News (Updated July 11, 2010)

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Antibodies That Prevent Most HIV Strains From Infecting Human Cells

11 Jul 2010   

Scientists have discovered two potent human antibodies that can stop more than 90 percent of known global HIV strains from infecting human cells in the laboratory, and have demonstrated how one of these disease-fighting proteins accomplishes this feat. According to the scientists, these antibodies could be used to design improved HIV vaccines, or could be further developed to prevent or treat HIV infection. Moreover, the method used to find these antibodies could be applied to isolate therapeutic antibodies for other infectious diseases as well.

"The discovery of these exceptionally broadly neutralizing antibodies to HIV and the structural analysis that explains how they work are exciting advances that will accelerate our efforts to find a preventive HIV vaccine for global use," says Anthony S. Fauci, M.D., director of the National Institute of Allergy and Infectious Diseases (NIAID), National Institutes of Health. "In addition, the technique the teams used to find the new antibodies represents a novel strategy that could be applied to vaccine design for many other infectious diseases."

Led by a team from the NIAID Vaccine Research Center (VRC), the scientists found two naturally occurring, powerful antibodies called VRC01 and VRC02 in an HIV-infected individual's blood. They found the antibodies using a novel molecular device they developed that homes in on the specific cells that make antibodies against HIV. The device is an HIV protein that the scientists modified so it would react only with antibodies specific to the site where the virus binds to cells it infects.

The scientists found that VRC01 and VRC02 neutralize more HIV strains with greater overall strength than previously known antibodies to the virus.

The researchers also determined the atomic-level structure of VRC01 when it is attaching to HIV. This has enabled the team to define how the antibody works and to precisely locate where it attaches to the virus. With this knowledge, they have begun to design components of a candidate vaccine that could teach the human immune system to make antibodies similar to VRC01 that might prevent infection by the vast majority of HIV strains worldwide.

NIAID scientists Peter D. Kwong, Ph.D., John R. Mascola, M.D., and Gary J. Nabel, M.D., Ph.D., led the two research teams. A pair of articles about these findings appears in the online edition of Science.

"We have used our knowledge of the structure of a virus - in this case, the outer surface of HIV - to refine molecular tools that pinpoint the vulnerable spot on the virus and guide us to antibodies that attach to this spot, blocking the virus from infecting cells," explains Dr. Nabel, the VRC director.

Finding individual antibodies that can neutralize HIV strains anywhere in the world has been difficult because the virus continuously changes its surface proteins to evade recognition by the immune system. As a consequence of these changes, an enormous number of HIV variants exist worldwide. Even so, scientists have identified a few areas on HIV's surface that remain nearly constant across all variants. One such area, located on the surface spikes used by HIV to attach to immune system cells and infect them, is called the CD4 binding site. VRC01 and VRC02 block HIV infection by attaching to the CD4 binding site, preventing the virus from latching onto immune cells.

"The antibodies attach to a virtually unchanging part of the virus, and this explains why they can neutralize such an extraordinary range of HIV strains," says Dr. Mascola, the deputy director of the VRC.

With these antibodies in hand, a team led by Dr. Kwong, chief of the structural biology section at the VRC, determined the atomic-level molecular structure of VRC01 when attached to the CD4 binding site. They then examined this structure in light of natural antibody development to ascertain the steps that would be needed to elicit a VRC01-like antibody through vaccination.

Antibody development begins with the mixing of genes into new combinations within the immune cells that make antibodies. Examination of the structure of VRC01 attached to HIV suggested that, from a genetic standpoint, the immune system likely could produce VRC01 precursors readily. The researchers also confirmed that VRC01 does not bind to human cells - a characteristic that might otherwise lead to its elimination during immune development, a natural mechanism the body employs to prevent autoimmune disease.

In the final stage of antibody development, antibody-producing B cells recognize specific parts of a pathogen and then mutate, or mature, so the antibody can bind to the pathogen more firmly. VRC01 precursors do not bind tightly to HIV, but rather mature extensively into more powerfully neutralizing forms. This extensive antibody maturation presents a challenge for vaccine design. In their paper, Dr. Kwong and colleagues explore how this challenge might be addressed by designing vaccine components that could guide the immune system through this stepwise maturation process and facilitate the generation of a VRC01-like antibody from its precursors. The scientists currently are performing research to identify these components.

The discoveries we have made may overcome the limitations that have long stymied antibody-based HIV vaccine design," says Dr. Kwong.

Notes:
The two research teams included NIAID scientists from the VRC, the Laboratory of Immunoregulation, and the Division of Clinical Research, all in Bethesda, Md.; as well as researchers from Beth Israel Deaconess Medical Center in Boston; Columbia University in New York; Harvard Medical School and Harvard School of Public Health in Boston; The Rockefeller University in New York City; and University of Washington in Seattle.

References:
Wu X et al. Rational design of envelope surface identifies broadly neutralizing human monoclonal antibodies to HIV-1. Science. 2010.
Zhou T et al. Structural basis for broad and potent neutralization of HIV-1 by antibody VRC01. Science. 2010.

 

The Use Of Zinc Finger Nucleases To Generate HIV Resistant Human Stem Cells

10 Jul 2010   

Sangamo BioSciences, Inc. (Nasdaq: SGMO) announced the publication of data demonstrating the preclinical efficacy of a human stem cell therapy for human immunodeficiency virus (HIV) based on its proprietary zinc finger DNA-binding protein nuclease (ZFN) technology. The ZFN approach enables the permanent disruption of the CCR5 gene, which encodes an important receptor for HIV infection, in all the cell types comprising the immune system that develop from hematopoietic stem cells (HSCs), and is the basis for a promising therapeutic strategy for the treatment of HIV/AIDS. Sangamo has two ongoing Phase 1 clinical trials to evaluate the safety and clinical efficacy of this approach in CD4+ T-cells.

The work, which was carried out in the laboratory of Paula Cannon, Ph.D., Associate Professor of Molecular Microbiology & Immunology at the Keck School of Medicine of the University of Southern California (USC), in collaboration with Sangamo scientists, was published on July 2, 2010, as an Advance Online Publication in Nature Biotechnology.

"These are very exciting data that provide proof of concept for a new approach to HIV treatment," said John Zaia, M.D., the Aaron D. and Edith Miller Chair in Gene Therapy and Chair of Virology, City of Hope . "The recent example of the 'Berlin Patient' who appears to have been cured of both his HIV and leukemia by receiving a bone marrow transplant (BMT) of stem cells from a donor that had a naturally occurring CCR5 mutation that makes them resistant to HIV infection, provided the model for this approach. However, the paucity of human donors with this natural CCR5 mutation and the risks of allogeneic BMT mean that we need a more practical solution to make this a therapeutic option. Modification of HSCs using ZFNs to recreate the CCR5 mutation is a potential solution."

Dr. Zaia is the leader of the recent $14.5 million Disease Team Research Award granted by the California Institute for Regenerative Medicine (CIRM) to a multidisciplinary team of investigators which includes City of Hope, Dr. Cannon and her colleagues at USC, and Sangamo scientists. The award funds the preclinical development of a ZFN CCR5-targeted approach which aims to complete an Investigational New Drug (IND) application to the U.S. Food and Drug Administration (FDA) for clinical testing of this ZFN method.

Sangamo's ZFNs are designed to permanently modify the DNA sequence encoding CCR5, a co-receptor that enables HIV to enter and infect cells of the immune system. Individuals carrying a naturally occurring mutation of their CCR5 gene, a variant known as CCR5-delta32, have been shown to be resistant to HIV infection. Building on this observation, a study published in the New England Journal of Medicine in 2009 reported a potential "cure" when an AIDS patient with leukemia received a bone marrow transplant from a "matched" donor with this delta-32 CCR5 mutation. This approach transferred the HSCs residing in the bone marrow from the delta-32 donor, and provided a self-renewable and potentially lifelong source of HIV-resistant immune cells. After transplantation, the patient was able to discontinue all anti-HIV drug treatments, CD4 counts increased, and viral load dropped to an undetectable level, demonstrating effective transplantation of protection from HIV infection. The data reported in the Nature Biotechnology publication replicate these findings for a ZFN-based treatment in a preclinical model.

"The data described in this paper are an important demonstration of the potential therapeutic possibilities of ZFN modification of human stem cells," commented Philip Gregory, D. Phil., Sangamo's vice president of research and chief scientific officer. "We have demonstrated efficient and specific modification of human hematopoietic stem cells, rendering them resistant to infection with HIV-1 while retaining their 'stemness' and ability to differentiate. These data pave the way for the use of this technology in other diseases for which HSC modification may be therapeutically useful."

Data Reported in the Nature Biotechnology Paper

The reported results demonstrate that a one-time exposure to CCR5-specific ZFNs resulted in the generation of an HIV-resistant population of human HSCs by the permanent genetic modification of the CCR5 gene. These ZFN-modified stem cells engrafted in NSG (NOD/SCID/IL2rγnull) mice, which lack a normal immune system and are able to tolerate engraftment of human cells and tissues. After 8-12 weeks the engrafted ZFN-modified human cells could be identified as different immune cell types in the peripheral blood, and various tissues of the mouse suggesting that they were functionally normal. Furthermore, the ZFN-modified HSCs produced progeny that could be harvested from one mouse and engrafted into a second animal, demonstrating that the modified HSCs retain their 'stemness' and ability to differentiate. In addition, the animals did not experience any obvious toxicity or ill-health. In HIV challenge experiments, researchers found that the ZFN-modified cells had a selective advantage over unmodified HSCs and not only survived infection but expanded and appeared to traffic normally to various tissues in the mouse. Moreover, the presence of ZFN-modified cells controlled HIV replication in the animals. These data suggest that human HSCs can be modified with ZFNs, expand and differentiate and have a selective advantage in the presence of HIV allowing them to evade infection and destruction leaving them able fight opportunistic infections and HIV itself.

About HIV/AIDS and CCR5

Human Immunodeficiency Virus (HIV) infection kills or impairs cells of the immune system, progressively destroying the body's ability to fight infections and certain cancers resulting in AIDS (Acquired Immune Deficiency Syndrome). Individuals diagnosed with AIDS are susceptible to life-threatening diseases called opportunistic infections, which are caused by microbes that usually do not cause illness in healthy people. According to UNAIDS/WHO, over 2.7 million people were newly infected with HIV in 2007. An estimated 2.0 million people died of AIDS in the same year. There are now over 33 million people living with HIV and AIDS worldwide. The CDC estimates that, in the United States alone, there were 1.2 million people living with HIV/AIDS, approximately 54,000 new infections and 23,000 deaths in 2007.

CCR5 is the chemokine receptor that HIV uses as a co-receptor to gain entry into immune cells. CCR5 is perhaps the most important of the known co-receptors for HIV, since the most commonly transmitted strains of HIV are strains that bind to CCR5 -- so-called "R5" strains. A small fraction of the population carries a mutation in their CCR5 gene, called the delta32 mutation. This mutated version of the gene results in a truncated CCR5 protein which cannot be used by HIV as a co-receptor. Individuals that have mutant delta 32 versions of both of their CCR5 genes are resistant to infection by R5 HIV strains.

About Sangamo

Sangamo BioSciences, Inc. is focused on the research and development of novel DNA-binding proteins for therapeutic gene regulation and modification. The most advanced ZFP Therapeutic™ development program is currently in a Phase 2b clinical trial for evaluation of safety and clinical effect in patients with diabetic neuropathy and a Phase 2 trial in ALS. Sangamo also has two Phase 1 clinical trials to evaluate safety and clinical effect of a treatment for HIV/AIDS and another Phase 1 trial to evaluate safety and clinical effect of a treatment for recurrent glioblastoma multiforme. Other therapeutic development programs are focused on neuropathic pain, nerve regeneration, Parkinson's disease and monogenic diseases. Sangamo's core competencies enable the engineering of a class of DNA-binding proteins known as zinc finger DNA-binding proteins (ZFPs). By engineering ZFPs that recognize a specific DNA sequence Sangamo has created ZFP transcription factors (ZFP TF) that can control gene expression and, consequently, cell function. Sangamo is also developing sequence-specific ZFP Nucleases (ZFN) for gene modification. Sangamo has established strategic partnerships with companies in non-therapeutic applications of its technology including Dow AgroSciences and Sigma-Aldrich Corporation.

 

Methadone 'improves survival rate'

July 8, 2010

The controversial heroin substitute methadone improves the long-term survival of drug abusers, according to academics.

A study showed the treatment reduced the frequency of drug use and led to a drop in the risk of death by 13% each year.

But the findings also showed the drug can prolong the number of years users continue to inject heroin.

Injecting can kill through overdose and the transmission of HIV and hepatitis. A recent outbreak of anthrax in Scotland caused 13 deaths among injecting users.

The research was carried out by the universities of Edinburgh , Bristol and Cambridge .

Roy Robertson, a GP who led the study at the University of Edinburgh , said: "Many injectors on a prescription will continue to occasionally inject, though may be reluctant to acknowledge this to their doctor for fear of a punitive response.

"Our research shows that despite this they still gain substantial health benefits from their prescription.

"Suggestions that methadone prescribing should be cut back or confined to the short-term are clearly misplaced and would lead to poorer health for drug injectors."

The research comes three months after a group of 40 experts from around the world said methadone should be "readily available" to addicts seeking help.

They argued that scrapping the treatment could lead to a rise in crime and drug deaths. But its use has been criticised by Scottish Conservatives, who claimed addicts are "parked" on methadone.

 

Diagnosing Latent TB: NICE Opens Public Consultation On The Use Of Interferon Gamma Tests

08 Jul 2010   

The National Institute for Health and Clinical Excellence (NICE) has today (8 July) published its draft clinical guideline on diagnosing latent TB in children and adults. A partial update of its 2006 guideline, the draft focuses on the diagnosis of latent TB using tuberculin skin tests (TST, also known as Mantoux and Heaf tests) and the newer interferon-gamma tests (IGT). The draft addresses which diagnostic strategy is most accurate in diagnosing latent TB in adults and children who are recent arrivals from countries where TB is highly prevalent; in adults and children who have been in close contact with patients with active TB, and in adults and children who are immunocompromised.

The original NICE guidance recognised that there was a lack of good quality evidence to show whether interferon-gamma tests are acceptable to patients and are more effective than tuberculin skin tests for predicting subsequent development of active TB, or diagnosing or ruling out current active TB. NICE therefore recommended further research to compare the strategies of skin test only, skin test then interferon gamma test if positive, and interferon gamma test only. Concern was also raised about the appropriateness of IGT use in current clinical practice during the planned review process for the original NICE guideline on the diagnosis and management of TB.

Based on a detailed analysis of this further research, the independent Guideline Development Group (GDG) has concluded that the relative benefit of IGT over TST in determining the need for treatment of latent TB infection is not certain - and in the case of younger children it feels that IGT may even perform less well. However, the GDG has made recommendations in populations where they considered IGT to be of clear benefit, especially in cases where IGT would reduce the uncertain diagnosis of TST. Recommendations in the draft guideline therefore include the following:

To diagnose latent TB in:

-- Household contacts 5 years and older, non household contacts and adult contacts:

- A Mantoux test should be performed. Those with positive results (or in whom Mantoux testing may be less reliable) should then be considered for IGT.

-- Recent arrivals from highly prevalent countries aged 5 - 34 years:

- Offer a Mantoux test followed by IGT if positive.

-- Recent arrivals from highly prevalent countries aged 16 years and above:

- An IGT test alone can be used.

-- Recent arrivals from highly prevalent countries aged under 5 years:

- Use Mantoux as the initial test. If positive, taking into account BCG history, undertake clinical assessment to exclude active disease and consider treatment of latent TB.

-- People who are immunocompromised:

- For people with HIV and CD4 counts (also called T-cells, these are types of cells that help protect the body from infection) of less that 200, perform both an IGT test and a TST. If either test is positive assess for active TB. Consider treatment of latent TB if active disease is excluded.

- For people with HIV and CD4 counts of 200-500, perform an IGT test alone or and IGT test with concurrent TST. If either test is positive, assess for active TB. Consider treatment of latent TB.if active disease is excluded.

Dr Fergus Macbeth, Director of the Centre for Clinical Practice at NICE said: "If TB is left untreated it can be very serious or fatal but antibiotic treatments are highly effective. Up to 15% of adults with latent TB will go on to develop active TB at some point in their lives and the risk in children may be much higher. In people who are immunocompromised - for example, if they are HIV positive - the chance of developing active TB within five years of infection is up to 50%. Detection of latent TB is therefore important in controlling the disease.

He continued: "The newer interferon gamma tests for latent TB may offer some advantages over the current internationally recognised standard test - the Mantoux test. However, despite the studies that have been carried out since the original NICE TB guidance was published, important questions remain unanswered about the potential role of these new tests in clinical practice in the UK . The draft guideline therefore adopts a cautious, pragmatic approach by recommending that for many cases the most effective way to diagnose latent TB infection is a Mantoux test followed, depending on the result, by an IGT test."

Stakeholders wishing to submit their comments on the draft guideline are invited to do so via the NICE website by 5 August 2010. NICE plans to publish its final guideline in December 2010.

Notes

About tuberculosis

1. Tuberculosis (TB) is an infectious disease caused by the bacterium Mycobacterium tuberculosis, also known as 'the tubercle bacillus'. TB commonly affects the lungs, but can also affect other parts of the body. The symptoms of TB are varied and depend on the site of infection. General symptoms may include fever, loss of appetite, weight loss, night sweats and tiredness.

2. TB is usually spread by coughs, but prolonged close contact with a person with TB is usually necessary for infection to be passed on. It can take many years for a person infected with M. Tuberculosis to develop active TB.

3. Latent TB is where the person has been exposed to the tubercle bacillus, which remains in the body, but where there are no symptoms of TB.

4. In the UK , although the introduction of the universal programme of BCG vaccination significantly reduced rates from around 50,000 cases in the 1950s, TB is still an important public health issue, with some 8500 cases each year.

5. Although rates of the disease are now very low in some parts of the country, in other areas, mainly cities, rates of the disease are higher and, in some cases, increasing. For example, two in every five cases of TB occur in London . Specific groups are disproportionately affected by TB, including the homeless and those in poor housing, new entrants, particularly those who come from areas that have a high prevalence of TB and those living in inner city areas.

6. Almost all cases of clinical TB in the UK contract the disease by breathing in infected respiratory droplets from a person with infectious respiratory active TB disease. The initial infection may either be eliminated, remain latent, or progress to active TB over the following weeks or months.

7. In people with latent TB, 10-15% of adults will go on to develop active TB at some point in their lives and the risk in children may be much higher. However, in people who are immunocompromised (for example, if they are HIV positive), the chance of developing active TB within 5 years of infection is up to 50%.

About the draft guideline

1. There is no gold-standard test for latent tuberculosis. Diagnosis has in the past relied on the TST but this has poor specificity if there has been BCG vaccination or exposure to environmental (non-tuberculous) mycobacteria, which can lead to false positive results. The test results have to be interpreted within a certain timescale, and patients who do not return, or delay returning, will have either no result or a possibly inaccurate one.

2. recently, selective immunological (interferon-gamma, or IGT) tests have been developed using two tuberculosis antigens, 'early secretion antigen target 6' (ESAT-6 ) and 'culture filtrate protein 10' (CFP-10), which are not present in BCG, and are found in only a few species of environmental mycobacteria. These tests can be done on either cells or cell products derived from whole blood. These tests aim to be more specific by removing false positive results, and to be better correlated with latent infection or dormant organisms.

3. NICE was asked by the Dept of Health to produce a short clinical guideline on interferon-gamma immunological testing for diagnosing latent TB (partial review of CG33) and make recommendations on:

- Which diagnostic strategy is most accurate in diagnosing latent tuberculosis in adults and children who are recent arrivals from highly prevalent countries?

- Which diagnostic strategy is most accurate in diagnosing latent tuberculosis in children?

- Which diagnostic strategy is most accurate in diagnosing latent tuberculosis in adults and children (children considered as a separate population) who have been in close contact with patients with active tuberculosis?

- Which diagnostic strategy is most accurate in diagnosing latent tuberculosis in immunocompromised patients?

4. The draft clinical guideline is available (from 8 July 2010) on the NICE website.

Source:
NICE

 

Viagra-popping seniors lead the pack for STDs

Wed, Jul 7 2010

By Frederik Joelving

NEW YORK (Reuters Health) - Even if you're past your prime and have a hard time getting an erection, you might still need to worry about unprotected sex, according to U.S. doctors.

In fact, they report in the Annals of Internal Medicine, the rate of sexually transmitted diseases (STDs) in older men taking erectile dysfunction drugs like Viagra is twice as high as in their non-medicated peers.

In both groups, however, the numbers are swelling. According to the Centers for Disease Control and Prevention, there were more than six new cases of STDs per 10,000 men over 40 in 2008, up almost 50 percent since 1996.

"Younger adults have far more STDs than older adults, but the rates are growing at far higher rates in older adults," said Dr. Anupam B. Jena of Massachusetts General Hospital in Boston , who led the study.

While the reasons for this development aren't well understood, he said more divorces and better health might have conspired to boost sexual prowess and activity among graying heads.

The problem, however, is that older adults appear to flout safe sex practices. For instance, the researchers note, 50-year-olds are six times less likely to use a condom than men in their 20s.

"We are typically unaccustomed to practice safe sex over the age of 50, because the risk of pregnancy is eliminated," Jena told Reuters Health.

To test whether the introduction of Viagra in 1998 might explain some of the STD surge, Jena and colleagues examined insurance records for more than 1.4 million U.S. men over 40. The average age in the study was about 60 years.

The most commonly found STD was HIV, followed by chlamydia, syphilis and gonorrhea.

Among the few percent of men who had filled prescriptions for erectile dysfunction drugs, more than two in a thousand had been treated for an STD in the year before they got the drug.

A year later, the number dropped to half that, suggesting that Viagra and its chemical cousins didn't fuel STDs.

However, the risk of contracting an STD turned out to be more than twice as high in men taking erectile dysfunction drugs compared with those who didn't.

"These users have a different sexual risk profile than non-users," said Jena , adding that the data didn't reveal any good explanation.

In an editorial, Dr. Thomas Fekete, of Temple University School of Medicine in Philadelphia , noted that it would have been valuable to know more about the frequency of sexual encounters, sexual partners and orientation.

He added that prevention strategies should still be directed at younger age groups, whose STD risk is at least 10 times higher than in middle-aged and older adults.

Still, he said, the authors remind us "that men older than 40 years remain sexually active, even if they need chemical assistance to do so. This study also serves as a reminder that sex after age 40 years is not necessarily safe."

Jena recommended that doctors take a few minutes to discuss safe sex with older men when they prescribe Viagra.

His advice? "Look, just realize that you are at higher risk for STDs, and try to be careful like you used to be 30 years ago."

SOURCE: www.annals.org/

Annals of Internal Medicine, online July 5, 2010.


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