BD BIOSCIENCES RESEARCH GRANTS

Fall 2013 Research Grant Recipients Talk About Their Research


Marco A. Biamonte, PhD
Chief Executive Officer

Abstract Title:
A Diagnostic to Aid the Elimination of All Filariases


BD: What is your educational background?

Marco Biamonte: I earned my BS in chemical engineering from the Swiss Institute of Technology, Lausanne, and my PhD in organic chemistry from the University of Cambridge, UK. My doctoral advisers were Drs. James Staunton and Ian Paterson. I then completed a postdoctoral fellowship in carbohydrate chemistry from the Swiss Institute of Technology, Zürich, under the direction of Prof. Andrea Vasella.

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

Marco Biamonte: I have always loved science: first the perfection of mathematics, then the beauty of galaxies, and later the equally awe-inspiring perfection of biological systems. I also love chemistry and organic synthesis. A multi-step synthesis feels like a chess game in which several moves (reaction conditions) are linked in non-obvious ways to reach the desired outcome. Looking into chemistry and biology simultaneously is even better. It is like being authorized to glimpse into the miracle of life with both eyes.

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

Marco Biamonte: I traveled quite a lot during my twenties, and I have seen injustices—lepers, for instance—that have shaped my vision of the world. Leprosy is an example of a disease that should not exist because it is curable for just a few dollars. My travels set in motion an intentional desire to do something practical, and to apply my skills to neglected diseases. I therefore trained myself in chemistry, and spent many years in the pharmaceutical industry learning the subtleties of drug discovery. I reached the point where it made sense for me to set up a non-profit organization with the mission to discover new treatments and diagnostics for diseases associated with poverty.

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

Marco Biamonte: Parasitic worms (helminths) infect hundreds of millions of people worldwide, predominantly in the poorest regions of the world. Although rarely lethal, helminth infections represent a major cause of disability, and are further associated with slower growth and lower cognitive functions in children.

Filariae are a subgroup of parasitic worms that can clog the lymphatic system (lymphatic filariasis), damage the optic nerve (river blindness), or accumulate under the skin (loiasis). Drugs exist for these conditions, but there are no rapid diagnostic tests against river blindness and loiasis. Such diagnostics would be very useful, especially in the context of mass drug administration (MDA) programs. For instance, the drug of choice against river blindness can cause severe side effects in those who have loiasis, so eliminating river blindness via MDAs is impossible in areas where loiasis is co-endemic.

Our goal is to deliver an inexpensive diagnostic that will simultaneously detect and distinguish the filariae implicated in these diseases. This will assist the World Health Organization and related organizations in targeting their MDA campaigns, and will assist doctors in prescribing optimal treatments.

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

Marco Biamonte: We are building a lateral flow assay in the same format as the consumer pregnancy assay, in which each worm gives rise to a different line. We anticipate a sensitivity tenfold higher than that of conventional lateral flow assays. We will design the diagnostic device to detect antigens in the blood and urine of infected patients, at a cost of less than one dollar per device.

Since each disease will give rise to a different "bar" on the readout, the result will be a "bar code" readable by the naked eye or by capturing its image with a smart phone camera. A reader "app" will quantify the signal from each bar, certify that the assay was reporting correct results via internal controls, and push the data into a Global Mapping and Monitoring Database to allow real-time analysis of disease distribution. Expert system analysis of the readout could even recommend treatment options for on-site healthcare workers.

Our short-term goal is to demonstrate that we can build a sensitive, proof-of-concept device. The mid-term goal is to build the accompanying app. The long-term goal is to optimize the device, initiate its production, and validate the tool in the field.

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

Marco Biamonte: Loiasis affects up to 40 million people worldwide, river blindness infects as many as 18 million, while lymphatic filariasis affects 120 million. Our proposed diagnostic tool will support MDA programs against filarial diseases. Once MDA programs are no longer necessary, the tool will still be useful for post-treatment surveillance and for individual case management. Ultimately, our goal is to contribute to eliminating all the filariae-related illnesses.

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

Marco Biamonte: We will use BD Biosciences antibodies to build early prototypes of the device. As a future development, it would be exciting to leverage the vast collection of BD Biosciences antibodies to expand the platform and diagnose certain cancers in low-to-middle income countries.


Kirstin M. Heutinck, PhD
Postdoctoral Fellow

Abstract Title:
Multiparameter Flow Cytometric Analysis of Kidney Resident T Cells: A Future Guide to Personalized Treatment of Injured Kidney Transplants


BD: What is your educational background?

Kirstin Heutinck: I earned my bachelors and masters degree from the University of Amsterdam, the Netherlands, where I subsequently spent eight months on a scientific internship in virology and molecular biology. I then came to the United States, where I completed a ten-month internship at the University of Virginia. There I studied the role of the thalamic mediodorsal nucleus in the generation of epileptic seizures. I then returned to the Netherlands, where I received my PhD in immunology at the Academic Medical Center, Amsterdam. My thesis work focused on innate immune responses in kidney transplantation. I remained at this institution for my post-doc.

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

Kirstin Heutinck: Since my mother was a biology teacher, I was exposed to science from childhood. During secondary school I focused on typical science and math subjects which, while interesting, did not directly inspire me to become a scientist. My "inspiration" did not occur until my undergraduate studies in medical biology. I enjoyed the practical labs and became fascinated with how the human body regulates such refined, diverse processes. After my masters studies, I decided I wanted to learn more and applied to a PhD program.

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

Kirstin Heutinck: As a masters student studying microbiology, I became interested in viral infections. Later, in graduate school, I studied how kidney cells responded to compounds of viruses, in particular viral RNA, increasing my knowledge about the interactions between host cell and virus. Working in the Department of Experimental Immunology, I became acquainted with other immunological topics, particularly adaptive immune responses. In my current position, I have the opportunity to combine the study of viruses and adaptive immunity, specifically how cells of the adaptive immune system, T cells, control viral infections in patients who have undergone kidney transplantation.

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

Kirstin Heutinck: To prevent rejection of kidney transplants, the immune system must be suppressed with a cocktail of immunosuppressive drugs. The downside of this treatment is an increased risk of infections. T cells, a type of immune cell, play a crucial role in both the control of viral infection and in rejection of transplants. Most studies have analyzed T cells isolated from blood. However, growing evidence indicates that T cells residing inside the tissue of an organ are significantly involved in local immune responses. Little is known about how T cells operate inside healthy and transplanted human kidneys. We found that human kidneys indeed possess a distinct subset of T cells. The aim of our project is to reveal the function of these kidney T cells, and investigate how the cells are recruited to and maintained inside the kidney. We hypothesize that the function of local T cells reflects the ongoing disease process within a transplanted kidney. For example, during rejection, T cells that recognize foreign cells will infiltrate the kidney transplant. Depending on the "killer" or "pacifying" function of these T cells, their presence will be harmful or beneficial for the function of the kidney transplant. We will use a wide array of functional markers, which can be visualized with fluorescent-labeled antibodies, to characterize T cells inside the healthy and injured kidneys.

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

Kirstin Heutinck: Short term, we hope to understand better the local immune response within healthy and transplanted kidneys, and determine if the function of local T cells reflects ongoing disease processes in kidney transplants. Long term, we hope to develop patient-specific treatment strategies based on the functional characterization of T cells isolated from kidney transplants.

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

Kirstin Heutinck: Approximately 70,000 kidney transplants occur worldwide each year. On average, these transplants last between ten and twelve years. I believe this could be improved significantly if the recipient immune responses were better understood. These studies will increase our knowledge about the function of T cells inside the human kidney and will reveal if the function of these T cells correlates to different causes of kidney transplant dysfunction, including transplant rejection and viral infections. We hope these insights will aid in developing personalized treatment strategies to delay or prevent organ rejection.

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

Kirstin Heutinck: I am planning to use fluorescent labeled monoclonal antibodies to perform extensive phenotypic and functional characterization of T cells that reside inside healthy and transplanted kidneys.


Aaruni Khanolkar, MBBS, PhD
Assistant Professor of Pathology

Abstract Title:
To Define a Cellular Signature that Predicts Disease Relapse in Pediatric Acute Lymphocytic Leukemia


BD: What is your educational background?

Aaruni Khanolkar: After receiving my medical degree from Jiwaji University, India, I moved to the US to pursue a PhD in microbiology and immunology from the University of Arkansas for Medical Sciences. I then undertook extensive post-doctoral training in microbiology and immunology at the University of Alabama at Birmingham, followed by the University of Iowa, where I worked in Dr. John T. Harty's laboratory. Thereafter I completed a clinical immunology fellowship in the Department of Pathology at the University of Utah and the affiliated ARUP National Reference Laboratory.

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

Aaruni Khanolkar: I was always fairly good at grasping facts, which helped me do well in scientific subjects, especially high school chemistry. Everyone in my circle of friends in grade school was interested in science. Deep down, I believe, we were aware of the harsh reality of our environment, which offered slim pickings in terms of stable career options outside science-related fields. I also kept my eyes and ears open to the regular news items about important scientific discoveries being made. It became obvious that the US was then, and still is, the science juggernaut of the world.

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

Aaruni Khanolkar: I was formally introduced to immunology, under the broader umbrella of pathology, during Phase 2 of my medical school curriculum. I performed well in the examination and although the immunology I learned then was rudimentary, it somehow stimulated my interest.

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

Aaruni Khanolkar: Between 75 and 80 percent of pediatric leukemias are of the acute lymphocytic variety (ALL), and they predominantly involve precursor B cells. Although a 90% cure rate is observed for pediatric B-cell ALL associated with favorable prognostic factors, almost one out of every four patients experiences front-line therapy failure. Accordingly, there is a need to identify novel actionable biomarkers that can help predict response to therapy and potentially identify targets for new therapeutic drugs. One factor that affects tumor cell behavior and response to therapy is the cell's ability to receive and process cues from their local microenvironment, a fact that might be used to advantage.

Our approach toward understanding factors involves analyzing signaling pathways in precursor B-ALL cells by flow cytometry following exposure to signals that these cells could potentially encounter in vivo. Our lab is in a unique position to conduct these studies because it is the sole CLIA and College of American Pathologists-certified laboratory in our institution that offers clinically validated multiparameter flow cytometric assessment of patient peripheral blood, bone marrow, and tissue samples for the diagnosis of blood disorders. We receive and analyze at least 15 pediatric leukemia/lymphoma samples each month, and over 90% of them are precursor B-cell ALL samples.

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

Aaruni Khanolkar: The overarching goal of this proposal is to test the hypothesis that a unique cellular and tumor cell-associated phosphoprotein signature identifies patients who are insensitive to standard front-line chemotherapy used in pediatric precursor B-cell ALL. We hope to identify a tumor cell specific signaling phosphoprotein signature at diagnosis that can identify patients at risk for front-line therapy failure.

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

Aaruni Khanolkar: Almost 3,800 children each year develop leukemias, which are the most common pediatric cancers in the US. This study will help identify, at the time of diagnosis, patients who will likely benefit from stem cell transplantation vs conventional chemotherapy. This decision is currently made by evaluating the persistence of leukemic blasts—minimal residual disease—four to six weeks after initiation of induction chemotherapy. For patients who would benefit from stem cell transplantation, this represents an unnecessary and potentially detrimental delay, and exposes them to unnecessary treatment.

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

Aaruni Khanolkar: We will be utilizing antibodies to examine phosphorylation status of a number of intracellular B-cell specific signaling molecules to identify a tumor cell-specific phosphoprotein signature.


Jianxun (Jim) Song, PhD
Assistant Professor

Abstract Title:
Directed, Antigen-Specific Induced Pluripotent Stem Cell-Derived Cytotoxic T Lymphocytes for Cell-based Therapies


BD: What is your educational background?

Jianxun Song: I received my undergraduate degree in biology from Chongqing Normal University, Chongqing, China. Afterwards, I entered the Third Military Medical University in China, where I earned my MSc in pathogenic biology, and my PhD in immunology. I was a postdoctoral fellow in immunology at the La Jolla Institute for Allergy & Immunology in California, where my mentor was Dr. Michael Croft.

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

Jianxun Song: I became interested in biology during my undergraduate studies, which is when I decided to pursue a career in science.

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

Jianxun Song: My research interests from before graduate school through today have involved T-cell activation and memory. Beginning with an etiology training program at the Department of Health in China, then during my time at the Third Military Medical University, I continued to study how signals from peripheral blood induce T-cell apoptosis. Then, while a postdoctoral fellow in the laboratory of Dr. Michael Croft at La Jolla, I decided to specialize in T-cell biology. After completing my postdoctoral training, I joined the Penn State faculty as a tenure-track assistant professor. I am now developing techniques and methods for producing T cells from pluripotent stem cells. This broadening of my skills will help me to understand the mechanisms by which T cells are activated, how their memory arises, and how to generate highly reactive T lymphocytes from induced pluripotent stem cells (iPSCs), specifically iPSC-T cells, for cell-based therapies.

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

Jianxun Song: Virus-specific T cells capable of controlling hepatitis B virus (HBV) and eliminating hepatocellular carcinoma expressing HBV antigen are deleted or dysfunctional in patients with chronic HBV infections. Current antiviral therapy, which targets the virus' reverse transcriptase, rarely establishes immunological control over HBV replication.

Adoptive cell transfer (ACT) of HBV-specific CD8+ cytotoxic T lymphocytes (CTLs) is a highly promising treatment for chronic HBV infection and related cancers. Naive or central memory T- cell–derived effector CTLs—the "right" or "highly reactive" CTLs—are optimal populations for ACT-based immunotherapy. These cells proliferate readily, are less prone to apoptosis than terminally differentiated cells, and respond more effectively to homeostatic cytokines. However, ACT often is not feasible due to difficulties in obtaining sufficient numbers of highly reactive cells from patients.

We propose to study the fundamental properties of HBV-specific iPSC-CTLs, genetically modified with HBV-specific T-cell receptors, as potential immunotherapies against chronic HBV infection and HBV-associated cancers. To define characteristics of HBV-specific iPSC-CTLs related to anti-HBV activity, we hypothesize that iPSCs genetically modified with antigen-specific T-cell receptors, followed by differentiation driven by Notch signaling, will enable pluripotent stem cells to pass hematopoietic and T-lineage differentiation checkpoints, resulting in naïve monoclonal antigen-specific CD8+ T cells. To test this hypothesis, we will study HBV-specific iPSC-CTLs with regard to phenotype, specificity, and function. Next, to determine the anti-HBV activity of HBV-specific iPSC-CTLs in HBV-bearing mice, we hypothesize that ACT of highly reactive HBV-specific iPSC-CTLs will generate functional CTL effectors that control chronic HBV infection and HBV-associated tumor growth. We plan to use well characterized murine HBV infection models to determine the efficacy of HBV-specific iPSC-CTLs for cell-based HBV therapies.

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

Jianxun Song: Short term, we hope to define the fundamental properties of HBV-specific iPSC-CTLs that relate to cytotoxic activity, and to determine the activity of HBV-specific iPSC-CTLs in hepatitis B-infected mice. Long-term, we hope to develop and optimize pluripotent stem cells expressing antigen-specific T-cell receptors as a source of highly reactive cytotoxic T lymphocytes for ACT-based immunotherapy.

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

Jianxun Song: Hepatitis B is a chronic viral infection affecting 240 million people worldwide, with approximately 600,000 deaths per year. Hepatitis B vaccination is the only preventive, but there is no effective treatment. Highly reactive hepatitis B virus specific cytotoxic T lymphocytes could potentially treat both HBV infection and HBV-associated liver cancer. This project is an early-stage attempt to develop an immunotherapeutic cell-based treatment for hepatitis B. The knowledge we gain may also provide insights into strategies for cell-based therapies of other infectious diseases and cancers.

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

Jianxun Song: We plan to use anti-mouse and anti-human antibodies, flow cytometry kits/cocktails, and immunology buffers. In addition, we anticipate purchasing cell-based assays, apoptosis assays, recombinant proteins, proliferation assays, and ELISA and ELISPOT kits. We rely heavily on magnetic cell separation, so we plan to acquire reagents for isolating human and mouse cells. Finally, we will employ transfer vectors and protein purification kits to monitor protein expression.


Saranya Sridhar, MBBS, DPhil
Research Associate

Abstract Title:
Identification of a Protective T-cell Signature Against Development of Symptomatic Pandemic Influenza Illness in Humans


BD: What is your educational background?

Saranya Sridhar: I completed my MBBS and internship at Nagpur University in India in 2003. Following my medical training, I received a Rhodes Scholarship to undertake a DPhil at the University of Oxford under Prof. Adrian Hill. My dissertation focused on the preclinical development of adenoviral vector vaccines against preerythrocytic-stage malaria, and demonstrated for the first time single-dose protection against murine malaria using a viral vector vaccine. In 2007, I received the NIH funded Fogarty International HIV/AIDS Training Fellowship to pursue a Masters in Epidemiology at the University of California, Berkeley. During this time I learned biostatistics, epidemiological study design, clinical epidemiology, and longitudinal data analysis. Since February 2010, I have been involved in postdoctoral work with Professor Ajit Lalvani at Imperial College London. My research combines my epidemiological and immunological training to investigate natural immunity to tuberculosis and influenza in humans.

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

Saranya Sridhar: I became interested in science at an early age, after reading Richard Feynman's book, Surely You're Joking, Mr. Feynman. The childlike excitement and curiosity Feynman projects for every natural phenomenon he discusses aroused my interest in science.

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

Saranya Sridhar: My interest in immunology began during my medical training, when I was awarded a studentship by the Indian Council of Medical Research. This award, which allowed me to undertake a laboratory-based project for three months, was a unique opportunity for a medical student. During this time I worked in the Department of Biochemistry isolating proteins from a worm that would help us to identify components of a potential vaccine against filariasis, a mosquito-borne disease endemic to that region of India. This project, my first insight into immunology, made me aware of the potential in understanding and exploiting the immune system to discover and develop new drugs or vaccines. Since then, I have focused on translational immunology.

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

Saranya Sridhar: One of the major barriers to influenza control is the constant evolution of the influenza virus, which enables it to evade the human immune system. This evolution leads to generation of new virus types that cause annual influenza epidemics or on occasion pandemics such as the 2009 swine flu H1N1(pH1N1) pandemic. During pandemics, limiting disease severity acquires great public health importance. Although an immune signature predicting asymptomatic versus symptomatic or severe illness following natural influenza infection is a global research priority, scientists have not yet identified relevant biomarkers.

Previous animal work and epidemiological evidence suggest that T cells may play a role in limiting severe illness. We propose to test this theory in our study. Addressing this fundamental question of protective T-cell correlates against pandemic influenza in humans requires a natural experiment involving emergence of an antigenically-shifted virus, a unique opportunity presented by the 2009 pH1N1 pandemic. We therefore recruited, collected samples from, and followed through the pandemic a cohort of healthy adults. Since investigating correlates of protection requires measuring these correlates before development of clinical influenza, collecting peripheral blood mononuclear cells from our cohort before clinical illness provided a unique opportunity to identify such correlates. The overall aim of this project is to identify the cellular immune signature prior to infection that is associated with limiting severity of influenza illness. We propose to phenotype immune cells collected from individuals using multicolor flow cytometry. This data will be analyzed in a hypothesis-driven and data-driven approach to identify the immune signature differentiating individuals developing asymptomatic influenza from those developing a more severe illness. We expect that CD8+ T cells capable of homing to the lung and secreting cytotoxic molecules will be associated with protection against severe influenza illness.

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

Saranya Sridhar: The short-term goal of this project is to identify a cellular immune correlate of protection against symptomatic pandemic influenza in naturally infected humans. Our long-term goal is to use the cellular immune signature to develop a universal influenza vaccine that protects against illness from new, antigenically shifted influenza viruses capable of causing pandemics.

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

Saranya Sridhar: Influenza is a major health problem, and despite the widespread deployment of vaccines, influenza annually causes an estimated 1 billion cases and between 250,000 and 500,000 deaths worldwide. An immune biomarker identifying those protected against symptomatic influenza would signal a paradigm shift in public health strategy against this disease. Our study could identify a cellular correlate of protection and provide the immunological basis for designing and evaluating influenza vaccines that induce protective cellular immunity against pandemic influenza.

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

Saranya Sridhar: We propose to use BD's flow cytometry antibodies against a variety of immune cell surface markers as well as intracellular cytokines to test the functionality of immune cells. We will conduct all flow cytometry work on the BD LSRFortessa™ cell analyzer. In addition, we propose to use the BD™ Cytometric Bead Array to test for different secreted chemokines and cytokines.


Amaya I. Wolf, PhD
Staff Scientist

Abstract Title:
The Role of MDSCs in Viral-bacterial Respiratory Co-infection


BD: What is your educational background?

Amaya Wolf: I studied biochemistry at the Free University of Berlin, Germany, where I specialized in immunology. I then worked on my masters degree in the laboratory of Dr. Ulrich von Andrian at the Center for Blood Research in Boston. I then joined Dr. Wolfgang Weninger's laboratory, at the Wistar Institute in Philadelphia, as a graduate student to study host immunity to influenza virus infection, specifically the role of plasmacytoid dendritic cells in anti-viral defense. As a postdoctoral fellow, I continued my work in the field of anti-viral immune responses with Dr. Jan Erikson at the Wistar Institute, investigating the regulation of B-cell–mediated immunity to influenza infection, vaccination, and viral-bacterial co-infection. I am interested in understanding the mechanisms underlying pathogenesis and protection during host-microbe interactions in the respiratory tract.

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

Amaya Wolf: I became interested in science because my sister had cystic fibrosis. I developed a strong desire to understand the biological causes of her disease and—in the end—to explore approaches to improved treatments for this disorder.

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

Amaya Wolf: Because cystic fibrosis causes chronic lung infections, my scientific interests and direction naturally evolved towards the study of immunity to respiratory infections, in particular to viruses and bacteria. My motivation for this field of study was further fueled when I became involved in a program project grant in viral immunity through my mentor, Dr. Jan Erikson. This project resulted in various exciting collaborative studies.

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

Amaya Wolf: We study the immune response to respiratory tract infections and co-infection with influenza virus and Streptococcus pneumoniae. Despite available vaccines, respiratory co-infections, in particular with these two pathogens, remain a major health problem worldwide and cause significant mortality. However, many individuals also carry Streptococcus pneumoniae and other microbes asymptomatically in the respiratory tract. The role of the respiratory tract microbiome in immunity to subsequent infections is unclear.

We have found that mice carrying Streptococcus pneumoniae harbor a myeloid population that appears to resemble myeloid-derived suppressor cells. We believe these cells are critical in regulating the response to a subsequent infection with influenza virus, that they carry out positive functions, and suppress disease. We have identified a virulence factor of Streptococcus pneumoniae that is fundamental for the induction of this distinct myeloid cell subset, and correlates with protection against influenza virus mediated disease. We expect our studies to reveal the mechanism by which the Strep-induced myeloid-derived suppressor cells prevent virus-induced lung pathology.

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

Amaya Wolf: Our short-term goal is to characterize myeloid populations in the lungs following exposure to Streptococcus pneumoniae. We propose to examine the role of distinct myeloid cell subsets in the regulation of immunopathology, in particular T-cell responses, in viral-bacterial respiratory co-infection. Our main objective is to understand better the molecular mechanism for the generation and the functions of myeloid-derived suppressor cells in lung inflammation during infection. Our long-term goal is to develop and utilize therapeutic strategies targeting myeloid cell subsets by specifically harnessing their immunosuppressive functions that can be beneficial for treatment of respiratory viral infection.

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

Amaya Wolf: Respiratory tract infections are a severe clinical problem, specifically those involving both influenza A viruses and Streptococcus pneumoniae. The order in which humans acquire these viral and bacterial respiratory pathogens remains unclear. Understanding how certain microbes in the respiratory tract modulate host immunity, for example through the activities of virulence factors that can promote pathogenesis but also trigger protective innate and adaptive immune responses, will help to improve current vaccine designs.

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

Amaya Wolf: Many of the proposed studies involve flow cytometry, cell sorting, and functional assays that allow us to identify, characterize, purify, and manipulate our cells of interest. All these approaches rely on BD fluorescent antibody conjugates (cell surface markers, intracellular cytokines, antibodies for BD Phosflow™), purified proteins (cytokines and growth factors for cell cultures and antibodies for in vivo treatment/depletion), ELISA assays, and BD™ Cytometric Bead Arrays to determine inflammatory/anti-inflammatory responses in lung lavages. Analysis will be performed on BD™ LSR II and BD FACSCalibur™ flow cytometers, and cell sorting on a BD FACSAria™ II cell sorter available in the Wistar Institute Flow Cytometry Core facility.


Shuang Zhang
Graduate Student

Abstract Title:
Testing the Therapeutic Potential of Enhancing Efferocytosis Efficiency to Prevent Heart Failure after Heart Attack


BD: What is your educational background?

Shuang Zhang: After studying for three years at Shandong Normal University in China, I entered East Tennessee State University, where I received my BS in biology. Since 2012 I have been at Northwestern University as a graduate student in the laboratory of Prof. Edward Thorp. I am studying inflammation-resolution in monocytes and macrophages after heart attack and during ischemia.

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

Shuang Zhang: My mom is a pediatrician. She influenced me quite a bit, which is why I have been interested in biomedical sciences since childhood. I was pretty good at biology in high school so I decided to major in bio at college. I didn't take any immunology courses, but I read a book called The Transformed Cell by Stephen Rosenberg, a Christmas gift from my undergrad PI Dr. Alok Agarawal. That amazing book stimulated my interests in immunology.

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

Shuang Zhang: As an undergrad I worked in an immunology lab, where I learned about macrophages in chronic inflammation diseases such as atherosclerosis. The father in my host family included a professor working on the adaptive immune system and hepatitis A virus. He told me about the importance of the adaptive immune system, and how little we know about it. As a result of those discussions, I began leaning towards studying immunology in grad school. I rotated in two immunology labs during my first year at Northwestern University. The experience of actually working in immunology solidified my interest in that field.

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

Shuang Zhang: Heart failure after myocardial infarction (MI) is a significant cause of illness and death worldwide. Though pharmacological advances in medicine have significantly reduced mortality, the residual risk of post MI heart failure remains high. Our work involves developing complementary approaches to preserve heart function.

After a heart attack, a burst of cardiomyocyte death triggers the recruitment of monocytes and macrophages that are responsible for clearing dead myocardial tissue. These innate immune cells execute their function via efferocytosis, the phagocytic clearance of apoptotic cells by macrophages. Efferocytosis is involved in inflammation resolution and homeostatic tissue remodeling via triggering anti-inflammatory pathways. Efficient efferocytosis of cardiomyocytes is a prerequisite for heart repair. However, studies indicate that clearance of dying heart tissue after heart attack is suboptimal.

My research focuses on targeting efferocytosis post MI to prevent heart failure. Specifically, this grant will provide funding towards the development of a reliable, accurate method for quantifying efferocytosis within the heart.

Effective efferocytosis requires the coupled presentation of "eat-me" signals with suppression of "don't-eat-me" signals on the surface of target cells. CD47 is a marker of cell viability and communicates "don't eat me" signals to phagocytes. The Weissman group at Stanford University has shown that blocking CD47 can enhance efferocytosis of tumor cells. However, the role of CD47 in regulating efferocytosis of cardiomyocytes is unclear. We hypothesize that blockade of CD47 therapeutically enhances myocardial efferocytosis, inflammation resolution, and heart repair.

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

Shuang Zhang: Our short-term goal is to study the function of CD47-blocking antibody in our in vivo mouse MI model to test the role of CD47 on cardiac function. As a long-term goal, we would like to translate results from this basic research into the development of novel therapeutics that enhance efferocytosis and resolve inflammation after MI. We expect these new therapies will complement current treatments.

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

Shuang Zhang: Heart failure after myocardial infarction (MI) is a significant cause of morbidity and mortality. Perhaps as many as 40% of individuals who suffer a heart attack experience heart failure, some quite soon after the MI. These individuals make up 20% of all heart failure cases. While mortality has fallen for these patients, the personal and economic costs of heart failure are quite high. A successful treatment regimen that reduces the incidence of post-MI heart failure could reduce suffering among these patients and their families, allow many to return to productive work, and save the national healthcare systems billions of dollars per year. I believe that strategies involving an anti-CD47 antibody to enhance efferocytosis and reduce inflammation could become a significant component of such a treatment strategy.

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

Shuang Zhang: We plan to use antibodies for flow cytometry to identify immune cell infiltration after myocardial infarction, and perhaps in novel in vivo efferocytosis assays. These antibodies include fluorescent conjugated anti-CD68, anti-desmin, anti-MerTK, anti-Ly6G, anti-Ly6C, and others, to study efferocytosis and inflammation after MI.