BD BIOSCIENCES RESEARCH GRANTS

Spring 2013 Research Grant Recipients Talk About Their Research


Ronald Gill, PhD
Professor of Surgery and Immunology/Scientific Director

Abstract Title:
Identifying Phenotypic Signature T Cells Targeting Islet Transplants in Autoimmune NOD Recipients


BD: What is your educational background?

Ronald Gill: I received my undergraduate degree in biology, and my PhD in biology and immunology from the University of California, Los Angeles. I then spent several years as a postdoctoral fellow at the University of Colorado, Health Sciences Center. As a postdoc I studied transplant immunology under the direction of Prof. Kevin Lafferty.

BD: How did you become interested in science?

Ronald Gill: I did not care much for science until my tenth grade biology class. That class was transformational for me and triggered a strong interest in the life sciences that has never diminished.

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

Ronald Gill: After deciding to major in biology I sought a wide range of experience in this field but increasingly gravitated towards cellular and molecular biology. During my senior year at UCLA I took a course on the biology of cancer and was given my first exposure to immunology. That experience almost instantly galvanized my interest in cellular immunology. I later undertook advanced lecture laboratory projects in immunology that sharpened my focus. I became intrigued with the concept of "self" versus "non-self" immune system discrimination and the related dilemmas it creates regarding pathogen reactivity, transplantation, and autoimmunity. My graduate research focused on basic issues of cellular cytotoxicity and cell membrane properties that affect the capacity of T cells to interact with and lyse specific cellular targets. Over time, my desire to perform more disease-related research increased, and I had persistent peripheral interests in both tissue and organ transplantation.

BD: Describe the project for which you received the BD grant.

Ronald Gill: Islet transplantation continues as a viable approach for restoring glucose homeostasis in Type 1 diabetes, especially for patients with hypoglycemia. While islet transplants restore insulin independence, durability of graft function is limited compared with most solid organ transplants. There is ongoing debate regarding the relative contribution of conventional transplant rejection versus the re-enactment of islet-specific autoimmunity in the destruction of islet transplants. Fortunately, we have a genetically autoimmune-prone animal model of diabetes with which to study spontaneous disease, the non-obese diabetic (NOD) mouse. Over many years we have explored several facets of how islet rejection occurs, both in this model and in non–autoimmune-prone mouse strains. Results indicate that either conventional transplant immunity or underlying islet-specific autoimmunity, is sufficient to destroy pancreatic islet transplants. However, the mechanisms by which these two immune processes result in islet injury appear to be different. Moreover, the combination of transplant immunity and autoimmune disease recurrence appear to synergize to inflict severe injury to islet allografts in the diabetic NOD recipient. So there is an ongoing need to identify the nature of both the specific immune response responsible for mediating islet graft rejection in diabetic recipients and to uncover immune pathways that appear to resist current treatment strategies.

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

Ronald Gill: Although there is ongoing debate on the relative contribution of autoreactive (islet-specific) versus general alloreactive T cells in islet graft injury in either type 1 diabetic patients or in diseased NOD mice, there has been very little direct analysis of such cells. We have the capacity to directly isolate islet graft-infiltrating T cells, and the major goals of this project are first to develop a phenotypic "signature" of T cells that infiltrate and destroy islet transplants in diabetic NOD recipients. Next, we hope to determine what T-cell subsets (autoreactive versus alloreactive) are resistant to tolerance-promoting therapies in diabetic NOD mice. It is not clear whether autoreactive, alloreactive, or both types of T cells are resistant to such transplant tolerance induction. Therefore, another important goal of this project is to directly assess the types of T cells that "break through" tolerance-promoting therapies and destroy islet transplants.

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

Ronald Gill: We know that autoreactive T lymphocytes injure insulin-producing islet beta cells, either during the initiation of type 1 diabetes or in the recurrence of disease in transplanted islets. Yet we have little understanding of the exact phenotypes of these islet-infiltrating T cells. Also, when islet transplants from unrelated donors are implanted, we do not know if the T cells that are autoreactive, alloreactive, or both predominate the response. Thus, the key implication of this project is to help identify the key immune barrier(s) to successful islet transplantation, and to understand the phenotypes of recipient cells found in recipients that are resistant to conventional therapies. Ultimately, the goal is to attenuate such graft-destructive immunity.

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

Ronald Gill: We will employ a variety of antibodies specific for both cell-surface and intracellular markers to identify the phenotype and functional state of our target immune system cells. Many BD reagents (such as the BD™ Cytometric Bead Array kits) will be used to determine cell surface phenotypes, effector molecules, and transcription factors characteristic of T cells that break through tolerance therapies and reject islet grafts in NOD mice.


Ana Gabriela Henriques, PhD
Neuroscience Laboratory Investigator

Abstract Title:
Profiling Neuroinflammatory Biomarkers in Alzheimer's Disease


BD: What is your educational background?

Ana Henriques: I received my undergraduate degree in biology from the University of Aveiro, which is in Portugal. During this time I received an internship in the Aveiro neuroscience laboratory, working with Profs. Odete da Cruz e Silva and Edgar da Cruz e Silva. I remained at the University of Aveiro in the da Cruz e Silva laboratory for my masters and doctoral degrees. My PhD thesis, in biochemistry, was titled Abeta Biology and its Contribution to Alzheimer´s Disease. I stayed at Aveiro for a postdoctoral fellowship in neuroscience, where I developed research in the identification and validation of biomarkers for early diagnosis of Alzheimer´s disease (AD).

BD: How did you become interested in science?

Ana Henriques: In the last year of the undergraduate degree, I undertook an internship in the neuroscience laboratory in the Center for Cell Biology at the University of Aveiro. During this period, I developed critical thinking and became aware of the key relevance of research in the advanced biomedicine. I was also moved by the enthusiasm and scientific contributions of both my mentors. The area of research in which I had the opportunity to participate, namely neuroscience, was an area recently introduced into the University. I thus had the opportunity to accompany the implementation of this new research area, which served to fuel my interest.

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

Ana Henriques: I've always been interested in areas related to medicine and society, in particular brain pathologies. As Alzheimer's disease is one of the most common forms of dementia, this field of study caught my attention early in my career. When I began working in neuroscience I knew that I had made the right choice.

BD: Describe the project for which you received the BD grant.

Ana Henriques: Dementia is characterized by a progressive deterioration in cognitive function that impairs performance of daily activities. Alzheimer's disease is a chronic, neurodegenerative disease and the most common cause of dementia in individuals older than 65 years. Differential diagnosis of AD is a challenge, in particular in the early stages or when clinical symptoms overlap with other types of dementia (eg, vascular dementia, dementia with Lewy bodies, and frontotemporal dementia). Moreover, with dementia progression, other factors besides a specific neuropathology might be at play, increasing the number of confounding factors that may mask an accurate diagnosis.

During my postdoctoral activity, and also as a consequence of my PhD work, I began to focus on Alzheimer's disease biomarker discovery. Other researchers have proposed that chronic inflammation has a key role in Alzheimer´s, as it can accelerate the disease or contribute to initiating it. Inflammatory cytokines may be produced by microglia, astrocytes, and macrophages and contribute to neuronal death and to disease pathogenesis. Also chemokines may be crucial elements in neuron-microglia communication. This project aims to identify potential inflammatory biomarkers by mimicking the disease conditions and by screening a large number of cytokines. The most relevant candidates will be evaluated in human Alzheimer´s samples from the Aveiro Cohort, a study of local Alzheimer´s patients and controls.

We plan to monitor and compare the levels of the candidate cytokine biomarkers in Alzheimer's and control patients. The study group will include patients evaluated by cognitive tests including Mini-Mental State Examination, Clock Test Scale Basic Activities of Daily Living, and the Scale of Instrumental Activities of Daily Living. Patients using NSAIDs will be excluded from the study, but one may also resort to this group for comparative purposes should the need arise.

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

Ana Henriques: The main goal of the project is to identify potential neuroinflammatory biomarker profiles for Alzheimer's disease. Successful completion of this project will contribute to appropriately diagnosing Alzheimer's at an early stage, but may also contribute to the diagnosis of other dementias.

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

Ana Henriques: The most significant contribution would be twofold. On one hand this is for the differential diagnosis of dementia but also this could be done using peripheral tissues in a non-invasive test. Additionally, targeting specific inflammatory biomarker candidates could potentially reveal new therapeutic avenues for the disease. Indeed, it has been reported that anti-inflammatory drugs can protect against and might diminish the risk for Alzheimer's disease.

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

Ana Henriques: We plan to use many BD reagents related to inflammation, including specific antibodies, purified proteins, ELISA assays, and inflammatory related BD™ Cytometric Bead Arrays that would guarantee the completion of the proposed project.


Christopher Jewell, PhD
Assistant Professor

Abstract Title:
Mapping Immune Signatures of Biomaterials


BD: What is your educational background?

Christopher Jewell: During my undergrad studies at Lehigh University I became interested in applying engineering to biological problems, so I completed a BS in chemical engineering and a BS in molecular biology. I earned my PhD in chemical engineering at the University of Wisconsin, Madison, in Dave Lynn's lab. My work focused on designing new polymer coatings for gene therapy. After that, I joined Prof. Darrell Irvine's lab at MIT as a Ragon Postdoctoral Fellow. My projects centered on developing biomaterial-based strategies to direct immune response. During my postdoc, I was also a visiting scientist at Harvard where I had an opportunity to interact with top immunologists. These experiences complemented my formal training in engineering. In autumn, 2012 I started my own lab in the Bioengineering Department at the University of Maryland.

BD: How did you become interested in science?

Christopher Jewell: When I was growing up my brother and I spent summers building with Legos and erector sets, and blowing up army men with chemistry sets in the back yard. I think that really got me interested in the fun and creativity of science. Over time, that kind of excitement evolved to study questions to which we do not yet have good solutions, or sometimes to find questions to apply the things we build to.

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

Christopher Jewell: Between graduate school and my postdoc, I worked in the healthcare practice of the Boston Consulting Group developing R&D strategy for biotech and vaccine companies. This experience exposed me to some of the challenges facing global health, and the amazing opportunities presented by new vaccines and immunotherapies. My excitement to work in this area led me to a postdoc in the Irvine lab at MIT, and ultimately to starting my own lab at the University of Maryland.

BD: Describe the project for which you received the BD grant.

Christopher Jewell: New vaccines and immunotherapies aimed at tough targets like malaria and cancer must carefully direct immune response to be effective. One way to achieve this goal is to design vaccines incorporating signals that activate specific parts of immunity or preferentially generate immune cells with the desired function. Delivering these signals at the right concentrations and combinations in animals or people is very challenging, so a lot of new vaccine candidates use polymers or other biomaterials that can control the release of several vaccine components over hours, days, or weeks. While these materials have a lot of potential for new vaccines, we don't fundamentally understand how the properties of these materials (eg, chemical modality, hydrophobicity) influence immune signaling and response. In fact, recent studies show that some common polymers inherently activate inflammatory "danger" or pathogen sensing pathways, even without foreign antigens or other adjuvants. The goal of this project is to link structural elements of vaccine carriers to activation of specific sets of immune signals or functions.

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

Christopher Jewell: In the short term, we will be creating polymer libraries with different chemical properties formulated as nanoparticles or microparticles. These vaccine carriers will be used to treat cells or mice, and then high-throughput tools (eg, cytokine arrays) will be used to evaluate how changing a particular polymer property impacts the resulting immune signaling in cells and animal models. Our first studies will focus on biomaterials without other immune signals, then we will incorporate common vaccine adjuvants/drugs and test how these additional signals impact immune signaling in cells or tissues when delivered in a controlled way from biomaterials (compared to conventional treatments involving soluble treatment without biomaterials). In the long term, we hope to establish patterns of immune signaling as a function of the properties of the polymers or other biomaterials. We hope ultimately to generate some design rules that could make developing the next generation of vaccines more efficient.

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

Christopher Jewell: An understanding of how biomaterials interact with and direct immune cells and tissues could allow new vaccines based on all-in-one materials that serve both as vaccine carriers and as agents actively direct or tune immune responses to treat a particular disease.

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

Christopher Jewell: Many of the proposed studies involve flow cytometry, so BD fluorescent antibody conjugates will be a fundamental type of reagent we use. Establishing cytokine signaling profiles as a function of biomaterial properties is a cornerstone of the project, and for this aspect we will use high-throughput BD™ Cytometric Bead Arrays (CBAs) to simultaneously analyze up to 30 analytes.


Yael Korin, PhD
Associate Researcher of Pathology and Laboratory Medicine

Abstract Title:
Multiparameter Immune Cell Flow Cytometry for the Characterization of MODS after MCSD


BD: What is your educational background?

Yael Korin: I received my undergraduate degree in genetics and zoology from the Hebrew University in Jerusalem. Afterward I studied for a master's degree in cell biology at California State University, Dominguez Hills, and for my PhD in virology and immunology at UCLA, where I remained for my postdoctoral fellowship and where I still work. Currently I am an associate researcher at the UCLA Immunogenetics Center, working under the advisory of Dr. Elaine Reed.

BD: How did you become interested in science?

Yael Korin: I grew up loving nature, animals, and plants. While still in high school I became attracted to biology as a formal discipline and began pursuing it. From that point forward I knew I wanted to be a biologist.

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

Yael Korin: I am mostly interested in studying the role and function of the immune system, particularly the T- and B-cell adaptive immunity, in responses to pathology and disease. My interest in HIV, during graduate school, related to how T cells interact with the virus, and the fate of infected cells. Now I'm focusing on T-cell immunity as it applies to organ transplantation, rejection, and tolerance.

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

Yael Korin: Heart failure affects more than 5 million people in the United States. Currently, Mechanical Circulatory Support Device (MCSD) therapy is the only alternative for patients with end-stage heart failure who are not candidates for transplantation. Patients undergoing MCSD therapy are at increased risk of multiple organ dysfunction syndrome (MODS), a life threatening condition caused by sustained activation of inflammatory responses. Risk evaluation for patients referred for MCSD is conducted, but no reliable methodology precisely assesses baseline immune system activation and predicts the risk of developing an uncontrollable systemic inflammatory response.

We will develop and evaluate the feasibility of multidimensional molecular biomarkers constructed by integrating peripheral blood mononuclear cell (PBMC) protein surface markers and mRNA gene expression in a prospective study, using sequential (time-dependent) blood samples obtained from patients undergoing MCSD.

We will collect peripheral blood from 15 study patients at days -1, 1, 3, 5, and 7 before and after MCSD surgery, and from 15 healthy volunteers at these five corresponding time points. Immunophenotyping will be performed using flow cytometry with antibodies against well-defined immune cell markers to determine the distribution of PBMC subsets, T- and B-cell memory and activation markers, homing markers, markers of immune senescence and exhaustion, markers to define viral clearance versus persistence, and cytotoxic markers.

We hypothesize that assessing simultaneously surface markers and gene expression in a time-dependent experiments will let us generate the necessary data to support our goal of developing integrative multilevel biomarkers.

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

Yael Korin: Our long-term goal is to develop and validate a reliable method that will afford us to precisely assess the baseline activation of the immune system and predicts the risk of developing an uncontrollable systemic inflammatory response that leads to multiple organ dysfunction syndrome. We hypothesize that multidimensional molecular biomarkers will achieve this goal. Thus, our short-term research goals are to develop and evaluate the feasibility of these biomarkers by analyzing sequential blood samples obtained from patients undergoing MCSD.

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

Yael Korin: What makes this study a very unique one is our ability to analyze the immunophenotype data together with the PBMC gene expression profiles obtained at the same time points. We will use our combined phenotype panel and gene expression profiles, along with powerful computational tools, to guide biomarker discovery and biomarker development. This is the first time that a simultaneous assessment of surface markers and gene expression in a time-dependent experiment is being conducted to generate the necessary data to support our goal of developing integrative multilevel biomarkers for the characterization of multiple organ dysfunction syndrome after beginning MCSD therapy.

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

Yael Korin: The UIC Immune Assessment Core will be developing an 8-template multiparameter flow cytometry panel comprised of 7, 10, and 11-color cocktails using PBMCs. These will be employed in a plate-based lyophilized format, conjugated to FITC, PE, PE-Cy™7, PerCP-Cy™5, APC, Alexa Fluor® 700, APC-H7, Pacific Blue™, BD Horizon™ Brilliant Violet™ 421/450, and BD Horizon™ PE-CF594. For cell fluorescence we will use the BD LSRFortessa™ flow cytometer from BD Biosciences and associated reagents.


Holly Rosenzweig, PhD
Assistant Professor

Abstract Title:
Interplay Between Neutrophils and T Cells in an Autoimmune Disease


BD: What is your educational background?

Holly Rosenzweig: I earned my BS in Molecular Biology from the University of Colorado and my PhD in immunology at Oregon Health and Science University, under the mentorship of Prof. Mary Stenzel-Poore. I then completed a postdoctoral fellowship at the Casey Eye Institute of the Oregon Health and Science University. There my mentor was Prof. James Rosenbaum. I'm currently Assistant Professor at the Oregon Health and Science University and the VA Medical Center.

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

Holly Rosenzweig: Science captured my interest from an early age. I received my first microscope when I was 6 years old.

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

Holly Rosenzweig: Most of my research interests (ie, PhD thesis work and postdoctoral fellowship) focused on innate immune responses. The cellular and signaling responses that shape innate immunity have fascinated me for a long time. With the recently defined "autoinflammatory diseases," the importance of the innate immune system in disease has been re-energized. This has transpired into the realization that innate immune mechanisms can also participate in previously defined autoimmune diseases. Historically, research efforts have focused on defined autoimmune responses—that is, antigen-specific T cells and autoantibodies—in disease, with little understanding of how such responses are orchestrated and how innate immunity is involved in this process.

BD: Describe the project for which you received the BD grant.

Holly Rosenzweig: The Th17 T-cell response has emerged as an important pathogenic response in many autoimmune disorders such as the spondyloarthropathies. Production of IL-17 from CD4+ T cells, and their participation has been a focus in spondyloarthritis research. However, recent findings highlight how IL-17 can be produced by innate cells such as neutrophils, which can play an important role in orchestration of T cells. Thus, the involvement of innate cells such as neutrophils could be a previously unconsidered and critical aspect involved in shaping the Th17 T-cell effector response.

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

Holly Rosenzweig: Using a previously established murine model of spondyloarthritis, wherein mice immunized with the cartilage constituent proteoglycan (PG) develop chronic progressive disease of the peripheral joints, spine, and uveitis (or ocular inflammatory disease), we propose to explore the interplay between neutrophils and antigen-specific T cells in development of disease and their participation in the Th17 response. To understand how the in vivo cellular interaction between neutrophils and T cells shapes "autoimmune" disease, we will use intravital videomicroscopy—an established technique in our lab that allows us to visualize in real time the cellular trafficking responses within the microvasculature and iris tissue of the eye.

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

Holly Rosenzweig: The Th17 response has emerged as a potential therapeutic target in many diseases including the spondyloarthropathies and their related diseases including psoriatic arthritis, psoriasis, inflammatory bowel disease, and uveitis. Understanding the cellular sources and signaling pathways could impact how we develop therapeutic targets.

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

Holly Rosenzweig: We will use antibodies for intracellular cytokine staining and flow cytometry, as well as fluorescent antibodies to label cells and visualize cell trafficking in vivo. Since we plan to measure cytokine production within the neutrophil population, we will run multiplex ELISAs.


Lynn Schnapp, MD
Professor of Medicine

Abstract Title:
Role of Lung Pericytes and Stromal Cells in the Pathogenesis of Pulmonary Fibrosis


BD: Tell us about your educational background.

Lynn Schnapp: I earned my Bachelor of Science degree in biology from the Massachusetts Institute of Technology, where I received the John Asinari award for undergraduate research. I then moved to the University of Pennsylvania, School of Medicine, where I received my MD and did a residency in internal medicine. I spent four years as a postdoctoral fellow in pulmonary and critical care medicine at the University of California, San Francisco. I have authored or co-authored nearly seventy journal articles and book chapters.

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

Lynn Schnapp: I was always what some people might call a science nerd, but my serious interest probably emerged during high school biology. On a recent visit to my parents, I rediscovered a certification I'd won in fourth grade from the Future Scientists and Engineers of America.

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

Lynn Schnapp: I'm a physician-scientist specializing in pulmonary and critical care medicine. I initially entered that field because I enjoy working in the intensive care unit. My exposure to this type of patent piqued my interest in research on lung injury and repair—what injures lungs, what gets them better, and the potential to apply the latest science to the patients I see. Cell biology provides deep insights into the real-world problems patients experience and can help drive further research activity that will eventually impact patient care.

BD: Describe the project for which you received the BD grant.

Lynn Schnapp: Acute lung injury and idiopathic pulmonary fibrosis both involve abnormal deposition of matrix proteins, for example collagens, in the lung. This scarring usually resolves after injury; in idiopathic pulmonary fibrosis its progression is relentless and fatal, and collagen production is out of control. Therapies, which include off-label immunosuppressants, steroids, and anti-oxidants, are ineffective, so there is a great deal of interest in developing new therapies. The key question we would like to answer is how and where the cells that make the scar-forming collagens arise. Figuring that out could lead to therapies targeting those cells, with the goal of reducing fibrosis and scar tissue formation.

We plan to use two mouse strains, each carrying fluorescently labeled cells that we believe are responsible for collagen deposition in the lung. We will also employ a cross between those mice, which will carry both labels. Our project involves characterizing those cell populations, and defining their distinctive behaviors with regard to fibrosis. We will grow those labeled cells in culture, separate them through flow cytometry, and determine their responses to stimulative cytokines, including their proliferation and generation of collagens.

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

Lynn Schnapp: Our research goal is to characterize these different cell populations, and understand their contributions to the abnormalities we observe in fibrosis. Ultimately, if the behaviors of these collagen-producing cells can be understood, it may be possible to develop targeted therapeutics against them. For example, if it turns out that FOXD1-derived cells are the major collagen-producing cell in fibrosis, one could design drugs that cause the cells to undergo apoptosis, or shut down their collagen expression.

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

Lynn Schnapp: More than 200,000 patients in the United States, and five million people worldwide, have idiopathic pulmonary fibrosis. Only 20% of patients survive five years post-diagnosis. Consequently, there is an urgent need for effective therapy. Experts estimate the potential market for treating pulmonary fibrosis at $2 billion per year. I believe that understanding the cellular mechanisms responsible for this disease will someday lead to effective therapies.

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

Lynn Schnapp: We plan to use recombinant proteins, including EGF, PDGF, aFGF, bFGF, and TGFbeta1, proliferation kits, multiplex bead-based assays for cytokine expression, ELISA kits, and antibodies for cell sorting and Western blot analysis.


Joseph Thome
Graduate Student

Abstract Title:
Analysis of Human Naïve T-Cell Persistence and Homeostasis in Lymphoid Tissue


BD: Tell us about your educational background.

Joseph Thome: I received my undergraduate degree in chemistry and biochemistry from Oberlin College, where I worked with Prof. Rebecca Whelan and Prof. William Fuchsman. Currently, I am a graduate student in the department of Microbiology and Immunology and the Columbia Center for Translational Immunology at Columbia University in the lab of Prof. Donna Farber.

BD: How did you become interested in science?

Joseph Thome: Volunteer and shadowing experiences at the local hospital when I was younger helped me develop an interest in medicine. From this, I became specifically curious about how invading pathogens affect the body and the possibility of advancing therapeutics through the lens of scientific discovery.

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

Joseph Thome: I began my research training in the field of bioanalytical chemistry, which sparked my enthusiasm to further work at the interface between experimental findings at the lab bench and clinical practice. Upon beginning my graduate studies, I knew that I wanted to develop a project where my results would be applicable to human health. Dr. Donna Farber's lab had a new development studying T-cell subset compartmentalization in human organ donor tissue sites around the body. The experimental findings show a diverse patterning of T-cell distribution in lymphoid and mucosal tissue sites that was remarkably consistent between donors. From this initial investigation, I became excited to work with this resource to expand knowledge in translational immunology and how the functional adaptive immune response varies with age.

BD: Describe the project for which you received the BD grant.

Joseph Thome: Naïve T cells are generated via thymopoiesis in early childhood followed by a gradual reduction in thymic function and an age-dependent involution of the functional organ volume beginning in puberty. This, combined with a decrease in naïve T-cell populations after pathogen exposure, begs questions into how an effective immune response can be generated with increasing age. Currently, it is not understood how and whether human naïve T cells are maintained in the context of decreasing thymic output throughout life, mainly because studies on human T-cell development have been limited to the sampling of peripheral blood.

We have established collaboration with the New York Organ Donor Network, which allows for the study of naïve T-cell maintenance beyond the blood and in specific organ sites where the majority of T-cell populations reside. Initial experiments show naïve T-cell persistence throughout the body in donor tissues analyzed from individuals in advanced age, long after the cessation of functional thymopoiesis. Additionally, preliminary data hint at a potential site of naïve T-cell maintenance and retention prior to further export into peripheral tissue sites, which needs elucidation. To test our initial hypotheses, we will use flow cytometry to characterize naïve T cells, specifically recent thymic emigrants (RTEs), using the markers CD31 and PTK7 alongside T-cell receptor excision circle measurements. From here we will assay the cytokine secretion profiles of the differential tissue environments where naïve T cells reside to understand the mechanisms of cell longevity.

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

Joseph Thome: The short-term goal of the project is to determine naïve T-cell longevity and the maintenance mechanisms of these immune cells in tissue sites around the body. We seek to uncover possible differential tissue cytokine environments that promote cell survival and identify a potential reservoir for naïve T-cell persistence in advanced age. From this, we will investigate whether naïve T cells keep their ability to respond to new pathogens as a function of age and tissue residence while also assessing the proliferative and repopulation capacity of these subsets compared to peripheral blood.

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

Joseph Thome: The results from this study will have implications in the immunology of aging, while also yielding new information on how naïve T cells are distributed throughout the body. Knowledge of naïve T-cell longevity will aid in mechanisms to promote immune responses to new pathogens while furthering the development of vaccines targeting organ-specific infectious agents. After identifying potential sites that are enriched for naïve T cells, these areas may prove useful in T-cell proliferation procedures to combat immunosenescence, elucidating the mechanisms of dysregulations that can occur with advanced age.

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

Joseph Thome: I will be using human antibodies for flow cytometry for most of the immune cell profiling in my work. However, I will also be using the BD™ Cytometric Bead Array kits to determine cytokine secretion environments in various lymphoid tissue sites.