Abstract: In various embodiments, the present invention is drawn to antibodies or antigen-binding fragments thereof that bind to a vertebrate high mobility group box (HMGB) polypeptide, methods of detecting and/or identifying an agent that binds to an HMGB polypeptide, methods of treating a condition in a subject characterized by activation of an inflammatory cytokine cascade and methods of detecting an HMGB polypeptide in a sample.

Abstract: Methods for the preparation of substantially purified populations of dendritic cells and monocytes from the peripheral blood of a mammal is described. Also described are vaccine compositions and methods for the treatment of certain diseases and medical conditions based on the substantially purified dendritic cells and monocytes.

Abstract: The invention consists in the use of a maturation agent comprising a mixture of ribosomal and/or membrane fractions for the preparation of mature dendritic cells from immature dendritic cells.

Abstract: Methods and compositions are provided for the persistent modification of cell membranes with exogenous proteins so as to alter the function of the cell to achieve effects similar to those of gene therapy, without the introduction of exogenous DNA. DNA sequences, the proteins and polypeptides embodying these sequences are disclosed for modulating the immune system. The modulations include down-regulation, up-regulation and apoptosis.

Abstract: The invention provides compositions and methods for cell separation. These reagents and techniques specifically agglutinate cells via surface antigen recognition and can be used to recover even rare cell types in high yield.

Abstract: Provided are methods for producing and screening proliferated populations of CD34-negative progenitor cells, mucosal mast cells, connective tissue-type mast cells and basophil cells. The methods generate uniform proliferated populations of cells. The proliferated populations contain a uniform population of a size suitable for use in high throughput screening methods, for example, screening for agents that alter exocytosis. The invention includes screening the proliferated populations with at least one candidate bioactive agent, and evaluating the cells to detect a cell with an altered phenotype. The invention also includes isolating a candidate bioactive agent that causes the altered phenotype. Additionally, cells formed according to the described methods are also encompassed by the invention.

Abstract: A method of stimulating an immune response in a human or animal subject, which method comprises administering to a subject in need thereof an effective amount of an attenuated herpes virus which: (i) lacks a functional vhs gene, or a functional equivalent thereof; (ii) lacks a functional ICP47 gene, or a functional equivalent thereof; and (iii) is incapable of expressing a substantial amount of functional ICP22, or a functional equivalent thereof, in mammalian dendritic cells.

Abstract: This invention is directed to a modified cyclosporin A and to a modified, genetically engineered version of its receptor, cyclophilin. This invention is further directed to a method for treating host versus graft disease following blood marrow transplantation by transfecting stem cells so that after introduction into a patient the stem cells will express the modified cyclophilin, and, as necessary, administer the modified cyclosporin A to the patient.

Abstract: The present invention relates to nonhuman transgenic animals in which the GP V gene has been modified. The invention is also useful for identifying agents that modulate the biological functions of GP V, including the screening and identification of potential therapeutic agents.

Abstract: The invention relates to the use of a secretor variant cell-line expressing the alpha moiety of human IgE binding protein to determine the allergic status of a given individual. Moreover, the cell-line is also used to provide an assay system for determining the allergenicity of substances and for subsequently providing therapeutic compositions which render said substances ineffective. In addition, the invention also relates to the use of said cell-line to determine the IgE independent irritancy of substances and compositions effective for attenuating the effects of said substances.

Abstract: The invention provides compositions and methods for cell separation. These reagents and techniques specifically agglutinate cells via surface antigen recognition and can be used to recover even rare cell types in high yield.

Abstract: A substantially enriched mammalian hematopoietic cell subpopulation is provided, which is characterized by progenitor cell activity for lymphoid lineages, but lacking the potential to differentiate into myeloid and erythroid lineages. Methods are provided for the isolation and culture of this common lymphoid progenitor cell (CLP). The cell enrichment methods employ reagents that specifically recognize CDw127 (IL-7 receptor ?); CD117 (c-kit) protein, in conjunction with other markers expressed on lineage committed cells. The murine cells are also characterized as expressing low levels of sca-1 (Ly-6E and Ly-6A). The CLPs are predominantly cycling, blast cells. These cells give rise to B cells, T cells and natural killer cells, as evidenced by their growth and differentiation in vitro and in vivo.

Abstract: The present invention provides tangible means and methods for stimulation of angiogenesis via enhanced endothelial expression of core proteins having a syndecan-4 cytoplasmic region intracellularly. The tangible means include a prepared DNA sequence fragment having separate and individual DNA sequenced portions coding for an heparan sulfate binding extracellular domain, a central transmembrane domain, and a cytoplasmic domain coding for the syndecan-4 polypeptide. The prepared DNA sequence unitary fragment may be delivered to endothelial cells in-situ, both under in-vivo and/or in-vitro conditions, using suitable expression vectors including plasmids and viruses. The resulting transfected endothelial cells overexpress heparan sulfate binding, core proteins; and the resulting overexpression of these proteoglycan entities causes stimulation of angiogenesis in-situ.

Abstract: A process is provided for expanding the population of endothelial cells obtained from peripheral blood which can be transformed with a vector comprising a DNA sequence encoding a preselected bioactive polypeptide. The resulting transgenic endothelial cells are useful to biocompatibilize implantable medical devices or can be used directly, as for gene therapy.

Abstract: The invention is directed to a purified population of mammalian endothelial, muscle, or neural stem cells. The invention further provides methods for isolating such populations of cells; methods for using such populations of cells for treating mammals; methods for making vectors for gene therapy; and methods for carrying out gene therapy with such vectors.

Abstract: The present invention provides methods for enhancing the development of APC from precursor cells by administering a combination of GM-CSF and IL-4. The precursor cells include: cells contained in peripheral blood, CD14+ cells and precursors in bone marrow. Thus, administration of GM-CSF and IL-4 can be used as a form of cytokine immunotherapy. One embodiment of the present invention involves systemic administration of GM-CSF and IL-4. In this embodiment, APC are required to directly access tumor antigens as they exist in vivo within the patient. A further embodiment of the present invention involves co-administration of a tumor-associated or tumor-specific antigen, with GM-CSF and IL-4, to induce antigen-specific immunity mediated by APC. Yet another embodiment of the present invention describes systemic administration of GM-CSF and IL-4 to achieve reduced tumor burden.

Abstract: A method for obtaining lineage committed human cells imbued with enhanced proliferative potential, biological function, or both, comprising culturing lineage committed human cells under physiologically acceptable liquid culture conditions, where the liquid culture medium is replaced at a rate and for a time sufficient to obtain the human lineage committed cells imbued with enhanced proliferative potential, biological function, or both; and isolating the cultured cells.

Abstract: The present invention relates to methods of preparing biological material, for use in various experimental, diagnostic or therapeutic uses, including immunotherapy treatment or prophylaxy of tumors. More particularly, the present invention relates to methods of preparing membrane vesicles (in particular exosomes) released by various types of mammalian cells, comprising diafiltration and/or density cushion centrifugation. The invention also provides novel methods for characterizing and analyzing exosome preparations, which can be used in quality control assay for the purpose of pharmaceutical product production. The invention is suitable to produce pharmaceutical grade preparations of such membrane vesicles and to fully characterize said preparations, for use in human beings.

Abstract: Disclosed and claimed are methods for the isolation and use of stem cell modulating factors for regulating stem cell cycle and for accelerating the post-chemotherapy peripheral blood cell recovery. Also disclosed and claimed are the inhibitors and stimulators of stem cell proliferation.

Abstract: A novel method has been developed for screening anti-scarring and anti-fibrotic agents. This method offers simplicity, it is reproducible and could be adopted to screen large numbers of new potential ant-fibrotic agents. This method has characteristics in common with the BAEC/BASMC co-culture system, but is more sensitive and does not require screening large number of clonal lines for developing an effective method. In this system, similar to the co-culture system, activation of L-TGF-&bgr;1 occurs by several independent mechanisms which involve binding of the latent complex to M6P/IGF-II receptors, thrombospondin and/or tissue type II transglutaminase. But, in contrast to the co-culture system, this macrophage-dependent system does not appear to involve plasmin.

Abstract: The present invention generally provides methods for modulating formation of new blood vessels. In one embodiment, the methods include administering to a mammal an effective amount of granulocyte macrophage-colony stimulating factor (GM-CSF) sufficient to form the new blood vessels. Additionally provided are methods for preventing or reducing the severity of blood vessel damage in a mammal which methods preferably include administering to the mammal an effective amount of GM-CSF. Provided also as part of this invention are pharmaceutical products and kits for inducing formation of new blood vessels in the mammal.

Abstract: Materials and methods are disclosed for regulation of biological events such as target gene transcription and growth, proliferation or differentiation of engineered cells.

Abstract: The present invention relates generally to a method to reduce, substantially reduce, inactivate or eliminate adventitious agents and/or toxins in a sample, particuarly in nutritive media, media supplements, media subgroups and buffer formulations. Specifically, the present invention provides powdered nutritive media, media supplements and media subgroups produced by the methods of the invention, particuarly cell culture media supplements (including powdered sera such as powdered fetal bovine serum (FBS)). The invention further provides powdered buffer formulations produced by the methods of the invention. The invention also provides kits and methods for cultivation of prokaryotic and eukaryotic cells, particularly bacterial cells, yeast cells, plant cells and animal cells (including human cells) using these nutritive media, media supplements, media subgroups and buffer formulations.

Abstract: A mouse expressing myc in B cells, because of defective function of one or more tumor suppressor genes, is useful for the testing of anti-lymphoma agents and for the testing of genes which may have an effect on the apoptotic pathway. Preferred embodiments include mice of genotypes E&mgr;-myc/p53+/−, E&mgr;-myc/Rb+/− and E&mgr;-myc/p16+/−, and cells derived from lymphomas arising in these mice, wherein the cells may have undergone further genetic alteration. Mouse strains, lymphoma cells and cell lines of the invention can be used in methods to discover new anti-lymphoma agents, methods to characterize tumors, and to characterize genes which may affect the development of resistance to anti-tumor agents. Such methods are also part of the invention.

Abstract: The invention relates to macrophages which have at least one of the following properties: their cytotoxic activity without IFN-7 is increased by about 20 to 30% with respect to standard macrophages, and is preferably of about 70%; their cytotoxic activity with IFN-&ggr; is increased by about 20 to about 40% with respect to standard macrophages, and is preferably of about 93%; the extension of the deactivation of the cytotoxic activity in reply to an activation of IFN-&ggr; is in a ratio such that after 60h of activation with IFN-&ggr;, the cytotoxic activity is higher than or equal to 30%, preferably of about 55%, compared to the maximum cytotoxic activity presented by the macrophages due to IFN-&ggr; activation, with said cytotoxic activity being measured as the percentage of inhibition of 3-H thymidine incorporation by target tumoral cells, particularly U 937 cells.

Abstract: The invention relates to the isolation and cloning of the VE-cadherin promoter. It also relates to transformed cells and transgenic animals containing the VE-cadherin promoter. The VE-cadherin promoter of the invention is particularly useful for the tissue-specific expression of a gene of interest in the vascular endothelium.

Abstract: A method for inducing differentiation of monocytes contained in an extracorporeal quantity of a subject's blood into functional dendritic antigen presenting cells is provided. The monocytes are first treated by exposure to physical perturbation, irradiation in the presence of a photoactivatable agent capable of forming photoadducts with cellular DNA components, and/or treatment with a DNA binding agent. The treated monocytes are then incubated for a period of time sufficient to maximize the number of functional dendritic cells in the treated cell population. Functional dendritic cells generated from induced monocytes are incubated together with disease effector agents to enhance the presentation of at least one disease-causing antigen expressed by the disease effector agents.

Abstract: The present invention relates to a method for producing dendritic cells from human hematopoietic progenitor cells by obtaining a cell sample that includes human progenitor cells and culturing the cell sample under plasma-free and serum-free conditions in the presence of a combination of cytokines to produce dendritic cells, where the combination of cytokines include TGF-&bgr;1.

Abstract: The invention relates to monocytes derived antigen presenting cells (MDAPCs) characterized in that they have the following properties: they present on their surface: antigen CD14 and CD64 with a mean intensity of about 5 to about 200; antigen (CD80 and CD86 with a mean intensity of about 20 to about 200; antigen CI)40 and mannose receptor with a mean intensity of 50 to 500; they are substantiallv devoid of the surface antigens CD1a and CD1c; they present a phagocytosis property; they have the property of stimulating the proliferation of allogenic lymphocytes.

Abstract: The present invention relates to antibody composition that are useful in preparing enriched cell preparations such as human hematopoietic progenitor cells and stem cells and non-hematopoietic tumor cells. The invention also relates to kits for carrying out the processes and to the cell preparations prepared by the processes.

Abstract: We teach a strategy to obtain large quantities of desired APCs, activated B cells, which are superior in their capacity to present tumor protein antigen in a multiadministration protocol. Human B cells can be obtained from peripheral blood in large numbers. These cells can be activated in vitro by coculture with CD40L (CD40-B cells) and an immunosuppressive agent such as cyclosporin A. They can expanded up to 1×103 to 1×104 fold in 2 weeks or 1×105 to 1×106 fold in 2 months. We demonstrate these cells are most efficient APCs comparable to DCs in stimulating allogeneic CD4+ CD45RA+, CD4+ CD45RO+, and CD8+ T cells. In contrast to DCs, CD40-B cells are fully functional even in the presence of immunosuppressive cytokines such as IL-10 and TGF&bgr;.

Abstract: A substantially enriched mammalian hematopoietic cell subpopulation is provided, which is characterized by progenitor cell activity for myeloid lineages, but lacking the potential to differentiate into lymphoid lineages. This population is further divided into specific myeloid progenitor subsets, including a common myeloid progenitor cells (CMP), megakaryocyte/erythroid progenitor cells (MEP) and granulocyte/monocyte lineage progenitor (GMP). Methods are provided for the isolation and culture of these subpopulations. The CMP population gives rise to all myeloid lineages, and can give rise to the two additional and isolatable progenitor populations that are exclusively committed to either the erythroid/megakaryocytic or myelomonocytic lineages. The cell enrichment methods employ reagents that specifically recognize CDw127 (IL-7 receptor &agr;); CD117 (c-kit) protein, in conjunction with other markers expressed on lineage committed cells.

Abstract: The present invention provides methods for generating antigen-reactive T cells in vitro comprising priming immune cells and incubating the primed immune cells in vitro with a non-covalent complex of an heat shock protein and an antigenic molecule. The present invention further relates to methods for generating antigen-reactive CD4+ T cells for immunotherapy. Methods and compositions are also disclosed for the treatment and prevention of cancer or infectious disease in a subject comprising administering to the subject MHC matched antigen-reactive T cells that are generated in vitro by the present methods.

Abstract: Described is an in vitro method for developing dendritic Langerhans type cells, for use in immunotherapy against cancer or infectious diseases and enhancement of graft survival in mammals. The method includes the steps of: culturing cells selected from peripheral blood monocytes and bone marrow cells in a medium containing platelets obtained from the same species or phylogenetically close species; and incubating the culture at 30 to 40° C for a period sufficient to enable formation of mature dendritic Langerhans type cells.

Abstract: A method for producing proliferating cultures of dendritic cell precursors is provided. Also provided is a method for producing mature dendritic cells in culture from the proliferating dendritic cell precursors. The cultures of mature dendritic cells provide an effective means of producing novel T cell dependent antigens comprised of dendritic cell modified antigens or dendritic cells pulsed with antigen, including particulates, which antigen is processed and expressed on the antigen-activated dendritic cell. The novel antigens of the invention may be used as immunogens for vaccines or for the treatment of disease. These antigens may also be used to treat autoimmune diseases such as juvenile diabetes and multiple sclerosis.

Abstract: The invention is directed to a purified population of mammalian endothelial stem cells. The invention further provides methods for isolating such populations of cells, methods for using such populations of cells for treating mammals in need of neovascularization and for making vectors for gene therapy, and methods for carrying out gene therapy with such vectors.

Abstract: Bacterial, yeast and animal cells and methods for overexpressing recombinant heparanase in cellular systems, methods of purifying recombinant heparanase therefrom and modified heparanase species which serve as precursors for generating highly active heparanase by proteolysis.

Abstract: The present invention relates to an antibody composition which contains antibodies directed to murine leukocyte and murine erythroid cells. This composition is used in a novel negative selection process to enrich for human hematopoietic cells from a sample from human-murine chimeric mice. The invention also relates to kits for carrying out this process.

Abstract: The present invention describes a simple technique that provides a biologically relevant measure of the inhibitory effect of cyclosporine in vivo. This ability to measure response to cyclosporine may improve prediction of the efficacy of immunosuppressive treatment in patients and may allow optimal immunosuppression in individual patients.

Abstract: Functional hepatic cells are generated from hematopoietic stem cells. In transplantation, populations of hematopoietic stem cells are shown to give rise to repopulating hepatocytes. The stem cells are obtained from a variety of sources, including fetal and adult tissues. The cells are useful in transplantation, for experimental evaluation, and as a source of lineage and cell specific products, including mRNA species useful in identifying genes specifically expressed in these cells, and as targets for the discovery of factors or molecules that can affect them.

Abstract: Methods for stimulating erythropoiesis using the hematopoietic protein thrombopoietin, optionally in combination with erythropoietin, are provided. The methods provided may be used to stimulate erythropoiesis in bone marrow and peripheral blood cells and in vitro and in vivo. In addition, methods for treatment of thrombocytopenia and anemia in patients are disclosed.

Abstract: This invention relates to transgenic non-human animals comprising a disrupted endothelial nitric oxide synthase gene. These animals exhibit abnormal wound-healing properties and hypertension. This invention also relates to methods of using the transgenic animals to screen for compounds having a potential therapeutic utility for vascular endothelial disorders, such as hypertension, cerebral ischemia or stroke, atherosclerosis and wound-healing activities. Moreover, this invention also relates to methods of treating a patient suffering from hypertension and wound-healing abnormalities with the compounds identified using the transgenic animals, and methods of making the transgenic animals. A method of treating a wound using nitroglycerin is also provided.

Abstract: Disclosed are cells and methods for treating or preventing tumor formation or infections with pathogens in a patient. The cells of the invention are antigen-presenting cells (e.g., dendritic cells or macrophage) that have been loaded with RNA derived from tumors or pathogens. By administering the RNA-loaded antigen-presenting cells to a patient, tumor formation or pathogen infections can be treated or prevented. Alternatively, the RNA-loaded cells can be used as stimulator cells in the ex vivo expansion of CTL. Such CTL can then be used in a variation of conventional adoptive immunotherapy techniques.

Abstract: The present invention provides methods of proliferating B cells as a means of obtaining large numbers of B cells. The present invention further provides methods of differentiating a proliferating B cell population to antibody producing cells.

Abstract: A new family of tumor rejection antigen precursors, and the nucleic acid molecules which code for them, are disclosed. These tumor rejection antigen precursors are referred to as DAGE tumor rejection antigen precursors, and the nucleic acid molecules which code for them are referred to as GAGE coding molecules. Various diagnostic and therapeutic uses of the coding sequences and the tumor rejection antigens, and their precursor molecules are described.

Abstract: Dendritic cells (DCs) are the primary antigen presenting cells during the initiation of T cell-dependent immune responses. The cells originate from the bone marrow and have been suggested to represent a distinct cell lineage. However, distinct DC precursors have not been identified in bone marrow, and mature monocytes can also give rise to DCs. The instant invention presents a distinct DC precursor among bone marrow CD34+ cells. The cells express high levels of the interleukin-3 receptor &agr; chain and CD4 and can be uniquely identified also in blood and lymphoid tissues by this phenotype.

Abstract: The present invention relates to ubiquitin-specific thiol proteases or deubiquitinating enzymes, referred to as DUB (DeUBiquitinating) enzymes, of eukaryotic origin which are members of a superfamily of deubiquitinating enzymes and which comprise a new subfamily of deubiquitinating enzymes which are similar in size and amino acid sequence to one another. DUB enzymes of the present invention are of eukaryotic origin, such as vertebrate origin, including mammalian (e.g., murine, human) origin, as well as yeast origin. All DUB enzymes of the present invention are inducible by at least one cytokine.

Abstract: This invention provides viral and retroviral vectors which comprises a nucleic acid molecule encoding a human cytosolic aldehyde dehydrogenase or a glutamylcysteine synthetase or combinations thereof. Further, this invention provides an isolated mammalian nucleic acid molecule encoding an cytosolic aldehyde dehydrogenase and glutamylcysteine synthetase. In addition, this invention provides a method for reducing the toxic effects of a cyclophosphamide in a subject which comprises replacing the subject's hematopoietic cells with hematopoietic cells of having the retroviral vector. Further, this invention provides a method for introducing a selectable marker into a mammalian cell which comprises transfecting the cell with a nucleic acid molecule encoding human cytosolic aldehyde dehydrogenase or glutamylcysteine synthetase. Lastly, this invention provides a method for selecting mammalian cells expressing protein of interest which comprises: a).

Abstract: A method of determining the presence of chronic volume dependent hypertension is provided wherein a determination is made as to whether there has been a substantial reduction in phosphorylation of the blood-derived protein or renal proximal brush border membrane protein and if such reduction exists concluding that chronic volume dependent hypertension exists in a patient. The method may advantageously be practiced by employing blood serum or blood plasma as the body specimen containing the protein in determining whether a patient has chronic volume dependent hypertension, a cellular component of the blood, such as a blood-derived protein coming from the plasma membrane of lymphocytes. The method may include subsequent therapeutic patient treatment. Related diagnostic apparatus is also provided.

Abstract: Transdominant repressors of viral gene phenotypic expression derived from the rev gene product of HIV-1 or the rex gene product of HTLV-1 and corresponding mutated genes are described, having the capability of repressing the Rev function in HIV-1 and/or the Rex function in HTLV-I and HTLV-II. Transient gene expression analysis of a series of missense and deletion mutants has been used. Sane of the mutants found repress both the Rev and the Rex function and are thus active in more than one viral species. Transdominant viral mutants represent a promising new class of anti-viral agents. Cellular expression of these transdominant inhibitors may be used in such therapeutic approaches as intracellular immunization in order to protect cells against the deleterious effects of viral, e.g. HIV-1 infection.