BD BIOSCIENCES RESEARCH GRANTS

Fall 2012 Research Grant Recipients Talk About Their Research


Henry Evans, PhD
Post-Doctoral Fellow

Abstract Title:
Multicolor Flow Cytometry to Parameterize Trogocytosis of HLA-G in Pregnancy and to Determine the Mechanisms Contributing to the Maintenance of a Tolerized Immune Compartment


BD: What is your educational background?

Henry Evans: I earned my BA in biological sciences from Oxford University in the UK, after which I received master's degrees in bioinformatics and in immunology and infection, both from Imperial College, London. I also completed my PhD in Immunology at Imperial College, in the lab of Prof. Dan Davis. I'm currently a postdoctoral research fellow at Harvard in Prof. Jack Strominger's lab in the Department of Stem Cell and Regenerative Biology. The overall theme of the lab is immunotolerance, with a specialty in human reproductive immunology.

BD: How and when did you become interested in science?

Henry Evans: My dad was an electrical engineer so I've always been interested in how things work. My interest in the life sciences was really stoked by my teacher for the last few years of high school. I can't understand why anyone wouldn't be interested in science!

BD: How did you become involved in your broad field of study?

Henry Evans: My PhD program included project rotations in different labs, one of which was in Prof. Davis' lab, which I later chose to join for my doctoral work. That is where I first became interested in immunology in general, and natural killer (NK) cell biology specifically. I have remained focused on natural killer cells during my postdoctoral work, but now I study the cells' role in pregnancy. Specifically, we study mechanisms that enable the maternal immune system to tolerate a fetus that is expressing paternal, and therefore foreign, antigens.

BD: Describe the project for which you were awarded the BD grant.

Henry Evans: I am studying natural killer cells because they are the dominant lymphocyte population in the decidua, which is the maternal side of the fetal-maternal interface. However, decidual natural killer cells do not exhibit the classical "killing" response of peripheral blood NK cells. Instead they are tolerized. HLA-G, a non-classical MHC class I molecule, is thought to play a role in establishing the tolerized decidual compartment. Healthy expression of HLA-G is limited to the fetal cells that invade the maternal decidua. This current project studies how HLA-G is physically distributed at the interactions between the decidual NK cells and the fetal EVT, and how this distribution impacts the functional capacity of the NK cells. The experiments will focus on imaging the interactions between these cell populations, analyzing the changes in phenotypes by flow cytometry, and assessing the impact on the functional responses of the NK cells. Functional studies will look at cytokine secretion and cytotoxicity, for which we have cell-based assays that exploit flow cytometry.

BD: What are the long- and short-term scientific goals of this project?

Henry Evans: The short-term goals are to understand whether protein transfer occurs between the fetus and mother, but more importantly, whether this contributes to the tolerized immune compartment at the interface. Beyond this, the goal is that if we can understand it, we can work on controlling it, and later exploiting it. What we are studying is how evolution solved the problem of tolerating a foreign body locally, while maintaining an otherwise active competent immune response systemically. We should learn from nature rather than designing similar therapeutics from scratch.

BD: What are the implications of your project for human health?

Henry Evans: While great advances have been made in improving the success rate of pregnancy, immune-related complications, such as preeclampsia, still affect more than 5% of all pregnancies. Understanding the pregnancy-related immune compartment thus remains an important aim in itself. However, in the broader medical context, it may be possible to exploit mechanisms that mediate maternal tolerance to the fetus in other situations that require modulating the immune response. For example, induction of a tolerized immune compartment would be beneficial for organ transplantation and autoimmune disorders.

BD: Which BD reagents do you plan to use, and for what purposes?

Henry Evans: Much of the work is flow cytometry based, so immune cell profiling antibodies are particularly useful. In addition, the BD™ cytometric bead array system and the BD Matrigel™ invasion chamber will be excellent for functional studies of the impact of HLA-G acquisition.


Mandy Jane McGeachy, PhD
Assistant Professor

Abstract Title:
Identifying IL-23 Driven Markers of Th17 Pathogenesis in Rheumatoid Arthritis Patients


BD: What is your educational background?

Mandy McGeachy: I received my bachelor's degree at the University of Glasgow, Scotland, with honors in immunology. I then entered Dr. Steve Anderton's lab at the University of Edinburgh for my doctorate. My PhD thesis focused on regulation of inflammation by regulatory T cells in central nervous system tissue, using the experimental autoimmune encephalomyelitis (EAE) model. After that, I joined the lab of Dr. Daniel Cua at DNAX/Schering Plough in Palo Alto, California. Continuing my interest in regulation of inflammation, I studied the role of IL-23 and other cytokines in promoting Th17 cell mediated inflammation. Being in an industry setting provided me with many interesting insights, resources, and challenges. In 2012 I had the opportunity to begin my own independent, academic lab at the University of Pittsburgh in the Department of Medicine.

BD: How and when did you become interested in science?

Mandy McGeachy: I always enjoyed biology and thought that I would be a veterinarian, but when I began university I realized that what really fascinated me was trying to understand the processes that cause and prevent disease. Spending summers in research labs working in the fields of microbiology and immunology confirmed my love for scientific thinking and discovery. On this basis I decided to pursue a PhD in immunology.

BD: How did you become interested in your broad field of study?

Mandy McGeachy: The precarious balance the immune system must achieve has always fascinated me. The immune system must be ready to tackle unpredictable invading pathogens quickly and effectively, but at the same time limit bystander damage and prevent immune attack on our own tissues. Hence, the mechanisms that regulate both pro-inflammatory and anti-inflammatory responses are sophisticated, complex, and dynamic. We still have much to learn, and my previous position working in industry gave me a better appreciation for just how important this knowledge is for developing better drugs to treat disease.

BD: Describe the project for which you were awarded the BD grant.

Mandy McGeachy: Rheumatoid arthritis (RA) is a chronic, debilitating autoimmune disease with serious health and economic consequences. Th17 cells have been identified as an inflammatory subset of cells that are involved in autoimmune inflammation, including RA, and we have shown that IL-23 drives proliferation of effector Th17 cells. We also recently found that IL-23 is required for activation of memory Th17 cells. The manuscript describing this work is in revision. During this study, we defined a set of genes that are expressed in Th17 cells in an IL-23–dependent manner in vivo. These studies were performed in mice, and the goals of this project are to test whether we see the same signature in human blood samples from patients with rheumatoid arthritis.

BD: What are the long- and short-term scientific goals of this project?

Mandy McGeachy: The long-term goal of my work is to elucidate the downstream effects of IL-23 signaling in mouse and human Th17 cells. We will test our findings on the functions of IL-23 in human RA samples, specifically to determine whether Th17 cells from RA patients show increased proliferative capacity by measuring markers of cell cycle ex vivo and after stimulation in vitro. We also hope to evaluate the IL-23 signature gene expression profile in Th17 cells isolated by fluorescence activated cell sorting from RA patients, and test expression of a gene list that we established in our mouse studies of IL-23 functions in inflammation.

BD: What are the implications of your project for human health?

Mandy McGeachy: Achieving the goals of this project will for the first time demonstrate the potential of IL-23–regulated genes both as disease activity biomarkers and as therapeutic targets in RA. These studies will also contribute to the development of personalized medicine approaches for individuals with RA. We have access to matched clinical data so are able to evaluate correlations between clinical treatment response and biological alterations in the IL-23 signature. While biologics such as TNF inhibitors are showing great benefit for some RA sufferers, around 40% of patients do not respond to these agents. Discovering novel drugs is imperative for preventing the long-term inflammation that leads to disability in these patients.

BD: Which BD reagents do you plan to use, and for what purposes?

Mandy McGeachy: This project relies heavily on flow cytometry, both for analysis and for sorting of T helper populations from patient and healthy control blood samples. We will use BD antibodies for cell surface markers, transcription factors, and cytokines, taking advantage of the new violet dyes and others becoming available to perform multicolor flow to optimize the data collected with each precious patient sample.


Vanesa Sanchez-Guajardo, PhD
Post-Doctoral Fellow

Abstract Title:
Treg Modulation of Microglia Activation in PD: Effect of Alpha-Synuclein Variants on Treg Function and their Role in Controlling Microglia Induced Neuroinflammation


BD: What is your educational background?

Vanesa Sanchez-Guajardo: I earned an Ms. Sc. degree from Aarhus University, Denmark, in molecular biology. I was then awarded a Danish Research Agency fellowship to pursue a PhD degree in immunology at the Université Pierre et Marie Curie, France. I remained in Paris for a short time as a postdoctoral fellow at the Curie Institute, where I studied TLR signaling with a grant from the Fondation pour la Recherche Médical. I then joined the research unit of Dr. Marina Romero Ramos, at Aarhus University, where with a personal fellowship from the Lundbeck Foundation I have been studying neuroinflammatory processes in Parkinson's disease (PD) for the past five years.

BD: How and when did you become interested in science?

Vanesa Sanchez-Guajardo: It was during secondary school, although I'm not sure why I became interested in science—perhaps because I enjoy being out in nature. I am extremely curious and observant.

BD: How did you become interested in your broad field of study?

Vanesa Sanchez-Guajardo: Serendipity! I am a trained immunologist, and I never considered studying the brain. But one day my husband and son went to the supermarket in Denmark and were speaking Spanish to each other. My current lab director, originally from Spain but based in Denmark, overheard and thought it odd that two very Nordic-looking men were conversing in Spanish. One thing led to another, and it turned out Dr. Ramos was looking for a postdoc to investigate neuroinflammation—and I was looking for a job. Five years later I have my own line of research within her group, studying neuroimmunology in degenerative diseases. It has been extremely exciting, as it is a totally new area of research with important implications for neurodegenerative diseases, especially Parkinson's and Alzheimer's diseases. It has been quite challenging to reconcile neuroscience (my mentor) with immunology (myself) and finding answers to questions nobody thought could actually apply to the brain.

BD: Describe the project for which you were awarded the BD grant.

Vanesa Sanchez-Guajardo: Our project involves the study of the regulative interactions between alpha-synuclein (aSyn) primed regulatory T cells (Tregs) and microglia. Our goal is to validate the use of antigen-specific Treg as an immunotherapy for modulating microglia's inflammatory response during Parkinson's disease. We postulate that the environment encountered by Tregs during PD could modify Treg function, in an as yet undefined manner, which could account for the adverse immune response observed in the brains of patients. We believe that the emergence of adequately primed Tregs in the periphery could exert a protective role in the brain, as these cells could then migrate to the brain and interact with microglia to modulate their activation response. Since HLA-DR (human leucocyte antigen-dopamine receptor) expression correlates to aSyn deposition in Parkinson's patients' brains, an aSyn variant seems an obvious candidate for Treg priming. We thus aim to study the effects of different aSyn variants on Treg function and how Tregs can modulate microglia activation profiles. We will achieve this by following two approaches. First, we will investigate the effects of aSyn variants on the immune system by exposing T cells/microglia to aSyn in vivo and in vitro, and then analyzing the influence of exposure on Tregs/microglia as determined by changes in surface markers and transcription factors. Secondly, aSyn primed Tregs will be co-cultured with primary microglia, to determine their potential modulatory role.

BD: What are the long- and short-term scientific goals of this project?

Vanesa Sanchez-Guajardo: In the short term we hope to determine how T cells and microglia interact, how peripheral immune events affect the brain immune cells (microglia), and determine which kind of peripheral stimuli make T cells migrate to the brain. Longer term we hope to be able to regulate microglia-T cell interactions, induce a specific microglia activation pattern via peripheral activation of T cells, and reduce the neuroinflammatory processes occurring during PD. It may also be possible, by priming microglia, to prevent or delay onset of Parkinson's.

BD: What are the implications of your project for human health?

Vanesa Sanchez-Guajardo: We would like to design a noninvasive, immune-based therapy to prevent and/or stop the neuroimmunological processes in Parkinson's. Indeed we believe that we can trigger the peripheral immune system to instruct microglia, the immune cells of the brain, into rescuing dying neurons—the dopaminergic neurons in the case of Parkinson's.

BD:Which BD reagents do you plan to use, and for what purposes?

Vanesa Sanchez-Guajardo: We will use fluorescently labeled antibodies for characterizing microglia and T-cell activation by 9-color flow cytometry. We will also employ antibodies for western blot, to determine transcription factors in T cells. Other reagents include BD IMag™ for concentrating T cells prior to sorting, the BD™ CBA (cytometric bead array) Th1/Th2/Th17 kit for fluorescence activated cell sorting determination of cytokines produced by cells, and BD Phosflow™ to determine STAT (signal transducers and activators of transcription) activation patterns in T cells.


Photo used with permission from Lars Kruse, AU Kommunikation

Adam Schrum, PhD
Assistant Professor

Abstract Title:
Human Protein-protein Interaction Networks Assessed by Flow Cytometry


BD: What is your educational background?

Adam Schrum: I received my PhD in immunology from the University of Pennsylvania, School of Medicine (now Perelman School of Medicine) in Philadelphia, Pennsylvania in 2002, having studied in the Transplantation Immunology laboratory of Larry Turka, MD. I then spent four years as a postdoctoral fellow in the Transplantation Nephrology laboratory of Ed Palmer, MD, PhD, at University Hospital Basel in Switzerland. Since 2007 I have been an assistant professor of Immunology at the Mayo Clinic in Rochester, Minnesota, where I focus on biochemical mechanisms of receptor biology and signal transduction as it impacts T-cell immunology.

BD: How and when did you become interested in science?

Adam Schrum: It's hard for me to answer this question. I became very interested in science as I became more and more aware of the creativity involved in setting up experimental models and testing them empirically. There is a very real artistic side to science that melds together with the quantitative, empirical, and engineering aspects. When they all come together, discovery can be very satisfying.

BD: How did you become interested in your broad field of study?

Adam Schrum: Before starting my PhD program, I worked as a summer intern at Chiron Viagene in La Jolla, California, where I was introduced to T-cell immunology. My project involved testing the cytotoxic T-lymphocyte activity from peripheral blood T cells of baboons that had been vaccinated against HIV. The positive control involved a xeno-reaction, where T cells from any baboon were capable of killing another species' cells, without any prior vaccination. I became obsessed with how that works, and I've been studying T-cell immunology ever since.

BD: Describe the project for which you were awarded the BD grant.

Adam Schrum: Protein-protein interactions (PPIs) are of outstanding interest as principal regulators of biochemical signal transduction, and as potential therapeutic drug targets. However, due to technological limitations, there is little capability for analyzing large collections of physiologic, human protein complexes. Our work attempts to address this problem using a new high sensitivity, flow cytometry-based method, Multiplex IP-FCM (immunoprecipitation-flow cytometry). The new method uses multicolor microbeads to capture protein complexes from human cells, flow cytometry to determine the proteins presenting together in molecular complexes, and computational and bioinformatics analyses to interpret the data. We are mounting the system for application to human T-cell signaling, and we have seen that the new data generates a novel class of biosignature, leading to unforeseen hypotheses regarding the biochemical signals that underlie states of health and disease.

The method is also extremely sensitive, capable of detecting PPI at 10–18 (attomolar) levels, without requiring epitope tagging, genetic engineering, or radioactivity. So this makes it compatible for analysis of healthy and diseased, clinically relevant human samples.

BD: What are the long- and short-term scientific goals of this project?

Adam Schrum: Currently we are focused on two goals: understanding the pattern of protein complexes that join together to transmit specific biochemical signals, and determining the protein complex signatures associated with specific states of health vs disease. These goals have us working not only with cell lines, but also with primary, rare, human T cells from healthy and patient samples. Although we are focused on T cells of the immune system, the technology will have general applicability to any protein assemblies relevant to many fields of study and classes of disease.

What are the implications of your project for human health?

Adam Schrum: This approach will reveal patterns of multiple protein interactions that previous to this new technology have not been observable from small samples such as those that originate from medical specimens. We envision Multiplex IP-FCM as a tool that will have clinical, diagnostic, pharmacologic, and immunologic applications, and its use in basic research will likely help identify specific protein complexes that could be targeted by medicines. The technology is also being mounted with an eye on its potential applications in medical diagnostics and drug discovery.

BD: Which BD reagents do you plan to use, and for what purposes?

Adam Schrum: We will use BD's rich inventory of antibodies, both unconjugated and fluorescent, to capture and analyze the protein complexes. Additionally, various cell isolation kits, cytokines, and other reagents will be used to grow specific cell types in tissue culture, prior to their analysis by Multiplex IP-FCM.


Peter Sieling, PhD
Assistant Director

Abstract Title:
Identification of T Cells Associated with Survival in Human Melanoma


BD: What is your educational background?

Peter Sieling: I earned my BS degree from UCLA in biology, and PhD from Wake Forest University in microbiology and immunology in the laboratory of Dr. Ivo van de Rijn. The title of my PhD thesis was "Purification and characterization of the Streptococcus adjacens (nutritionally variant Streptococcus serotype II) group antigen." Upon completion of my PhD studies I trained in the laboratory of Dr. Robert Modlin at the David Geffen School of Medicine at UCLA. My field of study at that time was T-cell immunity against intracellular pathogens, including tuberculosis and leprosy.

BD: How and when did you become interested in science?

Peter Sieling: The major influence on my interest in science was my father, a physician. Therefore, at a young age, probably as early as 10 or so, I was certain I wanted to be a scientist.

BD: How did you become interested in your broad field of study?

Peter Sieling: Having for many years studied T-cell immunity in the context of microbial infection, it was easy to transition to the field of T-cell immunity against tumors, my present field of study. The John Wayne Cancer Institute is a research institute dedicated to the understanding and curing of cancer. The institute has a vast repository of clinical specimens from cancer patients, creating an excellent opportunity to resolve the problem of deficient immunity leading to cancer progression.

BD: Describe the project for which you were awarded the BD grant.

Peter Sieling: Melanoma is an aggressive skin cancer that leads to 48,000 deaths annually worldwide. It is the most common cancer in young adults ages 20 to 30 and is the leading cause of cancer death in women ages 25 to 30. Immunosuppressed individuals have a three-fold greater risk of metastatic melanoma. At the same time immunomodulatory therapies can lead to prolonged remission of the disease, indicating a pivotal role for the immune system in fighting melanoma. Numerous studies establish a correlation between T cells at the site of disease and favorable outcome in cancer. Therefore, an examination of the underlying mechanisms that correlate with successful anti-tumor immune responses must be considered in improving therapies for melanoma.

BD: What are the long- and short-term scientific goals of this project?

Peter Sieling: The overall objective of this study is to gain insight into successful anti-tumor immunity by examining human T-cell responses in long-term survivors in melanoma.

One primary goal is to determine whether a distinct memory CD8 T-cell phenotype at the site of metastatic melanoma correlates with survival. We will investigate this using multicolor flow cytometry. Clinical observations and response to immunomodulatory therapies indicate an important role of memory CD8 T cells in the immune response of long-term survivors with advanced melanoma. We hypothesize that resident memory CD8 T cells contribute to melanoma survival by their presence at the site of disease. We propose to quantitatively define CD8 T-cell subsets from cells isolated at the site of disease from patients with metastatic melanoma.

Our other main objective is to define T-cell quality relevant to melanoma survival by measuring antigen-specific T cell responses in peripheral blood mononuclear cells (PBMCs) using intracellular flow cytometry. T cells producing multiple cytokines are associated with protective immune responses in microbial infection. However, there is limited evidence in human cancer for this phenomenon, also referred to as T-cell quality. We hypothesize that improved T-cell quality correlates with survival in melanoma. We propose to measure antigen-specific T-cell quality during the course of disease in melanoma, examining PBMCs collected at the time of staging and throughout the patients' therapy.

BD: What are the implications of your project for human health?

Peter Sieling: Identification of the distinct phenotypic and functional subset of T cells associated with improved survival in melanoma may allow one to predict survival, and identify novel therapies that elicit protective immune responses that may be applicable to other solid tumors.

BD: Which BD reagents do you plan to use, and for what purposes?

Peter Sieling: We will identify memory T-cell populations by flow cytometry using BD Biosciences antibodies, for which BD is world renown. T-cell quality will be determined by measuring cytokines and other inflammatory mediators in PBMCs from melanoma patients at several times before and during therapy using intracellular flow cytometry, also using BD Biosciences antibodies.


Zhihong Zeng, MD
Instructor

Abstract Title:
Investigating the Molecular Mechanisms of Bone Marrow Stromal-mediated Drug Resistance in Leukemia


BD: What is your educational background?

Zhihong Zeng: I received my MD degree from Jinan University Medical School in China. I came to the USA after receiving a fellowship in ophthalmology at the University of California, Irvine. I later received master's degrees in biochemistry and software engineering in computer science from Kansas State University.

BD: How and when did you become interested in science?

Zhihong Zeng: I became interested when I was taking the biochemistry class in medical school. I developed a great interest in the origins of disease, their mechanisms, how they develop, and of course possible treatments. My interest broadened further while I was undertaking my clinical studies.

BD: How did you become interested in your broad field of study?

Zhihong Zeng: After receiving my degrees from KSU, I had accepted a postdoctoral position at MD Anderson Cancer Center, studying leukemia. I was mentored by Drs. Andreeff and Konopleva, whose research bridged basic science directly with clinical applications, with an emphasis on utilizing advanced technologies to study the mechanisms of drug efficacy in the physiological tumor microenvironment. Working with this diverse and invigorating group of scientists helped me to continuously improve my skills and knowledge. I am grateful to have their guidance and support. This experience prepared me for my current position.

BD: Describe the project for which you were awarded the BD grant.

Zhihong Zeng: AML (acute myeloid leukemia) is a type of leukemia that often occurs in adults. This disease has a high relapse rate because drug-resistant cells, most likely leukemic stem cells (LSCs), repopulate after treatment. One focus of our research is to investigate how resistant cells develop in the bone marrow microenvironment. The major challenge in studying these cells is their rarity, which severely limits opportunities of studying these cells through conventional technologies such as Western blot, which requires relatively large numbers of cells. Multiparametric phospho-flow cytometry is an advanced technology that allows us to study both intracellular signaling within rare stem cells, and their interactions with stromal cells.

We will employ phospho-flow to study the interplay between AML stem cells and AML-derived stroma through their functional surface molecules. We will also characterize intracellular signaling in the LSCs and evaluate how they react to different inhibitors, and how these reactions can be altered by stroma, leading to emergence of a subset of drug-resistant stem cells. We will identify and profile stroma-mediated drug resistance in leukemic stem cells from AML patients at different stages of disease. Utilizing newly developed software, we will analyze all data in one platform, identifying the stroma modified pathway changes in LSCs and LSC subsets. The approach proposed here will improve our understanding of the stroma-regulated drug resistance in leukemic stem cells.

BD: What are your long- and short-term scientific goals?

Zhihong Zeng: Our short-term goal is to identify the unique signaling pathways related to stroma-mediated drug resistance in LSCs. A long-term goal is to identify druggable targets that will enable development of treatments to eradicate these cells.

BD: What are the implications of your project for human health?

Zhihong Zeng: Identifying resistance patterns and druggable targets is the critical step for the development of novel leukemia therapies targeting these cells. The ultimate goal is to reduce the frequency of relapse and increase AML cure rate. Our research may also benefit patients with other types of leukemias.

BD: Which BD reagents do you plan to use, and for what purposes?

Zhihong Zeng: We will use several BD products for phospho-flow experiments, including the BD Phosflow™ starter kit, several directly conjugated antibodies and paired isotype controls to detect surface markers and intracellular molecules, and to sort the identified cells. Our project will use BD LSRFortessa™ II flow equipment and the BD FACSAria™ cell sorter instrument.


Alessia Zoso, PhD
Scientist

Abstract Title:
New Approach to Effectively Modulate and Redirect the Autoimmune Response in T1D Patients: Phenotypical and Functional Analysis of the Components


BD: What is your educational background?

Alessia Zoso: After receiving my undergraduate degree in biology, and my PhD in oncology at the University of Padua, I completed the European Master of Advanced Studies of Immunology organized by the University of Milan-Bicocca and Institute Pasteur. I held a postdoctoral fellowship at the Johns Hopkins School of Medicine, where I worked on active anti-tumor immunotherapies. I completed a second postdoctoral fellowship at the University of Miami. My research focus is the induction of tolerance in autoimmune disease and graft-versus-host disease.

BD: How and when did you become interested in science?

Alessia Zoso: I had a great biology high school teacher who instilled in me a curiosity about the human body's finely tuned regulatory mechanisms and how it maintains equilibrium. Understanding how this equilibrium can be disrupted and re-established still represents for me an intriguing challenge.

BD: How did you become interested in your broad field of study?

Alessia Zoso: The immune system is complex and fascinating. Understanding the mechanisms that control the immune response against pathogens, while controlling the immune reaction against our own tissue, presents researchers with new and exciting challenges. Since I began working at the Diabetes Research Institute the main goal of my work is to identify novel strategies for inducing tolerance toward the transplanted islets and to control the autoimmune response accountable for the progression of type 1 diabetes. The application of the stem cells "educator" strategy represents a significant step for controlling progression of the disease, as well as the possibility to understand and unveil mechanisms that regulate the immunological response in other disorders.

BD: Describe the project for which you were awarded the BD grant.

Alessia Zoso: Autoimmunity in type 1 diabetes occurs through independent or concomitant factors including genetic predisposition, epigenetic factors, physical, social, and environmental conditions. A winning therapeutic approach for a cure should therefore address multiple factors. Although diabetes arises mainly through activation of autologous T cells against β cells, the disease is multifactorial and the entire immune system is dysfunctional. The only reasonable treatments for type 1 diabetes are daily insulin injections or transplantation of insulin-secreting tissues. Although insulin discovery transforms a deadly disease into a chronic one with quasi-normal life expectancy, many complications arise from poor control of blood glucose. Transplanted insulin-producing tissues, while clinically relevant, are subject to immune rejection. With this proposal we intend to establish a new strategy aimed at modulating the immune system and block the activated T cells at onset, thus potentially curing type 1 diabetes through transplantation. This approach takes advantage of the immunoregulatory property of umbilical cord-blood–derived stem cells. It involves the co-incubation of cord-blood–derived cells with peripheral blood mononuclear cells from diabetes patients, followed by their reinfusion. Pre-clinical and clinical data show that this approach significantly controls diabetes progression and induces generation of T-regulatory cells, with no graft-versus-host reactions or other significant adverse events.

Nevertheless, it is still unknown which cell populations are responsible for this therapeutic effect, and which mechanisms are involved in "re-education" of patients' T cells. With this award we will phenotypically and functionally characterize the cord-blood–derived populations and the changes they undergo through co-incubation with donor peripheral blood mononuclear cells.

BD: What are the long- and short-term scientific goals?

Alessia Zoso: Our primary goal is to understand the mechanisms and pathways activated during the co-culture of cord-blood–derived cells and mononuclear cells. We will focus on the suppressive capacity of the "educator" and "educated" cells, and how their function affects disease progression. In the long run, we aim to use the pathway we identify as a target for alternative and less-invasive therapies.

BD: What are the implications of your project for human health?

Alessia Zoso: The stem cells educator approach represents a breakthrough towards finding a cure for type 1 diabetes. Better knowledge of the mechanism would help us improve this strategy and extend its application to other autoimmune diseases. This strategy can significantly improve the quality of life of type 1 diabetics by reducing the daily management burden for glucose control and diet through prolonging pancreatic islet survival and postponing the need for transplantation.

BD: Which BD reagents do you plan to use, and for what purposes?

Alessia Zoso: BD Biosciences antibodies will be used for the analysis of both cord-blood–derived cells and mononuclear cells at different incubation times. We will also use the BD™ Cytometric Bead Array Kits to quantify cytokines in the culture media and sera of mononuclear cell donors.