Jeffrey A Bluestone, Professor And Director
University Of California San Francisco 3333 California St., Ste 315 San Francisco, Ca 941430962
Grant 1P30DK063720-019002 from National Institute Of Diabetes And Digestive And Kidney Diseases IRG: ZDK1
Abstract: The major objectives of this core are to produce and provide, to the UCSF DRTC community, pancreatic tissues and purified islets from mouse, non-human primates (NHP), and human donor pancreas. Our focus is to provide each investigator the best possible pancreatic tissue and islets, ensuring the quality and reproducibility of these samples. We will focus on developing new techniques that aim to minimize islet cell death, maximize islet yields and optimize islet function. We will work closely with experts in the islet, b-cell field to identify factors that will contribute to us reaching our goals for better new islet isolation techniques. The Core will continue to develop assays to better assess the quality of a pancreatic islet preparation. Investigate more informative quality control assays, which will prospectively predict in vivo islet function rather than the limited prospective studies relied upon today. The Core will work with the basic scientists, which we provide pancreas tissue and islets, to help us take full advantage of new research in islet cell expansion and neogenesis into large-scale islet transplantation settings. The Core will also maintain the state-of-the-art human islet isolation facilities meeting all FDA s cGMP regulations for human islet isolation and transplantation. The Core will also maintain the NHP and rodent pancreatic islet isolation facilities. The Core will focus on taking the positive aspects to what has been developed in the human islet isolation laboratory and apply these techniques and assays to both the NHP and rodent facilities and vise versa. The Islet Production Facility Core will play a critical role in the experimental and clinical programs of the DRTC and our objective is to provide the best possible tissues and to aid in the advancement of islet and beta-cells research.
Keywords: biomedical facility, pancreas, pancreatic islet, tissue /cell preparation, tissue resource /registry, cell biology, gene therapy, insulin dependent diabetes mellitus, noninsulin dependent diabetes mellitus, pancreatic islet transplantation, technology /technique development, Primate, human tissue, laboratory mouse
Project start date: 2002-09-01
Project end date: 2007-08-31
Sponsored Links Excellgen http://Excellgen.com
Diabetes Research And Training Center
Jeffrey A Bluestone, Professor And Director
University Of California San Francisco 3333 California St., Ste 315 San Francisco, Ca 941430962
Grant 1P30DK063720-01 from National Institute Of Diabetes And Digestive And Kidney Diseases IRG: ZDK1
Abstract: The University of California at San Francisco Diabetes Center, an organized research unit at UCSF, has functioned for more than a half-century as a basic and clinical research enterprise at the forefront of diabetes research. Historically, the program has had an ongoing enrichment program, Pilot and Feasibility studies, strong basic science and clinical research interface. The goal of the Center is to support a highly interactive team involved either directly or indirectly in Type 1 and Type 2 diabetes research to advance the study and treatment of the disease. In this application, the UCSF Diabetes Center proposes to establish a Diabetes Research and Training Center (DRTC) that will support Core Laboratories, an Enrichment Program and Pilot and Feasibility studies. The Center will encompass a broad range of intellectual and research expertise from over 12 departments and organized research units and three UCSF campuses focused on basic research with an eye towards clinical application. The center will combine immunology, metabolic research, cell biology and genetics in the field of diabetes to develop unique approaches to understand and treat this devastating disease. Investigators of the DRTC are organized in the following programmatic areas Cell Biology of Islets, Developmental Biology, Islet Transplantation and Immunology, Autoimmunity, Receptors and Signaling, and Obesity and Metabolism and Obesity, Complications of Diabetes. Seven Core facilities are designed to facilitate interdisciplinary investigations of these scientists ( Enrichment Core, Islet Production Facility Core, Microscopy and Cellular Imaging Core, Genomics and Bioinformatics Core, Mouse Genetics Core, Mouse Metabolism, and Human Metabolism Cores). A Pilot and Feasibility Grant Program serves to foster new initiatives in diabetes research primarily of junior faculty and those senior faculty from outside the diabetes focus area. An intensive academic enrichment program which organizes seminars and various symposia is designed to keep Center investigators abreast of the latest discoveries and to maintain the research program at this center at the forefront of biomedical science.
Keywords: biomedical facility, health science research, insulin dependent diabetes mellitus, noninsulin dependent diabetes mellitus, training, clinical research
Project start date: 2003-05-01
Project end date: 2008-01-31
1P30DK063720-01 (2003): $1178346
Jeffrey A Bluestone, Professor And Director
University Of California San Francisco 3333 California St., Ste 315 San Francisco, Ca 941430962
Grant 1P30DK063720-010001 from National Institute Of Diabetes And Digestive And Kidney Diseases IRG: ZDK1
Abstract: The UCSF Diabetes Center Pilot and Feasibility (P&F) Program was established 18 months ago to encourage the pursuit of highly novel research for understanding and treating type 1 and type 2 diabetes. The program has been targeted towards all members of the UCSF community - the existing faculty of the Diabetes Center as well as other investigators from other departments and institutes at UCSF - in order to capitalize on the many and varied opportunities within the institution that have pointed relevance to diabetes. The P&F program implemented at UCSF provides investigators with two years of support at $25,000 per year to support conceptually innovative studies. As importantly, these awards are hoped to provide seed funding that will generate sufficient data to pursue additional research efforts through other funding mechanisms. The DRTC P&F Program will endeavor to 1. facilitate and allow new investigators and junior faculty to establish research programs in diabetes; 2. facilitate and support established faculty wishing to transfer skills and research developments to the arena of diabetes research; 3. foster highly innovative directions in basic and clinical diabetes research, including clinical studies in minority and other underserved patient groups; and 4. foster new and innovative collaborations amongst DRTC UCSF Faculty. The Pilot and Feasibility Program committee is will be chaired by Dr. Douglas Hanahan and the Committee includes Drs. Gerold Grodsky, Michael German and Ira Goldfine. As a group this committee represents all the aspects of the Center to assist the director in the management of the program.
Keywords: health science research support, insulin dependent diabetes mellitus, noninsulin dependent diabetes mellitus, T cell receptor, T lymphocyte, autoimmunity, bioenergetics, cell differentiation, diabetes mellitus genetics, diabetes mellitus therapy, disease /disorder model, gene expression, genetic model, hematopoietic stem cell, immunity, leukocyte activation /transformation, natural killer cell, neuron, obesity, pancreatic islet, pathologic process, technology /technique development, NOD mouse, imaging /visualization /scanning, laboratory mouse
Project start date: 2002-09-01
Project end date: 2007-08-31
Grants awarded to Jeffrey A Bluestone
SPONTANEOUS AUTOIMMUNE MODEL OF PERIPHERAL NEUROPATHY
Jeffrey A Bluestone, Professor
University Of California San Francisco, 3333 California St., Ste 315, San Francisco, Ca 94143-0962
Grant 5R01AI050834-08 from National Institute Of Allergy And Infectious Diseases
Abstract: The treatment and cure of autoimmunity remains of paramount importance. The challenges to developing successful therapies are broad, ranging from complex genetics, similarities and differences among target tissues, differential pathogenic mechanisms and an incomplete knowledge of the target antigens. We have shown that the Non-Obese Diabetes (NOD) mouse strain can be used as a mouse model of multiple autoimmune disorders (AID). These other autoimmune diseases were most apparent when regulatory T cells (Tregs) were eliminated and co-stimulatory pathways altered. For instance, NOD mice develop a spontaneous autoimmune disease of the peripheral nervous system, termed Spontaneous Autoimmune Peripheral Polyneuropathy (SAPP), in the absence of CD28 interaction with B7-2. In addition, we observed that in the complete absence of CD28 signals, NOD mice were deficient in Tregs and developed SAPP, sialadenitis, autoimmune thyroiditis and a newly appreciated autoimmune exocrine disease similar to that observed in "fulminant type 1 diabetes" described in Japanese and some Australian patients. Significantly, these various autoimmune diseases could use different pathogenic and co-stimulatory pathways and result from the recognition of distinct as well as potentially overlapping self-antigen specificities. These results have led to the conclusion that the NOD mouse represents a unique model for studying multi-organ autoimmunity. The combination of genetic propensity for autoimmunity and the tools that we have developed in this mouse strain will be exploited to address several key questions. Do unique and/or overlapping antigen specificities distinguish/link these diseases? Are the pathogenic pathways evident for one disease critical for the manifestation of others? Are there common co-stimulation pathways that control the susceptibility and progression of these distinct autoimmune diseases? The following aims are proposed to address these questions Specific Aim #1 To generate tissue antigen-specific effector and regulatory T cell TCR Tg mice based on candidate antigens. Specific Aim #2 To generate tissue antigen-specific effector and regulatory T cell TCR Tg mice using T cell hybridomas and mimotopes. Specific Aim #3 To determine the effector and regulatory pathways and the role of co-stimulation in the distinct autoimmune diseases in NOD mice. Specific Aim #4 To develop green fluorescence protein (GFP)-specific systems to study autoimmunity in NOD mice. Together, the results of these studies will test the hypothesis that the phenotypic manifestation of multi- organ autoimmune diseases is regulated by a coalescence of common and tissue-specific pathways. Moreover these common and distinct pathways are critical for understanding of the immunopathology of these different autoimmune diseases and development of novel therapies
Keywords: APC; ATGN; Address; Animals; Antigen Targeting; Antigen-Presenting Cells; Antigens; Autoantigens; Autoimmune; Autoimmune Diseases; Autoimmune Process; Autoimmune Status; Autoimmune thyroiditis; Autoimmunity; Autologous Antigens; Axon; B7-1; BB1; Body Tissues; CD28; CD28 gene; CD28LG; CD28LG1; CD80; CD80 gene; Cell Communication and Signaling; Cell Signaling; Cells; Characteristics; Chronic Lymphocytic Thyroiditis; Complex; Critical Paths; Critical Pathways; Development; Diabetes Mellitus; Diabetes Mellitus, Brittle; Diabetes Mellitus, Insulin-Dependent; Diabetes Mellitus, Juvenile-Onset; Diabetes Mellitus, Ketosis-Prone; Diabetes Mellitus, Sudden-Onset; Diabetes Mellitus, Type 1; Diabetes Mellitus, Type I; Disease; Disorder; Exocrine pancreas; Fluorescence; Genetic; Goals; Grant; Hybridomas; IDD; IDDM; ITX; Image; Immunologic Accessory Cells; Immunologically Directed Therapy; Immunotherapy; In Vitro; Individual; Inflammatory; Insulin-Dependent Diabetes Mellitus; Intracellular Communication and Signaling; Investigators; Japanese; Japanese Population; Knowledge; LAB7; Lead; Lesion; Link; Lytotoxicity; MHC Receptor; Major Histocompatibility Complex Receptor; Mammals, Mice; Mice; Modeling; Molecular; Monocytes / Macrophages / APC; Mouse Protein; Mouse Strains; Murine; Mus; Nervous; Neurilemma Cell; Neurilemmal Cell; Neuropathy; Non obese; Nonobese; Organ; PNS Diseases; Pathogenicity; Pathway interactions; Patients; Pb element; Peripheral; Peripheral Nerve Diseases; Peripheral Nervous System; Peripheral Nervous System Diseases; Peripheral Nervous System Disorders; Peripheral Neuropathy; Phenotype; Polyneuropathy; Predisposition; Production; Promoter; Promoters (Genetics); Promotor; Promotor (Genetics); Proteins; Reagent; Receptors, Antigen, T-Cell; Regulation; Regulatory Pathway; Reporter; Research Personnel; Researchers; Role; Schwann Cells; Self-Antigens; Sialadenitis; Sialoadenitis; Signal Transduction; Signal Transduction Systems; Signaling; Specificity; Study models; Susceptibility; System; System, LOINC Axis 4; T-Cell Receptor; T-Cells; T-Lymphocyte; T1 diabetes; T1D; T1DM; T44; Testing; Thymus-Dependent Lymphocytes; Thyroiditis; Thyroiditis, Lymphocytic; Thyroiditis, Lymphomatous; Tissues; Transgenic Mice; Transgenic Organisms; Type 1 diabetes; accessory cell; autoimmune disorder; autoreactive T cell; base; biological signal transduction; cytokine; cytotoxicity; diabetes; disease/disorder; gene product; heavy metal Pb; heavy metal lead; imaging; immune therapy; immunogen; immunopathology; in vivo; insulin dependent diabetes; islet; juvenile diabetes; juvenile diabetes mellitus; ketosis prone diabetes; mouse model; neural; neuropathic; novel; oncogene protein pp60(v-src) (137-157); pathway; peptide A; relating to nervous system; self recognition (immune); social role; thymus derived lymphocyte; tool; transgenic; type I diabetes
Project start date: 2001-12-15
Project end date: 2012-05-31
Budget start date: 1-JUN-2009
Budget end date: 31-MAY-2010
5R01AI050834-08 (2009): $341020
2R01AI050834-06A1 (2007): $346406
COSTIMULATION IN PERIPHERAL T CELL TOLERANCE AND ANERGY
Jeffrey A Bluestone, Professor And Director
Medicineuniversity Of California San Francisco
3333 California St., Ste 315
san Francisco, Ca 941430962
Grant 7P01AI035294-08 from National Institute Of Allergy And Infectious Diseases IRG: AITC
Abstract: The overall theme of this Program Project is to understand the mechanisms which regulate peripheral tolerance and to facilitate the implementation of scientific advances gained using in vitro model systems to the establishment of peripheral tolerance in vivo. Initiation and maintenance of peripheral T cell tolerance relies on a complex set of events dependent on the responding T cell subpopulation, the antigen and the antigen-presenting cells (APC), expression of co- stimulatory molecules and cytokines, and the genes expressed during the early signalling cascade following exposure to antigen. The overall goals are to dissect the mechanisms which lead to peripheral T cell tolerance by examining the molecular, biochemical, and immunobiological consequences of T cell interactions with antigen presenting cells. To achieve these general goals, three complementary projects will be undertaken 1. To understand the interactions between receptor-mediated signal transduction pathways that regulate the functions of different T lymphocyte subsets. The Pis will further characterize the signaling block in anergic T cells to produce IL-2. They also will characterize the CD28-mediated signaling events that promote augmented cytokine gene expression and that prevent energy induction, focussing on serine/threonine kinase intermediates that may lead to activation of Jnk-1/Jnk-2. They will identify additional costimulatory molecules that regulate the growth of Th2 cells. 2. To define the molecular basis of for the activity of co-stimulatory molecules. The PI will continue to define the molecular basis of the opposing effects of CTLA-4 and CD28 on the immune response in collaboration with the other Program Project investigators. In addition, Dr. Thompson will study the role CD30, 4-1-BB, and OX40 as co-stimulatory molecules, emphasizing the role of the TRAF family of molecules in mediating their effects. He also will continue his studies of the role of co-stimulation in preventing apoptosis through the bcl-xl and Fas pathways. 3. To determine the mechanisms of tolerance induction and the individual roles of CD28/CTLA-4 and B7- 1/B7-2. The PI will determine the individual roles of B7-1 and B7-2 on donor and host tissues during the initiation and progression of transplant rejection. He also will further characterize the biological effects of CTLA4 engagement and blockade on exposure to antigen in vitro and in vivo, determining the importance of the stage of the immune response, the tissue transplanted, and the APC targeted. These projects should provide information on the mechanism of peripheral T cell tolerance and help develop new therapeutic approaches for autoimmunity and graft rejection
Keywords: T lymphocyte, anergy, antigen presenting cell, immune tolerance /unresponsiveness, leukocyte activation /transformation
Project start date: 1993-09-01
Project end date: 2001-08-31
7P01AI035294-08 (2000): $749572
5P01AI035294-06 (1998): $707900
2P01AI035294-05 (1997): $681856
IMMUNOMODULATION OF TRANSPLANT REJECTION
Jeffrey A Bluestone, Professor And Director
Ben May Instituteuniversity Of Chicago
5801 S Ellis Ave
chicago, Il 60637
Grant 5P01AI029531-05 from National Institute Of Allergy And Infectious Diseases IRG: SRC
Abstract: The overall goals of this Transplantation Program Project are to dissect the mechanisms which lead to allograft rejection and to facilitate the implementation of scientific insights gained in the laboratory as treatment modalities in the clinic. To attain these general goals, four complementary projects have been proposed 1. A hamster mAb anti-murine CD3 mAb will be used to evaluate systematically the use of anti-CD3 to suppress transplantation responses in vivo in a well-defined, small animal model. The anti-CD3 and combined mAbs therapies will be exploited to more efficiently suppress transplantation responses in vivo. Other T cell subset-specific mAbs and antigens including staphylococcal toxins will be examined for their ability to selectively suppress T cell subsets which may play a predominant role in graft rejection. These studies will hopefully lead to the development of more sophisticated immunosuppressive regimens that selectively alters immunity towards the organ transplant. 2. The kinds of T cells involved in rejection of allografts which differ from the host in specific, defined regions of the MHC complex as well as the mechanisms by which these T cells cause graft rejection will be defined. Alloreactive T cells from unprimed spleen and lymph nodes, from the lymph nodes draining the site of a rejecting skin allograft, and from rejecting skin allografts will be characterized in terms of the array of lymphokines produced, the cell surface molecules expressed, and the specific epitopes with which these clones react. 3. The human/SCID mouse model will be developed to study the human cell interaction involved in graft rejection. This model will aid in the analyses of novel therapeutic approaches to overcome rejection of human allografts. 4. New modalities in man based on the basic research performed in the small animal and human/SCID mouse model systems will be directly analyzed at the humoral, cellular and molecular levels. Together, the four projects should provide information as to the mechanism of graft rejection and should help to develop new therapeutic modalities that will be directly implemented in the clinic
Keywords: T lymphocyte, cellular immunity, histocompatibility, homologous transplantation, immunosuppression, immunotherapy, monoclonal antibody, transplant rejection
Project start date: 1990-06-01
Project end date: 1995-09-29
5P01AI029531-05 (1994): $707142
5P01AI029531-04 (1993): $590411
5P01AI029531-03 (1992): $579734
Jeffrey A Bluestone, Professor And Director
Institution:
Grant 2P30CA014599-230004 from National Cancer Institute
Abstract: The overall goal of the UCCRC program in Immunology and Cancer is to dissect the multiple closely interrelated factors that determine immune responses to conventional and tumor antigens. This program is closely related to the Committee on Immunology which has been highly successful in developing interactive research and educational efforts to enhance the scientific endeavors of the UCCRC and the University. One example of the interactive nature of this program is the ongoing USPHS-NCI Program Project "Immunity and Cancer" H. Schreiber, Program Director. The overall goals of the UCCRC Program in Immunology and Cancer are (1) to enhance the scientific interactions among the various immunology researchers interested in cancer; (2) to bring together individuals interested in the most basic elements of the immune response to coordinate efforts towards a better understanding of tumor immunology; (3) to develop organized educational programs for predoctoral and postdoctoral students in order to foster increased interest in cancer research and (4) to manipulate the immune system in the treatment of cancer. Ultimately, these activities will lead to developing preclinical models to test novel immunotherapies for cancer and clinical studies through collaborative efforts with the Developmental Therapeutics Program of the UCCRC.
Keywords: neoplasm /cancer immunology, antigen presentation, cytokine, immunoglobulin, neoplasm /cancer education, protein transport, tissue /cell culture
Sponsored Links Excellgen http://Excellgen.com
Jeffrey A Bluestone, Professor And Director
University Of Chicago 5801 S Ellis Ave Chicago, Il 60637
Grant 5P01DK049799-059001 from National Institute Of Diabetes And Digestive And Kidney Diseases
Abstract: The immune system is a highly complex network of interacting cells, receptors and factor that combine to yield a potent immune response. Over the last decade, major advances in the study of the immune system has depended on our ability to examine individual cells of the immune network. Automated flow cytometry using fluorescence activated cell sorters (FACS) has enabled immunologists to analyze cell surface as well as intracellular proteins and DNA on as few as 1% of the cells. More recently, FACS has become an even more powerful tool capable of examining rapid intracellular events such as apoptosis, cell cycle events and cell signalling (Ca++flux) in individual cells of multicell conjugates. Equally important is the preparative sorting capability of these machines. These instruments an separate, with extremely high efficiency, individual subpopulations of cells and yield tens of millions of cells with a purity of greater than 99%. Access to these instruments is critical to the Program Project members. Every project depends on both analytical and preparative flow cytometry. Drs. Herold and Bluestone will use the facility to analyze the cell surface phenotype of APCs, activated T cells and islets from normal NOD and tolerant mice. They will prepare purified T cells from NOD and TCR transgenic mice for adoptive transfer studies. Dr Sant will use flow cytometry to analyze class II transfectants, newly-derived transgenic mice and natural APC populations. Dr. Miller will depend on flow cytometry to analyze TCR transgenic mice and select for new monoclonal antibodies specific for novel co-stimulatory molecules. Dr. Thompson has developed a highly sensitive method for examine apoptosis using flow cytometry to analyze individual cells. The cell sorter facility is, therefore, an essential resource for the research proposed in the program project.
Keywords: biomedical facility, flow cytometry, T lymphocyte, antigen presenting cell, pancreatic islet
Project start date: 1999-06-21
Project end date: 2000-05-31
Jeffrey A Bluestone, Professor And Director
University Of Chicago
5801 S Ellis Ave
chicago, Il 60637
Grant 5P01CA019266-169002 from National Cancer Institute
Abstract: The immune system is a high complex network of interacting cells, receptors and factors that combine to yield a potent immune response. aver the last decade, major advances in the study of the immune system had depended on our ability to examine individual cells of the immune network. Automated flow microfluorometry using fluorescence activated cell sorters has enabled immunologists to analyze cell surface as well as intracellular proteins and DNA. The instruments allows detailed analysis of subpopulations represented by as few as 1% of the cells. It allows highly efficient preparative separations of individual cell populations and can analyze intracellular events such as calcium fluxes. All of the members of the program project need and use the cell sorter facility. Each of the projects depends on the multifaceted capabilities of these instruments to assure completion of the proposed studies. The cell sorter facility is, therefore, an essential resource for the research proposed in the program project
Keywords: biomedical facility, fluorescence activated cell sorter T lymphocyte, calcium flux, cell population study, cellular immunity, intracellular transport, neoplasm /cancer immunology
Jeffrey A Bluestone, Professor And Director
University Of Chicago 5801 S Ellis Ave Chicago, Il 60637
Grant 5P30CA014599-209008 from National Cancer Institute
Abstract: The goals of the University of Chicago Cancer Research Center Animal Barrier Facility are multifold. The Facility provides to any investigator the means to perform animal studies using specific pathogen-free mice and rats. In addition, the Facility provides expertise in viral analyses of biological materials that are introduced into the animals a5P30CA014599-030028
Keywords: 1976
Sponsored Links Excellgen http://Excellgen.com
Jeffrey A Bluestone, Professor
University Of California San Francisco, 3333 California St., Ste 315, San Francisco, Ca 94143-0962
Abstract: The Flow Cytometry Core will meet the special needs of the Program Project members. The availability of quality sorting and analysis capabilities is essential for the success of the research efforts proposed in this Program Project Grant. We will be active users of both the MoFlo cell sorting and LSR II fluorescence analyzer in the studies dealing with sorting of small numbers of cells based on either cell surface or transduced gene markers; and multiparameter analyses to detect biochemical and biological changes in rare populations of lymphocytes as well as whole populations transduced with various genes. In many cases, the individual program directors will need access to machines that will require triple or even quadruple staining for the sophisticated experiments outlined in the proposal. The specific aims of this core are 1. To provide high speed cell sorting to isolate cells that can be used for functional and biochemical studies; 2. To provide analytical flow cytometry using a state-of-the-art LSR II to perform multiparameter analyses and 3. To provide innovative approaches to study immune tolerance using flow microscopy. The availability of the core ensures that the PPG investigators will have access to this essential facility on the UCSF campus
Keywords: Arts; Autoimmune Status; Autoimmunity; Biochemical; Biological; Cell Count; Cell Isolation; Cell Number; Cell Segregation; Cell Separation; Cell Separation Technology; Cell surface; Cells; Cytofluorometry, Flow; Ensure; Flow Cytofluorometries; Flow Cytometry; Flow Microfluorimetry; Fluorescence; Genes; Immune Tolerance; Immunologic Tolerance; Individual; Investigators; Lymphocyte; Lymphocytic; Microfluorometry, Flow; Microscopy; P01 Mechanism; P01 Program; Population; Program Project Grant; Program Research Project Grants; Programs (PT); Programs [Publication Type]; Research; Research Personnel; Research Program Projects; Researchers; Sorting - Cell Movement; Speed; Speed (motion); Staining method; Stainings; Stains; base; cell sorting; experiment; experimental research; experimental study; flow cytophotometry; immune system tolerance; immune unresponsiveness; immunological paralysis; innovate; innovation; innovative; lymph cell; meetings; member; programs; research study; self recognition (immune); sorting; success
Budget start date: 1-AUG-2009
Budget end date: 31-JUL-2010
5P01AI035297-18_9002 (2009): $227347
Jeffrey A Bluestone, Professor And Director
University Of California San Francisco 3333 California St., Ste 315 San Francisco, Ca 941430962
Grant 5P01AI035297-119002 from National Institute Of Allergy And Infectious Diseases IRG: ZAI1
Abstract: The Flow Cytometry Core will meet the special needs of the Program Project members. The availability of quality sorting and analysis capabilities is essential for the success of the research efforts proposed in this Program Project Grant. All the program members will be active users of both the MoFlo cell sorting and LSR fluorescence analyzer in the studies dealing from sorting of small numbers of cells based on either cell surface or transduced gene markers; and multiparameter analyses to detect biochemical and biological changes in rare populations of lymphocytes as well as whole populations transduced with various genes. In many cases, the studies proposed in the individual programs will require triple or even quadruple staining combined with state of the art cell sorting for cell surface and/or intracellular proteins. Thus, the FIRST SPECIFIC AIM of this Core is to provide access to a newly purchased high-speed cell sorter, MoFlo. The SECOND SPECIFIC AIM of this core is to provide access to the flow analyzers including the multiparameter LSR.
Keywords: biomedical equipment resource, biomedical facility, cell sorting, flow cytometry, immunofluorescence technique
IN VIVO MODELS OF PERIPHERAL TOLERANCE AND ANERGY
Jeffrey A Bluestone, Professor And Director
University Of Chicago 5801 S Ellis Ave Chicago, Il 60637
Grant 2P01AI035294-050004 from National Institute Of Allergy And Infectious Diseases
Abstract: The ability of T cells to distinguish self from non-self remains one of the central paradigms of the immune system. This specialized capability begins early in T cell development and under normal circumstances is maintained throughout the life of an individual. Self- tolerance is maintained by peripheral as well as central (thymic) mechanisms. The primary goal of this proposal is to study the role of the CD28/CTLA-4/B7 family of co-stimulatory molecules in the regulation of peripheral tolerance. Studies performed during the previous program project grant identified the B7-1 and B7-2 molecules as major co-stimulatory ligands; demonstrated that CD28 signaling regulates sustained T cell proliferative responses and Th1/Th2 differentiation in naive T cells. Of equal importance, we demonstrated that CTLA-4/B7 interactions profoundly down-regulate T cell responses. This application proposes to take advantage of in vitro and in vivo model systems to define the roles of B7-1 and B7-2 in the regulation of both CD28 and CTLA-4-mediated functions; to study the biochemical basis of CTLA-4-mediated down-regulation of immunity; and to develop novel regulatory drugs to manipulate the immunological activities of these receptors. In order to accomplish these goals, the following specific aims are proposed. 1. Determine the mechanism of tolerance induction and the individual roles of B7-1 and B7-2 on donor and host tissues during the initiation and progression of transplant rejection; 2. Determine the biological effects of CTLA-4 engagement and blockade on exposure to antigen in vitro and in vivo; and 3. Determine the molecular and biochemical basis of CTLA-4-mediated suppression of T cell function in vitro. The results of these studies will provide important insights into the mechanisms of CD28/CTLA-4 regulation of immune responses and may lead to novel immunotherapeutic approaches to the induction and maintenance of peripheral tolerance.
Keywords: CD28 molecule, T lymphocyte, anergy, immune tolerance /unresponsiveness, leukocyte activation /transformation, CD3 molecule, T cell receptor, antigen presentation, biological signal transduction, pancreas transplantation, transplant rejection, laboratory mouse, tissue /cell culture, transgenic animal
FASEB SUMMER CONFERENCE--LYMPHOCYTES AND ANTIBODIES
Jeffrey A Bluestone, Professor And Director
Federation Of Amer Soc For Exper Biology
9650 Rockville Pike
bethesda, Md 208143998
Grant 1R13AI047114-01 from National Institute Of Allergy And Infectious Diseases IRG: AITC
Abstract: The 2000 FASEB summer research conference on Lymphocytes and Antibodies, will be a small meeting, held from June 24 to June 29, 2000 at Saxtons River, Vermont. The planned subjects to be covered include areas in which major new advances are underway and have potential relevance to human health and disease. The meeting is structured to be highly interactive, promoting maximal scientific interchange and discussion among participants. There are no other competing conferences of similar format or size in the months before or after this conference
Keywords: antibody, lymphocyte, meeting /conference /symposium biological signal transduction, developmental immunology, immune tolerance /unresponsiveness, programmed cell death, receptor, transplantation immunology, virus travel
Project start date: 2000-06-01
Project end date: 2001-05-31
1R13AI047114-01 (2000): $8000
Sponsored Links Excellgen http://Excellgen.com
Jeffrey A Bluestone, Professor And Director
Diabetes Centeruniversity Of California San Francisco
3333 California St., Ste 315
san Francisco, Ca 941430962
Grant 5R01DK055351-04 from National Institute Of Diabetes And Digestive And Kidney Diseases IRG: ZDK1
Abstract: Recent technical advances have made pancreatic islet cell transplantation a feasible approach to the treatment of insulin- dependent diabetes mellitus (IDDM). However, the need for continuous, often debilitating, immunosuppressive drug therapy in transplant recipients makes the use of this therapy in diabetic patients most problematic. Thus, the development of long term donor-specific tolerance remains a primary goal of transplant biology. Activation of graft-specific pathogenic T-lymphocytes requires a first signal originating from the ligation of the TCR complex and an additional second co-stimulatory signal. However, novel immunotherapies successfully developed in small animal models to block costimulation often fail to induce the desired immunologic effects in large animals. It is critical to move these therapies forward towards the clinic using primate models that lack the confounding factors present in clinical transplantation. In order to accomplish this goal, the following specific aims are proposed. Specific Aim number 1. To determine the effect of blockade of the CD28/B7 co-stimulatory pathway on the survival of allogeneic islet cells transplanted into insulin- dependent Cynomolgus monkeys. In this study, we propose to administer CD28/B7 antagonists short-term in the absence of other immunosuppressive drugs, to prolong pancreatic islet allograft survival, prevent the development of donor specific antibodies and promote tolerance in a non-human primate model of islet transplantation; Specific Aim number 2. To integrate costimulation inhibition into novel drug combination treatment strategies in the non-human primate in order to establish consistent long term donor-specific tolerance. Antagonism of CD28 co-stimulation, even utilizing optimal reagents and dosing, may not completely inhibit T cell activation and differentiation. We hypothesize that additional treatments (anti-IL-2R, anti-ICAM- 1, 4-1BBFc) aimed at preventing the development of proinflammatory cell-mediated anti-donor responses will facilitate engraftment and induce a consistent donor specific tolerance in a non-human primate transplant model. Together the experiments proposed above will provide a detailed study of co- stimulation blockade in an islet transplant model in non-human primates. The immunologic, biologic and physiologic studies will afford us an opportunity to explore the basis of T cell activation and tolerance induction. The outcome of these studies will have important implications for islet cell transplantation for the treatment of IDDM
Keywords: diabetes mellitus therapy, immune tolerance /unresponsiveness, immunotherapy, inhibitor /antagonist, insulin dependent diabetes mellitus, pancreatic islet transplantation, technology /technique development CD28 molecule, T lymphocyte, antibody, antireceptor antibody, cell adhesion molecule, cytokine receptor, disease /disorder model, interleukin 2, leukocyte activation /transformation, model design /development, tumor necrosis factor alpha Macaca fascicularis, immunocytochemistry, in situ hybridization, laboratory mouse, tissue /cell culture
Project start date: 1999-06-15
Project end date: 2003-05-31
5R01DK055351-04 (2001): $325869
5R01DK055351-02 (2000): $455668
Role Of Notch 1 In Immune Tolerance
Jeffrey A Bluestone, Professor And Director
University Of California San Francisco 3333 California St., Ste 315 San Francisco, Ca 941430962
Grant 5R21AI059979-02 from National Institute Of Allergy And Infectious Diseases IRG: ZAI1
Abstract: Recent advances in the understanding of T-cell activation have led to new therapeutic approaches in the treatment of immunological disorders. One attractive target of intervention has been the blockade of T cell-mediated co-stimulatory pathways, which result in more selective effects on only those T-cells that have encountered specific antigen. In fact, in some instances, CD28/B7 co-stimulatory pathway antagonists can induce antigen-specific tolerance that prevents the progression of autoimmune diseases and organ graft rejection. However, many other cell surface molecules exist on naive T cells that may play an important role in initiating immunity from the quiescent state. Moreover, it is clear that quiescent T cells can also function as regulatory T cells to promote immune tolerance by an as yet unknown mechanism suggesting that some of the cell surface proteins may provide functional activities beyond just determining whether cells effectively go into cell cycle. Notch, Serrate, and Delta are type 1 integral membrane proteins involved in cell-cell signaling crucial to normal cellular differentiation and in organogenesis. In the immune system, Notch signaling is critical for normal T cell development. Recently, a role for Notch signaling in mature T cells has been proposed based on the observation that engagement of Notch by Serrate could induce a profound state of unresponsiveness in peripheral T cells in vivo. Most interesting was the observation that these "unresponsive" T cells possessed regulatory abilities that on several levels appear similar to the CD25+ regulatory cells that are able to regulate autoimmunity. Our preliminary studies suggest that Notch-1 is an important regulator of T cell signaling in resting T cells and may be involved in tolerance. We hypothesize that Notch-1 plays a critical role in the intrinsic regulation of T cell responses by influencing the primary signaling pathways critical for activating naive T cells. This results in T cell inactivation or altered differentiation. Moreover, we hypothesize that manipulation of immune responses through Notch-1 can be used to alter autoimmunity and transplantation tolerance. In this application we propose to explore the mechanisms of Notch signaling in T cells and its application as follows Specific Aim #1. To Identify the biochemical, cellular and genetic effects of Notch signaling in T cells; and Specific Aim #2. In vivo application of Notch signaling for the establishment of peripheral T cell tolerance.
Keywords: T cell receptor, biological signal transduction, immunologic receptor, leukocyte activation /transformation, CD28 molecule, cellular immunity, chimeric protein, dendritic cell, diabetes mellitus, genetic transcription, immune tolerance /unresponsiveness, pancreatic islet transplantation, NOD mouse, SCID mouse, genetically modified animal, laboratory mouse, transfection
Project start date: 2004-04-01
Project end date: 2007-03-31
5R21AI059979-02 (2005): $227250
CTLA-4-based Tolerance In Autoimmune Diabetes
Jeffrey A Bluestone, Professor And Director
University Of California San Francisco 3333 California St., Ste 315 San Francisco, Ca 941430962
Grant 5P01AI035297-140006 from National Institute Of Allergy And Infectious Diseases IRG: ZAI1
Abstract: Recent advances in the understanding of T-cell activation have led to new therapeutic approaches in the treatment of immunological disorders. One attractive target of intervention has been the blockade of T cell-mediated co-stimulatory pathways, which result in more selective effects on only those T-cells that have encountered specific antigen. In fact, in some instances, CD28/B7 co-stimulatory pathway antagonists can induce antigen-specific tolerance that prevents the progression of autoimmune diseases and organ graft rejection. However, the negative regulatory role of CTLA-4 in autoimmune diseases and graft rejection supported a dynamic but complex process of immune regulation prominent in the control of self-reactivity. As an example, CD28 and CTLA-4 can regulate pathogenic as well as suppressor cell development and function resulting, paradoxically, in opposite function consequences of receptor-ligand interactions. Finally, unlike CD28, the biochemical signals initiated through the CTLA-4 pathway are biochemically-linked to the T cell receptor signaling-complex. Thus, we hypothesize that CTLA-4 plays a critical role in intrinsic regulation of the ongoing autoimmune response by influencing the activation threshold of the T cells resulting in inactivation or altered differentiation. Moreover, manipulation of immune responses through CTLA-4 will depend on effective co-ligation of the TCR and CTLA-4. In order to test this hypothesis, the following specific aims are proposed. 1. TO DETERMINE THE MOLECULAR AND BIOCHEMICAL BASIS OF CTLA-4-MEDIATED T CELL FUNCTION IN IMMUNE TOLERANCE; 2. TO DETERMINE THE ROLE OF CTLA-4/B7 INTERACTIONS IN REGULATION OF PATHOGENESIS AND TOLERANCE INDUCTION AND MAINTENANCE IN NOD DIABETES; 3. TO DETERMINE THE ROLE OF CTLA-4 ON REGULATORY T CELLS THAT SUPPRESS NOD DIABETES; AND 4. TO USE A SINGLE CHAIN ANTI-CTLA-4 AND pepMHC-IG DIMER TO PRODUCE A NOVEL DIMERIC pepMHC-IG/ANTI-CTLA-4 HETEROCONJUGATE TO SPECIFICALLY TOLERIZE AUTOREACTIVE T CELLS. The studies proposed will be complimented by the other projects within this program project grant. We will work closely with Dr. Abbas to understand the role of CTLA-4 engagement at different stages of the immune response and solicit the collaborative efforts of Drs. Weiss and DeFranco to elucidate the detailed biochemical events that result from CTLA-4 engagement at the TCR complex. The results of these studies will provide important insights into the mechanism of CD28/CTLA-4 regulation of immune responses and may lead to novel immunotherapeutic approaches to the induction and maintenance of peripheral tolerance.
Keywords: CD28 molecule, autoimmune disorder, diabetes mellitus, immune tolerance /unresponsiveness, immunoregulation, leukocyte activation /transformation, MHC class I antigen, T cell receptor, antigen antibody reaction, autoantigen, biological signal transduction, cytogenetics, immunogenetics, pathologic process, receptor binding, NOD mouse, immunoprecipitation, microarray technology, polymerase chain reaction, transfection, western blotting
Role Of Notch 1 In Immune Tolerance
Jeffrey A Bluestone, Professor And Director
University Of California San Francisco 3333 California St., Ste 315 San Francisco, Ca 941430962
Grant 1R21AI059979-01 from National Institute Of Allergy And Infectious Diseases IRG: ZAI1
Abstract: Recent advances in the understanding of T-cell activation have led to new therapeutic approaches in the treatment of immunological disorders. One attractive target of intervention has been the blockade of T cell-mediated co-stimulatory pathways, which result in more selective effects on only those T-cells that have encountered specific antigen. In fact, in some instances, CD28/B7 co-stimulatory pathway antagonists can induce antigen-specific tolerance that prevents the progression of autoimmune diseases and organ graft rejection. However, many other cell surface molecules exist on naive T cells that may play an important role in initiating immunity from the quiescent state. Moreover, it is clear that quiescent T cells can also function as regulatory T cells to promote immune tolerance by an as yet unknown mechanism suggesting that some of the cell surface proteins may provide functional activities beyond just determining whether cells effectively go into cell cycle. Notch, Serrate, and Delta are type 1 integral membrane proteins involved in cell-cell signaling crucial to normal cellular differentiation and in organogenesis. In the immune system, Notch signaling is critical for normal T cell development. Recently, a role for Notch signaling in mature T cells has been proposed based on the observation that engagement of Notch by Serrate could induce a profound state of unresponsiveness in peripheral T cells in vivo. Most interesting was the observation that these "unresponsive" T cells possessed regulatory abilities that on several levels appear similar to the CD25+ regulatory cells that are able to regulate autoimmunity. Our preliminary studies suggest that Notch-1 is an important regulator of T cell signaling in resting T cells and may be involved in tolerance. We hypothesize that Notch-1 plays a critical role in the intrinsic regulation of T cell responses by influencing the primary signaling pathways critical for activating naive T cells. This results in T cell inactivation or altered differentiation. Moreover, we hypothesize that manipulation of immune responses through Notch-1 can be used to alter autoimmunity and transplantation tolerance. In this application we propose to explore the mechanisms of Notch signaling in T cells and its application as follows Specific Aim #1. To Identify the biochemical, cellular and genetic effects of Notch signaling in T cells; and Specific Aim #2. In vivo application of Notch signaling for the establishment of peripheral T cell tolerance.
Keywords: T cell receptor, biological signal transduction, immunologic receptor, leukocyte activation /transformation, CD28 molecule, cellular immunity, chimeric protein, dendritic cell, diabetes mellitus, genetic transcription, immune tolerance /unresponsiveness, pancreatic islet transplantation, NOD mouse, SCID mouse, genetically modified animal, laboratory mouse, transfection
Project start date: 2004-04-01
Project end date: 2006-03-31
1R21AI059979-01 (2004): $227250
CTLA-4-based Tolerance In Autoimmune Diabetes
Jeffrey A Bluestone, Professor And Director
University Of California San Francisco 3333 California St., Ste 315 San Francisco, Ca 941430962
Grant 5P01AI035297-130006 from National Institute Of Allergy And Infectious Diseases IRG: ZAI1
Abstract: Recent advances in the understanding of T-cell activation have led to new therapeutic approaches in the treatment of immunological disorders. One attractive target of intervention has been the blockade of T cell-mediated co-stimulatory pathways, which result in more selective effects on only those T-cells that have encountered specific antigen. In fact, in some instances, CD28/B7 co-stimulatory pathway antagonists can induce antigen-specific tolerance that prevents the progression of autoimmune diseases and organ graft rejection. However, the negative regulatory role of CTLA-4 in autoimmune diseases and graft rejection supported a dynamic but complex process of immune regulation prominent in the control of self-reactivity. As an example, CD28 and CTLA-4 can regulate pathogenic as well as suppressor cell development and function resulting, paradoxically, in opposite function consequences of receptor-ligand interactions. Finally, unlike CD28, the biochemical signals initiated through the CTLA-4 pathway are biochemically-linked to the T cell receptor signaling-complex. Thus, we hypothesize that CTLA-4 plays a critical role in intrinsic regulation of the ongoing autoimmune response by influencing the activation threshold of the T cells resulting in inactivation or altered differentiation. Moreover, manipulation of immune responses through CTLA-4 will depend on effective co-ligation of the TCR and CTLA-4. In order to test this hypothesis, the following specific aims are proposed. 1. TO DETERMINE THE MOLECULAR AND BIOCHEMICAL BASIS OF CTLA-4-MEDIATED T CELL FUNCTION IN IMMUNE TOLERANCE; 2. TO DETERMINE THE ROLE OF CTLA-4/B7 INTERACTIONS IN REGULATION OF PATHOGENESIS AND TOLERANCE INDUCTION AND MAINTENANCE IN NOD DIABETES; 3. TO DETERMINE THE ROLE OF CTLA-4 ON REGULATORY T CELLS THAT SUPPRESS NOD DIABETES; AND 4. TO USE A SINGLE CHAIN ANTI-CTLA-4 AND pepMHC-IG DIMER TO PRODUCE A NOVEL DIMERIC pepMHC-IG/ANTI-CTLA-4 HETEROCONJUGATE TO SPECIFICALLY TOLERIZE AUTOREACTIVE T CELLS. The studies proposed will be complimented by the other projects within this program project grant. We will work closely with Dr. Abbas to understand the role of CTLA-4 engagement at different stages of the immune response and solicit the collaborative efforts of Drs. Weiss and DeFranco to elucidate the detailed biochemical events that result from CTLA-4 engagement at the TCR complex. The results of these studies will provide important insights into the mechanism of CD28/CTLA-4 regulation of immune responses and may lead to novel immunotherapeutic approaches to the induction and maintenance of peripheral tolerance.
Keywords: CD28 molecule, autoimmune disorder, diabetes mellitus, immune tolerance /unresponsiveness, immunoregulation, leukocyte activation /transformation, MHC class I antigen, T cell receptor, antigen antibody reaction, autoantigen, biological signal transduction, cytogenetics, immunogenetics, pathologic process, receptor binding, NOD mouse, immunoprecipitation, microarray technology, polymerase chain reaction, transfection, western blotting
Spontaneous Autoimmune Model Of Peripheral Neuropathy
Jeffrey A Bluestone, Professor And Director
University Of California San Francisco 3333 California St., Ste 315 San Francisco, Ca 941430962
Grant 5R01AI050834-05 from National Institute Of Allergy And Infectious Diseases IRG: IMS
Abstract: provided by investigator) Guillain-Barre Syndrome (GBS) and Chronic Inflammatory Demyelinating Polyradiculoneuropathy (CIDP) are autoimmune diseases characterized by inflammatory demyelinating lesions of the peripheral nervous system that cause devastating neurological deficits and paralysis. We have developed the first model of spontaneous autoimmune disease of the peripheral nervous system, called Spontaneous Autoimmune Peripheral Polyneuropathy (SAPP). NOD mice deficient in the co-stimulatory molecule B7-2 exhibit a progressive and generalized limb paralysis associated with severe demyelination and axonal damage due to massive inflammation of the peripheral nervous system. Adoptive transfer experiments showed that the disease is mediated, at least in part, by CD4+ T cells. The course of the disease and its pathophysiological features are strikingly similar to human GBS and CIDP. SAPP is observed in mice of the autoimmune-prone NOD background, which has been widely recognized as a valuable animal model of autoimmunity in humans. Furthermore, B7-2-deficient NOD mice do not develop diabetes. Thus, this animal model will provide insight into the influence of the co-stimulatory milieu on the polarization of autoimmunity towards different tissues and the development of distinct diseases in individuals presenting a general susceptibility to autoimmunity. The following specific aims are proposed to study this disease Specific aim #1 To determine the antigen(s) targeted by autoreactive T cells in the peripheral nerves. Specific aim #2 To understand the influence of B7 costimulation on the polarization of autoimmunity in nod mice. Specific aim #3 To determine the immunopathology of SAPP, particularly the cell subsets and the effector mechanisms involved in the disease. Specific aim #4 To analyze the genetic control of SAPP. We propose to study the hypothesis that SAPP shares a subset of immunological and genetic attributes similar to those associated with the other autoimmune syndromes in the NOD strain. Our results will allow us to define genetic and immunological characteristics that determine the general susceptibility of individuals to autoimmunity and identify distinct mechanisms that lead to spontaneous destructive immune response in the nervous system versus the pancreas.
Keywords: autoimmune disorder, pathologic process, polyneuritis, T lymphocyte, autoantigen, biological signal transduction, cytokine, disease /disorder model, leukocyte activation /transformation, NOD mouse, genetically modified animal
Project start date: 2001-12-15
Project end date: 2007-05-31
5R01AI050834-05 (2006): $258895
Sponsored Links Excellgen http://Excellgen.com
5R01AI050834-04 (2005): $265125
5R01AI050834-03 (2004): $265125
5R01AI050834-02 (2003): $263446
1R01AI050834-01 (2002): $289325
IMMUNOSUPPRESSIVE EFFECTS OF ANTI-T CELL MABS
Jeffrey A Bluestone, Professor And Director
Institution:
Grant 5P01AI029531-070001 from National Institute Of Allergy And Infectious Diseases
Abstract: OKT3 is a murine monoclonal antibody directed against the CD3epsilon chain of the T cell receptor complex on human T-cells. OKT3 has been widely used over the past decade in transplantation to treat corticosteroid- resistant organ graft rejections, and more recently, as prophylaxis of rejection. This mAb induces an efficient and rapid immunosuppression, at least in part due to, the prevention of allorecognition as a result of modulation of the T cell receptor (TCR) from the surface of T lymphocytes and clearance of T cells from the circulation. However, there are several clinical and scientific issues that remain unresolved regarding anti-CD3 therapy. First, a variety of side effects are associated with OKT3 that limits its usefulness in other diseases such as autoimmunity and bone marrow transplantation. These limitations are, in large part, a result of the first dose toxicity as a consequence of T cell activation and the subsequent massive cytokine release. In addition, the therapy has been limited by the generation of a potent humoral antibody response against the murine mAb limiting the potential for retreatment. During the past 4 years, Dr. Bluestone s laboratory has systematically evaluated the use of anti-CD3 to suppress transplantation responses in vivo first in a well- defined, small animal model and then in the hu-SPL-SCID model in collaboration with Dr. Thistlethwaite. The toxic effects of the mAb were defined and largely eliminated, immunosuppressive agents such as deoxyspergualin and CTLA4Ig were used in conjunction with anti-CD3 to potentiate immunosuppression and block the humoral response, and genetically-engineered monoclonal anti-CD3 antibodies were developed to examine the role of T cell activation and Co-stimulation in immune suppression. However, many questions remain including the relative efficacy of "activating" and "non-activating" anti-CD3 mAbs; the signal transducing ability of "non-activating" anti-CD3 mAbs and how it regulates T cell activation; the role of co-stimulatory in the activating and toleragenic effects of anti-CD3; and the role of other inhibitors of T cell signalling, such as cyclosporin A in the efficacy of anti-CD3 therapy. In order to answer these questions the following specific aims are proposed 1. To study the in vivo effects of non-mitogenic anti-CD3 mAbs alone and in conjunction with donor antigen; 2. To determine the role of CD28/B7 interactions in anti-CD3-mediated immunosuppression; 3. To determine the molecular mechanism(s) controlling apoptosis and anergy in mice treated with mitogenic anti-CD3. The results of these studies will provide important insights into the effects of anti-CD3 and the CD28/B7 family of cell surface molecules in the regulation of transplant rejection that may translate into clinical application.
Keywords: CD3 molecule, antireceptor antibody, artificial immunosuppression, monoclonal antibody, transplant rejection, CD antigen, T lymphocyte, anergy, biological signal transduction, cell cycle protein, cell migration, cyclosporine, etoposide, interleukin 2, isoantigen, pancreatic islet transplantation, programmed cell death, sirolimus, skin transplantation, surface antigen, laboratory mouse, tissue /cell culture
IMMUNOBIOLOGY OF TCR GAMMA/DELTA T-CELLS
Jeffrey A Bluestone, Professor And Director
University Of Chicago 5801 S Ellis Ave Chicago, Il 60637
Grant 5R01AI026847-08 from National Institute Of Allergy And Infectious Diseases IRG: EI
Abstract: More than half a decade after their identification, the biological role of TCRgammadelta cells remains uncertain. While TCRgammadelta cells are clearly dominant in certain pathologic conditions, their lack of significant involvement in most classical T cell responses suggests that they perform a unique and special role in host immune defense. In fact, TCRgammadelta cells have several unique features that distinguish them from TCRalphabeta cells including (a) preferential expression in epithelial tissue; (b) a broad repertoire skewed towards non-classical major histocompatibility complex (MHC) class I antigens, heat shock proteins, and non-processes antigens; and (c) substantial expansion during a variety of inflammatory diseases thus constituting a major arm of the immune system. The major objective of the previous grant was to dissect and functionally characterize this unique T cell subset. During this time, we identified TCRgammadelta cells showing that TCRgammadelta cells can be MHC-specific (classical class I-, class II-, and TL-specific) and nominal antigen- specific. The antigen-specific TCRgammadelta cells were used 1) To demonstrate that the TCRgammadelta repertoire is diverse; 2) To create TCRgammadelta transgenic mice that establish positive and negative selection in TCRgammadelta development; 3) To produce multiple antisera and mABs that have been widely used to characterize TCRgammadelta subsets and examine the functional potential (i.e. lymphokine production and cytolytic activity) of this novel T cell population in normal and transgenic mice. During the next grant period, we will use the mABs, T cell clones and transgenic mice to define TCRgammadelta cell activation and intracellular signal transduction pathways, the physiologic processes that affect positive and negative selection, and the genetic elements that control antigen recognition by this eclectic population. Specifically, we will 1) Study the precise antigen specificity and physiologic role of herpesvirus-specific and TL-specific TCRgammadelta cells. We will examine the role of antigen processing and the requirement for peptide- presentation in the specificity of these T cells; 2) Examine the biochemical and molecular events associated with TCRgammadelta maturation and selection. We will examine directly the TCR complex and early signal transduction pathways in developing TCRgammadelta cells and during antigen-driven activation; and 3) Determine the genetic elements that control TCRgammadelta repertoire development. We will identify the genes and antigens that control strain-specific repertoire differences; These studies will help to define the role of TCRgammadelta cells in immune responses.
Keywords: T lymphocyte, biological signal transduction, cell differentiation, genetic regulation, leukocyte activation /transformation, MHC class I antigen, T cell receptor, antigen presentation, gene mutation, genetic transcription, herpes simplex virus 1, interleukin 2, receptor binding, hamster, immunoprecipitation, laboratory mouse, monoclonal antibody, tissue /cell culture, transfection, transgenic animal
Project start date: 1988-09-01
Project end date: 1998-08-31
5R01AI026847-08 (1995): $182105
5R01AI026847-07 (1994): $181719
5R01AI026847-10 (1997): $196597
IMMUNOSUPPRESSIVE EFFECTS OF ANTI-T CELL MABS
Jeffrey A Bluestone, Professor And Director
University Of Chicago 5801 S Ellis Ave Chicago, Il 60637
Grant 5P01AI029531-050001 from National Institute Of Allergy And Infectious Diseases
Abstract: Monoclonal antibody (mAb) treatment of organ transplant recipients has been a major development in the treatment of allograft rejection. The advantages of mAbs include their known specificity, consistent biological activity, and ease of administration. Of all mAbs clinically and experimentally tested in man, the most efficacious is the OKT3 mAb which recognizes the T cells receptor (TCR) complex on the surface of alloreactive cells. However, several limitations to the use of the OKT3 reagent persists. These include the significant adverse reactions following the initial dose, production by the recipient of antibodies to the OKT3 antibody, and the broad but transient nature of the induced immunosuppression. Our laboratory has recently developed an anti-murine CD3 mAb that reacts with all mouse T cells expressing a TCR complex. Initial studies in the laboratory suggest that treatment with the mAb in vivo has profound long term immunosuppressive effects including prolonged skin graft survival and depressed cellular immunity. Although the initial immunosuppression is a consequence of T cell depletion and receptor blockade, T cells that survive treatment continue to be hyporesponsive. Thus, this hamster mAb provides the first anti-CD3 that can be used in a well-defined small animal model to provide a systematic evaluation of the use of anti-CD3 in vivo to suppress transplantation responses. Since anti-CD3 suppresses the immune response by affecting all T cells, the potential for generalized immunosuppression and susceptibility to opportunistic infections remains a great risk. Therefore, efforts will be designed to focus immunosuppression specifically on T cell subsets critical in allorecognition. Thus, subset-specific, mAbs and other T cell reagents will be used to suppress individual V(beta) expressing T cells which predominate in allorecognition. In addition, anti-CD3 will be coupled to anti-CD4 and anti-CD8 to suppress these subsets of T cells. Thus, the more specific mAb treatment should lead to a more sophisticated immunosuppressive regimen that selectively alters immunity towards the organ transplant.
Keywords: CD3 molecule, cellular immunity, histocompatibility, homologous transplantation, immunosuppressive antileukocyte serum, monoclonal antibody, CD4 molecule, CD8 molecule, T cell receptor, T lymphocyte, cell type, diaziquone, disease model, histocompatibility antigen, immune tolerance /unresponsiveness, immunoglobulin, immunoglobulin gene, immunotherapy, leukocyte activation disorder, major histocompatibility complex, staphylococcal enterotoxin, athymic mouse, bone marrow transplantation, drug administration route, flow cytometry, laboratory mouse, pancreas transplantation, skin transplantation, thymectomy, tissue /cell culture
Sponsored Links Excellgen http://Excellgen.com