Efficient gene transfer into primary lymphocytes obviating the n

Details Efficient gene transfer into primary lymphocytes obviating the n Efficient gene transfer into primary lymphocytes obviating the n

1995-06-07

1997-09-16

Guzo, David

Chemistry: molecular biology and microbiology

Treatment of micro-organisms or enzymes with electrical or...

Modification of viruses

4353201, 435355, 435325, C12N 510, C12N 1564

Patent

active

056679980

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.7 units/ml; (b) isolating a lymphoid cell subpopulation which can repopulate a specific lymphoid lineage or is a long-lived population by treating a suspension of lymphoid cells with a monoclonal antibody which removes undesired lymphoid cells to obtain an enriched lymphoid subpopulation; (c) culturing the enriched lymphoid subpopulation from step (b) with growth factors specific to the lymphoid subpopulation; (d) co-cultivating the lymphoid subpopulation from step (c) with a lawn of irradiated virus-producing cell line from step (a) to produce an infected lymphoid subpopulation; and (e) harvesting the infected lymphoid subpopulation. The invention further relates to a population of transfected lymphocytes, in which greater than about 90% of the lymphocytes contain a provirus.

REFERENCES:
patent: 4650764 (1987-03-01), Temin et al.
patent: 4980289 (1990-12-01), Temin et al.
patent: 5124263 (1992-06-01), Temin et al.
patent: 5399346 (1995-03-01), Anderson et al.
A. Dusty Miller, "Human Gene Therapy Comes of Age", Nature, vol. 357, pp. 455-460 Jun. 1992.
Finer et al., "kat: A High-Efficiency Retroviral Transduction Sysytem for Primary Human T Lymphocytes", Blood, vol. 83, No. 1, pp. 43-50 Jan. 1994.
Sutkowski et al., 1994, Proc. Natl. Acad. Sci. USA 91:8875-79.
Sprent and Tough, 1994, Science 265: 1395-99.
Tough and Sprent, 1994, J. Exp. Med. 179: 1127-35.
Kuo et al., 1993, Blood 82:845-52.
Sutkowski et al., 1993, J. Cell. Biochem. Suppl. 17B: 166.
Braakman et al., 1992, Eur. J. Immunol. 22: 63-69.
van Beusechem et al., 1992, Proc. Natl. Acad. Sci. USA 89: 7640-44.
Malim et al., 1992, J. Exp. Med. 176: 1197-1201.
Riddell et al., 1992, Science 257: 238-41.
Zimmermann et al., 1992, Human Gene Therapy 3: 155-61.
Culver et al., 1991, Human Gene Therapy 2: 107-109.
Culver et al., 1991, Proc. Natl. Acad. Sci. USA 88: 3155-59.
Fauser, 1991, J. Cell. Bioch. 45:353-58.
Ferrari et al., 1991, Science 251: 1363-66.
Greenberg, 1991, Adv. Immunol. 49: 281-355.
Krauss et al., 1991, Human Gene Therapy 2:221-28.
Lupton et al., 1991, Mol. Cell. Biol. 11: 3374-78.
Sprent et al., 1991, J. Exp. Med. 174: 717-28.
Kasid et al., 1990, Proc. Natl. Acad. Sci. USA 87: 473-77.
Wilson et al., 1990, Proc. Natl. Acad. Sci. USA 87: 439-43.
Bender et al., 1989, Mol. Cell. Biol. 9: 1426-34.
Bodine et al., 1989, Proc. Natl. Acad. Sci. USA 86: 8897-8901.
Lim et al., 1989, Proc. Natl. Acad. Sci. USA 86:8892-96.
Szilvassy et al., 1989, Proc. Natl. Acad. Sci. USA 86:8798-8802.
Markowitz et al., 1988, J. Virol. 62: 1120-24.
Lemischka et al., 1986, Cell 45: 917-27.
Reimann et al., 1986, J. Immunol. Methods 89: 93-101.
Uchida et al., 1986, J. Immunol. 136: 1876-79.
Kantoff et al., 1986, Proc. Natl. Acad. Sci USA 83: 6563-67.
Richter et al., 1984, Mol. Cell. Biol. 4: 151-59.
Mann et al., 1983, Cell 33: 153-59.

Affiliated with

Also associated with

Rating

Efficient gene transfer into primary lymphocytes obviating the n does not yet have a rating. At this time, there are no reviews or comments for this patent.

If you have personal experience with Efficient gene transfer into primary lymphocytes obviating the n, we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Efficient gene transfer into primary lymphocytes obviating the n will most certainly appreciate the feedback.

Rate now

Comments { 0 }

Profile ID: LFUS-PAI-O-217567