The Rochester Human Immunology Center (RHIC) has been awarded a $4 million renewal of its grant from the National Institute of Allergy and Infection Illness (NIAID). The renewal enables RHIC to continue leading the field of immunology in a worldwide effort to standardize how researchers use complex technologies like flow cytometry which are central to the discovery of new treatments.
When standardization is achieved, researchers is going to be better able to compare data collected worldwide, with the outcomes of numerous studies combined into massive datasets to guide the construction of hyper-accurate computer models of the mechanisms of disease. Such simulations will yield scientific conclusions which are dramatically far more valid and reproducible in areas like infectious disease, transplantation and cancer, researchers said. The resulting sophistication in understanding of the human immune system, and of how it responds to influenza, HIV or smallpox for instance, will guide the far more precise design of near-future vaccines.
Since its founding four years ago, the RHIC has proven that it can drive the development of standard operating procedures, having helped a lot more than 39 investigators and teams standardize their testing, and lending expertise that helped result in the winning of 22 major grants. As an internationally identified resource, the RHIC is part of working groups seeking to standardize methods for analyzing flu viruses and HIV in partnership with the Division of Microbiology and Infectious Illnesses at NIAID, the HIV Vaccine Trials Network and also the New York Influenza Center of Excellence.
“Standardizing test methods to achieve comparable outcomes goes far beyond getting able to compare apples to apples,” stated Sally Quataert, Ph.D., director of Core Facilities for the University of Rochester School of Medicine and Dentistry, and co-principle investigator for the grant renewal. “Validated methods assure researchers that their experimental outcomes are good science, and truly meaningful. Standard methods also directly contribute to translational science, where the results of studies are far more readily affirmed by regulatory bodies, and with fewer conflicting studies.”
Prior to 2004, Quataert was director of Immunobiological Laboratory Services at Wyeth Vaccines Investigation in Rochester for seven years. Her industry background prepared her to apply Good Laboratory Practice (GLP) standards to academic studies, making them much more likely to withstand scrutiny by the U.S. Food and Drug Administration as sound evidence that new treatments are safe and effective. If applied internationally, such standards would make drug candidates discovered at universities much easier to license to industry, with possible to improve the number of new drugs becoming accessible.
Also recently joining the RHIC executive committee was Tim Bushnell, Ph.D., director of the Flow Cytometry Core in the University of Rochester Medical Center. Bushnell’s active role inside the International Society of Analytical Cytology (ISAC) has positioned RHIC, not just to create expertise, but also to take a leading role in the groups that are shaping related standards worldwide.
Flow cytometry is actually a method of counting and sorting cells that have already been labeled by fluorescent markers which correspond with physical and chemical qualities of the cell. Modern flow cytometers can analyze thousands of cells in real time to unravel the complicated interactions driving illness processes. The devices beam laser light into a stream of liquid which carries the cells to be analyzed. Detectors catch the patterns of fluorescent light that bounce off single cells as they pass through the laser. The cells have been prepared with fluorescent dyes that absorb the laser light and emit light at lower frequencies in scatter patterns that reveal details about the cell.
Modern instruments have numerous lasers and fluorescence detectors, with 18 fluorescence detectors now commonplace in larger analysis institutions. Increasing the number of lasers and detectors enables a lot more precise identification of target cell populations by their characteristics. Four years ago, the RHIC had eight-color flow cytometry, but has since upgraded to 11- and 18-color cytometers with support from the Medical Center and its Clinical Translational Science Institute (CTSI). Researchers receive training on flow cytometry along with other key discovery techniques and gain access to a growing suite of RHIC equipment. Using the renewal of the NIAID grant, the center will continue to create and standardize immunological methods.
Beyond general leadership and standard setting, the center will take the lead inside the next five years in helping the field to make better use of three cutting-edge technologies “of immense importance.” The goal is to advance technologies to maturity so that they may be applied widely, using the new standards communicated through publications, the RHIC Web site and symposia.
The first technology is arrayed image reflectometry, which promises to be extremely useful in large-scale analysis of immune responses against many disease-causing proteins like those encountered throughout influenza outbreaks. Immune system proteins may be coated having a thin film that prevents light from reflecting off of them, but that turn out to be a lot more reflective as the proteins bind to disease-related proteins. When several proteins to be tested are attached to a biochip, a great many proteins could be analyzed for little cost.
The grant renewal will also bring together RHIC experts in cytometry and in a second new technology: quantum dot nanostructures. These microscopic machines have optical qualities that could be harnessed to measure cellular qualities via flow cytometry with far more precision than fluorescent dyes. Like dyes, the dots could be designed to inhabit certain regions of the cell, or cling to certain proteins, so that their number and characteristics could be measured. Dyes are an older technology, limited in number and with overlapping emission spectra, which indicates they interfere with every single other. RHIC teams will seek to establish new Q-dot production methods to replace and extend dye capability. As RHIC machinery gets far more complex, it will turn into increasingly in a position to pick apart and to build accurate immune cell models.
Thirdly, the team will seek to design better “gating” procedures. Flow cytometry separates cell types by brightness into groups, and gating is the process by which researchers select which cells go into every single group. The present process in a lot of labs is arbitrary, tedious and has poor reproducibility in between operators because study teams select patterns of cell characteristics “by eye.” The field of flow cytometry is in desperate need of high-speed automation to process the “mountains of data” generated by the tests, and when gating the outcomes, researchers said. The existing process is greatly slowing the pace of discovery as investigation teams do double duty with lab function and time-consuming computational function.
One unique capability that drove the RHIC renewal was that the Medical Center has a world-leading core of biostatisticians along with leading immunologists. Led by Hulin Wu, Ph.D., chief of the Department of Biostatistics and Computational Biology, the biostatistics team within RHIC will seek to establish statistically-rigorous, automated gating protocols for flow cytometry analysis that will greatly enhance the reproducibility of data and also the speed of flow cytometry lab by lab.
“The NIAID reviewers that voted to renew the RHIC grant seem to have validated the fundamental idea of our center – that centralizing the development and deployment of sophisticated technologies increases the efficiency of numerous other studies,” stated Tim Mosmann, Ph.D., principal investigator of the RHIC grant, and director and Michael and Angela Pichichero Director’s Endowed Chair of the David H. Smith Center for Vaccine Biology and Immunology in the Medical Center. “Although investigation core facilities can give centralized expertise on a fee-for-service basis, the RHIC grant allows us to move beyond this model by actively pursuing new technologies and creating promising methods via direct funding, and by providing expertise in human immunology on a collaborative basis. Secondly, the RHIC has a strong training function, so that expertise developed within the RHIC spreads rapidly through the research community. We are delighted that several researchers see the RHIC as an asset, and it’s their achievements, publications and grant awards that had been a major reason for the RHIC grant renewal.”
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Source: Greg Williams
University of Rochester Medical Center