heroic, and sometimes tragic -- depended in large part on the institutions' preparedness training, planning and forethought in calmer times.

When bad weather strikes or illness invades, zoos and aquariums are among the most vulnerable facilities affected, said University of Illinois veterinarian Yvette Johnson-Walker, a clinical epidemiologist who contributes to emergency response training efforts at animal exhibitor institutions. She is a clinical instructor in the department of veterinary clinical medicine at Illinois, and lead author of a new paper on emergency preparedness at zoos and aquariums in the journal Homeland Security & Emergency Management.

Some animals are likely to suffer if the electricity goes out for long, she said. Others are large, skittish and dangerous under normal conditions.

Training caretakers and keepers to minimize their own risks while attending to their animals in an emergency is a challenge, but leads to the best outcomes, she said.

In 2012, Johnson-Walker joined forces with Yvonne Nadler, a project manager with the Zoo and Aquarium All Hazards Preparedness Response and Recovery Center, to bring vital emergency training to accredited animal exhibitor institutions in Illinois, Indiana and Missouri. This effort, funded by the U.S. Department of Agriculture and supported by the Association of Zoos & Aquariums, has since expanded, providing training to staff from zoos and aquariums in 23 states.

The trainings, dubbed "Flu at the Zoo," focus on avian influenza, a viral disease that spread in the 2000s among wild and captive birds and also infected hundreds of people, primarily in Asia, Africa and the Middle East. Bird flu serves as a useful model scenario to help train participants in basic preparedness skills.

One such skill is familiarity with the Incident Command System (ICS), a framework developed by firefighters and adopted by the Federal Emergency Management Agency (FEMA) that allows first responders to quickly set up their emergency response operation and assign vital tasks. The ICS has long been used by public safety, law enforcement and public health entities involved in emergency response.

"We wanted zoos and aquariums to have a seat at the table when there's planning for how we're going to respond to emergencies, and to be able to fit into the system, know who to talk to and how to communicate," Johnson-Walker said.

It's also important to recognize the other responders and understand their roles, she said. If the event involves a disease like bird flu, the USDA, FEMA, National Institutes of Health, state veterinarian, state and federal wildlife services, public health authorities, veterinary organizations, police, hospitals and perhaps even local poultry operations will be involved in the response. Knowing who does what can speed communication in a crisis.

Planning also helps managers make best use of the limited supplies or equipment they have on hand, Nadler said.

"There are certain types of livestock trailers, for example, that can be adapted to moving big cats," she said. "Is that your preferred method of movement? Of course it isn't, but in an emergency that might be your only option."

One beneficiary of the emergency training, Melinda Arnold, knows firsthand the value of preparedness. Arnold is public relations director for Friends of the Zoo, affiliated with Dickerson Park Zoo in Springfield, Missouri. The zoo suffered a blackout during a 2007 ice storm that shut off power for most of the city for several days.

"We did have backup generators," Arnold said. "The greatest problem with the generators was that those fueling stations in town that did have gas didn't have power, so they couldn't pump the gas."

Zoo staff had to travel many miles outside of the affected area with gas cans to collect gas to run the backup generators, she said.

"Now we have some propane-powered backups," Arnold said.

A more recent incident at the zoo, the accidental death of a zookeeper in 2013, caused Dickerson Park Zoo officials to re-evaluate all of their safety protocols. Even though the zookeeper had decades of experience and was guarded by a protective barrier, a skittish elephant rushed him at an unguarded moment, and he fell and was trampled to death.
"It made us step back, not only in our elephant management but in all areas of the zoo, and look at our safety procedures and points of contact with dangerous animals and evaluate those safety conditions and make improvements," Arnold said.

The preparedness plans, drills, discussions and training all help zoos and aquariums reassess their procedures, even those that seem to be safe after decades of operations and no major incidents, she said.

Source: University of Illinois at Urbana-Champaign

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