Showing posts with label virus. Show all posts
Showing posts with label virus. Show all posts

Friday, 16 March 2012

The Future of Personalized Medicine: First-ever Integrative 'Omics' Profile

Stanford scientist discovers and tracks his diabetes onset  
Friday, 16 March 2012

Geneticist Michael Snyder, PhD, has almost no privacy. For more than two years, he and his lab members at the Stanford University School of Medicine pored over his body's most intimate secrets: the sequence of his DNA, the RNA and proteins produced by his cells, the metabolites and signaling molecules wafting through his blood. They spied on his immune system as it battled viral infections.

Finally, to his shock, they discovered that he was predisposed to type-2 diabetes and then watched his blood sugar shoot upward as he developed the condition during the study. It's the first eyewitness account — viewed on a molecular level — of the birth of a disease that affects millions of Americans. It's also an important milestone in the realization of the promise of truly personalized medicine, or tailoring health care to each individual's unique circumstances.

The researchers call the unprecedented analysis, which relies on collecting and analyzing billions of individual bits of data, an integrative Personal "Omics" Profile, or iPOP. The word "omics" indicates the study of a body of information, such as the genome (which is all DNA in a cell), or the proteome (which is all the proteins). Snyder's iPOP also included his metabolome (metabolites), his transcriptome (RNA transcripts) and autoantibody profiles, among other things.

The researchers say that Snyder's diabetes is but one of myriad problems the iPOP can identify and predict, and that such dynamic monitoring will soon become commonplace.

"This is the first time that anyone has used such detailed information to proactively manage their own health," said Snyder.

"It's a level of understanding of health at the molecular level that has never before been achieved."

The research will be published in the March 16 issue of Cell. Snyder, who chairs the Department of Genetics, is the senior author. Postdoctoral scholars Rui Chen, PhD, George Mias, PhD, Jennifer Li-Pook-Than, PhD, and research associate Lihua Jiang, PhD, are co-first authors of the study, which involved a large team of investigators.

The study provides a glimpse into the future of medicine — peppered with untold data-management hurdles and fraught with a degree of self-examination and awareness few of us have ever imagined. And, despite the challenges, the potential payoff is great.

"I was not aware of any type-2 diabetes in my family and had no significant risk factors," said Snyder, "but we learned through genomic sequencing that I have a genetic predisposition to the condition. Therefore, we measured my blood glucose levels and were able to watch them shoot up after a nasty viral infection during the course of the study."

As a result, he was able to immediately modify his diet and exercise to gradually bring his levels back into the normal range and prevent the ongoing tissue damage that would have occurred had the disease gone undiagnosed.

Snyder provided about 20 blood samples (about once every two months while healthy, and more frequently during periods of illness) for analysis over the course of the study. Each was analyzed with a variety of assays for tens of thousands of biological variables, generating a staggering amount of information.

The exercise was in stark contrast to the cursory workup most of us receive when we go to the doctor for our regular physical exam.

"Currently, we routinely measure fewer than 20 variables in a standard laboratory blood test," said Snyder, who is also the Stanford W. Ascherman, MD, FACS, Professor in Genetics.

"We could, and should, be measuring many, many thousands."

For Snyder, one set of measurements was particularly telling. On day 301, about 12 days after a viral infection, his glucose regulation appeared to be abnormal. Shortly thereafter his glucose levels became elevated, prompting him to visit his primary care physician. On day 369, he was diagnosed with type-2 diabetes.

"We are all responsible for our own health," said Snyder.

"Normally, I go for a physical exam about once every two or three years. So, under normal circumstances, my diabetes wouldn't have been diagnosed for one or two years. But with this real-time information, I was able to make diet and exercise changes that brought my blood sugar down and allowed me to avoid diabetes medication."

Snyder started his study in the months after arriving at Stanford in 2009, when whole-genome sequencing of individuals was just becoming a reality. Stephen Quake, PhD, who is Stanford's Lee Otterson Professor of Bioengineering, had recently completed the complete sequencing of his own genome and was working to use the information to predict his risk for dozens of diseases.

But while the predictive power in genomic information is due in part to its static nature — because it doesn't change over time, a one-time analysis can hint at future events — our bodies are dynamic. They use our DNA blueprints to churn out RNA and protein molecules in varying amounts and types precisely calibrated to respond to the changing conditions in which we live. The result is an exquisitely crafted machine that turns on a dime to metabolize food, flex our muscles, breathe air, fight off infections and make all the other little adjustments that keep us healthy. A misstep can lead to disease or illness.

To generate Snyder's iPOP, he first had his complete genome sequenced at a level of accuracy that has not been achieved previously. Then, with each sample, the researchers took dozens of molecular snapshots, using a variety of different techniques, of thousands of variables and then compared them over time. The composite result was a dynamic picture of how his body responded to illness and disease.

A number of molecular cues led to the discovery of Snyder's diabetes. His genomic sequence suggested he had an increased risk for high cholesterol, coronary artery disease (which he knew already), as well as basal cell carcinoma and type-2 diabetes, which was unexpected. Conversely, the sequence predicts his risk for hypertension, obesity and prostate cancer is lower than that of other men his age (54 when the study started). A check of his triglyceride levels at the start of the study confirmed that they were high: 321 mg/dL. Snyder took the cholesterol-lowering drug simvastatin, and his levels dropped dramatically to 81-116 mg/dL. Based on the type-2 diabetes prediction, the team decided to also monitor Snyder's blood sugar levels, which were normal when the study began.

Snyder, who has two small children, experienced two viral infections during the course of the study: one with rhinovirus (at day 0), and one with respiratory syncytial virus (beginning at day 289). Each time, his immune system reacted by increasing the blood levels of pro-inflammatory cytokines — secreted proteins that cells use to communicate and coordinate their responses to external events such as an infection. Snyder also exhibited increased levels of auto-antibodies, or antibodies that reacted with his own proteins, after viral infection. Although auto-antibody production can be a normal, temporary reaction to illness, the researchers were interested to note that one in particular targeted an insulin receptor binding protein.

The researchers also sequenced the RNA transcripts present in Snyder's cells during infection at an unheard-of level of detail.

"We generated 2.67 billion individual reads of the transcriptome, which gave us a degree of analysis that has never been achieved before," said Snyder.

"This enabled us to see some very different processing and editing behaviors that no one had suspected. We also have two copies of each of our genes and we discovered they often behave differently during infection."

Overall, the researchers tracked nearly 20,000 distinct transcripts coding for 12,000 genes and measured the relative levels of more than 6,000 proteins and 1,000 metabolites in Snyder's blood.

In Snyder's case, the researchers observed unexpected relationships and pathways between viral infection and type-2 diabetes by comparing the results of a variety of "omics" studies.

"This study opens the door to better understanding this concerted regulation, how our bodies interact with the environment and how we can best target treatment for many other complex diseases at a truly personal level," said Li-Pook-Than.

The researchers identified about 2,000 genes that were expressed at higher levels during infection, including some involved in immune processes and the engulfment of infected cells, and about 2,200 genes that were expressed at lower levels, including some involved in insulin signaling and response.

"We were looking for common pathways that were changing in response to infection," said Snyder.

"In a study like this, you are your own best control. You compare your altered, or infected, states with the values you see when you are healthy."

Snyder's iPOP is a proof of principle that the researchers hope will lead to a more-streamlined, less-complex version for regular use in the clinic.

"In the future, we may not need to follow 40,000 variables," said Snyder.

"It's possible that only a subset of them will be truly predictive of future health. But studies like these are important to know which are important and which don't add much to our understanding.”

"Right now, this type of analysis is very expensive. But we have to expect that, like whole-genome sequencing, it will get much cheaper. And we also have to consider the savings to society from preventing disease."

Contact: Krista Conger
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Tuesday, 11 May 2010

Viruses Against Cancer

Complete regression of malignant brain tumours after treatment with parvoviruses
Tuesday, 11 May 2010

Particular parvoviruses normally infect rodents, but they are also infectious for human cells. However, they do not cause any disease symptoms in humans. Most importantly, these viruses have an astonishing property: They kill infected tumours cells without causing any damage to healthy tissue. Therefore, scientists in the teams of Jean Rommelaere and Jörg Schlehofer at the German Cancer Research Center (Deutsches Krebsforschungszentrum, DKFZ) have been investigating over the past years whether these viruses can be used as weapons against cancer.

Many different viruses have been tested before in cancer therapy, particularly for treating those types of cancer for which there are no effective established treatment methods. The DKFZ researchers realized early on that parvovirus H-1 has important advantages over other viruses. Now they have been the first to prove that malignant glioblastomas regress completely as a result of treatment with these viruses.

The treatment experiments were conducted in rats who had received brain tumours cells by implantation. Once the resulting brain tumours had reached a specified size, the animals were given parvoviruses, either by direct injection into the tumour or via the blood stream. In those animals in which the viruses had been injected directly into the tumour, the tumours shrank visibly after only three days and even disappeared completely in eight of twelve animals treated. The rodents survived without any symptoms, while untreated control animals suffered from severe disease symptoms within three weeks following tumour cell implantation. In the intravenously treated group, tumours regressed completely in six of nine animals. The animals have survived for more than one year now without any symptoms or late side effects of therapy.

The researchers found no infection-related damage in the nervous tissue surrounding the tumour. The viruses did not spread to the whole organism. Although parvovirus DNA was detectable in all organs after several days following virus transfer, this was only for a short time. The viruses had infected healthy cells, but these did not produce a new virus generation. However, in the tumour tissue, the viruses reproduced and viral protein production was detected only in these cells. In rats that did not bear tumours, the viruses did not reproduce. Thus, it appears that the presence of cancer cells is a necessary condition for the parvoviruses to reproduce.

After the positive results of these experiments the DKFZ researchers are convinced that parvoviruses are suitable candidates for use in cancer treatment.

Professor Jean Rommelaere summarizes the reasons why:

"Parvovirus H-1 does not cause any disease symptoms in humans. Since we are normally not immune against rodent viruses, it is not immediately eliminated by the human immune system after injection. Parvoviruses kill tumours due to natural properties so that their genetic material does not need to be genetically manipulated like herpes viruses, polio viruses or adenoviruses, which have been used in other studies. Moreover, they do not incorporate their genetic material into the host cell's genome, so we need not fear that they might 'accidentally' boost growth-promoting genes."

Rommelaere's colleague, Jörg Schlehofer, adds two more qualities that could be decisive for therapy of glioblastomas, in particular:

"Parvoviruses pass the blood brain barrier so that they can be administered via the blood stream. In addition, they reproduce in cancer cells, which is particularly important for successful treatment of glioblastoma with its diffuse growth. Thus, the second generation viruses reach and eliminate even those cancer cells that have already settled at some distance from the primary tumour."

Parvovirus therapy to be tested in clinical trial
The promising results in the animal model have encouraged the DKFZ scientists, jointly with Dr. Karsten Geletneky of the Neurosurgery Department of Heidelberg University, to plan a clinical trial on the treatment of advanced glioblastomas. Glioblastoma is considered the most threatening type of brain tumour; only about half of those affected survive the first year after diagnosis. Even innovative drugs that have been made available recently can prolong survival only marginally. Therefore, new treatment approaches for this type of cancer are urgently needed.

Preparing such a trial is a tremendous effort. Thus, large amounts of virus have to be produced under controlled conditions for toxicological tests. Therefore, even a large institute like DKFZ could not afford financing a transfer of these results into clinical practice. Continuation of viral therapy development was made possible only by funds from Munich-based company Oryx. The company aims to provide funds for the development of therapeutically effective substances into clinically applicable drugs in an early stage.

Many of the required toxicological data have already been obtained and submitted to the drug approval authority by the researchers so that they expect to be able to admit the first patients to the trial by the end of the year. In addition, DKFZ and Oryx have recently signed another agreement: Oryx will also get involved in the development of a parvovirus therapy against pancreatic cancer.

Reference:
Regression of advanced rat and human gliomas by local or systemic treatment with oncolytic parvovirus H-1 in rat models
Karsten Geletneky, Irina Kiprianova, Ali Ayache, Regina Koch, Marta Herrero y Calle, Laurent Deleu, Clemens Sommer, Nadja Thomas, Jean Rommelaere and Jörg R. Schlehofer
Neuro-Oncology 2010, DOI: 10.1093/neuonc/noq023
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Wednesday, 13 January 2010

The Viruses Within and What Keeps Them There

Biologists wake dormant viruses and uncover mechanism for survival Wednesday, 13 January 2010 This is Professor Trono with co-author Helen Rowe in their laboratory. Credit: EPFL.It is known that viral "squatters" comprise nearly half of our genetic code. These genomic invaders inserted their DNA into our own millions of years ago when they infected our ancestors. But just how we keep them quiet and prevent them from attack was more of a mystery until EPFL researchers revived them. The reason we survive the presence of these endogenous retroviruses — viruses that attack and are passed on through germ cells, the cells that give rise to eggs and sperm — is because something keeps the killers silent. Now, publishing in the journal Nature, Didier Trono and his team from EPFL, in Switzerland, describe the mechanism. Their results provide insights into evolution and suggest potential new therapies in fighting another retrovirus — HIV. This shows the functioning of Kap1 protein in mouse embryonic stem cells cells. Credit: Pascal Coderay, pascal@salut.ch.By analysing embryonic stem cells in mice within the first few days of life, Trono and team discovered that mouse DNA codes for an army of auxiliary proteins that recognize the numerous viral sequences littering the genome. The researchers also demonstrated that a master regulatory protein called KAP1 appears to orchestrate these inhibitory proteins in silencing would-be viruses. When KAP1 is removed, for example, the viral DNA "wakes up," multiplies, induces innumerable mutations, and the embryo soon dies. Because retroviruses tend to mutate their host's DNA, they have an immense power and potential to alter genes. And during ancient pandemics, some individuals managed to silence the retrovirus involved and therefore survived to pass on the ability. Trono explains that the great waves of endogenous retrovirus appearance coincide with times when evolution seemed to leap ahead. "In our genome we find traces of the last two major waves. The first took place 100 million years ago, at the time when mammals started to develop, and the second about fifty million years ago, just before the first anthropoid primates," he says. The discovery of the KAP1 mechanism could be of interest in the search for new therapeutic approaches to combat AIDS. The virus that causes AIDS can lie dormant in the red blood cells it infects, keeping it hidden from potential treatments. Waking the virus up could expose it to attack. To view a YouTube video related to this release, please visit The Viruses Within - Interview with Didier Trono, EPFL. Reference: KAP1 controls endogenous retroviruses in embryonic stem cells Helen M. Rowe, Johan Jakobsson, Daniel Mesnard, Jacques Rougemont, Séverine Reynard, Tugce Aktas, Pierre V. Maillard, Hillary Layard-Liesching, Sonia Verp, Julien Marquis, François Spitz, Daniel B. Constam & Didier Trono Nature 463, 237-240 (14 January 2010), doi:10.1038/nature08674 ......... ZenMaster


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Tuesday, 8 December 2009

Stem Cells Can be Engineered to Kill HIV

Innovative strategy could be effective against other chronic viral diseases Tuesday, 08 December 2009 Researchers from the UCLA AIDS Institute and colleagues have for the first time demonstrated that human blood stem cells can be engineered into cells that can target and kill HIV-infected cells — a process that potentially could be used against a range of chronic viral diseases. The study, published Dec. 7 in the-peer reviewed online journal PLoS ONE, provides proof-of-principle — that is, a demonstration of feasibility — that human stem cells can be engineered into the equivalent of a genetic vaccine. "We have demonstrated in this proof-of-principle study that this type of approach can be used to engineer the human immune system, particularly the T-cell response, to specifically target HIV-infected cells," said lead investigator Scott G. Kitchen, assistant professor of medicine in the division of haematology and oncology at the David Geffen School of Medicine at UCLA and a member of the UCLA AIDS Institute. "These studies lay the foundation for further therapeutic development that involves restoring damaged or defective immune responses toward a variety of viruses that cause chronic disease, or even different types of tumours." Taking CD8 cytotoxic T lymphocytes — the "killer" T cells that help fight infection — from an HIV-infected individual, the researchers identified the molecule known as the T-cell receptor, which guides the T cell in recognizing and killing HIV-infected cells. These cells, while able to destroy HIV-infected cells, do not exist in enough quantities to clear the virus from the body. Therefore, the researchers cloned the receptor and genetically engineered human blood stem cells, then placed the stem cells into human thymus tissue that had been implanted in mice, allowing them to study the reaction in a living organism. The engineered stem cells developed into a large population of mature, multifunctional HIV-specific CD8 cells that could specifically target cells containing HIV proteins. The researchers also found that HIV-specific T-cell receptors have to be matched to an individual in much the same way that an organ is matched to a transplant patient. The next step is to test this strategy in a more advanced model to determine if it would work in the human body, said co-author Jerome A. Zack, UCLA professor of medicine in the division of haematology and oncology and associate director of the UCLA AIDS Institute. The researchers also hope to expand the range of viruses against which this approach could be used. However, the results of the study suggest that this strategy could be an effective weapon in the fight against AIDS and other viral diseases. "This approach could be used to combat a variety of chronic viral diseases," said Zack, who is also a professor of microbiology, immunology and molecular genetics. "It's like a genetic vaccine." ......... ZenMaster


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Monday, 11 May 2009

Swine Flu II

Early findings about pandemic potential reported in new study Monday, 11 May 2009 Early findings about the emerging pandemic of a new strain of influenza A (H1N1) in Mexico are published today in Science. Researchers from the MRC Centre for Outbreak Analysis and Modelling at Imperial College London, working in collaboration with the World Health Organisation and public health agencies in Mexico, have assessed the epidemic using data to the end of April. Their key findings are as follows:

  • The data so far is very consistent with what researchers would expect to find in the early stages of a pandemic.
  • The researchers' best estimate is that in Mexico, influenza A (H1N1) is fatal in around 4 in 1,000 cases, which would make this strain of influenza as lethal as the one found in the 1957 pandemic. The researchers stress that healthcare has greatly improved in most countries since 1957 and the world is now better prepared.
  • The epidemic of influenza A (H1N1) is thought to have started in Mexico on 15 February 2009. The data suggests that by the end of April, around 23,000 people were infected with the virus in Mexico and 91 of these died as a result of infection. However, the figures are uncertain – for example, some mild cases may have gone unreported. The numbers infected could be as low as 6,000 people or as high as 32,000 people.
  • The uncertainty around the numbers of people who have been infected with influenza A (H1N1) in Mexico means that the case fatality ratio (CFR) of 0.4% (4 deaths per 1000) cannot be definitely established. The CFR is in the range of 0.3% to 1.5%, but at this stage the researchers believe that 0.4% is the most likely.
  • For every person infected, it is likely that there will be between 1.2 and 1.6 secondary cases. This is high compared to normal seasonal influenza, where around 10-15 percent of the population are likely to become infected. However, it is lower than would be expected for pandemic influenza, where 20-30 percent of the population are likely to become infected.
  • In an outbreak in an isolated village called La Gloria, Mexico, children were twice as likely to become infected as adults, with 61% of those aged under 15 becoming infected, compared with 29% of those over 15. This may suggest that adults have some degree of immunity against infection, because of having been previously infected with a related strain of influenza, or it may mean that children are more susceptible to infection because they interact much more closely together, for example in school, than adults.

Professor Neil Ferguson, the corresponding author of today's research from the MRC Centre for Outbreak Analysis and Modelling at Imperial College London, said: "Our study shows that this virus is spreading just as we would expect for the early stages of a flu pandemic. So far, it has been following a very similar pattern to the flu pandemic in 1957, in terms of the proportion of people who are becoming infected and the percentage of potentially fatal cases that we are seeing.” "What we're seeing is not the same as seasonal flu and there is still cause for concern – we would expect this pandemic to at least double the burden on our healthcare systems. However, this initial modelling suggests that the H1N1 virus is not as easily transmitted or as lethal as that found in the flu pandemic in 1918," added Professor Ferguson. Reference: Pandemic Potential of a Strain of Influenza A (H1N1): Early Findings The WHO Rapid Pandemic Assessment Collaboration Science Published Online May 11, 2009, DOI: 10.1126/science.1176062 ......... ZenMaster


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Swine Flu I

What does it do to pigs? Monday, 11 May 2009 The effects of H1N1 swine flu have been investigated in a group of piglets. Scientists writing in BioMed Central's open access Virology Journal studied the pathology of the virus, finding that all infected animals showed flu-like symptoms between one and four days after infection and were shedding virus two days after infection. Roongroje Thanawongnuwech led a team of researchers from Chulalongkorn University, Bangkok, who infected 22-day old pigs with both the H1N1 strain of swine flu and the less dangerous H3N2 subtype. "The results demonstrated that both swine flu subtypes were able to induce flu-like symptoms and lung lesions in weanling pigs. However the severity of the disease with regard to both gross and microscopic lung lesions was greater in the H1N1-infected pigs", he said. All infected pigs developed respiratory symptoms such as nasal discharge, coughing, sneezing and conjunctivitis. Upon pathological examination, lung lesions large enough to be seen by the naked eye were observed. "These lesions were characterized by dark plum-coloured, consolidated areas on lung lobes and were most severe two days after infection, especially in the H1N1-infected pigs, where approximately a third of the lung was covered", according to Thanawongnuwech. The course of infection was limited to less than a week and none of the animals died. Reference: Pathogenesis of swine influenza virus (Thai isolates) in weanling pigs: an experimental trial Donruethai Sreta, Roongtham Kedkovid, Sophon Tuamsang, Pravina Kitikoon and Roongroje Thanawongnuwech Virology Journal 2009, 6:34 doi:10.1186/1743-422X-6-34 ......... ZenMaster


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Saturday, 7 March 2009

Virus-free Embryonic-like Stem Cells Made from Skin

Breakthrough produces Parkinson's patient-specific stem cells free of harmful reprogramming genes Saturday, 07 March 2009 Researchers reporting in the March 6th issue of the journal Cell have developed a new way to produce human embryonic-like stem cells that are free of the viruses used to insert the key ingredients. They showed they could make those embryonic-like cells by reprogramming cells taken from people with unexplained (or idiopathic) Parkinson's disease. This marks the first time researchers have generated human iPS cells that have maintained their embryonic stem-cell-like properties after the removal of reprogramming genes. "Until this point, it was not completely clear that when you take out the reprogramming genes from human cells, the reprogrammed cells would actually maintain the iPS state and be self-perpetuating," says Frank Soldner, a postdoctoral researcher in Whitehead Member Rudolf Jaenisch's laboratory and co-author of the article. "We used a modified virus you can excise," said Rudolf Jaenisch of The Whitehead Institute and Massachusetts Institute of Technology. "After they've done their job, you can get rid of them." That is important, he explained, because the use of viruses encoding the reprogramming factors represents a major limitation of the current technology. Even low activity of the virus-inserted genes may alter the potential for these human embryonic-like cells, or induced pluripotent stem (iPS) cells, to differentiate into other cell types or to cause cancer. (Pluripotent refers to the ability of these cells to differentiate into most other cell types). In the new study, the team also converted these iPS cells into the neurons that are lost in patients with Parkinson's disease. The Parkinson's disease patient-specific iPS cells and neurons made from them offer a powerful new way to study the disease. The demonstration that this conversion from iPS cells to neurons is possible also represents another step toward the ultimate goal of using such iPS cell-derived cells in replacement therapies. A 2006 report, also in the journal Cell, showed that the introduction of four factors (Oct4, Sox2, c-Myc and Klf4) could transform differentiated cells taken from adult mice into iPS cells with the physical, growth, and genetic characteristics typical of embryonic stem cells. The same recipe was later shown to also work with human skin cells. Last month, a Cell report showed for the first time that adult neural stem cells can take on the characteristics of embryonic stem cells with the addition of just one factor. In each case, the scientists used viral vectors to insert the critical genes that encode the factors. In recent months, other methods to reprogram stem cells without the use of viruses have been developed, but those techniques have so far proved to be very inefficient, Jaenisch added. It also remains unclear whether they could be made to work in human cells. Now, Jaenisch and his colleagues show that fibroblasts from the skin of five patients with idiopathic Parkinson's disease can be efficiently reprogrammed and subsequently differentiated into dopaminergic neurons using "Cre-recombinase excisable viruses" that could be inserted and then removed. In this method, Whitehead researchers used viruses to transfer the four reprogramming genes and a gene coding for the enzyme Cre into skin cells from Parkinson's disease patients. The reprogramming genes were bracketed by short DNA sequences, called loxP, which are recognized by the enzyme Cre. After the skin cells were reprogrammed to iPS cells, the researchers introduced the Cre enzyme into the cells, which removed the DNA between the two loxP sites, thereby deleting the reprogramming genes from the cells. The result is a collection of iPS cells with genomes virtually identical to those of the Parkinson's disease patients from whom original skin cells came. The resulting factor-free iPSCs maintained their ability to differentiate into other cell types, they found. The cells also showed a global gene expression profile more closely related to human embryonic stem cells than to human iPSCs still carrying the virally inserted transgenes. "People had worried that when you have viral vectors, there can still be viral gene expression," Jaenisch said. "We didn't know if it was functionally important or not." The new findings show that indeed it is, a result that Jaenisch said he found rather surprising. "The vector-free cells are much more closely related to embryonic stem cells than to the parental cells [they are derived from]. It argues that even low vector expression somehow changes the transcriptional profile of cells." Those differences may have real implications for the differentiation of virus-carrying iPSCs into other cell types and for their use in transplantation, he said. "The reprogramming factors are known to bind to and affect the expression of 3,000 genes in the entire genome, so having artificial expression of those genes will change the cell's overall gene expression," Dirk Hockemeyer, who is also a co-author of the Cell article. "That's why the four reprogramming genes can mess up the system so much. From now on, it will be tough for researchers to leave the reprogramming genes in iPS cells." Jaenisch's new method for producing virus-free cells represents an important breakthrough for scientists aiming to better understand the causes and consequences of Parkinson's and other diseases. The advance is also good news for those in search of potential new drug therapies as well as those who hope ultimately to use a patient's own cells to replace neurons lost to the degenerative disease. "Such patient-specific cells will provide, for the first time, a system to investigate the proposed molecular and cellular mechanisms of sporadic Parkinson's disease, such as protein aggregation, mitochondrial dysfunction, oxidative stress and altered kinase activity," the researchers noted. Due to the relatively short lifespan of cultured neurons – which survive for a matter of weeks – studies of the typically late-onset disease may require methods to accelerate the development of symptoms, for instance by challenging the cells with oxidative stresses, neurotoxins, or by increasing the activity of known Parkinson's disease-related genes. "Such in vitro models could be utilized for large-scale genetic or drug-based screens since large numbers of hiPSCs can be generated and robustly differentiated into dopaminergic neurons," the researchers wrote. "Furthermore our finding that dopaminergic neurons from Parkinson's disease-patients can be derived regardless of the underlying disease or the age of the donor substantiates the idea that hiPSC-based cell replacement could become a feasible therapeutic option for Parkinson's disease in the future." Because the cells reside in the patients' brains, researchers cannot easily access them to investigate how the disease progresses at the cellular level, what kills the cells, or what might prevent cellular damage. Therefore, the ability to create patient-specific iPS cells, derive the dopamine-producing cells, and study those patient-specific cells in the lab could be a great advantage for Parkinson's disease researchers. Although the initial results are extremely promising, Jaenisch acknowledges that the process is far from over. "The next step is to use these iPS-derived cells as disease models, and that's a high bar, a real challenge. I think a lot of work has to go into that." Reference: Parkinson's disease patient-derived induced pluripotent stem cells free of viral reprogramming factors Frank Soldner, Dirk Hockemeyer, Caroline Beard, Qing Gao, George W. Bell, Elizabeth G. Cook, Gunnar Hargus, Alexandra Blak, Oliver Cooper, Maisam Mitalipova, Ole Isacson, Rudolf Jaenisch Cell, Volume 136, Issue 5, 964-977, 6 March 2009, doi:10.1016/j.cell.2009.02.013 ......... ZenMaster


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Thursday, 8 January 2009

Converting Adult Somatic Cells to Pluripotent Stem Cells Using a Single Virus

A single lentiviral vector of the expression of a 'stem cell cassette' dramatically boosts reprogramming efficiency and puts iPS technology one step closer toward human clinical trials. Thursday, 08 January 2009 A Boston University School of Medicine-led research team has discovered a more efficient way to create induced pluripotent stem (iPS) cells, derived from mouse fibroblasts, by using a single virus vector instead of multiple viruses in the reprogramming process. The result is a powerful laboratory tool and a significant step toward the application of embryonic stem cell-like cells for clinical purposes such as the regeneration of organs damaged by inherited or degenerative diseases, including emphysema, diabetes, inflammatory bowel disease, and Alzheimer's Disease. Their research titled "iPS Cell Generation Using a Single Lentiviral Stem Cell Cassette" appears on line in the journal Stem Cells. Prior research studies have required multiple retroviral vectors for reprogramming — steps that depended on four different viruses to transfer genes into the cells' DNA – essentially a separate virus for each reprogramming gene (Oct4. Klf4, Sox2 and c-Myc). Upon activation, these genes convert the cells from their adult, differentiated status to what amounts to an embryonic-like state. However, the high number of genomic integrations — 15 to 20 — that typically occurs when multiple viruses are used for reprogramming, poses a safety risk in humans, as some of these genes (i.e. c-Myc) can cause cancer. In addition, the viruses can integrate in cell locations turning on potential oncogenes. The major milestone the six-member research team, led by Gustavo Mostoslavsky, Boston University Assistant Professor of Medicine in the Gastroenterology Section, achieved was combining the four vectors into a single "stem cell cassette" containing all four genes. The cassette (named STEMCCA) is comprised of a single multi-cistronic mRNA encoding the four transcription factors using a combination of 2A peptide technology and an internal ribosomal entry site (IRES). With the STEMCCA vector, the researchers were able to generate iPS cells more efficiently — 10 times higher than previously reported studies. "The use of a single lentiviral vector for the derivation of iPS cells will help reduce the variability in efficiency that has been observed between different laboratories, thus enabling more consistent genetic and biochemical characterizations of iPS cells and the reprogramming process," the researchers concluded. "We believe that the specific design of the cassette together with the fact that all four genes are expressed from the same transcript could account for the high efficiency we obtained" commented Cesar A. Sommer, first author in the paper and a postdoctoral fellow at Boston University Medical School's Gastroenterology Section. Most importantly, several iPS clones were generated with a single viral integration, a major advance compared to the multiple integrations observed in other studies. "Now we could move forward toward the elimination of the whole cassette using recombination technologies", noted Mostoslavsky. Darrell N. Kotton, another co-author on the paper and an Assistant Professor at Boston University Medical School's Pulmonary Section mentioned that preliminary studies already confirmed that the STEMCCA vector works with high efficiency for the reprogramming of human cells. Reference: iPS Cell Generation Using a Single Lentiviral Stem Cell Cassette Cesar A. Sommer, Matthias Stadtfeld, George J. Murphy, Konrad Hochedlinger, Darrell N. Kotton, Gustavo Mostoslavsky Stem Cells, December 18, 2008; doi:10.1634/stemcells.2008-1075 ......... ZenMaster


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Tuesday, 30 December 2008

Genes That Made 1918 Flu Lethal Isolated

Genes That Made 1918 Flu Lethal Isolated Tuesday, 30 December 2008 By mixing and matching a contemporary flu virus with the "Spanish flu" — a virus that killed between 20 and 50 million people 90 years ago in history's most devastating outbreak of infectious disease — researchers have identified a set of three genes that helped underpin the extraordinary virulence of the 1918 virus. Writing today (Dec. 29) in the Proceedings of the National Academy of Sciences, a team led by University of Wisconsin-Madison virologists Yoshihiro Kawaoka and Tokiko Watanabe identifies genes that gave the 1918 virus the capacity to reproduce in lung tissue, a hallmark of the pathogen that claimed more lives than all the battles of World War I combined. "Conventional flu viruses replicate mainly in the upper respiratory tract: the mouth, nose and throat. The 1918 virus replicates in the upper respiratory tract, but also in the lungs," causing primary pneumonia among its victims, says Kawaoka, an internationally recognized expert on influenza and a professor of pathobiological sciences in the UW-Madison School of Veterinary Medicine. "We wanted to know why the 1918 flu caused severe pneumonia." Autopsies of 1918 flu victims often revealed fluid-filled lungs severely damaged by massive haemorrhaging. Scientists assumed that the ability of the virus to take over the lungs is associated with the pathogen's high level of virulence, but the genes that conferred that ability were unknown. Discovery of the complex and its role in orchestrating infection in the lungs is important because it could provide a way to quickly identify the potential virulence factors in new pandemic strains of influenza, Kawaoka says. The complex could also become a target for a new class of antiviral drugs, which is urgently needed, as vaccines are unlikely to be produced fast enough at the outset of a pandemic to blunt its spread. To find the gene or genes that enabled the virus to invade the lungs, Kawaoka and his group blended genetic elements from the 1918 flu virus with those of a currently circulating avian influenza virus and tested the variants on ferrets, an animal that mimics human flu infection. Substituting single genes from the 1918 virus onto the template of a much more benign contemporary virus yielded, for the most part, agents that could only replicate in the upper respiratory tract. One exception, however, included a complex of three genes that, acting in concert with another key gene, allowed the virus to efficiently colonize lung cells and make RNA polymerase, a protein necessary for the virus to reproduce. "The RNA polymerase is used to make new copies of the virus," Kawaoka explains. Without the protein, the virus is unable to make new virus particles and spread infection to nearby cells. In the late 1990s, scientists were able to recover genes from the 1918 virus by looking in the preserved lung tissue of some of the pandemic's victims. Using the relic genes, Kawaoka's group was able to generate viruses that carry different combinations of the 1918 virus and modern seasonal influenza virus. When tested, most of the hybrid viruses only infected the nasal passages of ferrets and did not cause pneumonia. However, one did infect the lungs, and it carried the RNA polymerase genes from the 1918 virus that allowed the virus to make the key step of synthesizing its proteins. In 2004, Kawaoka and his team identified another key gene from the 1918 virus that enhanced the pathogen's virulence in mice. That gene makes haemagglutinin, a protein found on the surface of the virus and that confers on viral particles the ability to attach to host cells. "Here, I think we are talking about another mechanism," Kawaoka says. “The RNA polymerase is used to make copies of the virus once it has entered a host cell. The role of haemagglutinin is to help the virus gain access to cells.” Reference: Viral RNA polymerase complex promotes optimal growth of 1918 virus in the lower respiratory tract of ferrets Tokiko Watanabe, Shinji Watanabe, Kyoko Shinya, Jin Hyun Kim, Masato Hatta, and Yoshihiro Kawaoka PNAS, December 29, 2008, doi: 10.1073/pnas.0806959106 ......... ZenMaster


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Friday, 5 December 2008

Gene Therapy Corrects Sickle Cell Disease in Laboratory

New St. Jude treatment alleviates long-term anaemia and organ damage in mice and paves the way for human applications Friday, 05 December 2008 Using a harmless virus to insert a corrective gene into mouse blood cells, scientists St. Jude Children's Research Hospital have alleviated sickle cell disease pathology. In their studies, the researchers found that the treated mice showed essentially no difference from normal mice. Although the scientists caution that applying the gene therapy to humans presents significant technical obstacles, they believe that the new therapy will become an important treatment for the disease. Sickle cell disease, which affects millions of people worldwide, arises because of a tiny genetic defect in the gene for beta-globin, a protein component of haemoglobin. This defect causes haemoglobin-containing red blood cells to tend to deform, clump and break apart. The resulting clogged blood vessels can lead to cognitive dysfunction by causing small strokes in the brain and cause damage to kidneys, liver, spleen and lungs. The only permanent cure for the disease is a bone marrow transplant to give recipients blood-forming cells that will form normal beta-globin. However, such transplants are rare because of the lack of compatible donors. Researchers have long known that symptoms of the disease could be alleviated by persistence in the blood of an immature foetal form of haemoglobin in red blood cells. This immature haemoglobin, which usually disappears after birth, does not contain beta-globin, but another form called gamma-globin. St. Jude researchers had found that treating patients with the drug hydroxyurea encourages the formation of foetal haemoglobin and alleviates disease symptoms. "While this is a very useful treatment for the disease, our studies indicated that it might be possible to cure the disorder if we could use gene transfer to permanently increase foetal haemoglobin levels," said Derek Persons, M.D., Ph.D., assistant member in the St. Jude Department of Hematology. He and his colleagues developed a technique to insert the gene for gamma-globin into blood-forming cells using a harmless viral carrier. The researchers extracted the blood-forming cells, performed the viral gene insertion in a culture dish and then re-introduced the altered blood-forming cells into the body. The hope was that those cells would permanently generate red blood cells containing foetal haemoglobin, alleviating the disease. In the experiments, reported in the advanced, online issue of the journal Molecular Therapy, the researchers used a strain of mouse with basically the same genetic defect and symptoms as humans with sickle cell disease. The scientists introduced the gene for gamma-globin into the mice's blood-forming cells and then introduced those altered cells into the mice. The investigators found that months after they introduced the altered blood-forming cells, the mice continued to produce gamma-globin in their red blood cells. "When we examined the treated mice, we could detect little, if any, disease using our methods," said Persons, the paper's senior author. "The mice showed no anaemia, and their organ function was essentially normal." The researchers also transplanted the altered blood-forming cells from the original treated mice into a second generation of sickle cell mice to show that the gamma-globin gene had incorporated itself permanently into the blood-forming cells. Five months after that transplantation, the second generation of mice also showed production of foetal haemoglobin and correction of their disease. "We are very encouraged by our results," Persons said. "They demonstrate for the first time that it is possible to correct sickle cell disease with genetic therapy to produce foetal haemoglobin. We think that increased foetal haemoglobin expression in patients will be well tolerated and the immune system would not reject the haemoglobin, in comparison to other approaches." While Persons believes that the mouse experiments will lead to treatments in humans, he cautioned that technical barriers still need to be overcome. "It is far easier to achieve high levels of gene insertion into mouse cells than into human cells," he said. "In our mouse experiments, we routinely saw one or two copies of the gamma-globin gene inserted into each cell. However, in humans this insertion rate is at least a hundred-fold less." Persons' laboratory is currently working with other animal and human cells to develop methods to achieve a high enough gene insertion rate to make the gene therapy clinically useful. About St. Jude Children's Research Hospital St. Jude Children's Research Hospital is internationally recognized for its pioneering work in finding cures and saving children with cancer and other catastrophic diseases. Founded by late entertainer Danny Thomas and based in Memphis, Tenn., St. Jude freely shares its discoveries with scientific and medical communities around the world. No family ever pays for treatments not covered by insurance, and families without insurance are never asked to pay. St. Jude is financially supported by ALSAC, its fundraising organization. ......... ZenMaster


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Tuesday, 4 November 2008

Effective HPV Vaccine Needed for Both Men and Women

Effective HPV Vaccine Needed for Both Men and Women Monday, 03 November 2008 A call to explore a broader use of HPV (human papillomavirus) vaccines and the validation of a simple oral screening test for HPV-caused oral cancers are reported in two studies by a Johns Hopkins Kimmel Cancer Center investigator. Leading HPV expert Maura Gillison, M.D., Ph.D., reports her latest work in the November 3, 2008, journal Clinical Cancer Research and in a Centers for Disease Control and Prevention (CDC) monograph. The CDC report on HPV-associated cancers appears on line November 3 and in the November 15, 2008, supplement edition of Cancer. She was the first to identify HPV infection as the cause of certain oral cancers and who identified multiple sex partners as the most important risk factor for these cancers. In the CDC report, believed to be the first and most comprehensive assessment of HPV-associated cancer data in the United States, investigators analyzed cancer registry data from 1998-2003 and found 25,000 cancer cases each year occurred at cancer sites associated with HPV infection. In additional analysis, Gillison and colleagues at the National Cancer Institute identified HPV infection as the underlying cause of approximately 20,000 of these cancers. Gillison and team found approximately 20,000 cases of cancer in the United States each year are caused by HPV infection. Oral cancers are the second most common type of HPV-associated cancers and are increasing in incidence in the U.S., particularly among men. Add to that anal, penile, vaginal, and vulvar cancers that are also linked to HPV infection, and Gillison says these cancers, when combined, equal the number of cervical cancers, the most common and well known of the cancers caused by HPV. While about one-quarter of HPV-linked cancers occur in men, vaccines are currently approved only for use in girls and young women for cervical cancer prevention. "We need to have a more comprehensive discussion of the potential impact the HPV vaccine could have on cancer rates among men and women in this country," says Gillison, associate professor of oncology. "Currently available HPV vaccines have the potential to reduce the rates of HPV-associated cancers, like oral and anal cancers, that are currently on the rise and for which there no effective or widely-applied screening programs." Gillison notes, however, that studies are needed to confirm that the vaccine effectively prevents HPV infections that lead to oral and anal cancers. Gillison's findings were part of a project known as ABHACUS (Assessing the Burden of Human Papillomavirus-Associated Cancers). The data studied came from the CDC's National Program of Cancer Registries and the National Cancer Institute's Surveillance, Epidemiology, and End Results program. More than 80 investigators from across the country participated in the project, which addressed a variety of HPV-cancer associated issues, including racial disparity, economic impact, behavioural risk factors, and cancer mortality. Other than prevention, early detection is held by cancer experts as the best way to control cancer. In the Clinical Cancer Research study, the first to track the disease and related oral infections over an extended period, Gillison found that simple "swish and spit" oral rinses can successfully track oral HPV infection over time. These findings open the door to a potential, non-invasive screening test to detect the disease and monitor for tumour recurrence. Head and neck cancer is the broad term for a variety of cancers of the oral cavity, including the tonsils, base of the tongue, and the side and back wall of the throat. The study found that oral rinses successfully detected high-risk HPV infections in patients with HPV 16-positive head and neck cancers for up to five years after treatment for their cancer. Gillison says the findings indicate a high rate of persistent infection and reaffirms the connection between high-risk types of HPV and HPV-positive head and neck cancers. In the study, the researchers used oral rinses to collect cells shed from inside the mouths of 135 head and neck cancer patients. The researchers genetically sequenced the DNA obtained from the rinses and tumour samples to identify those with HPV-positive cancers and determine the HPV type. There are approximately 120 types of HPV, but HPV 16 is one of the two most common associated with cancer. The analysis revealed 44 patients with HPV 16-positive tumours and found that these patients were more likely to have continuing oral HPV 16 infections both before and after cancer treatment. While this study did not link the continued post-treatment infections to tumour recurrence, it was noted that patients with high-risk oral HPV infections prior to therapy, maintained high rates of infection after completing therapy. The team plans further, long-term research to determine if this continued infection leads to cancer recurrence. In 2000, Gillison identified HPV-positive head and neck cancer as a distinct subtype of the disease and linked it to improved survival. "There is no question of cause," says Gillison. "It has now become a question of tracking the infection over time to identify those at risk of developing cancer or cancer recurrence." See also: CDC Releases First Estimate of Human Papillomavirus-Associated Cancer Data CDC News - November 3, 2008 ......... ZenMaster


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Monday, 20 October 2008

Is A Universal Bird Flu Vaccine Achievable?

H9N2 virus ‘core’ bird flu vaccine protects both people and pets Monday, 20 October 2008 A single vaccine could be used to protect chickens, cats and humans against deadly flu pandemics, according to an article published in the November issue of the Journal of General Virology. The vaccine protects birds and mammals against different flu strains and can even be given to birds while they are still in their eggs, allowing the mass vaccination of wild birds. The emergence of bird flu has posed a major challenge to scientists designing vaccines as it can infect a number of different animals, including birds, pets and people. Now, researchers in the USA have discovered that a vaccine based on a bird flu virus could be used to protect several species against different influenza viruses. "The world is experiencing a pandemic of influenza in birds caused by an H5N1 virus. Although it has been restricted to Eurasia and some countries in Africa, there is a risk that this virus may spread worldwide," said Professor Daniel Perez from the University of Maryland, USA. "The H5N1 virus also has an unusual expanded host range: not only birds and humans have been infected but also cats, which are usually resistant to influenza. To prepare for a pandemic, it would be ideal to have a vaccine that could be used in multiple animal species." The researchers found that the central genes or 'backbone' of the H9N2 virus that infects guinea fowl can protect birds and mice against highly pathogenic strains of influenza. They modified the virus to make it less pathogenic and then used it to vaccinate mice. Three weeks after being vaccinated, the mice were infected with the potentially lethal H1N1 virus – the same virus that caused the 1918 Spanish flu pandemic. All the vaccinated mice survived with no signs of disease. Vaccinated mice also survived infection with the deadly H5N1 bird flu virus, again showing no signs of disease. "Our results show that the H9N2 backbone vaccine can be used to protect mice against two different, highly pathogenic strains of influenza. We chose genes from H9N2 influenza for the vaccine because the virus can infect many different animals, including chickens, mice and pigs," said Professor Perez. "A very important limitation in the current design of flu vaccines is that they are usually species specific. Our approach involves a universal backbone that can be used in several different species, including humans." More importantly, this live attenuated virus provided effective protection when it was administered to birds before they had hatched. By vaccinating eggs against influenza, we could protect wild bird species as well as domestic chickens against pandemic flu strains, limiting the spread of disease to humans. "If an emerging strain of bird flu spreads among a broad range of animal species, we should expect major health, economic and ecological consequences," said Professor Perez. "It is unrealistic to consider preparing different vaccines specifically tailored to different animal species in this situation. An influenza vaccine that could protect different species would save valuable time during a pandemic." ......... ZenMaster


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Friday, 10 October 2008

New Technique to Reprogram Stem Cells

Yamanaka eliminates viral vector in stem cell reprogramming, improving safety Friday, 10 October 2008 Shinya Yamanaka MD, PhD, of Kyoto University and the Gladstone Institutes of Cardiovascular Disease (GICD) has taken another step forward in improving the possibilities for the practical application of induced pluripotent stem (iPS) cell technology. Shinya Yamanaka MD, PhD.. Courtesy of the Gladstone Institutes.Previously, Dr. Yamanaka had shown that adult cells can be reprogrammed to become embryonic stem cell–like using a cancer-causing oncogene as one of the four genes required to reprogram the cells, and a virus to transfer the genes into the cells. In the last year, Dr. Yamanaka and other labs showed that the oncogene, c-Myc, is not needed. However the use of viruses that integrate into the genome prohibit use of iPS cells for regenerative medicine because of safety concerns: its integration into the cell's genome might activate or inactivate critical host genes. Now Dr. Yamanaka's laboratory in Kyoto has eliminated the need for the virus. In a report published this week in Science, they showed that the critical genes can be effectively introduced without using a virus. The ability to reprogram adult cells into iPS cells without viral integration into the genome also lays to rest concerns that the reprogramming event might be dependent upon viral integration into specific genomic loci that could mediate the genetic switch. "The iPS field and stem cell research in general is progressing rapidly," said GICD Director Deepak Srivastava, MD. "But, as Shinya has shown, each step forward reveals a new set of challenges." Dr. Yamanaka's team began this series of experiments by replacing the retrovirus with an adenoviral vector. While transfections with the genes on separate vectors didn't work, they did work when the genes were arranged in a specific order on a single vector. The same arrangement worked when the genes were incorporated into a plasmid. To determine if the plasmid-mediated reprogrammed cells were pluripotent, the scientists transplanted the cells under the skin of immunocompromised mice. The resulting tumours contained a wide variety of cell types from all three germ layers. iPS cells injected into embryos resulted in chimeric mice with the injected cells contributing to almost all cell types. Still, other problems remain to be solved. The efficiency of the gene transfer with the plasmid was lower than with the retrovirus. Nevertheless, this significant step moves us closer to realizing the promise of stem cells in the understanding and eventual cure of diseases. About the Gladstone Institutes The J. David Gladstone Institutes, affiliated with the University of California, San Francisco (UCSF), is dedicated to the health and welfare of humankind through research into the causes and prevention of some of the world's most devastating diseases. Gladstone is comprised of the Gladstone Institute of Cardiovascular Disease, the Gladstone Institute of Virology and Immunology and the Gladstone Institute of Neurological Disease. Reference: Generation of Mouse Induced Pluripotent Stem Cells without Viral Vectors Okita K, Nakagawa M, Hyenjong H, Ichisada T, Yamanaka S. Science, Published Online October 9, 2008, DOI: 10.1126/science.1164270 ......... ZenMaster


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Monday, 6 October 2008

Virus Work Awarded in 2008 Nobel Prize in Physiology or Medicine

HPV and HIV virus work avarded this year Monday, 06 October 2008 Harald zur Hausen in his lab.Harald zur Hausen, at the German Cancer Research Centre in Heidelberg, Germany get half of the Nobel Prize in Physiology or Medicine 2008: "for his discovery of human papilloma viruses causing cervical cancer". See also in CellNEWS: Vaccine Makes Cervical Cancer Control Feasible Thursday, 28 August 2008 Luc MontagnierFrançoise Barré-SinoussiThe other half is scared by Françoise Barré-Sinoussi and Luc Montagnier, Institut Pasteur Paris, France, for their discovery of HIV/AIDS virus: "for their discovery of human immunodeficiency virus" .

See also: Dispute behind Nobel Prize for HIV research French researchers win for virus discovery; controversial scientist shunned MSNBC - October 6, 2008 This year's Nobel Prize awards discoveries of two viruses causing severe human diseases. Harald zur Hausen went against current dogma and postulated that oncogenic human papilloma virus (HPV) caused cervical cancer, the second most common cancer among women. He realized that HPV-DNA could exist in a non-productive state in the tumours, and should be detectable by specific searches for viral DNA. He found HPV to be a heterogeneous family of viruses. Only some HPV types cause cancer. His discovery has led to characterization of the natural history of HPV infection, an understanding of mechanisms of HPV-induced carcinogenesis and the development of prophylactic vaccines against HPV acquisition. Françoise Barré-Sinoussi and Luc Montagnier discovered human immunodeficiency virus (HIV). Virus production was identified in lymphocytes from patients with enlarged lymph nodes in early stages of acquired immunodeficiency, and in blood from patients with late stage disease. They characterized this retrovirus as the first known human lentivirus based on its morphological, biochemical and immunological properties. HIV impaired the immune system because of massive virus replication and cell damage to lymphocytes. The discovery was one prerequisite for the current understanding of the biology of the disease and its antiretroviral treatment. Discovery of human papilloma virus causing cervical cancer Against the prevailing view during the 1970s, Harald zur Hausen postulated a role for human papilloma virus (HPV) in cervical cancer. He assumed that the tumour cells, if they contained an oncogenic virus, should harbour viral DNA integrated into their genomes. The HPV genes promoting cell proliferation should therefore be detectable by specifically searching tumour cells for such viral DNA. Harald zur Hausen pursued this idea for over 10 years by searching for different HPV types, a search made difficult by the fact that only parts of the viral DNA were integrated into the host genome. He found novel HPV-DNA in cervix cancer biopsies, and thus discovered the new, tumourigenic HPV16 type in 1983. In 1984, he cloned HPV16 and 18 from patients with cervical cancer. The HPV types 16 and 18 were consistently found in about 70% of cervical cancer biopsies throughout the world. Importance of the HPV discovery The global public health burden attributable to human papilloma viruses is considerable. More than 5% of all cancers worldwide are caused by persistent infection with this virus. Infection by the human papilloma virus is the most common sexually transmitted agent, afflicting 50-80% of the population. Of the more than 100 HPV types known, about 40 infect the genital tract, and 15 of these put women at high risk for cervical cancer. In addition, HPV is found in some vulval, penile, oral and other cancers. Human papilloma virus can be detected in 99.7% of women with histologically confirmed cervical cancer, affecting some 500,000 women per year. Harald zur Hausen demonstrated novel properties of HPV that have led to an understanding of mechanisms for papilloma virus-induced carcinogenesis and the predisposing factors for viral persistence and cellular transformation. He made HPV16 and 18 available to the scientific community. Vaccines were ultimately developed that provide ≥95 % protection from infection by the high risk HPV16 and 18 types. The vaccines may also reduce the need for surgery and the global burden of cervical cancer.


Nobel Prize in Physiology or Medicine 2008
Discovery of HIV Following medical reports of a novel immunodeficiency syndrome in 1981, the search for a causative agent was on. Françoise Barré-Sinoussi and Luc Montagnier isolated and cultured lymph node cells from patients that had swollen lymph nodes characteristic of the early stage of acquired immune deficiency. They detected activity of the retroviral enzyme reverse transcriptase, a direct sign of retrovirus replication. They also found retroviral particles budding from the infected cells. Isolated virus infected and killed lymphocytes from both diseased and healthy donors, and reacted with antibodies from infected patients. In contrast to previously characterized human oncogenic retroviruses, the novel retrovirus they had discovered, now known as human immunodeficiency virus (HIV), did not induce uncontrolled cell growth. Instead, the virus required cell activation for replication and mediated cell fusion of T lymphocytes. This partly explained how HIV impairs the immune system since the T cells are essential for immune defence. By 1984, Barré-Sinoussi and Montagnier had obtained several isolates of the novel human retrovirus, which they identified as a lentivirus, from sexually infected individuals, haemophiliacs, mother to infant transmissions and transfused patients. The significance of their achievements should be viewed in the context of a global ubiquitous epidemic affecting close to 1% of the population. Importance of the HIV discovery Soon after the discovery of the virus, several groups contributed to the definitive demonstration of HIV as the cause of acquired human immunodeficiency syndrome (AIDS). Barré-Sinoussi and Montagnier's discovery made rapid cloning of the HIV-1 genome possible. This has allowed identification of important details in its replication cycle and how the virus interacts with its host. Furthermore, it led to development of methods to diagnose infected patients and to screen blood products, which has limited the spread of the pandemic. The unprecedented development of several classes of new antiviral drugs is also a result of knowledge of the details of the viral replication cycle. The combination of prevention and treatment has substantially decreased spread of the disease and dramatically increased life expectancy among treated patients. The cloning of HIV enabled studies of its origin and evolution. The virus was probably passed to humans from chimpanzees in West Africa early in the 20th century, but it is still unclear why the epidemic spread so dramatically from 1970 and onwards. Identification of virus−host interactions has provided information on how HIV evades the host’s immune system by impairing lymphocyte function, by constantly changing and by hiding its genome in the host lymphocyte DNA, making its eradication in the infected host difficult even after long-term antiviral treatment. Extensive knowledge about these unique viral host interactions has, however, generated results that can provide ideas for future vaccine development as well as for therapeutic approaches targeting viral latency. HIV has generated a novel pandemic. Never before has science and medicine been so quick to discover, identify the origin and provide treatment for a new disease entity. Successful anti-retroviral therapy results in life expectancies for persons with HIV infection now reaching levels similar to those of uninfected people.
  • Harald zur Hausen, born 1936 in Germany, German citizen, MD at University of Düsseldorf, Germany. Professor emeritus and former Chairman and Scientific Director, German Cancer Research Centre, Heidelberg, Germany.
  • Françoise Barré-Sinoussi, born 1947 in France, French citizen, PhD in virology, Institut Pasteur, Garches, France. Professor and Director, Regulation of Retroviral Infections Unit, Virology Department, Institut Pasteur, Paris, France.
  • Luc Montagnier, born 1932 in France, French citizen, PhD in virology, University of Paris, Paris, France. Professor emeritus and Director, World Foundation for AIDS Research and Prevention, Paris, France.

......... ZenMaster


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