TO GET MORE...   If you are interested in publishing any of the releases in this media report or if would like to schedule an interview or get high resolution photos related to the story, please contact Claudia Costabile at ccostab1@jhmi.edu. Let us know your opinion or special request here.	NEW ALLERGY VACCINE PROVIDES LONG-LASTING HAY FEVER RELIEF AFTER JUST SIX WEEKS OF SHOTS   Researchers at The Johns Hopkins University School of Medicine have successfully used an experimental DNA-based vaccine to protect against ragweed allergies, commonly known as hay fever, after just six injections. Patients receiving the vaccine showed an average 60 percent reduction in allergy symptoms compared to those receiving a placebo. The experimental therapy holds the promise of one day eliminating the need for traditional allergy medicines targeting allergy symptoms, such as nasal steroids and antihistamines, and providing a safer, faster replacement for immunotherapy regimens, which are costly and take years to work, the researchers say. During the test period, allergic symptoms were monitored and recorded, right down to how often volunteers' noses ran and how many times they sneezed. Compared to the placebo group, those who received the vaccine exhibited a 60 percent reduction in all of their allergy symptoms, including sneezing, runny nose, watery eyes and itching. Relief from allergic symptoms was as pronounced in the second year as in the first, even though no more vaccine was administered. Lead investigator Peter Creticos, M.D., medical director of the Johns Hopkins Asthma and Allergy Center in Baltimore, explained that such prolonged relief is an important part of his team's findings because it appears that the vaccine's efficacy doesn't wear off quickly. A new study, currently under way, will further examine the drug's lasting effects in a larger group of participants. "This therapeutic intervention heralds a major advance in the treatment of allergic rhinitis," says Creticos. "Long-lasting relief can be achieved with a concise, six-week injection regimen, as opposed to the current, tedious, four- to five-year course of treatment with allergen immunotherapy." Investigators at the University of California, San Diego (UCSD) had previously observed that a particular sequence of DNA, derived from bacteria, shuts down a T-helper cell (Th2) involved in the body's inflammatory response. Creticos and his team, recognizing that allergic disease is driven by Th2 inflammation, then embarked on a series of studies to evaluate the effectiveness of using this approach to treat allergies. The central question that they sought to answer was: What would happen if the DNA strand was linked to the most allergenic portion of the ragweed pollen protein, which is the No. 1 cause of seasonal allergies in North America? Dynavax Technologies Corp., of Berkeley, Calif., developed the vaccine and funded several of Creticos' early safety studies. Creticos was a paid consultant of Dynavax during that period. The current study was sponsored by the Immune Tolerance Network, which receives its funding from the National Institute of Allergy and Infectious Diseases and the National Institute of Diabetes and Digestive and Kidney Diseases. Creticos explained that the vaccine works in two ways: by suppressing acute allergic reactions (like sneezing) and by helping the body better regulate chronic inflammation (like itchy eyes and a runny nose). "We are turning off an inappropriate or abnormal allergic response and returning the body to normalcy," says Creticos. "Our hope is that we can one day provide a long-term cure for hay fever and other chronic inflammatory diseases."   LEADING REASON FOR CORNEAL TRANSPLANTS COMES INTO FOCUS   Guided by families with an unusual number of cases, scientists at Johns Hopkins have discovered the genetic origins of at least one form of Fuchs corneal dystrophy, FCD, the leading reason for corneal transplantation in the United States. In one form or another, FCD's trademark deterioration of the cells covering the clear, outermost lens of the eye affects more than 4 percent of the population over 40. Late in life, the dystrophy causes swelling of the cornea and can severely affect vision, making it impossible to see well even with glasses or contact lenses. It's believed that various forms of FCD are due to multiple gene mutations. A team led by Hopkins ophthalmologist John Gottsch, M.D., says they were able to map a common form of Fuchs, found most often in women, to chromosome 18. "Finding this chromosomal locus is putting us in the right neighborhood to find culprit genes," says Gottsch. "Now we have to start knocking on every door." Gottsch is heartened by success with earlier Fuchs gene hunting studies. The Hopkins group tracked down its first FCD related gene in a Virginia family with multiple, early onset cases. That gene, labeled COL8A2, was mapped to chromosome 1. Prior to that, a large Indiana family with FCD, including a boy of 10, led the team to yet another gene variant, on chromosome 13. The scientists use linkage analysis, a process of elimination gene hunting technique that analyzes inheritance patterns in families with relatively large numbers of affected individuals and trace genetic traits co inherited or "linked" with the disorder. Researchers search for a common location for all the linked traits until they wind up with a single chromosome address. In his latest study, Gottsch used not only linkage analysis but also a method of identifying variations in DNA sequences to examine three FCD families in which 43 members had the disease and 33 did not. He and his coworkers were able to narrow down the linked traits in all three families to the short arm of chromosome 18 (the whole "address" is 8q21.2 q21.32). "Because the same location popped up for three different families with similar forms of Fuchs dystrophy, we believe we have the chromosome locus for the most common genetic mutation resulting in Fuchs," he said. "It's a painstaking process of elimination, but now we are closing in on the gene that causes what we believe is the most widespread form of Fuchs, not just the rare types in individual families. Our methods have clearly shown that Fuchs is not just one disease, but rather a disorder with several genetic flavors." Gottsch became interested in FCD more than six years ago when he treated a woman with a corneal dystrophy of unknown origin that looked remarkably similar to FCD despite slightly different symptoms. "I knew it wasn't classic Fuchs, but rather something new," remembers Gottsch. "In the end, it wasn't Fuchs at all, but a sort of mutated distant cousin of the disease. It made me wonder, however, if there were more genetic variants of the disease out there." He stopped wondering a few years later when he examined the Indiana family and discovered what he thought at the time was the youngest case of FCD ever described in the scientific literature, the 10 year old boy. However, literature published 25 years ago described a Virginia family with a 3 year old girl with the disease. Gottsch contacted the original investigator and was able to reexamine the family. He was then able to determine the gene that afflicted this family and that it resulted in a unique and severe form of FCD. "I knew Fuchs was not one but several diseases with multiple genes involved when I started comparing the symptoms of the two families," Gottsch says. "The Indiana family members developed their disease late in life, women were mostly affected, and those who were had large bumps, or guttae, on the back of the cornea. The Virginia family had severe early disease with men and women equally affected, and the cornea had smaller, finer bumps." Gottsch is looking to one day use various gene therapies to suppress the mutations that he has found. It is hoped that someday the millions of people who suffer from FCD in the United States will eventually benefit from having the genetic basis of their disease identified and gene therapies developed. At present, corneal transplantation is the only solution for those severely affected.   MOUSE TESTS PREDICT DRUG RESPONSE IN RELAPSING PANCREATIC CANCER PATIENTS   By slicing up bits of patient tumors and grafting them into mice, Johns Hopkins Kimmel Cancer Center specialists have figured out how to accurately "test drive" chemotherapy drugs to learn in advance which drug treatments offer each individual pancreatic cancer patient the best therapeutic journey. Although "xenografting" with either cells or fresh tissue is already used widely to test cancer therapies, the Johns Hopkins design is personalized to each patient who has relapsed after an initial course of chemotherapy. "Eventually our approach offers a promising way to individualize therapy earlier in treatment instead of first giving everyone the standard drug gemcitabine, which has a success rate of less than 10 percent," says Antonio Jimeno, M.D., instructor in oncology at the Johns Hopkins Kimmel Cancer Center. Results of preliminary tests of the Johns Hopkins method in 14 patient samples taken after surgery shows that each xenografts' genetic profile remained stable through three and four generations of mice so that "test drives" would accurately represent a patient's tumor. The scientists also found they could build xenografts in 80 percent of their pancreatic patients, a success rate higher than efforts with colon cancer patients, for which rates are typically less at about 50 percent. Reporting on their work in a recent issue of Clinical Cancer Research and at the September meeting of the American Association for Cancer Research in Chicago, the Johns Hopkins team said it took tiny bits of a patient's tumor removed after surgery, and implanted them into one or two mice. After letting the resulting tumor grow for several months, they removed the mass and cut it into pieces to implant into additional mice, eventually creating 20 animals containing matching samples of a single patient's tumor. "By scaling up this way, we got enough tumor samples to randomize mice into groups for testing candidate drugs," says Manuel Hidalgo, M.D., Ph.D., associate professor at Johns Hopkins' Kimmel Cancer Center, who says the process currently requires about six months to get information on which drugs work best. "In the meantime, most patients are receiving their first rounds of chemotherapy and radiation. Initially, xenograft information can guide therapy once patients relapse, which is generally in nine to 12 months with pancreatic cancer." The Johns Hopkins group is conducting a clinical trial of the xenograft model in 40 patients undergoing surgery at Johns Hopkins for non-metastatic pancreas cancer. In the trial, a portion of each patient's tumor is shuttled directly from the pathologist to the Johns Hopkins' laboratory where the first mice are implanted and the 20 mice "built" to test the 20 or so drugs currently available against pancreatic cancer. Says Jimeno, "If this model works, then we'll need to develop ways to apply it to a broader population of pancreatic cancer patients since there are significant laboratory resources necessary for each patient." Information from the study also may reveal new biomarkers that predict drug response and data on how certain therapies act within the body. Ultimately, they hope to broaden use of xenografting to tumor samples that can be accessed via biopsy through fine needle aspiration. Pancreatic cancer accounts for more than 33,000 new cases in the United States and almost as many deaths. It is one of the deadliest cancers, with less than five percent of patients living beyond five years. This study was funded by the Goldman and Lee families, the Viragh Foundation for Cancer Research, and the National Institutes of Health/National Cancer Institute. Additional authors include Belen Rubio-Viqueria, George Cusatis, Xianfeng Zhang, Christine Iacobuzio-Donahue, Collins Karikari, Chanjusn Shi, Nadia Bouraoud, Maria L. Amador, Soner Altiok, Piotr Kulesza, Charles Yeo, Wells Messersmith, James Eshleman, Ralph H. Hruban and Anirban Maitra from Johns Hopkins; Kathleen Danenberg from Response Genetics, Inc.; Peter V. Danenberg, Hidekazu Kuramochi, and Koji Tanaka from the University of Southern California; and Sharat Singh and Hossein Salimi-Moosavi from Monogram Bioscience, Inc.   HOPKINS JOINS UGANDAN RESEARCHERS TO STUDY PEDIATRIC AIDS VACCINE   Scientists at Makerere University, in Kampala, Uganda, along with scientists from Johns Hopkins and other institutions worldwide, have begun the first clinical safety trial in Africa of a vaccine to prevent mother-to-child transmission of HIV through breastfeeding. Breast milk is a leading route of infection in the developing world, according to the United Nations World Health Organization, which estimates that each day 1,800 newborns are infected with the AIDS virus, 30 percent to 40 percent by virus carried in their mother's milk. Enrollment of the first newborn took place at Mulago Hospital in Kampala. The so-called phase I study is designed to test the safety of injecting newborns with the vaccine, formally known as ALVAC-HIV (vCP1521). If the vaccine is found to be safe in this study, and if it is later shown to be effective in reducing the chance of infants' becoming infected during breastfeeding, researchers estimate that it could potentially stop up to 8,000 of Uganda's 22,000 infections a year in children. Initial results are expected by mid-2007. "A vaccine is the easiest way to help prevent mother-to-child transmission of the disease, as healthy alternatives to breastfeeding, such as infant formula, are not available or affordable to most new mothers in the developing world, many of whom do not know they are HIV positive," says study protocol chair and pediatric infectious disease specialist Laura Guay, M.D., who will lead Johns Hopkins' efforts. "Vaccines often involve several injections over a short period of time, whereas other drug therapies that might prevent transmission are less convenient and must be taken daily over a longer period of time and potentially have more side effects," says Guay, an associate professor at the Johns Hopkins University School of Medicine. Guay notes that discouraging breastfeeding altogether is not a practical option for many women because the lack of safe alternatives increases five- to sevenfold a newborn's chances of dying of pneumonia or diarrhea, illnesses which breastfeeding can help prevent through nutrition and ingestion of the mother's antibodies. Moreover, she says, it would be potentially harmful to deny this key means of nutrition to the 80,000 newborns in Uganda who do not contract HIV each year from their infected mothers, either from pregnancy, labor or delivery, or breast milk. Indeed, Guay points out that the Ugandan Ministry of Health has identified development of a vaccine as a key priority for reducing the country's high HIV transmission rates. A group of local community members provided advice on how the study was to be carried out and participated in advance in educational seminars. The goal of the Johns Hopkins team is to eventually find a vaccine that will allow infants to develop immunity to HIV just as they would to polio, diphtheria and hepatitis B after vaccination for those disorders. Many of these vaccines, researchers point out, are already combined into a single vaccination. The goal is to one day provide an AIDS vaccine as part of a child's regular vaccination program. Previous research using an ALVAC-HIV vaccine in adults in Uganda showed it to be safe, but it is not yet known if it is effective in preventing infection. In 2005, a phase I study done in newborns in the United States using a similar ALVAC-HIV vaccine found that the vaccine was very safe. Related clinical research from other institutions using the same vaccine is under way in Thailand, involving 16,000 participants, a much larger sample because researchers there are testing the vaccine's broad effectiveness rather than its initial safety. However, research in monkeys has shown that ALVAC-SIV vaccine was successful in preventing oral transmission through milk of simian immunodeficiency virus, or SIV, in 11 of 17 newborns given the vaccine. The ALVAC-HIV vaccine is one of at least five HIV vaccines under study in Africa, all of which are being studied in adults. It is manufactured by extracting cell cultures that have been grown in embryonated chicken eggs. The cell cultures contain live and weakened canarypox virus, which does not infect humans, but has had genetic material from specific strains of HIV inserted into it. The theory, according to researchers, is that the human body could develop a cellular immunity to HIV by developing immunity to its genetic components in the non-harmful canarypox virus. More than 20 other test vaccines against the disease are in various stages of early development. Studies are proceeding in multiple countries to assess other vaccines' safety against all subtypes and various cross-mixes of the virus. "A major advantage to this kind of research is that it opens up access to better AIDS care, including medications, regular checkups and home care, to the people of sub-Saharan Africa, where the need is great," notes study co-investigator Brooks Jackson, M.D., M.B.A., professor and director of pathology at Johns Hopkins. "We hope this study will lead to more effective research and treatment and more vaccine trials in infants in Africa." Jackson, a pathologist and virologist, has studied HIV disease in Africa for the past 16 years. According to the latest statistics from the United States Centers for Disease Control and Prevention, in 2004, more than 1 million Americans currently live with HIV, the virus that causes AIDS. However, the advent of antiretroviral therapy and better understanding of how to prevent transmission from mother to child have dropped the annual number of pediatric cases from nearly 2,000 in the early 1990s to under 200 in recent years, mostly to mothers who did not know they were HIV positive. Funding for the study is provided by the National Institute of Allergy and Infectious Diseases, part of the U.S. National Institutes of Health. Vaccine is being supplied by the manufacturer, sanofi pasteur, a division of Sanofi-Aventis. Neither Guay nor Jackson receives any financial benefit from the manufacturer for their participation.   KEY TO LUNG CANCER CHEMO RESISTANCE REVEALED   Scientists at Johns Hopkins have discovered how taking the brakes off a "detox" gene causes chemotherapy resistance in a common form of lung cancer. Products made by a gene called NRF2 normally protect cells from environmental pollutants like cigarette smoke and diesel exhaust by absorbing the materials and pumping them out of the cell. Another gene called KEAP1 encodes products that stop this cleansing process. But lung cancer cells sabotage the expression of these same genes to block assault from chemotherapy drugs. "What we're seeing is that lung cancer cells recruit and distort NRF2 and KEAP1 expression to help tumor cells evade the toxic effects of chemotherapy," says Shyam Biswal, Ph.D., associate professor at the Johns Hopkins Bloomberg School of Public Health and Kimmel Cancer Center, who published results of cell culture studies in the October 3, 2006 issue of PLoS Medicine. Past studies have shown that NRF2 detoxifies cells by directing proteins to absorb and pump out pollutants and chemicals. The NRF2 gene makes a "trigger" protein which starts the production of other proteins and enzymes that sweep the cell clear of toxins. To halt the detox process, proteins manufactured by KEAP1 bind to the NRF2 triggers tagging them for destruction. In cancer cells, NRF2 activity runs amok, sweeping away all cellular toxins, including chemotherapy agents. Biswal says that blocking NRF2 activity could improve the effectiveness of standard chemotherapy drugs, particularly platinum based compounds widely used for lung cancer. In Biswal's study, half of 12 lung cancer cell lines and 10 of 54 tissue samples from non small cell lung cancer patients had mutations in the KEAP1 gene rendering it inactive and unable to keep NRF2 activity in check. In addition, half of the tissue samples were missing one copy of the KEAP1 gene - cells usually have two copies of each gene. No missing genes or mutations were observed in normal lung tissues from the same patients. NRF2 activity along with its cleansing proteins and enzymes were higher in tumor samples than normal cells, according to the researchers. Their cell culture tests also show that cancer cells with KEAP1 mutations are more resistant to chemotherapy drugs than normal lung cells. Tumor samples with normal KEAP1 genes also show increased levels of NRF2 and its enzymes, suggesting other ways of dismantling KEAP1, such as splicing the gene to make a shortened, ineffective protein, he said. The researchers plan to confirm their findings with a larger set of samples and then to screen for appropriate drugs. Funding for the study was provided by the National Cancer Institute Lung SPORE (Specialized Program of Research Excellence), National Heart Lung and Blood Institute, National Institute of Environmental Health Sciences Center, National Institute of Health, and the Flight Attendant Medical Research Institution. Co authors include Anju Singh, Vikas Misra, Rajesh K Thimmulappa, Hannah Lee, Stephen Ames, Mohammad O. Hoque, James G. Herman, Stephen B. Baylin, David Sidransky, Edward Gabrielson and Malcolm Brock from Johns Hopkins.   HOPKINS RESEARCHERS UNCOVER CRITICAL PLAYER IN CELL COMMUNICATION   Johns Hopkins researchers have teased out the function of a protein implicated in Williams Beuren syndrome, a rare cognitive disorder associated with overly social behavior and lack of spatial awareness. Called TFII I, or TF "two eye," the protein long known to help control a cell's genes also controls how much calcium a cell takes in, a function critical for all cells, including nerves in the brain. The study will be published in Science. "While the previously described function of TFII I very well also could contribute to the cognitive defects of Williams Beuren syndrome, its role controlling calcium makes much more sense," says Stephen Desiderio, M.D., Ph.D., a professor of molecular biology and genetics and director of the Institute of Basic Biomedical Sciences at Hopkins. And, says Desiderio, others have shown that defects in a cell's ability to take in calcium can lead to other neurological and behavioral conditions. Williams Beuren syndrome is associated with craniofacial defects, problems with the aorta and a very specific mental retardation that causes those affected to be talkative, sociable and empathetic but at the same time have significant spatial learning defects. Those affected are highly expressive, have exceptionally strong language abilities and "can talk up a storm," for example. But at the same time, they are poor at global organization, having problems re creating patterns in drawings. The syndrome occurs in roughly one in 25,000 births and is caused by a deletion of a small section of chromosome 7 that contains several genes, including the gene that encodes the TFII I protein. The discovery came after Desiderio and his team used biochemical "bait" to fish for candidate proteins that physically bind to TFII I. The fishing expedition returned one protein known to control when and how much calcium a cell takes in. "The partner we found in the fishing experiment and the abundance of TFII I outside the cell nucleus led us to suspect that this protein must be doing more than regulate gene expression," says Desiderio. Under normal conditions, calcium does not flow freely into and out of cells until a demand for it - such as a muscle contraction or nerve function - triggers cells to take up the free floating element from their surroundings. Cells store calcium until still other signals occur to release it again. "The finding was stunning to us because calcium is one of the most important messengers in cells," says Desiderio," "and both it and TFII I are in every cell. That affirmed our suspicion that TFII I could be doing something important with calcium signaling." In one experiment, the Hopkins team knocked down the amount of TFII I in lab grown cells and looked for changes in calcium flow under a high power microscope using a dye that glows when it comes in contact with calcium. A camera attached to the microscope recorded the brightness of the glow and fed that measure into a computer that calculates the amount of calcium. Knocking down TFII I and separately assaulting the cells with chemicals caused the cells to take up more calcium than usual. The researchers realized that when they depleted the cells of TFII I, the cell responded by installing more calcium channels in their surfaces that allow calcium and only calcium to enter the cell. "We think TFII I must control calcium entry into the cell by somehow limiting the number of calcium channels at the cell's surface," says Desiderio. "There's good evidence suggesting that the frequency and intensity of this ebb and flow of calcium can determine a cell's response to external cues," says Desiderio. "TFII I may be a universal player in communication between cells, in the brain, the immune system and elsewhere." The researchers were funded by the Howard Hughes Medical Institute, the National Cancer Institute and the Institute for Cell Engineering at Hopkins. Authors on this paper are Gabriela Caraveo, Damian van Rossum, Randen Patterson, Solomon Snyder and Desiderio, all of Hopkins.