Abstract: The present invention relates to a method of gene or DNA targeting in cells of vertebrate, particularly mammalian, origin. That is, it relates to a method of introducing DNA into primary or secondary cells of vertebrate origin through homologous recombination or targeting of the DNA, which is introduced into genomic DNA of the primary or secondary cells at a preselected site. The present invention further relates to primary or secondary cells, referred to as homologously recombinant (HR) primary or secondary cells, produced by the present method and to uses of the homologously recombinant primary or secondary cells. The present invention also relates to a method of turning on a gene present in primary cells, secondary cells or immortalized cells of vertebrate origin, which is normally not expressed in the cells or is not expressed at significant levels in the cells.

Abstract: The present invention relates to the discovery, identification and characterization of nucleic acids that encode a novel protein differentially expressed within the TH2 cell subpopulation (hereinafter referred to as STIF).

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: Dimerization and oligomerization of proteins are general biological control mechanisms that contribute to the activation of cell membrane receptors, transcription factors, vesicle fusion proteins, and other classes of intra- and extracellular proteins. We have developed a general procedure for the regulated (inducible) dimerization or oligomerization of intracellular proteins. In principle, any two target proteins can be induced to associate by treating the cells or organisms that harbor them with cell permeable, synthetic ligands. To illustrate the practice of this invention, we have induced: (1) the intracellular aggregation of the cytoplasmic tail of the .zeta.

Abstract: Mammalian stem cells are obtained and maintained in vitro whose genome has at least one nucleic acid construct encoding an antibiotic resistance gene operatively linked to a promoter specific for mammalian stem cells. The preferential expression of the antibiotic resistance gene in the stem cells results in the preferential survival of the stem cells in the presence of antibiotic.

Abstract: The present invention relates, first, to the identification of novel nucleic acid molecules, termed RATH genes and RATH gene products encoded by such nucleic acid molecules, or degenerate variants thereof, that participate in the regulation, control and/or modulation of G-protein-mediated signal transduction involved in T cell activation, including, but not limited to T helper (TH) cell and TH cell subpopulation activation. Specifically, the nucleic acid molecules of the present invention include the genes corresponding to the mammalian RATH genes, including the RATH1.1 genes. Sequence analysis indicates that the RATH genes are novel genes belonging to the RGS ("regulator of G-protein signalling") gene family, a gene family which encodes gene products involved in G-protein-mediated signal transduction.

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: Pyruvate kinase deficiency is a blood disease characterized by hemolytic anemia. PCR analysis of cDNA leukocytic extracts of patients with hereditary pyruvate kinase deficiency anemia revealed two overexpressed markers when compared to extracts from healthy individuals. These two markers, proposed as human markers A and B and associated with hereditary hemolytic anemia, have been cloned (HUMDNAMA and HUMDNAMB, Gen Bank Association numbers M64700 and M64701). Our attention has focused on human DNA marker B, a 451 bp open-reading frame. The marker has been cloned in the bacteria vector pGEX-2TK and the expressed protein, called HMF (hemolytic anemia related factor), has been tested for biological activity. It has been observed that the HMF-glutathione transferase fusion protein, added to freshly prepared leukocytes cultures, has an immediate cytotoxic effect and a delayed lymphoblastic effect. The lymphoblastic effect is greatly enhanced when erythrocytes are added to the leucocytes.

Abstract: Transgenic animals expressing Fc receptors and uses for the animals in testing the efficacy of human antibodies and in generating novel antibodies are described.

Abstract: A single laser flow cytometric method for the enumeration of micronuclei in erythrocyte populations, wherein a sample of peripheral blood or bone marrow is obtained and the cell populations in the sample are fixed. Reticulocytes in the fixed samples are treated simultaneously with RNAse and with a fluorescent labeled antibody having binding specificity for a surface marker for erythroblasts/reticulocytes. The erythrocyte populations are then stained with a nucleic acid stain which stains DNA representing micronuclei, if present. The stained and/or labeled erythrocyte populations are then exposed to a laser beam of appropriate excitation wavelength for both the nucleic acid staining dye and the fluorescent label to produce fluorescent emission. The fluorescent emission and light scatter produced by the erythrocyte populations are detected by the flow cytometer from which is calculated the number of specific erythrocyte populations in said sample.

Abstract: Mice which produce measurable levels of glucose-6-phosphate dehydrogenase (G6PD) deficient red blood cells can be used as animal models to evaluate new drugs for risk of inducing hemolytic anemia in G6PD-deficient individuals, and for pre-clinical evaluation of gene therapy protocols to correct G6PD deficiency. Deficient and wild-type cells can be distinguished using a tetrazolium dye, and the numbers of each type of cell counted before and after exposure of the cells to the drug or therapy.

Abstract: A monoclonal preparation of lymphocytes, their production and use.

Abstract: The invention relates to macrophages which have at least one of the following properties: their cytotoxic activity without IFN-.gamma. is increased by about 20 to 30% with respect to standard macrophages, and is preferably of about 70%; their cytotoxic activity with IFN-.gamma. 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-.gamma. is in a ratio such that after 60 h of activation with IFN-.gamma., 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-.gamma. 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 monoclonal antibodies against a tumor-associated antigen which is mainly derived from tumors from the group of carcinomas of the breast, ovaries and prostate, as well as adenocarcinomas of the lung, which additionally react with polymorphic epithelial mucin (PEM), to the preparation and use thereof and to the use of the epitope defined by the antibody for diagnosis and therapy.

Abstract: Hairpin ribozyme lacking a substrate moiety, comprising atleast six bases in helix 2 and able to base-pair with a separate substrate RNA, wherein the said ribozyme comprises one or more bases 3' of helix 3 able to base-pair with the said substrate RNA to form a helix 5 and wherein the said ribozyme can cleave and/or ligate said separate RNA(s) in trans.

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 a chain and CD4 and can be uniquely identified also in blood and lymphoid tissues by this phenotype.

Abstract: Switch regions derived from an immunoglobulin (Ig) gene are used to direct recombination between a targeting construct containing a promoter, a switch region (S.sub.1), and 2) a target locus minimally containing a promoter, a switch region (S.sub.2), and a target sequence.

Abstract: Methods and compositions are provided for expression of mammalian genes in culture. An amplifiable gene is introduced by homologous recombination in juxtaposition to a target gene, the resulting combination of amplifiable gene and target gene transferred to a convenient host and the target gene amplified by means of the amplifiable gene. The resulting expression host may then be grown in culture with enhanced expression of the target gene.

Abstract: Disclosed are methods, compositions and uses of bone precursor cells. Bone precursor cells are cells which are not hematopietic and which can differentiate into osteoblasts upon exposure to a bone growth factor and deposit calcium into the extracellular matrix. In addition, methods of differentiating bone precursor cells into osteoblasts are disclosed. Bone precursor cells are useful in the treatment of certain bone related disorders and diseases such as, promoting fracture repair.

Abstract: The present invention relates generally to the control of body weight of animals including mammals and humans, and more particularly to materials identified herein as modulators of weight, and to the diagnostic and therapeutic uses to which such modulators may be put. In its broadest aspect, the present invention relates to the elucidation and discovery of nucleotide sequences, and proteins putatively expressed by such nucleotides or degenerate variations thereof, that demonstrate the ability to participate in the control of mammalian body weight. The nucleotide sequences in object represent the genes corresponding to the murine and human ob gene, that have been postulated to play a critical role in the regulation of body weight and adiposity. Preliminary data, presented herein, suggests that the polypeptide product of the gene in question functions as a hormone.

Abstract: The present invention relates to monoclonal antibodies specific for a cell receptor specific for human stem cell factor (hSCF) as well as pharmaceutical compositions containing such monoclonal antibodies and uses of such monoclonal antibodies.

Abstract: A monoclonal antibody, which has reactivity with human von Willebrand factor, which has action to inhibit RIPA (ristocetin-induced platelet aggregation), BIPA (botrocetin-induced platelet aggregation), and SIPA (shear stress-induced platelet aggregation) of human platelet, and which does not express bleeding action in an medicinally effective dose to exhibit antithrombotic action, is used as an active ingredient of an antithrombotic agent.

Abstract: The present invention pertains to a method for efficiently introducing exogenous genes into primary lymphoid cells without drug selection which comprises the steps (a) deriving a retroviral vector and a helper cell combination that will yield a level of virus production in the range from 5.times.10.sup.6 to 5.times.10.sup.7 units/ml by transfecting a vector into a helper cell followed by selection, isolation of cell clones, and determination of viral titers to identify which virus-producing cell lines produce a virus titer in the range from 5.times.10.sup.6 to 5.times.10.sup.

Abstract: Process for the in vitro production of non-immortalised haematopoietic progenitor cells of the erythroid lineage, in which a population of erythroid progenitor cells is exposed to a combination of growth factors containing a glucocorticoid and an oestrogen and at least one ligand of a tyrosine kinase receptor at least until the cells begin to renew themselves.

Abstract: The present invention relates to novel immortalized precursor cell populations derived from embryonic stem cell populations and methods to produce such cell populations. Also disclosed is an assay to identify regulatory compounds capable of controlling cell growth for therapeutic and experimental use.

Abstract: The present invention provides compositions of purified cells that are selectively enriched for proliferating hematopoietic progenitor cells. These cells are both CD34 positive and express a galactose-binding surface structure. Methods for the purification and use of these cells, for example, in improving transplantations and reducing graft-versus-host disease also are provided.

Abstract: A vector for the integration of a gene into the genetic material of a mammalian host cell such that the gene may be expressed by the host cell. The vector comprises a promoter and the gene and an immunoglobulin dominant control region derived from the mouse .lambda. immunoglobulin gene locus capable of eliciting host cell-type restricted, integration site independent, copy number dependent expression of the said gene. The DNaseI super hypersensitive site exemplified are i) about 2.35 kb upstream of the CAP site of the rearranged .lambda..sub.1 gene, ii) about 2.5 kb upstream of the genomic V.lambda..sub.2 segment or iii) about 30 kb downstream of the rearranged .lambda..sub.1 gene. Mammalian host cells transformed with the vector are disclosed as are transgenic mammals transformed with the vector and a method of producing a polypeptide comprising culturing a transformed mammalian cell.

Abstract: Immunogenic chimeric proteins comprising an endoplasmic reticulum signal sequence and at least one other peptide are disclosed. The invention relates to the design of vaccinia virus constructs capable of directing host organism synthesis of immunogenic chimeric proteins which can be used as immunogens, as vaccines, or in methods of treatment for cancer, infectious diseases, or autoimmune diseases.

Abstract: A gene expression system for producing galactosyltransferase comprises an NSO cell recombinantly modified with a polynucleotide encoding galactosyltransferase or a portion thereof which catalyzes the reaction:UDP-D-galactose+N-acetylglucosamine.fwdarw.UDP+D-galactosyl-N-acetyl-D-gluc osamineFurther provided is a method for producing galactosyltransferase by culturing the gene expression system.

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: A process for production of a desired protein comprising the steps of:a culturing animal cells capable of producing the desired protein in a medium containing trichostatin compounds; andrecovering the desired protein from the culture.

Abstract: The present invention relates to a growth factor preparation derived from a mixed lymphocyte culture, and a process for its production. The invention also relates to a cell culture medium containing said growth factor preparation. The invention further relates to a process for culturing plasma cells and producing antibodies by using said cell culture medium.

Abstract: The present invention provides improved methods for the amplification and expression of recombinant genes in cells. The methods of the present invention permit the isolation of cell lines which have co-amplified input recombinant sequences which encode an amplifiable marker, one or more expression vectors encoding a protein of interest and optionally a selectable marker. The present methods allow the efficient isolation of amplified cell lines which express the protein(s) of interest in a relatively short period of time. The present invention also provides compositions comprising amplified T lymphoid cell lines.

Abstract: The present invention relates to mammalian hematopoietic facilitatory cells (FC). In particular, it relates to the isolation, characterization and uses of the FC. The FC of the present invention can be distinguished from all other known bone marrow cells by their morphology, cell surface phenotype and in vivo function. It has now been established that purified hematopoietic stem cells alone or bone marrow cells depleted of FC do not readily engraft in a recipient. When co-administered with other bone marrow cells, especially the hematopoietic stem cells into a recipient, the FC enhance their engraftment, without apparent adverse biologic activities. In fact, the ability of the FC to enhance the engraftment of bone marrow cells in esablishing lymphohematopoietic chimerism without producing graft versus host disease also induces donor-specific tolerance to permit the permanent acceptance of donor's cells, tissues and organs.

Abstract: The sequence, molecular structure and expression of a cDNA clone, denoted D4, of human and murine origin, preferentially expressed in hematopoietic cells is described herein. The human cDNA clone has been expressed in bacteria and the predicted 24 Kd protein purified. The protein has been used in studies of its biochemical function. As predicted on the basis of sequence, D4 can function as a GDP-dissociation inhibitor of at least several small GTP-binding proteins (CDC42 and rac). The D4 protein was used to generate a polyclonal antibody specific for the protein. The human cDNA was used to obtain several full length murine genomic clones. A clone has been analyzed and sequenced to use for the construction of a gene-targeting vector to produce animals deficient in D4 through disruption of the gene by homologous recombination. These animals can then be used as models for fundamental and applied research on the GTP-binding proteins.

Abstract: A procedure for carrying out T cell differentiation of CD34+ stem cells in an in vitro culture of thymic epithelial fragments whereby the differentiated T cells achieve full immunocompetence. The invention also includes the procedure for differentiation of stem cells from HIV seropositive individuals or genetically modified stem cells. The invention broadly relates to the culture of cultured thymic epithelial fragments and provides procedures for verifying true immunocompetence of the resulting T cells and for analyzing the effects of various compounds on the differentiation process. The invention also comprises several novel applications for utilizing the procedure of the invention, including grafting fortified cultured thymic epithelial fragments and infusing immunocompetent T cells into patients with compromised immune systems.

Abstract: Vectors for the integration of a gene into the genetic material of a mammalian host cell such that the gene may be expressed by the host cell comprise a promoter and the gene and include a dominant activator sequence capable of eliciting host cell-type restricted, integration site independent, copy number dependent expression of the gene.

Abstract: The present invention provides compositions and methods useful for isolating calcineurin as well as inhibiting calcineurin activity. The compositions are peptides that contain regions that are homologous to calcineurin-binding regions of AKAP 79. Also provided are methods for determining if a cell contains a calcineurin-binding and PKA-binding anchoring protein that are useful for identifying additional proteins that bind both calcineurin and PKA. Another aspect of the present invention is methods for enhancing expression of interleukin 2 by T cells.

Abstract: Induction of an antigen-specific T-lymphocyte response in a T-lymphocyte culture, e.g. a primary cytotoxic T-lymphocyte (CTL) response, by loading antigen-presenting vehicles which carry empty MHC molecules with an antigen-derived T-cell-immunogenic MHC-binding peptide, culturing T-lymphocytes in the presence of the peptide-loaded antigen-presenting vehicles under specific T-lymphocyte response-inducing conditions. Optionally, an antigen-specific T-lymphocyte is isolated from the resulting culture and cultured. The process can be used for preparing CTL which are specific for viral or other foreign antigens, or CTL which are specific for autologous peptides. The process can also be used for the identification of peptides that are capable of binding to MHC and inducing a T cell response.

Abstract: The invention relates to monoclonal antibodies against a tumor-associated antigen which is mainly derived from tumors from the group of carcinomas of the breast, ovaries and prostate, as well as adenocarcinomas of the lung, which additionally react with polymorphic epithelial mucin (PEM), to the preparation and use thereof and to the use of the epitope defined by the antibody for diagnosis and therapy.

Abstract: The invention involves a nonapeptide derived from the tumor rejection antigen precursor encoded by gene MAGE-3. This nonapeptide is presented by HLA molecules HLA-A1. The resulting complexes are identified by cytolytic T cells. Such recognition may be used in diagnostics, or therapeutically.

Abstract: The present invention pertains to a method for efficiently introducing exogenous genes into primary lymphoid cells without drug selection which comprises the steps (a) deriving a retroviral vector and a helper cell combination that will yield a level of virus production in the range from 5.times.10.sup.6 to 5.times.10.sup.7 units/ml by transfecting a vector into a helper cell followed by selection, isolation of cell clones, and determination of viral titers to identify which virus-producing cell lines produce a virus titer in the range from 5.times.10.sup.6 to 5.times.10.sup.

Abstract: Disclosed are vectors for achieving high level expression in eucaryotic cells. The vectors include an expressible gene encoding a protein product of interest, an expressible gene encoding a marker protein which permits selection of useful transformants, and an enhancer element, preferably a cellular enhancer element, which functions to increase the level of transcription of genes disposed on its 3' and 5' sides. A blocking element is interposed between the enhancer element and the marker gene which shields the promoter of the marker gene from the transcription-stimulating function of the enhancer, thereby limiting the effect of the enhancer to transcriptions of the DNA encoding the protein product of interest. Use of the vectors permits isolation of viable clones characterized by a very high level of expression of the protein of interest.

Abstract: Macrophages have been developed, which possess at least one of the following properties:their cytotoxic activity without IFN-.gamma. is increased by about 20 to 30% with respect to standard macrophages, and is preferably of about 70%;their cytotoxic activity with IFN-.gamma. is increased by about 20 to about 40% with respect to standard macrophages, and is preferably of about 93%;deactivation of the cytotoxic activity following activation of IFN-.gamma. is such that sixty hours after activation with IFN-.gamma., the residual cytotoxic activity is at least 30%, preferably about 55%, of the maximum cytotoxic activity presented by the macrophages due to IFN-.gamma. activation, with said cytotoxic activity being measured as a percentage of the inhibition of 3-H thymidine incorporation by target tumoral cells, particularly U 937 cells. The macrophages are prepared by culturing healthy human monocytes and lymphocytes in a culture medium containing 1,25-dihydroxy vitamin D.sub.3 and GM-CSF.

Abstract: The present invention relates to a population of blood borne mammalian cells that express a unique profile of surface markers that includes certain markers typical of connective tissue fibroblasts, and are referred to herein as "blood-borne mesenchymal cells." In particular, it relates to the isolation, characterization and uses of such blood-borne mesenchymal cells. The cells of the present invention can be distinguished from peripheral blood leukocytes by their distinct size, morphology, cell surface phenotype and biologic activities, and are likewise distinguishable from connective tissue fibroblasts by other surface phenotypic markers. These cells proliferate in culture, and in vivo, as demonstrated in animal models, are capable of migrating into wound sites from the blood.

Abstract: The present invention provides an immature dendritic cell line derived from p53 growth suppressor gene deficient animals. The immature dendritic cell line may be induced to become an activated dendritic cell line that will stimulate T-cells to proliferate. The cell line is useful for presentation of antigens involved in autoimmune disease and analysis of peptides that produce a T-cell response.

Abstract: A transformant, which harbors a recombinant vector containing a DNA which codes for human nerve growth factor 2 and a DNA which codes for the pro-region of a nerve growth factor at 5'-terminal of said DNA, produces human nerve factor 2 in stable and large amount in a culture medium.

Abstract: Positive-negative selector (PNS) vectors are provided for modifying a target DNA sequence contained in the genome of a target cell capable of homologous recombination. The vector comprises a first DNA sequence which contains at least one sequence portion which is substantially homologous to a portion of a first region of a target DNA sequence. The vector also includes a second DNA sequence containing at least one sequence portion which is substantially homologous to another portion of a second region of a target DNA sequence. A third DNA sequence is positioned between the first and second DNA sequences and encodes a positive selection marker which when expressed is functional in the target cell in which the vector is used. A fourth DNA sequence encoding a negative selection marker, also functional in the target cell, is positioned 5' to the first or 3' to the second DNA sequence and is substantially incapable of homologous recombination with the target DNA sequence.

Abstract: Disclosed are (1) a DNA sequence encoding a mutant L3T4 protein which, when expressed on the surface of a cell, is capable of facilitating infection of the cell by human immunodeficiency virus; the mutant protein includes at least one amino acid residue substitution or deletion in a segment corresponding to the gp120 binding epitope of a native L3T4 protein so as to increase homology between that segment and its counterpart in a CD4 protein; (2) a murine cell line or strain transfected with such a DNA sequence; and (3) a transgenic rodent susceptible to infection by human immunodeficiency virus.

Abstract: Mice lacking expression of CD28 or particular CD45 isoforms in certain cells of the immune system are provided. Also provided are methods of using these mice.