Mutant cell lines unresponsive to interleukin 1

Details Mutant cell lines unresponsive to interleukin 1 Mutant cell lines unresponsive to interleukin 1

1998-06-10

2000-10-03

Mertz, Prema

Chemistry: molecular biology and microbiology

Animal cell, per se ; composition thereof; process of...

435366, 435369, 4352523, C12N 510, C12N 516, C12N 522

Patent

active

061271764

ABSTRACT:
Mutant cell lines which have lost their ability to respond to IL-1 are provided. In one embodiment, the mutant cell line lacks or is essentially free of IL-1 receptor associate kinase (IRAK), an IL-1 signaling pathway component. The present invention also provides a method for making mutant cell lines that are unresponsive to IL-1, TNF, or to both cytokines. The method comprises the steps of: transfecting cells with a Herpes Simplex Virus thymidine kinase (HSV-TK) gene and a second gene for positive selection, each of said genes being operatively linked to an IL-1 inducible promoter or a TNF inducible promoter; selecting for transfected cells that express HSV thymidine kinase and the product of the positive selection gene in response to IL-1 or TNF; determining a gancyclovir concentration which kills the selected cells in the presence of IL-1 or TNF and which does not kill the selected cells in the absence of IL-1 or TNF; mutagenizing the selected cells using a chemical mutagenizing agent; and treating the mutagenized cells with the determined concentration of gancyclovir and IL-1 or TNF. The present invention also relates to a method of identifying domains or amino acids in IRAK that are essential for IRAK to function in the IL-1 signaling pathway.

REFERENCES:
Altmeyer et al. Cellular Immunology, vol. 138(1), pp. 94-107, Oct. 1991.
"Jak-STAT Pathways and Transcriptional Activation in Response to IFNs and Other Extracellular Signaling Proteins" by Darnell, et al., Scientce, vol. 264, Jun. 3, 1994, pp. 1415-1421.
"Use of a Selectable Marker Regulated by Alpha Interferon To Obtain Mutations in the Signaling Pathway" by Pelligrini, et al., Molecular and Cellular Biology, vol. 9, No. 11, Nov. 1989, pp. 4605-4612.
"RIP mediates tumor necrosis factor receptor 1 activation of NF-.kappa.B but not Fas/APO-1 initiated apoptosis" by Ting, et al., The EMBO Journal, vol. 15, No. 22, 1996, pp. 6189-6196.
"Early Lethality, Functional NF-.kappa.B Activation, and Increased Sensitivity to TNF-Inducesd Cell Death in TRAF2-Deficient Mice" by Yeh, et al., Immunity, vol. 7, Nov. 1997, pp. 715-725.
MyD88: An Adapter That Recruits IRAK to the IL-1 Receptor Complex by Wesche, et al., Immunity, vol. 7, Dec. 1997, pp. 837-847.
"A cytokine-responsive I.kappa.B kinase that activates the transcription factor NF-.kappa.B" by DiDonato, et al., Nature, vol. 388/7, Aug. 1997, pp. 548-554.
"Identification and Characterization of an I.kappa.B Kinase" by Regnier, et al., Cell, vol. 90, Jul. 25, 1997, pp. 373-383.
"Complementation Cloning of NEMO, a Component of the I.kappa.B Kinase Complex Essential for NF-.kappa.B Activation" by Yamaoka, et al., Cell, vol. 93, Jun. 26, 1998, pp. 1231-1240.
"IRAK: A Kinase Associated with the Interleukin-1 Receptor" by Cao, et al., Science, vol. 271, Feb. 23, 1996, pp. 1128-1131.

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