MOLECULAR MECHANISMS UNDERLYING ACAID-INDUCTION
Sharmila Masli
Schepens Eye Research Institute, 20 Staniford St, Boston, Ma 02114
Grant 5R01EY015472-07 from National Eye Institute
Abstract: Thrombospondin (TSP-1) is essential for maintaining ocular immune privilege and preventing potentially blinding effects of ocular inflammation. An extracellular matrix protein TSP-1 is synthesized by several ocular epithelia as well as resident antigen presenting cells (APCs), which contribute to the immune privilege status of the eye. The anti-inflammatory effects of TSP-1 in ocular inflammation are apparent from the spontaneous development of chronic ocular surface inflammation in TSP-1 deficient mice. The studies described in this proposal seek to elucidate immunologic mechanisms underlying such inflammatory responses noted in the absence of TSP-1. The first specific aim seeks to determine the significance of thrombospondin-1 (TSP-1) in the prevention of ocular inflammation. Experiments in this aim will allow us to characterize immune effectors developed in the absence of TSP-1 that lead to ocular surface inflammation. The second aim seeks to investigate the specific contribution of local TSP-1 in the lacrimal glands towards altering immune effectors. Considering the multidomain structure of a large molecule like TSP-1 with multiple cell type specific biological effects, these investigations will clarify mechanisms by which TSP-1 contributes to the prevention of chronic ocular inflammation. Since TSP-1 is known to be important in maintaining immune privilege these studies can build further on the existing knowledge of ocular immune responses and provide insights into novel potential therapeutic strategies
Keywords: APC; ATGN; Address; Anterior Chamber; Anterior Chamber of the Eye; Anterior chamber of eye structure; Anti-Inflammatories; Anti-Inflammatory Agents; Anti-inflammatory; Antigen-Presenting Cells; Antigens; Antiinflammatories; Antiinflammatory Agents; Autoimmune; Autoimmune Diseases; Autoimmune Process; Biological; CD120b; Candidate Disease Gene; Candidate Gene; Cell Communication; Cell Interaction; Cell-to-Cell Interaction; Chronic; Delayed Hypersensitivity; Delayed-Type Hypersensitivity; Development; Differential Display; Disease; Disorder; Displays, mRNA Differential; Dryness of eye; Environment; Exposure to; Eye; Eyeball; Gene Expression; Genes; Goals; Graft Rejection; INFLM; Immune; Immune Tolerance; Immune response; Immunologic Accessory Cells; Immunologic Tolerance; Immunologic, Immunochemical; Immunologics; Inflammation; Inflammatory; Inflammatory Response; Investigation; Knowledge; Mammals, Mice; Mediating; Mice; Microarray Analysis; Microarray-Based Analysis; Molecular; Molecular Analysis; Monocytes / Macrophages / APC; Murine; Mus; Ocular Pathology; P75TNFR; Peptides; Peripheral; Play; Population; Prevention; Proteins; Receptor Protein; Regulation; Regulatory T-Lymphocyte; Reticuloendothelial System, Spleen; Role; Sight; Sjogren`s Syndrome; Sjogrens; Spleen; Structure; T-Cells; T-Lymphocyte; TBPII; THBS1; THBS1 gene; TNF-R-II; TNF-R75; TNFBR; TNFR2; TNFR80; TNFRSF1B; TNFRSF1B gene; TSP; TSP Gene; TSP-1; TSP1; TSP1 Gene; Testing; Therapeutic; Therapeutic Intervention; Thrombospondin 1; Thrombospondin-1 Gene; Thymus-Dependent Lymphocytes; Transplant Rejection; Transplantation; Transplantation Rejection; Tuberculin-Type Hypersensitivity; Type IV Hypersensitivity; Vision; Visual; accessory cell; anterior chamber; autoimmune disorder; autoimmune uveitis; base; cell mediated hypersensitivity; cell type; delayed-type hypersensitivity response; disease/disorder; dry eye; experiment; experimental research; experimental study; eye dryness; gene product; host response; immune system tolerance; immune unresponsiveness; immunogen; immunological paralysis; immunoresponse; improved; insight; intervention therapy; mRNA Differential Displays; microarray technology; novel; ocular surface; peripheral tolerance; prevent; preventing; public health relevance; receptor; research study; response; social role; thymus derived lymphocyte; transplant
Relevance: Immune privilege of the eye is an evolutionary phenomenon that protects the eye from potential vision-compromising inflammatory immune response. Understanding immunologic mechanisms underlying this phenomenon can help improve current therapeutic approaches used to treat ocular inflammation. The long-term goal of these investigations is to develop novel anti-inflammatory therapeutic strategies to counter inflammatory conditions such as autoimmune diseases and transplant rejections
Project start date: 2004-09-01
Project end date: 2011-08-31
Budget start date: 1-SEP-2010
Budget end date: 31-AUG-2011
PFA/PA: PA-07-070
5R01EY015472-07 (2010): $487500
Sponsored Links Excellgen http://Excellgen.com
MOLECULAR MECHANISMS UNDERLYING ACAID INDUCTION
Sharmila Masli
Schepens Eye Research Institute
20 Staniford St
boston, Ma 02114
Grant 5R01EY015472-05 from National Eye Institute IRG: ZRG1
Abstract: Immune response to ocular antigens is unique in that it is selectively deficient in delayed type hypersensitivity (DTH) mediating Th1 effectors and complement fixing immunoglobulins, while non-complement fixing immunglobulins and cytotoxic T cells remain detectable. Such deviation from a conventional immune response is referred to as Anterior Chamber Associated Immune Deviation (ACAID). This immune response is known to contribute to the immune privilege status of the eye. Resident ocular antigen presenting cells (APCs) are endowed with the ability to induce this response in an antigen specific manner by migrating out to the recipient spleen and presenting antigen to the responders T cells in a manner that generates regulatory cells. These cells in turn are known to prevent DTH responses. The ACAID-inducing ability of ocular APCs is mainly attributed to their exposure to TGFbeta in the eye. Conventional APCs derived from non-ocular sources are also known to acquire such unique ACAID-inducing properties when exposed to TGFbeta. Understanding molecular mechanisms underlying this functional transformation in APCs offers potential to develop therapeutic strategies to prevent ocular inflammation as well as to generate regulatory APCs ex vivo to be used for various clinical applications that require regulation of an undesirable immune response. To this end we have attempted identification of genes that are selectively expressed in TGFbeta-treated APCs. Applying differential analysis of genes (RAP-PCR and Genechip Microarrays) to RNA isolated from untreated APCs and TGFbeta-treated APCs, we isolated several differentially expressed genes that can be relevant in ACAID-inducing properties of APCs. Such an approach has allowed us to select potentially significant genes that have been unsuspected, so far, to be relevant in ACAID-induction. To begin with, we have chosen genes based on their known properties and as they relate to existing information available regarding various aspects of ACAID and APCs involved in its induction as well as the unique ocular microenvironment. We now propose to analyze the participation of these gene products (thrombospondin, TNFR II and IkappaBetaalpha) and their role in ACAID induction by TGFbeta-treated APCs. First, significance of these molecules in development of various aspects of ACAID will be evaluated followed by analysis of individual mechanisms regulated by these gene products. By comparing the effects of these genes and assessing interrelated mechanisms we propose to test a possibility to effectively use TGFbeta-treated APCs to prevent experimental autoimmune uveitis (EAU) and skin allograft rejection
Keywords: antigen presenting cell, cytokine, eye disorder, immunopathology, molecular pathology, transforming growth factor CD antigen, I kappa B beta, biological signal transduction, cytokine receptor, cytotoxic T lymphocyte, delayed hypersensitivity, interferon gamma, interleukin 12, nuclear factor kappa beta, protein biosynthesis, tumor necrosis factor alpha laboratory mouse
Project start date: 2004-09-01
Project end date: 2009-08-31
5R01EY015472-05 (2008): $373031
5R01EY015472-04 (2007): $380644
5R01EY015472-03 (2006): $382788
5R01EY015472-02 (2005): $392000
Grants awarded to Sharmila Masli
MOLECULAR MECHANISMS UNDERLYING ACAID-INDUCTION
Sharmila Masli, Assistant Scientist
Schepens Eye Research Institute, 20 Staniford St, Boston, Ma 02114
Grant 2R01EY015472-06 from National Eye Institute
Abstract: Thrombospondin (TSP-1) is essential for maintaining ocular immune privilege and preventing potentially blinding effects of ocular inflammation. An extracellular matrix protein TSP-1 is synthesized by several ocular epithelia as well as resident antigen presenting cells (APCs), which contribute to the immune privilege status of the eye. The anti-inflammatory effects of TSP-1 in ocular inflammation are apparent from the spontaneous development of chronic ocular surface inflammation in TSP-1 deficient mice. The studies described in this proposal seek to elucidate immunologic mechanisms underlying such inflammatory responses noted in the absence of TSP-1. The first specific aim seeks to determine the significance of thrombospondin-1 (TSP-1) in the prevention of ocular inflammation. Experiments in this aim will allow us to characterize immune effectors developed in the absence of TSP-1 that lead to ocular surface inflammation. The second aim seeks to investigate the specific contribution of local TSP-1 in the lacrimal glands towards altering immune effectors. Considering the multidomain structure of a large molecule like TSP-1 with multiple cell type specific biological effects, these investigations will clarify mechanisms by which TSP-1 contributes to the prevention of chronic ocular inflammation. Since TSP-1 is known to be important in maintaining immune privilege these studies can build further on the existing knowledge of ocular immune responses and provide insights into novel potential therapeutic strategies
Keywords: APC; Address; Adoptive Transfer; Angiogenesis Antagonists; Angiogenesis Blockers; Angiogenesis Inhibitors; Angiogenetic Antagonists; Angiogenic Antagonists; Angiostatic Agents; Anti-Angiogenetic Agents; Anti-Angiogenic Agents; Anti-Angiogenic Drugs; Anti-Inflammatories; Anti-Inflammatory Agents; Anti-inflammatory; Antiangiogenesis Agents; Antiangiogenic Agents; Antigen-Presenting Cells; Antiinflammatories; Antiinflammatory Agents; Autoimmune Diseases; Biological; Chronic; Cornea; Development; Dysfunction; Epithelium; Epithelium, Anterior Corneal; Epithelium, Corneal; Extracellular Matrix Proteins; Eye; Eyeball; Functional disorder; Goals; Graft Rejection; INFLM; Immune; Immune response; Immunologic Accessory Cells; Immunologic, Immunochemical; Immunologics; Inflammation; Inflammatory; Inflammatory Response; Inhibitors, Angiogenetic; Inhibitors, Angiogenic; Investigation; Iris; Iris (Eye); Knowledge; Lacrimal Glands; Lacrimal gland structure; Lead; Maintenance; Maintenances; Mammals, Mice; Mediating; Mice; Molecular; Monocytes / Macrophages / APC; Murine; Mus; Neovascularization Inhibitors; Pb element; Peptides; Physiopathology; Play; Prevention; Process; Property; Property, LOINC Axis 2; Regulation; Role; Sight; Structure; Structure of corneal epithelium; THBS1; TSP; TSP-1; TSP1; Testing; Therapeutic; Therapeutic Intervention; Thrombospondin 1; Transplant Rejection; Transplantation Rejection; Vision; Work; Wound Healing; Wound Repair; accessory cell; angiogenesis; antiangiogenic; autoimmune disorder; cell type; corneal; corneal epithelial; corneal epithelium; design; designing; experiment; experimental research; experimental study; heavy metal Pb; heavy metal lead; host response; immunoresponse; improved; insight; intervention therapy; novel; ocular surface; pathophysiology; prevent; preventing; reconstitute; reconstitution; research study; social role; tissue repair
Relevance: Immune privilege of the eye is an evolutionary phenomenon that protects the eye from potential vision-compromising inflammatory immune response. Understanding immunologic mechanisms underlying this phenomenon can help improve current therapeutic approaches used to treat ocular inflammation. The long-term goal of these investigations is to develop novel anti-inflammatory therapeutic strategies to counter inflammatory conditions such as autoimmune diseases and transplant rejections
Project start date: 2004-09-01
Project end date: 2011-08-31
Budget start date: 30-SEP-2009
Budget end date: 31-AUG-2010
PFA/PA: PA-07-070
2R01EY015472-06 (2009): $489375
GENES THAT PROMOTE ACAID AND INHIBIT OCULAR INFLAMMATION
Sharmila Masli
Schepens Eye Research Institute 20 Staniford St Boston, Ma 02114
Grant 5R03EY013775-03 from National Eye Institute IRG: ZEY1
Abstract: The unique nature of ocular immunity is characterized by anterior chamber associated immune deviation (ACAID), an unusual immune response that is eye characterized by suppressed DH induced to antigens introduced in the anterior chamber. Such an immune response is systemic in nature. Indigenous ocular antigen presenting cells are known to induce a unique type of antigen-specific regulatory cell population in the spleen which is believed to be responsible for suppressed DH responses in ACAID. While it is known that various factors in the ocular environment influence antigen presenting cells to modify their function such that these cells acquire the ability to induce ACAID, the exact mechanisms by which these APCs acquire and express this unique ability is not dearly understood. A cytokine found in abundance in the ocular environment, TGFP, has been demonstrated to play a significant role in functional modification of APCs thereby conferring upon them the ability to induce ACAID. We propose to study molecular mechanisms underlying ACAID induction by examining changes induced within these cells at genetic levels, by TGF[3, changes which in turn lead to functional alterations of an APC. Taking advantage of the latest advances in molecular biology techniques we have successfully compared the transcriptional program of conventional APCs with that of ACAID-inducing APCs. Applying differential display analysis (using RAP-PCR) we have identified genes uniquely expressed by the APCs that functionally resemble eye- derived APCs. Furthermore, we have recently employed DNA microarray technology to accomplish a more comprehensive analysis of expression profiles of these APCs. We now propose to first determine which of the differentially expresssed genes are reasonable candidates for ACAID induction based on our current knowledge of the immune response in ACAID and the available literature. We then adopt a systematic approach, employing various strategies (such as antisense inhibition, antibody or antagonist mediated inhibition, use of transgenic mice selectively deficient in certain genes, induction by chemical or pharmacological . agents or gene transfection) to study the functional relevance to ACAID of the selected candidate genes. Such an approach to examine involvement of specific gene products in conferring unique functional capabilities on eye-derived APCs will not only enhance our understanding of carefully regulated ocular immune responses but give rise to suggested therapies by which the ocular microenvironment can be selectively manipulated.
Keywords: antigen presenting cell, eye disorder, gene expression, gene induction /repression, immune response gene, inflammation, transforming growth factor, immunogenetics, immunoregulation, antisense nucleic acid, enzyme linked immunosorbent assay, hybridoma, laboratory mouse, microarray technology, polymerase chain reaction, tissue /cell culture, transgenic animal
Project start date: 2001-09-01
Project end date: 2004-08-31
5R03EY013775-03 (2003): $186000
1R03EY013775-01 (2001): $174000
MOLECULAR MECHANISMS UNDERLYING ACAID INDUCTION
Sharmila Masli
Schepens Eye Research Institute
20 Staniford St
boston, Ma 02114
Grant 3R01EY015472-05S1 from National Eye Institute IRG: ZRG1
Abstract: Immune response to ocular antigens is unique in that it is selectively deficient in delayed type hypersensitivity (DTH) mediating Th1 effectors and complement fixing immunoglobulins, while non-complement fixing immunglobulins and cytotoxic T cells remain detectable. Such deviation from a conventional immune response is referred to as Anterior Chamber Associated Immune Deviation (ACAID). This immune response is known to contribute to the immune privilege status of the eye. Resident ocular antigen presenting cells (APCs) are endowed with the ability to induce this response in an antigen specific manner by migrating out to the recipient spleen and presenting antigen to the responders T cells in a manner that generates regulatory cells. These cells in turn are known to prevent DTH responses. The ACAID-inducing ability of ocular APCs is mainly attributed to their exposure to TGFbeta in the eye. Conventional APCs derived from non-ocular sources are also known to acquire such unique ACAID-inducing properties when exposed to TGFbeta. Understanding molecular mechanisms underlying this functional transformation in APCs offers potential to develop therapeutic strategies to prevent ocular inflammation as well as to generate regulatory APCs ex vivo to be used for various clinical applications that require regulation of an undesirable immune response. To this end we have attempted identification of genes that are selectively expressed in TGFbeta-treated APCs. Applying differential analysis of genes (RAP-PCR and Genechip Microarrays) to RNA isolated from untreated APCs and TGFbeta-treated APCs, we isolated several differentially expressed genes that can be relevant in ACAID-inducing properties of APCs. Such an approach has allowed us to select potentially significant genes that have been unsuspected, so far, to be relevant in ACAID-induction. To begin with, we have chosen genes based on their known properties and as they relate to existing information available regarding various aspects of ACAID and APCs involved in its induction as well as the unique ocular microenvironment. We now propose to analyze the participation of these gene products (thrombospondin, TNFR II and IkappaBetaalpha) and their role in ACAID induction by TGFbeta-treated APCs. First, significance of these molecules in development of various aspects of ACAID will be evaluated followed by analysis of individual mechanisms regulated by these gene products. By comparing the effects of these genes and assessing interrelated mechanisms we propose to test a possibility to effectively use TGFbeta-treated APCs to prevent experimental autoimmune uveitis (EAU) and skin allograft rejection
Project start date: 2004-09-01
Project end date: 2009-08-31
3R01EY015472-05S1 (2008): $48125
1R01EY015472-01 (2004): $392000