Efficient gene transfer into primary murine lymphocytes obviatin

Details Efficient gene transfer into primary murine lymphocytes obviatin Efficient gene transfer into primary murine lymphocytes obviatin

1996-03-19

1999-05-25

Guzo, David

Chemistry: molecular biology and microbiology

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

Modification of viruses

4353201, 435355, 435357, 435372, 4353722, 4353723, 435373, C12N 1510, C12N 1563, C12N 1586, C12N 510

Patent

active

059069287

DESCRIPTION:

BRIEF SUMMARY
FIELD OF THE INVENTION

The present invention relates to a method for efficiently introducing exogenous genes into lymphoid cells using retroviral vectors and helper cells. The gene transfer method of the present invention is achieved without drug selection and is potentially important for somatic cell gene therapy and for studying lymphocyte biology.


BACKGROUND OF THE INVENTION

The disclosures referred to herein to illustrate the background of the invention and to provide additional detail with respect to its practice are incorporated herein by reference. For convenience, the disclosures are referenced in the following text and respectively grouped in the appended bibliography.
Retroviral vectors are the most efficient tools for the stable introduction of genes into vertebrate cells. Clinical experiments have been conducted to use retrovirus vectors to cure a genetic disease in humans (adenosine deaminase (ADA) deficiency). Besides correcting inborn errors of metabolism, gene therapy is also being tested in clinical trials to cure cancer and various other diseases (Science 1992, Vol. 258, pp. 744-746).
Retroviral vectors are basically retroviral particles that contain a crippled viral genome in which all viral protein coding sequences have been replaced with the gene(s) of interest. As a result, such viruses cannot further replicate after one round of infection without the help of a helper virus. Retroviral vector particles are produced by helper cells. Such helper cells contain plasmid constructs which express all retroviral proteins necessary for particle production and replication. After the introduction (transfection) of the retroviral vector genome into such helper cells, the vector genome (an RNA genome) is encapsulated into virus particles (due to the presence of specific encapsidation sequences). Virus particles are released from the helper cell carrying a genome containing only the gene(s) of interest. After infection of a fresh target cell, the RNA genome is reverse transcribed into DNA and the DNA copy is integrated into the host genome. The integrated viral DNA is called the provirus. In the last decade, several retroviral vector systems, derived from chicken or murine retroviruses, have been developed for the expression of various genes (for reviews see Temin, 1987; Gilboa, 1990).


Efficient Gene Transfer into Primary Murine Lymphocytes

Much attention has been focused upon experimental animal models aimed at the efficient introduction of exogenous genes into hematopoietic stem cells using retroviral vectors (Williams et al., 1984, Nature 310:476; Dzierzak et al., 1989, Nature 331:35; Bender et al., 1989, Mol. Cell Biol. 9:1426). The ability of pluripotential stem cells to repopulate all hematopoietic cell lineages, and their capacity for self-renewal (Williams et al., 1984, Nature 310:476; Lemischka et al., 1986, Cell 45:917) make them attractive target cells to correct genetic defects affecting hematopoietic cells (Parkman, 1986, Science 232:1373). Technically, however, this has proven somewhat difficult, primarily because whole bone marrow contains very few pluripotential stem cells and as yet, no unique cell surface markers have been identified on these cells making it difficult to purify sufficient quantities for detailed analysis. Thus, gene transfer into these cells has been inefficient (Szilvassy et al., 1989, Proc. Natl. Acad. Sci. USA 86:8798) and even when transfer has been achieved, appropriate expression of the transferred gene has often been problematic, possibly because pluripotential stem cells undergo many differentiation steps before reaching maturity which might interfere with the proper expression of the introduced gene. It has therefore been suggested that for certain diseases which effect the lymphoid compartment, or which might be treated by augmenting immune responses, gene transfer into primary lymphocytes might be helpful, and in some cases preferred (Culver et al., 1991, Proc. Natl. Acad. Sci. USA 88:3155; Culver et al., 1991, Human Gene Therapy 2:107). Primary

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.
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.
Miller, Nature, vol. 357, Jun. 11, 1992, pp. 455-460.
Finer et al., Blood, vol. 83, No. 1, Jan. 1, 1994, pp. 43-50.

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