Abstract: The present invention relates to diagnostic applications. For autoimmune diseases more particularly, it is demonstrated herein that individuals with SLE, APLA, MCDS and PSS have antibodies that are specific for SR proteins. Thus, in particular aspects the present invention provides methods and compositions for diagnosing autoimmune disease using SR proteins and antibodies to detect the presence of SR protein-specific antibodies in an individual suspected of having autoimmune disease, wherein the presence of such antibodies is indicative of said individual suffering from autoimmune disease.

Abstract: The present invention demonstrates that JSRV envelope protein (Env) can be used to transduce human and other mammalian cells. Hybrid retrovirus packaging cells have been constructed that express the JSRV Env and retrovirus Gag-Pol proteins, and can produce JSRV-pseudotype vectors at high titers. Using high-titer virus the host range for JSRV has been established, and included sheep, human, monkey, bovine and dog cells, but not murine, rat or hamster cells. Retroviral packaging cell lines comprising the JSRV envelope protein, and receptor binding fragments thereof, are provided which transiently and stably produce high titers of recombinant retrovirus particles which can be used to transfer a heterologous gene to a eukaryotic cell.

Abstract: Immortalized human stromal cell lines sustain and expand human hematopoietic precursor cells. The precursor cells are obtained from a blood product and inoculated into a culture medium conditioned by exposure to a human stromal cell line. Preferred human stromal cell lines secrete SCF, LIF, MIP1&agr;, and IL-6, as exemplified by a human stromal cell line designated HS-1. The conditioned culture medium may be supplemented with additional growth factors, such as interleukin-3. After expansion the human hematopoietic precursor cells are harvested and returned to a patient or frozen and stored. The immortalized human stromal cell lines can also be used as feeder layers in ex vivo bone marrow cultures or in colony forming assays.

Abstract: Transgenic non-human animals one having a general deletor construct and a second having a general reporter construct are described. The general deletor animals express a heterologous recombinase under the control of an ubiquitously expressed endogenous promoter. Specifically, the Cre recombinase is inserted into the ROSA26 locus of the mouse. The general reporter animals have a gene which is desired to remove flanked by sites recognized by the is heterologous recombinase. This flanked sequence is operatively associated with a marker gene such that when the gene sequence flanked by sites recognized by the heterologous recombinase is excised, the reporter gene is expressed. When the general deletor mouse is crossed with the general reporter mouse the heterologous recombinase is expressed in essentially all cells of the resultant descendants under the control of the ubiquitous promoter. Expression of the recombinase results in the excision of the desired gene in essentially all cells of the descendant animals.

Abstract: The present invention provides methods for identifying targets of a drug in a cell by comparing (i) the effects of the drug on a wild-type cell, (ii) the effects on a wild-type cell of modifications to a putative target of the drug, and (iii) the effects of the drug on a wild-type cell which has had the putative target modified of the drug. In various embodiments, the effects on the cell can be determined by measuring gene expression, protein abundances, protein activities, or a combination of such measurements. In various embodiments, modifications to a putative target in the cell can be made by modifications to the genes encoding the target, modification to abundances of RNAs encoding the target, modifications to abundances of target proteins, or modifications to activities of the target proteins. The present invention also provides methods for drug development based on the methods for identifying drug targets.

Abstract: Provided are methods of classifying and prognosticating human neuroectodermal tumors by analyzing a sample of the tumor to determine whether various bHLH proteins are detectably expressed in the sample. In the methods disclosed herein, expression of the bHLH genes is assessed by measuring transcripts or proteins expressed from the genes.

Abstract: A family of compounds effective in suppressing lymphocyte activation is comprised of 7-(oxoalkyl)1,3-dialkyl xanthines, other than denbufylline, of the formula in which R1 and R2 are the same or different and are selected from the group consisting of straight-chain or branched alkyl radicals with 2 to 6 carbon atoms, cyclohexyl, alkoxyalkyl and hydroxyalkyl radicals, and A represents a hydrocarbon radical with up to 4 carbon atoms which can be substituted by a methyl group.

Abstract: The invention provides HSV antigens that are useful for the prevention and treatment of HSV infection. Disclosed herein are epitopes confirmed to be recognized by T-cells derived from herpetic lesions. T-cells having specificity for antigens of the invention have demonstrated cytotoxic activity against cells loaded with virally-encoded peptide epitopes, and in many cases, against cells infected with HSV. The identification of immunogenic antigens responsible for T-cell specificity provides improved anti-viral therapeutic and prophylactic strategies. Compositions containing antigens or polynucleotides encoding antigens of the invention provide effectively targeted vaccines for prevention and treatment of HSV infection.

Abstract: The present invention relates to a method of identifying one or more secondary drug targets and their use in the identification of drug or drug candidates, particularly for the treatment of cancer. The yeast-based synthetic lethal screens were used to functionally identify and validate new gene targets to kill tumor cells with defects in cell cycle checkpoints and damage response pathways. These newly identified gene targets can be used to develop new cancer chemotherapeutics.

Abstract: rDNA promoter constructs useful in plasmids and vectors directing transcription of RNAs in a Pol I-specific manner constructed of four elements in a serial array: a first nucleotide sequence, capable of hybridizing under stringent conditions to an rDNA promoter element; a second nucleotide sequence, capable of hybridizing under stringent conditions to an internal ribosome entry signal (IRES); a third nucleotide sequence containing a coding region interest; a fourth nucleotide sequence containing a polyadenylation (polyA) signal sequence; a fifth rDNA enhancer element may be positioned upstream of the serial array. Also recombinant permissive cells and genetically engineered stable cell lines that contain the subject constructs.

Abstract: An isolated protein designated p27 is disclosed. The p27 protein has an apparent molecular weight of about 27 kD, and is capable of binding to and inhibiting the activation of a cyclin E-Cdk2 complex. A nucleic acid sequence encoding p27 protein is disclosed, as well as a method for producing p27 in cultured cells. In vitro assays for discovering agents which affect the activity of p27 are also provided. Methods of diagnosing and treating hypoproliferative disorders are provided.

Abstract: Retroviral vectors for producing coordinately expressed polycistronic mRNA in transfected host cells. A representative retroviral construct capable of forming a proviral genome in a host cell contains a first nucleotide coding sequence, a second nucleotide coding sequence, and a third nucleotide sequence capable of hybridizing under stringent conditions to a 5′ nontranslated region (NTR) of a picornavirus RNA or its complementary RNA strand. The first, second, and third nucleotide sequences are operably linked such that transcription of the proviral genome gives rise to a messenger RNA molecule containing transcripts of the first, second, and third nucleotide sequences. The transcript of the third nucleotide sequence in the messenger RNA molecule contains a nucleic acid capable of forming a regulatory stem-loop nucleic acid structure followed by at least one operable AUG start codon.

Abstract: The subject invention is directed to the discovery of a protein involved in regulation of cell-cycle progression, and includes reagents and methods related thereto.

Abstract: Therapeutic hybrid cytokines, having a size ranging from about 10 to about 30 kDa, comprise portions of cytokines: leukemia inhibitory factor (LIF), granulocyte-colony stimulating factor (G-CSF), interleukin-6 (IL-6), interleukin-11 (IL-11), oncostatin-M (OSM), and ciliaryneurotrophic factor (CNTF). Hybrid cytokines comprise three or four &agr;-helical sequences selected from &agr;-helical sequences of IL-6, G-CSF, LIF, IL-11, CNTF and OSM and linking sequences of 5-40 amino acids in length, selected from the linking sequences of IL-6, G-CSF, LIF, IL-11, CNTF and OSM or other, desirable linking sequences. In the hybrid cytokines, at least one &agr;-helical sequence is derived from a different cytokine than at least one other &agr;-helical sequence; or, at least one linking sequence of a cytokine differentiates the hybrid cytokine from a corresponding cytokine.

Abstract: The present invention provides methods for identifying targets of a drug in a cell by comparing (i) the effects of the drug on a wild-type cell, (ii) the effects on a wild-type cell of modifications to a putative target of the drug, and (iii) the effects of the drug on a wild-type cell which has had the putative target modified of the drug. In various embodiments, the effects on the cell can be determined by measuring gene expression, protein abundances, protein activities, or a combination of such measurements. In various embodiments, modifications to a putative target in the cell can be made by modifications to the genes encoding the target, modification to abundances of RNAs encoding the target, modifications to abundances of target proteins, or modifications to activities of the target proteins. The present invention also provides methods for drug development based on the methods for identifying drug targets.

Abstract: Retroviral packaging cells produce replication-defective retroviral particles capable of binding to Mus dunni endogenous virus retroviral receptors on target cells and are useful in gene transfer and gene therapy. The packaging cell employs a vector encoding a M. dunni retroviral Env protein and produces the retroviral particles at high titer.

Abstract: A therapeutic method of modulating the immune response, by administering to a patient an amount of IL-4 effective to promote peripheral blood lymphocyte adhesion to microvascular endothelial cells in lymphoid organs. The IL-4 is preferably coadministered with IL-1.beta..An improved method of screening a cell line for the production of a binding partner that binds with a cell adhesion molecule, by contacting the binding partner with IL4-activated and nonactivated microvascular endothelial cells, and selecting binding partners that bind to the IL4-activated microvascular endothelial cells but not to the nonactivated microvascular endothelial cells. The selected binding partners may thereafter be tested for the ability to block lymphocyte binding to cytokine-activated endothelial cells. The binding partners are preferably also characterized by binding to human VCAM-1 and to IL4- or TNF.alpha.-activated bone marrow stromal cells. A representative embodiment is mAb 6G10 produced by hybridoma ATTC No. HB 10519.

Abstract: Nucleic acid sequences are disclosed encoding an E170 epithelial ligand and capable of hybridizing under stringent conditions to the nucleotide sequences derived from cDNA clones shown in the figure. Also disclosed are vectors containing the nucleic acid sequences, and cells transformed with the vectors. Methods are given for purifying and utilizing epiligrin, an epithelial glycoprotein complex, and its component glycoproteins, and for raising antibodies against components of this complex. Assay methods are further provided for identification of functional epiligrin in tissues.

Abstract: Immortalized human stromal cell lines sustain and expand human hematopoietic precursor cells. The precursor cells are obtained from a blood product and inoculated into a culture medium conditioned by exposure to a human stromal cell line. Preferred human stromal cell lines secrete SCF, LIF, MIP1.alpha., and IL-6, as exemplified by a human stromal cell line designated HS-1. The conditioned culture medium may be supplemented with additional growth factors, such as interleukin-3. After expansion the human hematopoietic precursor cells are harvested and returned to a patient or frozen and stored. The immortalized human stromal cell lines can also be used as feeder layers in ex vivo bone marrow cultures or in colony forming assays.

Abstract: The present invention provides a rapid expansion method (termed "REM"), for quickly generating large numbers of T lymphocytes, including cytolytic and helper T lymphocytes. REM involves culturing the T cells in association with a disproportionately large concentration of nondividing feeder cells, preferably .gamma.-irradiated peripheral blood mononuclear cells ("PBMC") present at an excess of at least 40-fold (relative to the number of target T cells), more preferably at an excess of at least about 200-fold. Cultures grown under REM exhibit dramatically enhanced expansion rates that can be even further elevated by the use of appropriate concentrations of an additional feeder cell, an anti-CD3 monoclonal antibody and IL-2, as described herein. Clonal expansions in the range of 500-fold to 3000-fold can be achieved within a single stimulation cycle of about 10-13 days, which is more than 100-fold more efficient than currently employed methods of culturing human T cell clones.