Chemistry: molecular biology and microbiology – Process of mutation – cell fusion – or genetic modification – Introduction of a polynucleotide molecule into or...
Reexamination Certificate
1995-06-07
2004-02-17
Nguyen, Dave T. (Department: 1632)
Chemistry: molecular biology and microbiology
Process of mutation, cell fusion, or genetic modification
Introduction of a polynucleotide molecule into or...
C435S320100, C424S093210
Reexamination Certificate
active
06692964
ABSTRACT:
BACKGROUND
The expression of exogenous DNA in eukaryotic cells permits the study of a broad array of biological topics ranging from the regulation of gene expression to the treatment of disease by gene transfer-based therapies. A number of methods for gene transfer into mammalian cells have evolved. These include in vivo and in vitro infection with cloned retroviral vectors (Shimotohno, K., and Temin, H. M. (1981)
Cell
26:67-77; Cone, R. D., and Mulligan, R. C. (1984)
Proc. Natl. Acad. Sci. USA
81:6349-6353; Dubensky, T. W., Campbell, B. A., and Villareal, L. P. (1984)
Proc. Natl. Acad. Sci. USA
81:7529-7533; Seeger, C., Ganem, D. and Varmus, H. E. (1984)
Proc. Natl. Acad. Sci. USA
81:5849-5852), coprecipitation of DNA with calcium phosphate (Chu, G., and Sharp, P. (1981)
Gene
13:197-202; Benvenisty, N., and Reshef, L. (1986)
Proc. Natl. Acad. Sci. USA
83:9551-9555), encapsulation of DNA in liposomes (Felgner, P. L., and Ringold, G. M. (1989)
Nature
337:387-388; Kaneda, Y., Iwai, K., and Uchida, T. (1989)
Science
243:375-378), direct injection of plasmid DNA (Wolff, J. A., Malone, R. W., Williams, P., Chong, W., Acsadi, G., Jani, A., and Felgner, P. L. (1990)
Science
247:1465-1468), DEAE-dextran (McCutchan, J. H., and Pagano, J. S. (1968)
J. Natl. Cancer Inst
. 41:351-357), electroporation (Neumann, E., Schaefer-Ridder, M., Wang, Y., and Hofschneider, P. H. (1982)
EMBO J
. 1:841-845; Cann, A. J., Koyanagi, Y., and Chen, I. S. Y. (1988)
Oncogene
3:123-128), and DNA-coated particle bombardment of cells and tissues (Yang, N-S., Burkholder, J., Roberts, B., Martinell, B., and McCabe, D. (1990)
Proc. Natl. Acad. Sci. USA
87:9568-9572).
Although transfection of numerous cell types with an exogenous nucleic acid molecule containing a gene results in efficient expression of the exogenous gene, primary T lymphocytes, e.g. peripheral blood T lymphocytes obtained from an individual, have been found to be refractory to transfection and expression of exogenous DNA. Primary T lymphocytes also have been found to be refractory to expression of the introduced nucleic acid when first stimulated to proliferate. Thus, a system that allows for efficient introduction of exogenous DNA into primary T cells and expression of the exogenous DNA in the T cell is still needed.
SUMMARY
The present invention provides an improved method for transfecting T cells with a nucleic acid molecule containing a gene such that expression of the gene in the T cells is enhanced as compared to classic transfection techniques. The method of the invention is particularly useful for transfecting primary T cells which are refractory to classical transfection techniques. The method involves contacting a proliferating T cell with one or more agents which stimulate the proliferating T cell prior to introducing the nucleic acid molecule into the T cell. In one embodiment of the invention, the T cell is stimulated with a combination of a first agent which provides a primary activation signal to the T cell and a second agent which provides a costimulatory signal. The method of the invention has numerous applications, in particular for gene therapy.
REFERENCES:
patent: 5272082 (1993-12-01), Santoli et al.
patent: 5399346 (1995-03-01), Anderson et al.
patent: 5443983 (1995-08-01), Ochoa et al.
patent: 5733543 (1998-03-01), Nabel et al.
patent: 5738852 (1998-04-01), Robinson et al.
patent: 5827642 (1998-10-01), Riddell et al.
patent: 5858358 (1999-01-01), June et al.
patent: WO 90/05541 (1990-05-01), None
patent: WO 94/12196 (1994-06-01), None
patent: WO 94/29436 (1994-12-01), None
patent: WO 95/03408 (1995-02-01), None
patent: WO 95/33823 (1995-12-01), None
patent: WO 95/34320 (1995-12-01), None
patent: WO 96/01122 (1996-01-01), None
patent: WO 96/02646 (1996-02-01), None
patent: WO 96/06942 (1996-03-01), None
Fundamental Immunology, Second Edition, edited by WE Paul, Raven Press Ltd., New York ch. 13 and 15, 1989.*
Pullen, AM. Superantigens. in: Encyclopedia of Immunology, IM Roitt, ed. Academic Press, New York, 1993.*
Crystal, Science, vol. 270, 404-410, 1995.*
Coghlan, New Scientist, vol. 148 :14-15, 1995.*
Gunzburg et al., Molecular Medicine Today , 1/9, pp. 410-427, 1995.*
Mastrangelo et al., Seminars In Oncology, vol. 23, 1:4-21, 1996.*
Guba, S.C. et al., “Interleukin-3 Gene Induction in Normal Human T Cells Via Stimulation of CD3 and CD28,”Clinical Research, vol. 37, No. 4, (1989).
Cann, A.J., et al, “High Efficiency Transfection of Primary Human Lymphocytes and Studies of Gene Expression,”Oncogene, vol. 3, 123-128 (1988).
Fauser, A.A., “Long-Term Expression of Gene Introduction into Normal Human T-Lymphocytes by Retroviral-Mediated Gene Transfer,”Journal of Cellular Biochemistry, vol. 45, 353-358 (1991).
Finer, M., et al., “kat: A High-Effeciency Retroviral Transduction System for Primary Human T Lymphocytes,”Blood, vol. 83, No. 1, 43-50 (1994).
Lowenthal, J., et al., “Tumor Necrosis Factor &agr; Induces Proteins that Bind Specifically to kB-like Enhancer Elements and Regulate Interleukin 2 Receptor &agr;-chain Gene Expression in Primary Human T Lymphocytes,”Proc. Natl. Acad. Sci. USA, vol. 86, 2331-2335 (1989).
Novak, T., et al., “In Vitro Transfection of Fresh Thymocytes and T Cells Shows Subset-Specific Expression of Viral Promoters,”Molecular and Cellular Biology, vol. 12, 1515-1527 (1992).
Park, J. et al., “Transcriptional Regulation of Interleukin 3 (IL3) in Primary Human T Lymphocytes,”The Journal of Biological Chemistry, vol. 268, No. 9, 6299-6308 (1993).
Philip, R., et al., “Effecient and Sustained Gene Expression in Primary T Lymphocytes and Primary and Cultured Tumor Cells Mediated by Adeno-Associated Virus Plasmid DNA Complexed to Cationic Liposomes,”Molecular and Cellular Biology, vol. 14, 2411-2418 (1994).
Tiberghien, P., et al., “Ganciclovir Treatment of Herpes simplex Thymidine Kinase-Transduced Primary T Lymphocytes: An Approach for Specific In Vivo Donor T-Cell Depletion After Bone Marrow Transplantation?,”Blood, vol. 84, No. 4, 1333-1341 (1994).
June Carl H.
Kim Suil
Thompson Craig B.
Hale and Dorr LLP
Nguyen Dave T.
The United States of America as represented by the Secretary of
LandOfFree
Methods for transfecting T cells does not yet have a rating. At this time, there are no reviews or comments for this patent.
If you have personal experience with Methods for transfecting T cells, we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Methods for transfecting T cells will most certainly appreciate the feedback.
Profile ID: LFUS-PAI-O-3277210