1985 — 1988 |
Rodriguez, Moses |
K07Activity Code Description: To create and encourage a stimulating approach to disease curricula that will attract high quality students, foster academic career development of promising young teacher-investigators, develop and implement excellent multidisciplinary curricula through interchange of ideas and enable the grantee institution to strengthen its existing teaching program. |
Pathogenesis of Viral Induced Demyelination @ Mayo Clinic Coll of Medicine, Rochester
Mechanisms of demyelination are studied in two virus induced demyelinating diseases in mice by means of electron microscopy, ultrastructural immunolabeling techniques, and assays of cellular and humoral immunity which use infected glial cells in tissue culture as targets. The experimental models are induced by Theiler's murine encephalomyelitis virus (TMEV) or the temperature sensitive mutant 472 of the Chandipura virus (CV). The objectives in studying TMEV are 1) to determine definitively wheter TMEV demyelination is immune mediated by transferring spleen cells from mice during the development of the white matter lesion and searching for histologic lesions in recipient mice, using as positive controls the transfer of lymphoid cells from mice afflicted with experimental autoimmune encephalomyelitis, 2) to determine if demyelination is the result of an autoimmune process directed against myelin by desensitizing mice with myelin components prior to infection with TMEV, 3) to determine if demyelination is caused by neutral proteases released by macrophages by attempting to prevent demyelination with neutral protease inhibitors and macrophage depletion, 4) to determine if demyelination is caused by an immune response directed against persistently infected oligodendroglial cells by using persistently infected primary glial cultures as targets to study lymphocytotoxicity. The objectives in studying the demyelination associated with infective Chandipura virus are to determine 1) the ultrastructural morphologic features of demyelination, 2) the ultrastructural characteristics of the cells infected during various stages of infection, 3) ultrastructural evidence of viral antigens expressed on the surface of infected cells by using monospecific antibodies to the structural proteins of TS472CV as probes to label antigens. The findings are expected to contribute to the understanding of human demyelinating diseases, such as multiple sclerosis.
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1987 — 2008 |
Rodriguez, Moses |
R01Activity Code Description: To support a discrete, specified, circumscribed project to be performed by the named investigator(s) in an area representing his or her specific interest and competencies. |
Immune Promotion of Remyelination @ Mayo Clinic Coll of Medicine, Rochester
myelination; psychoneuroimmunology;
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1994 — 1998 |
Rodriguez, Moses |
R01Activity Code Description: To support a discrete, specified, circumscribed project to be performed by the named investigator(s) in an area representing his or her specific interest and competencies. |
T-Cell Function in a Murine Model of Multiple Sclerosis @ Mayo Clinic Coll of Medicine, Rochester
The etiologies of primary demyelinating diseases such as multiple sclerosis (MS) are unknown. One testable hypothesis is that destruction of myelin and oligodendrocyte results from an immune attack directed against antigen triggered by virus infection. We have used infection to mice with Theiler's murine encephalomyelitis virus (TMEV), a picornavirus, to study the role of the immune response in demyelination and in neurologic disease. The long term goal of these experiments is to reveal the immune mechanisms of demyelination and neurologic disease with the hope that this will provide new insights into the treatment of MS. There are two major specific aims in the proposed experiments. First, in order to determine which viral genes are involved in inducing a protective immune response and which genes are targeted in the pathogenic phase of demyelinating disease, we have divided the TMEV genome into three regions so that they can be expressed individually in target cells in vitro and as transgenes in vivo. Fibroblasts transfected with the TMEV coding blocks will be used to determine the location of the genes recognized by T cells appearing early during the protective host response to viral infection and late during the pathogenic phase of disease. The same coding blocks expressed as transgenes in resistant and susceptible strains will presumably induce tolerance to sets of viral antigens, permitting to assess the significance of immune recognition of these antigens in resistance to viral infection and in the pathogenesis of demyelination. In the second specific aim we will utilize beta2- macroglobulin deficient (-/-) mice which when infected with TMEV develop prominent demyelination but no neurologic deficits. These experiments have direct relevance to MS, in which frequently there is a discrepancy between demyelination provides a unique opportunity to dissect those components of the immune response important in demyelination versus those important in neurologic deficits. We will first determine the nature of the immune response (CTL, Th1, and Th2) in CNS of infected Beta-2m (+/+) and Beta-2m (-/-) of susceptible and resistant haplotypes. We will then take advantage of the Beta2-m (-/-) model to establish the immunologic basis of neuronal injury in the demyelinated host. Using a new technique in the mouse to measure motor and sensory spinal cord conduction in vivo, we will determine those components of the immune response responsible for neurophysiologic abnormalities. These experiments have the potential to elucidate new strategies for the treatment of human CNS demyelinating disorders.
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1999 — 2003 |
Rodriguez, Moses |
R01Activity Code Description: To support a discrete, specified, circumscribed project to be performed by the named investigator(s) in an area representing his or her specific interest and competencies. |
T Cell Function in a Murine Model of Multiple Sclerosis @ Mayo Clinic Coll of Medicine, Rochester
A major question in multiple sclerosis research is the mechanism responsible for the development of neurologic deficits following demyelination. The goal of this proposal is to test the hypothesis that effector molecules released by CD8+ T cells cause neurologic deficits following demyelination by destroying or disrupting the function of axons and neurons. We have used the Theiler's virus model of demyelination to dissect the components of the MHC-restricted class I (CD8 T cells) versus MHC-restricted class II (CD4 T cells) responses to neurologic dysfunction following demyelination. Infection of class I- deficient mice results in prominent demyelination in the absence of neurologic deficits, whereas infection of class 11-deficient mice of identical genotype results in demyelination with severe neurologic dysfunction. Our first specific aim is to determine the effector molecules produced by cytotoxic T cells (IFN-gamma, Fas, TNF, and perforin) critical for induction of neurologic deficits by examining neurologic function and electrophysiology in immune knockout mice (IFN- gamma-/-, IFN-gammaR -/-, TNF -/-, TNFR -/-, PFP -/-, Fas -/-, FasL -/- )in mice of genotype resistant or susceptible to demyelination. We will also do adoptive transfer experiments into immunodeficient RAG -/- mice to determine the contribution of effector molecules with CD8 and CD4 cells to demyelination and neurologic deficit. Our specific second aim is to define the morphologic basis for functional deficits by determining whether axons are destroyed or injured in mice with immune effector deficiency following virus infection. We will assess degree of axonal dropout in normal and demyelinated white matter, perform neurofilament staining by immunofluorescence, count neurons in nuclei by retrograde labeling, and assess sodium channel distribution by [H3] saxitoxin autoradiography and fluorescence immunocytochemistry. These experiments are expected to provide new insights into the effector molecules released by the inflammatory response responsible for neurologic deficits in demyelinating disease.
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2000 — 2002 |
Rodriguez, Moses |
P01Activity Code Description: For the support of a broadly based, multidisciplinary, often long-term research program which has a specific major objective or a basic theme. A program project generally involves the organized efforts of relatively large groups, members of which are conducting research projects designed to elucidate the various aspects or components of this objective. Each research project is usually under the leadership of an established investigator. The grant can provide support for certain basic resources used by these groups in the program, including clinical components, the sharing of which facilitates the total research effort. A program project is directed toward a range of problems having a central research focus, in contrast to the usually narrower thrust of the traditional research project. Each project supported through this mechanism should contribute or be directly related to the common theme of the total research effort. These scientifically meritorious projects should demonstrate an essential element of unity and interdependence, i.e., a system of research activities and projects directed toward a well-defined research program goal. |
Core--Pathology @ Mayo Clinic Coll of Medicine, Rochester
The Pathology Core is responsible for all the morphological assessments of various strains of mice injected with Theiler's virus, or immunized with myelin antigens for development of experimental autoimmune encephalomyelitis. The Pathology Core is housed in Dr. Rodriguez's research laboratory. The major focus of the Pathology Core is primarily on plastic-embedded sections embedded in either glycol methacrylate or Araldite. These are the standard assays for assessment of demyelination, remyelination, spinal cord atrophy, and axonal loss. In addition, brain and spinal cord sections are embedded in paraffin for immunocytochemistry and in situ hybridization to detect virus replication. Frozen sections are used for immunofluorescence staining for neurofilament proteins. T cell and cytokine expression and [3H] saxitoxin-labeling to assess axonal integrity.
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2000 — 2004 |
Rodriguez, Moses |
P01Activity Code Description: For the support of a broadly based, multidisciplinary, often long-term research program which has a specific major objective or a basic theme. A program project generally involves the organized efforts of relatively large groups, members of which are conducting research projects designed to elucidate the various aspects or components of this objective. Each research project is usually under the leadership of an established investigator. The grant can provide support for certain basic resources used by these groups in the program, including clinical components, the sharing of which facilitates the total research effort. A program project is directed toward a range of problems having a central research focus, in contrast to the usually narrower thrust of the traditional research project. Each project supported through this mechanism should contribute or be directly related to the common theme of the total research effort. These scientifically meritorious projects should demonstrate an essential element of unity and interdependence, i.e., a system of research activities and projects directed toward a well-defined research program goal. |
Immunogenetics of Demyelination @ Mayo Clinic Coll of Medicine, Rochester
The major goal of this program project is to investigate the immunogenetic basis of demyelination and neurological deficits. The experiments will utilize two excellent models of human multiple sclerosis, i.e. Theiler's virus infection and experimental autoimmune encephalomyelitis. The program project is under the direction of Dr. Moses Rodriguez Professor of Neurology and Immunology of the Mayo Medical School who has ha a long interest in the pathogenesis of demyelination and remyelination in human multiple sclerosis. Members of the Program Project (Drs. Chella S. David, Dr. Larry R. Pease, and Dr. Moses Rodriguez) are well established investigators and Professors of Immunology in the Mayo Medical School. This group has an impressive record of investigations on the immunogenetics in murine models of human disease. Project 1: (Dr. Rodriguez, Principal Investigator) will investigate transgenic expression of Theiler's virus genomes in mice. The experiments will also investigate the mechanisms of decreased demyelinating disease observed in human HLA transgenic mice and address the specificity of the pathogenic immune response important in development of functional neurological deficits following demyelination. Project 2: (Dr. Pease, Principal Investigator) will investigate the basis for H-2K vs. H-2D polarity of anti-TMEV lymphocyte responses in the central nervous system. Experiments will identify the importance of virus-specific CD8+ T cells in the resistance to persistence virus infection and in neuropathology. Project 3: (Dr. David, Principal Investigator) will investigate experimental autoimmune encephalomyelitis in HLA expressing HLA-DR or DQ haplotypes. Experiments will examine the presentation of putative autoantigens linked to MS by HLA-DR and DQ molecules. This Program Project focusing on the immunogenetic basis of demyelination by a group of highly interactive investigators is expected to provide new insights into the pathogenesis of myelin and neural injury in multiple sclerosis.
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2000 — 2002 |
Rodriguez, Moses |
P01Activity Code Description: For the support of a broadly based, multidisciplinary, often long-term research program which has a specific major objective or a basic theme. A program project generally involves the organized efforts of relatively large groups, members of which are conducting research projects designed to elucidate the various aspects or components of this objective. Each research project is usually under the leadership of an established investigator. The grant can provide support for certain basic resources used by these groups in the program, including clinical components, the sharing of which facilitates the total research effort. A program project is directed toward a range of problems having a central research focus, in contrast to the usually narrower thrust of the traditional research project. Each project supported through this mechanism should contribute or be directly related to the common theme of the total research effort. These scientifically meritorious projects should demonstrate an essential element of unity and interdependence, i.e., a system of research activities and projects directed toward a well-defined research program goal. |
Transgenic Expression of Theiler's Virus Encoded Regions @ Mayo Clinic Coll of Medicine, Rochester
The goal of this project is to investigate the nature and specificity of the immune response that protects resistance mice from Theiler's murine encephalomyelitis virus (TMEV) persistent infection but which may also contribute in susceptible mice to demyelination and neurologic deficits. In this murine model of multiple sclerosis the immune system functions both to clear virus infection but also to exacerbate the pathogenic response which mediates myelin and axonal injury. The hypothesis to be tested is that antigens encoded by the TMEV genome are critical for protective immunity (resistance) but possibility may also contribute to immunopathology (susceptibility). The experiments will utilize a series of transgenic mice expressing independent three continuous regions of the TMEV genome. Transgenic mice have been created under control of a class I continuous regions of the TMEV genome. Transgenic mice have been created under control of a class I promoter expressing region I coding sequence 5' of VP1 (L, VP4, VP2, and VP3), region II (VP1 coding block), and region III coding sequence 3' of VP1 92A, 2B, 2C, 3A, 3B, 3C, and 3D). By challenging mice expressing TMEV transgenes with infectious virus, we will be able to address the role of immune response to TMEV coding regions in vivo. These experiments will also evaluate demyelination and neurologic deficits in transgenic mice expressing human class II MHC genes infected with TMEV. Finally we will study the phenotype and specificity of the immune response contributing to neurologic deficits utilizing adoptive transfer experiments with perforin deficient mice when injected with TMEV show demyelination but fail to show neurologic deficits. The experiments are expected to provide unique insights into the mechanisms of myelin injury and neurologic deficits with relevance to human multiple sclerosis.
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2003 — 2006 |
Rodriguez, Moses |
T32Activity Code Description: To enable institutions to make National Research Service Awards to individuals selected by them for predoctoral and postdoctoral research training in specified shortage areas. |
Medical Scientist Training Program At Mayo Clinic
DSCRIPTION: (provided by applicant): A critical requirement for the future of outstanding health care in the United States is the training of physician-scientists who will investigate fundamental basic science questions which ultimately relate to clinical problems. The mission of the M.D./Ph.D. program of Mayo Medical and Graduate Schools is to train a small number of gifted students in the methods, tools and disciplines of the laboratory basic scientist, while also helping them develop skills for relieving human suffering. The faculty for the Mayo MSTP will include 57 of the most highly productive, well-funded research scientists at the Mayo Clinic with demonstrated abilities for mentoring young scientists. The seamless integration of the M.D./Ph.D. program between Mayo Medical School and Mayo Graduate School parallels the complete integration of basic research and clinical investigation at Mayo. These collaborations among Ph.D. scientists and physician scientists model the future careers for M.D./Ph.D. students. The Mayo M.D./Ph.D. program enrolls approximately 6 new students each year in a medical school class o 42. M.D./Ph.D. students are selected from an annual applicant pool of about 150 of the most talented future physician scientists in the country. During their M.D./Ph.D. training they mature into highly productive basic scientists. The integrated basic science core curriculum and programmatic approach to research allows flexible access to an outstanding selection of research faculty as mentors for Mayo MSTP students. Although this is a relatively young program, 39 students have graduated and 36 are currently enrolled. Graduates are continuing on toward successful careers and continuing to make significant scientific contributions. As an NIGMS-designated MSTP the Mayo program will be able to further improve and increase its impact on the training of much-needed physician scientists prepared to apply the explosive growth in biomedical information to important clinical problems.
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2006 — 2010 |
Rodriguez, Moses |
R01Activity Code Description: To support a discrete, specified, circumscribed project to be performed by the named investigator(s) in an area representing his or her specific interest and competencies. |
T Cell Function in Murine Model of Multiple Sclerosis
[unreadable] DESCRIPTION (provided by applicant): There is strong evidence that chronic permanent neurologic impairment in multiple sclerosis is the result of injury and potential loss of axons. We have shown in a viral model of multiple sclerosis induced by Theiler's virus that the immune response contributes to neurologic deficits and axonal dropout months following demyelination of the spinal cord. In this set of experiments we will test the hypothesis that axonal loss which follows demyelination can be the result of immune-mediated damage to axons. Although we have striking data demonstrating the expression of Class I molecules on neurons and axons under pathologic conditions, there is still significant controversy regarding the expression of MHC on CNS cells. Our hypothesis depends in part on the expression of these proteins in the CNS during chronic demyelination. Therefore the first goal of this project is to examine, using double and triple labeling con-focal immunofluorescence, the expression of MHC Class I and MHC Class II on CNS cells with a particular focus on neurons and axons. We will also examine the interaction of MHC molecules on CNS cells with CD8+ T cells, CD4+T cells, NK cells, macrophages, and perforin in the spinal cords of animals with chronic demyelination induced by Theiler's virus infection and in myelin mutant mice with denuded axons. The second goal is to quantify immune mediated axonal loss and changes in neurologic function by adoptive transfer experiments using purified populations of antigen-specific and non antigen-specific immune cells into mice with chronic demyelination either induced by virus or mutations that lead to spontaneous dysmyelination. The third goal is to develop transgenic mouse models in which neurons are specifically deficient in MHC class I function or in which MHC class I function is only expressed on neurons, and to characterize axonal injury following Theiler's virus infection in these animals. These experiments will specifically address whether immune cells require the direct engagement of MHC molecules for induction of axonal injury. The findings are expected to provide new insights into the pathogenesis of axonal and neuronal injury in demyelinating disorders such as multiple sclerosis. [unreadable] [unreadable]
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2011 — 2012 |
Rodriguez, Moses |
R21Activity Code Description: To encourage the development of new research activities in categorical program areas. (Support generally is restricted in level of support and in time.) |
Clinical Translation of a Neuron Protective Recombinant Human Antibody
DESCRIPTION (provided by applicant): There are currently no effective treatments to prevent or reverse neurologic deficits in MS. All available drugs target the immune system. Long term CNS repair may require a paradigm shift to focus on drugs that target the nervous system. We have identified a human monoclonal IgM that binds to the surface of living neurons, including those of human origin. The IgM promotes neurite extension even in the presence of normally inhibitory CNS myelin and protects neurons in culture from cell death. The IgM was isolated from the serum of an individual who carried it at high levels for many years without adverse effects and has shown no signs of toxicity to cells or animals. In vivo the IgM improves spinal cord axon health in a virus (Theiler's murine encephalomyelitis virus, TMEV) mediated mouse model of chronic progressive MS. A recombinant form of the IgM, called rHIgM12, was constructed, a research production cell line established, certified for GMP production and banked at a vendor. We have manufactured and purified over 200 mg of rHIgM12 using a procedure appropriate for GLP and GMP production. The efficacy of a single i.v. dose of rHIgM12 (25 mg/kg) has been demonstrated to protect axons and preserve neurologic deficits in the TMEV model of MS. Over the time frame of functional improvement, the number of myelinated axons in the spinal cord are preserved, and N-acetyl aspartate (NAA) concentrations in the brain stem increase, as measured by magnetic resonance spectroscopy (MRS). We have validated the use of NAA in the brain stem as a surrogate marker of axon preservation throughout the spinal cord (37). This MRS based assay is easily applicable to human studies and may become a valid clinical trial endpoint. After peripheral administration rHIgM12 accumulates within spinal cord lesions, co-localized with the axon marker, neurofilament. Our data supports a mechanism of action in which the IgM clusters neuron membrane domains by binding to gangliosides, activating tubulin and initiating signals that result in axon protection and process extension. This project is designed to generate sufficient safety and dose response data for rHIgM12 to support a larger translational program. 1) Efficacy and safety studies in an autoimmune mediated model of MS (EAE) will address concerns that administering a CNS binding Ab in the face of active autoimmunity may exacerbate disease. 2) There is clear efficacy in protecting axons in the TMEV model. A rigorous dose response study in this model will further define the minimum effective dose and guide safety studies. 3) Studies to measure the ability of rHIgM12 to cross the blood brain barrier will strengthen the data supporting direct signaling within the CNS and tissue binding studies across species using rHIgM12 will justify species selection in safety studies.
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