Chemistry: molecular biology and microbiology – Animal cell – per se ; composition thereof; process of... – Animal cell – per se – expressing immunoglobulin – antibody – or...
Patent
1995-03-30
1998-12-22
Stucker, Jeffrey
Chemistry: molecular biology and microbiology
Animal cell, per se ; composition thereof; process of...
Animal cell, per se, expressing immunoglobulin, antibody, or...
435325, 435326, 435330, 435333, 435339, 4353391, 435366, 435372, 435419, 424577, 424578, 935 70, 935 71, 935 95, 935 99, 935100, 935101, 935102, 935107, C12N 1500
Patent
active
058518290
DESCRIPTION:
BRIEF SUMMARY
The present invention is directed to a method for intracellular binding of specific molecules, preferably proteins. More specfically, this method involves the intracellular expression and subsequent use of antibodies specific for a desired molecule.
Various abnormalities appear to be the result of the undesired expression of a particular molecule such as a protein. For example, many tumors are believed to be the result of the overexpression of cellular oncogenes, such as neu, myc, abl, etc. Other malignancies are believed to be the result of expression of an altered receptor. Certain illnesses are caused by the undesired cellular expression of viral proteins. For example, the human immunodeficiency virus (HIV) uses mammalian cells for the preparation of viral encoded proteins including structrual proteins and regulatory enzymes. Human T-cell Leukemia virus type 1 or 2, (HTLV-1 or 2) produce tumors in infected individuals as a result of viral expression. Such viral encoded proteins can result in the assembly of virions which can in turn infect other cells.
Therapeutic strategies have included the development of drugs to target the undesired proteins, means of intercellular blocking of such proteins, for example, soluble CD4, and the use of drugs which will selectively kill cells expressing the undesired proteins.
Another method of treatment that has been suggested is the transfer of genetic materials into cell. For example, by receptor mediated gene delivery, transkaryotic implantation and viral shuttle vectors such as retroviral gene transfer. In such methods, broadly referred to as gene therapy, cells which are either deficient in a protein or produce a dysfunctional protein are hoped to be mended by introducing into the cell DNA coding for the normal gene product.
In vivo gene expression has been reported following direct injection of non-infectious, non-oncogenic plasma DNA encapsulated in lyposomes (1987)! and in a liposome/red blood cell membrane hybrid (Kaneda, Y., et al., Science 243:375 (1989)!. Expression from a variety of calcium phosphate-precipitated gene sequences has been reported following direct 83:9551 (1986); Feigner, P. L., et al., Nature 349:351 (1991)! or 242:714 (1987)!. In vivo gene targeting has also been accomplished by receptor mediated gene delivery in which a complex between an asialoorosomucoid/polysine conjugate and plasmid receptor genes have been used to target expression exclusively to the liver, following intravenous Retroviral gene transfer is reported to offer high efficiency of F., Science 226:401 (1984)!. In vivo gene therapy has been initiated in patients with ADA deficiency who have had reinfused into their blood, autologous lymphocytes carrying the ADA gene and in cancer patients with advanced melanoma who have had reinfused tumor infiltrating lymphocytes A., et al., N. Eng. J. Med. 323:570 (1990) all of these articles are specifically incorporated herein by reference!.
Gene modification of cells which continually express a viral inhibitor and result in the inhibition of viral infection have been proposed and 335:395-196 (1988)!. Towards this goal, several approaches have been Lisziewicz, J., et al., VII Internat'l. Conf. AIDS 2:28 (1991)!, dominant (1990); Hasseloff, J., et al., Nature 334:585-591 (1988); VanderKrol, A. R., et al., BioTechniques 6:958-976 (1988); Malim, M. H., et al., Cell 58:205-214 (1989); and Trono, D., et al., Cell 59:113-120 (1989)!. A major impediment to the development of effective gene inhibition protocols using such antisense RNA or ribozymes is the ability to achieve a high level of expression of the inhibitor encoding DNA template in the transformed cells and this may also be a potential problem for using dominant negative mutants because of the competitive nature of the inhibition.
It would be desirable to have a method which can be used to achieve a high level of expression of an inhibitor to the desired molecule.
It would be desirable to have a method which can specifically target these undesired molecules and which has wide applicabi
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Haseltine William A.
Marasco Wayne A.
Conlin David G.
Dana-Farber Cancer Institute
Eisenstein Ronald I.
Resnick David S.
Stucker Jeffrey
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