Abstract: The present invention provides human corneal epithelial cell lines with extended lifespan. The cell lines are useful as an in vitro model of the human ocular surface. Methods for making and using the cell lines are also provided.

Abstract: Genetically engineered or transduced hepatocytes which express genetic material of interest introduced or incorporated into them, as well as methods of producing, transplanting and using the genetically engineered hepatocytes. The genetic material of interest can be incorporated through use of a vector, such as a recombinant retrovirus, which contains the genetic material of interest, or by other means.

Abstract: Coronaviruses can be a significant factor in bovine shipping fever. A new human rectal tumor cell line, HRT-18G, is suitable as a host cell line for the propagation of these bovine respiratory coronavirus-shipping fever viruses, and also is well suited for the propagation of other bovine coronaviruses.

Abstract: Disclosed herein is a transgenic mouse carrying a transgene which expresses non-infectious HIV ribonucleic acid, and the complementary proteins thereof. The transgenic mouse is useful as a source for obtaining the complementary proteins, and as an animal model to study HIV host cell interactions and to evaluate anti-HIV drugs.

Abstract: The invention concerns certain insecticidal compositions of ingestible insecticides selected from the group consisting of DNA viruses, RNA viruses and bacteria of the order Bacillus such as, for example, Bacillus thuringiensis var. israelensis entrapped by a suitable charged polymer. The invention also concerns a process for the preparation of and the use of such insecticidal compositions.

Abstract: Disclosed is a gel of microbially-produced cellulose, characterized in that the microbially-produced cellulose is modified by (1) physically or chemically bonding an animal cell adhesive protein to the cellulose, and/or (2) substituting hydrogen atoms of at least parts of hydroxyl groups of the cellulose with a positively or negatively charged organic group. This gel is valuable as a carrier for mass culture of animal cells or as a medical vulnerary cover.

Abstract: All lines of mammalian origin which have been stably transformed with a chimeric toxin gene expressed under the regulatory control of HIV cis-acting sequences and HIV trans-acting factors are provided by the present invention. HIV infection of a cell of such a transformed cell line results in the death of that cell due to the specific induction of toxin gene expression within the cell. As specifically exemplified, the toxin gene is the diphtheria toxin fragment A gene or a tox176 fragment A chain gene. Also provided by the present invention are recombinant nucleic acid molecules suitable for the stable transformation of a mammalian cell line to produce a transformed cell which will effectively commit suicide in response to HIV infection due to induction of toxin gene expression.

Abstract: Strains of HIV-2 capable of infecting humans and non-human primates such as baboons and causing an immune system disease are disclosed. The HIV-2 strain are used to infect non-human primates which infected primates present symptoms of a disease of the immune system and are used to test drugs, e.g., anti-virals and/or vaccines for their ability to treat and/or prevent lentiviral infections such as HIV infections. Useful strains are HIV-2.sub.UC2, HIV-2.sub.UC12 and HIV-2.sub.UC14 and preferred non-human primates are baboons.

Abstract: Genetically engineered or transduced hepatocytes which express genetic material of interest introduced or incorporated into them, as well as methods of producing, transplanting and using the genetically engineered hepatocytes are disclosed. The genetic material of interest can be incorporated through the use of a vector, such as a recombinant retrovirus, which contains the genetic material of interest, or by other means.

Abstract: The present invention provides a replicatable and hybridizable recombinant single-stranded RNA probe molecule comprising: a recognition sequence for the binding of an RNA-directed RNA polymerase; a sequence required for the initiation of product strand synthesis by the polymerase; and a heterologus RNA sequence inserted at a specific site in the internal region of the recombinant molecule and complementary to an oligo or polynucleotide of interest. This invention also provides methods for determining the presence of concentration of an oligo- or polynucleotide of interest in a sample and for simultaneously determining the presence or concentration of several different oligo- or polynucleotides of interest in a sample.

Abstract: Purified trypsin-sensitive agent or analog thereof, able to downregulate collagenase, PAI-1 and .beta.APP expression in senescent cells isolatable from culture medium having growing fibroblasts or U937 promyelocytic cells.

Abstract: A live or inactivated canine corona virus vaccine is provided which is derived from a virus of the novel antigenic type of the canine corona virus strain I-743 (CNCM, Institut Pasteur, Paris). A method for the preparation of this vaccine and the use of said vaccine in protecting susceptible animals against canine corona virus injection are also disclosed.

Abstract: The present invention relates to a ruminant immortalized mammary epithelial cell line which has normal physiological responses in that it produces milk constituents which comprises .alpha. and .beta.-casein and lactose. There is provided, using the cell line of the present invention a method in vitro studying lactation. There is provided a method of in vitro screening for gene expression of DNA constructs for transgenic ruminant animals. The cell line can be further used in a method for expressing foreign genes. One cell line of the present invention has been deposited at the ATCC under the accession number CRL10274.

Abstract: An insoluble mammalian protein is extracted from transformed bacteria expressing the mammalian protein while avoiding irreversible insolubilization of bacterial host proteins by homogenizing the fermentation broth, centrifuging the homogenized broth and removing the supernatant liquid for the inclusion body containing pellet. In another embodiment, the pH of the homogenized broth is adjusted to 2.0 prior to centrifugation. The acidified broth is then centrifuged, and the pellet is resuspended in buffer, homogenized again, and the inclusion body is isolated by centrifugation.

Abstract: An inventive bioassay method can be used to determine the presence in a fluid sample of a toxin having sodium channel-affecting activity. The method includes the steps of incubating a plurality of cultures of cells which are responsive in a dose-dependent manner to sodium channel-affecting toxins with (i) a medium comprising a solution of ouabain and veratridine and (ii) a portion of the fluid sample, each culture being incubated with a different concentration of the fluid sample; removing the medium and fluid sample from the cultures; incubating the cultures with a medium comprising an indicator which is acted upon by living cells to form a measurable product; measuring the amounts of product formed during the preceding step; and relating the amounts of product measured to a standard calibration curve to determine the presence of the toxin in the sample. The method is readily embodied in kit form and is amenable to automation.

Abstract: Disclosed is an isolated and purified feline immunodeficiency virus (FIV) culture having the identifying characteristics of FIV isolate NCSU.sub.1. A biologically pure culture of host cells containing a FIV having the identifying characteristics of FIV isolate NCSU.sub.1 is also disclosed, along with isolated and purified DNA coding for (a) an FIV having the identifying characteristics of FIV isolate NCSU.sub.1, or (b) an antigenic fragment of an FIV having the identifying characteristics of FIV isolate NCSU.sub.1. Various vaccine formulations containing active agents derived from the foregoing FIV virus, DNA encoding the virus, and DNA encoding antigenic fragments of the virus are also disclosed herein.Also disclosed are immunodeficient mice containing feline tissue, which feline tissue is capable of infection with a feline immunodeficiency virus such as (but not limited to) FIV isolate NCSU.sub.1.

Abstract: A method of producing active immunity against a viral disease in an animal subject comprises administering to the subject a vaccine complex consisting essentially of a live virus and a neutralizing factor bound to the live virus. The neutralizing factor is selected from the group consisting of antibodies and antibody fragments. The live virus is one capable of producing disease in the subject, and the antibody or antibody fragment is one capable of neutralizing the live virus. Preferred subjects are birds, a preferred virus is Infectious Bursal Disease Virus, and a preferred route of administration to birds is by in ovo administration.

Abstract: A method of producing active immunity against infectious bursal disease virus in an avian subject comprises administering to the subject a vaccine complex consisting essentially of a live virus and a neutralizing factor bound to the live virus. The neutralizing factor is selected from the group consisting of antibodies and antibody fragments. The live virus is one capable of producing disease in the subject, and the antibody or antibody fragment is one capable of neutralizing the live virus. A preferred route of administration to birds is by in ovo administration.

Abstract: An attenuated pseudorabies virus (PRV) having a reduced ability to reactivate from latency is produced by introducing (1) a genomic modification in the early protein 0 (EP0) gene whereby said virus is characterized by the inability to express the early protein 0; or (2) a genomic modification in the large latency transcript (LLT) gene whereby said virus is characterized by disruption of the synthesis of said large latency transcript; or (3) the genomic modifications described in both (1) and (2). The attenuated virus is useful in a vaccine for psuedorabies-susceptible animals, particularly swine. Swine vaccinated with a deletion mutant in the EP0/LLT overlap region displayed reduced virus shedding and fewer clinical signs than animals inoculated with a wild type virus. The deletion mutant-vaccinated swine also harbored less PRV DNA in the nervous tissue and showed reduced ability to reactivate the virus.

Abstract: The present invention relates to cell medium developed to support long term multiplication and permanent establishment of a cell line of human liver epithelial cells. The medium may contain an effective cell growth promoting amount of calcium ions; an effective cell growth promoting amount of glucose; an effective amount of insulin to aid cells in glucose uptake; an effective cell growth promoting amount of hydrocortisone; an effective amount of epidermal growth factor to bind epidermal growth factor receptors on cells; an effective amount of transferrin to increase DNA synthesis in cells; an effective amount of cholera toxin to increase DNA synthesis in cells; an effective amount of triiodothyronine to increase DNA synthesis in cells; and an effective growth promoting amount of mammalian hormones and mitogenic factors, including lipoprotein, cholesterol, phospholipids and fatty acids.

Abstract: The present invention relates to a process, a cell culture medium, and a kit for the rapid, high-efficiency transfection of mammalian cells with exogenous DNA. The process comprises incubating a cell culture in the presence of a transfection medium comprising a serum that is different from the serum in the normal growth medium used to grow the cells, and DNA to produce transfected cells. In a preferred embodiment the normal growth medium comprises fetal bovine serum and the transfection medium comprises a serum such as human, calf, horse, lamb, or pig serum. The transfection medium may further comprise an hydroxylated sterol such as 25-hydroxycholesterol.

Abstract: An assay for detecting molecules and compounds which specifically bind to sites which regulate cellular potassium channels, for use, inter alia, as a method for identifying drugs with activity specific for modulating K.sup.+ channels.

Abstract: A method for producing cells having at least one common optical property, electromagnetic property, or biological property. The cells derived from selected living cells located at particular hole positions on an apertured carrier. The selected living cells are separated from all other cells on the carrier either by removing the selected cells from the carrier, or by removing undesired living cells from the carrier, or by killing undesired living cells on the carrier. The selected cells are growing either on the carrier or after having been removed therefrom.

Abstract: The invention is a method for selectively isolating cells which secrete higher levels of monoclonal antibody than did the parental culture from which they were derived. The method is based on the membrane characteristics of the cell and requires the use of a cell sorter. At present, the invention's best mode utilizes fluorescent probes in conjunction with a fluorescence activated cell sorter. The method is based on the correlation of a high level of cell surface immunoglobulin with high monoclonal antibody secretion rates. The invention provides a predictable and rapid process for selecting high monoclonal antibody producing cells.

Abstract: The present invention relates to a method of preparing efficiently producing antibody avian cell clones. The method generally involves infecting a B-lymphocyte population with a v-rel gene using transfeThe government may have certain rights in the invention pursuant to grant No. CA 41450 from the National Institutes of Health.

Abstract: Novel live, cold-adapted temperature-sensitive (CaTs), attenuated Newcastle disease virus vaccines are provided which are effectively immunogenic and contain a mutant of a Hitchner B.sub.1 parent strain of Newcastle disease virus.

Abstract: The present invention relates to a bovine immortalized mammary epithelial cell line which has normal physiological responses in that it produces milk constituents which comprises .alpha.- and .beta.-casein and lactose. There is provided, using the cell line of the present invention a method of `in vitro` studying lactation. There is provided a method of `in vitro` screening for gene expression of DNA constructs for transgenic cows, since the cell line of the present invention is a bovine one. The cell line can be further used in a method for indefinitely expressing foreign genes. The cell line of the present invention has been deposited at the ATCC under the accession number CRL 10274.

Abstract: A transgenic mouse offspring produced by the mating of a first transgenic mouse carrying a transresponder transgene whose expression is regulated by a viral gene product of HSV-1 and a second transgenic mouse carrying a transactivator transgene. A process for expressing a gene of interest which comprises the mating of a first transgenic mouse carrying a transresponder transgene whose expression is regulated by a viral gene product of HSV-1 and a second transgenic mouse carrying a transactivator transgene.

Abstract: A method for producing a recombinant mammal in vivo is disclosed. The mammal has a mammary gland, with mammary epithelial cells lining a duct in that gland One increases the rate of mitosis of the mammary epithelial cells (e.g. by using perphenazine or a mitogen), and one then exposes the mammary epithelial cells in vivo in the gland to a recombinant vector (e.g. a retrovirus vector). The mammary epithelial cells can be pretreated (or simultaneously treated) with polybrene to facilitate infection by virus vectors, and the vector can have a signal sequence which causes foreign proteinaceous material to be secreted into the milk produced by the mammary gland. Also disclosed are recombinant non-human mammals produced by such methods and novel milk products that they produce.

Abstract: The present invention provides a dye other than red and purple which is obtained from cultured cells of Euphorbia milli. The present invention also provides cultured cells containing quercetin glucuronide in a large amount, derived from tissues or cells of Euphorbia milli.

Abstract: An improved method for producing Factor VIII:c is disclosed. The method involves culturing mammalian cells which contain DNA encoding Factor VIII:c and which are capable of expressing Factor VIII:c. In accordance with this invention the cells are cultured in a medium containing an effective amount of a Factor VIII:c-stabilizing substance comprising (a) von Willebrand Factor (VWF), (b) a phospholipid or phospholipid mixture, or a mixture of (a) and (b).

Abstract: Methods for amplifying and detecting a predetermined target nucleic acid in a biological specimen are accomplished even where there is a mismatch in a single position between a primer and the target nucleic acid. The mismatch is located at or near the 3' end of the primer. Such a mismatch is overcome using a primer having a nucleotide with a thymine base at the position of the mismatch. The use of such primers is most likely to prime the target and form primer extension products. This method is particularly useful for detection of a nucleic acid sequence which is not fully known, or where there is considerable heterogeneity in DNA target from patient samples.

Abstract: Methods for identifying individuals at increased risk of diabetes are disclosed. The methods disclosed utilize the discovery of the DQw3.2 variant, which identifies a specific allelic polymorphism at a sinle gene locus. One preferred method utilizes a labeled probe to detect the DQw3.2 allele. This method involves estimating the size of the hybridizable DNA fragment generated by a specific restriction endonuclease and therefrom determining the presence of the allele. A second preferred method involves the serologic detection of the DQw3.2 allele. Within this method, immunocomplexes formed between two different MAb's and separate portions of a cell collection are detected and the presence or absence of the allele determined.

Abstract: A cell line transformed by EBV and infected with a virus, e.g., HIV, which cell line does not release extracellular virus, but which forms syncytia when 1) these cells are co-cultivated with lymphocytes permissive for HIV infection and 2) antibodies to HIV are not present (syncytia formation is specifically inhibited by antibodies by HIV).Syncytial formation induced by cell lines not producing detectable HIV provides a unique and less hazardous test for the detection of non-productive HIV infection as well as for the determination of antibody response to HIV vaccines.

Abstract: The present invention relates to a novel tumor-associated antigen that is a cell-surface glycoprotein having a molecular weight in the range of 110,000-140,000 daltons that is present in a variety of carcinomas, including squamous cell carcinomas and adenocarcinomas. The invention also comprises antibodies reactive with the antigen, hybridoma cell lines that produce the antibodies of the invention, and methods for using the antibodies in the diagnosis and treatment of cancer.

Abstract: A novel retrovirus isolated from human lymphoma cells is disclosed. The retrovirus is characterized by a C-type retroviral particle of approximately 100 nm diameter; and an approximately 27,000 molecular weight p24 core protein. Also disclosed are cell lines from which the virus can be obtained and screening methods for detecting the presence of the virus in human sera.

Abstract: A method for introducing a replication-defective retroviral vector into pluripotent stem cells of embryos of an avian species, including chickens, turkeys, quails or ducks. The method is useful for transferring nucleic acid sequences into embryonic avian cells which may differentiate into somatic or germ cells. Transfer into germ cells has been achieved to produce transgenic animals. The replication-defective retroviral vector used for transfer may be a recombinant retroviral vector containing both a retroviral derived nucleic acid sequence and a non-retroviral derived nucleic acid sequence. Examples of non-retroviral nucleic acid sequences are a neomycin resistance gene from the bacterial transposon Tn5, a herpes simplex virus thymidine kinase gene and a chicken growth hormone gene, however, any prokaryotic or eukaryotic nucleic acid sequence of interest may be used. Transgenic chickens have been produced whose cells contain and express a replication-defective retroviral vector nucleic acid sequence.

Abstract: The present invention discloses a biologically active basement membrane composition. When polymerized under physiogical conditions, the composition forms gel-like structures whose ultrastructure resembles interconnected thin sheets of the lamina densa zone of basement membrane. The major components of the composition include laminin, type IV collagen, heparan sulfate proteoglycan, entactin and nidogen. These components polymerize in constant proportions when redissolved and allowed to reconstitute. Molecular sieve studies on the soluble extract demonstrate that laminin, entactin and nidogen are associated in a large but dissociable complex. The reconstituted matrix is biologically active and stimulates the growth and differentiation of a variety of cells, including epithelial cells, nerve cells, hair follicles and the like. The reconstituted matrix can also be used for determining metastatic potential of tumor cells and for isolating metastatic tumor cells.

Abstract: Monoclonal antibodies having binding specificity to human prostate tumor-associated antigens but not to prostate-specific antigen (PSA) or prostatic acid phosphatase (PAP); and methods of diagnosis and treatment employing the same.

Abstract: Multiple copies of a foreign DNA I coding for the production of a protein may be introduced into eucaryotic cells by cotransforming suitable cells with foreign DNA I and with foreign DNA II which includes a functionally deficient, amplifiable gene coding for a selectable or identifiable trait, preferably carried on the same DNA molecule as a foreign DNA III, which includes a functional, amplifiable gene coding for another selectable or identifiable trait. The cotransformation is carried out under suitable conditions permitting selection or identification of cells expressing the gene on DNA I or that on DNA III, but not that on DNA II. The cotransformed cells so identified or selected are recovered and cloned under conditions where the functionally deficient gene on DNA II is expressed. Cells expressing the gene on DNA II are recovered. They contain multiple copies of DNA I. This method can be used to produce mRNA transcripts or protein products such as human and animal growth hormone, insulin and the like.

Abstract: A method of preserving live viruses comprises subjecting an aqueous system containing the virus to drying either in the frozen state or at ambient temperature, in the presence of trehalose.

Abstract: The invention relates to a live virus vaccine against Newcastle Disease containing at least .sup.10 log 5.5 EID.sub.50 per dose of the strain NDW, deposited at the CNCM of Institut Pasteur in Paris under accession no. I-781.

Abstract: The present invention provides general protocols by which a substance may be identified and characterized as an androgen agonist and/or an androgen antagonist. The described protocols are in-vitro methods which utilize androgen dependent cells in culture and a medium comprising an inhibitor endogenous to the sera of humans and animals which is able to prevent the proliferation of these cultured cells in-vitro. The methodology is rapid, reproducible, and accurate; and provides the major advantage of being able to test large numbers of unevaluated substances for their androgen agonistic and/or antagonistic properties as primary properties or secondary side-effects.

Abstract: A method for diagnosing a neuronal abnormality in a person includes providing living non-neural somatic cells from the person, maintaining the cells in culture under conditions in which the cells become neuronally differentiated, and detecting in the culture a metabolic indicator associated with a symptom of the neuronal abnormality. Also, a method for assaying the effectiveness of a potential therapeutic agent for treatment of a symptom of a neuronal abnormality in a patient, the symptom being associated in the neuronal abnormality with a metabolic indicator, includes treating neuronally differentiated somatic cells with the potential therapeutic agent, and detecting the metabolic indicator in the culture.

Abstract: A kit for testing materno-fetal immunoincompatibility in women, particularly in women whose fetuses are in danger, comprises two containers containing an identical amoung of HLA-D antigen-containing material and one container containing male serum as control. In the test, maternal serum of the patient is added to one container of antigen while the control serum is mixed with the other container of antigen. The amount of expressed HLA-D antigen remaining in each case is established by conventional HLA-D directed monoclonal antibody immunoassay techniques such as conventional radio-immunoassay. The maternal serum assay divided by the control serum assay gives an immunoincompatibility quotient (IQ). The IQ is from 40 to 50% in healthy pregnant women and is about 70% in pathological cases. The test provides reliable results since the antigenic portion is non-living and therefore does not offer the inconstances of living test reagents.

Abstract: Factor-dependent cell line, HU-01, provides the first in vitro system in which human CD34 positive hematopoietic cells have been induced to undergo complete differentiation into enucleated hemoglobin-containing erythrocytes. HU-01 cells may provide a source of monoclonal antibodies useful for early diagnosis and possible treatment of human leukemia conditions, as well as of regulatory factors having therapeutic utility.

Abstract: A novel cell line for use as a parent for hybridoma preparation is provided. This cell line is derived from tumor cells, in particular human tumor cells, other than those originating from bone marrow cells. A method for establishing this cell line is also provided. By fusing this cell line with cells producing useful physiologically active substances, such as B cells immunized with an antigen, a hybridoma capable of active growth can be prepared. This hybridoma can be cultivated for the production of useful physiologically active substances.

Abstract: An improved helper cell for growing up stocks of replication incompetent retrovirus vectors is disclosed. The helper cell resists recombination events due to the fact that natural promoters and poly(A) sequences in the helper sequences have been replaced with foreign promoters and poly(A) sequences bearing little or no homology to the vectors. Plasmids containing these modified sequences can still create a helper cell with resulting expression of the needed helper proteins, yet there is much less risk of recombination events in the helper cell. Also disclosed are vectors produced by such helper cells, target cells infected by such vectors, and helper cells which convert vectors so that they can be used with hosts from different species.

Abstract: A monoclonal antibody which binds preferentially to a subset of the human CD4+ lymphocyte population whereby to positively and precisely distinguish between helper-inducer and suppressor-inducer cells in the CD4+ cell population. The monoclonal antibody recognizes a novel antigen on the CD4+ lymphocytes by means of which it can bind CD4+ cells which express the antigen on the surface of CD4+ cells. The CD4+ subset cell population to which this antibody preferentially binds is the CD4+ helper-inducer population. This selectivity of the monoclonal antibody enables cell sorting, diagnostic and possible therapeutic applications thereof to be realized. The monoclonal antibody also reacts with CD8 cells, B cells and macrophages.

Abstract: Compositions for the prevention of, and for the treatment of autoimmune diseases which comprise as active ingredient membrane material shed from autoimmune T lymphocytes, or T lymphocytes activated by a pressure application and release process. There is also provided a process for obtaining such active materials and for preparing pharmaceutical compositions for these.