Immortalization of splenic and peripheral blood macrophages using a multi-cistronic v-raf/v-myc lentivirus
AssigneeThe Henry M. Jackson Foundation for the Advancement of Military Medicine, Inc.
InventorsBrian C. Schaefer, Chelsi Beauregard
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Abstract
Vectors and methods are disclosed for immortalizing mammalian cells by co-expression of v-raf and v-myc proteins. A replication-defective viral vector is used for improved safety. The vector comprises an optional marker gene, and is especially useful for producing an immortalized macrophage by a method that involves contacting the vector with a monocyte, proliferatively growing the monocyte, growing the monocytic cell on a solid surface, and then growing the monocytic cell on a porous surface. An immortalized macrophage is also disclosed.
Claims9
A replication-deficient viral vector for immortalizing mammalian cells, wherein the replication-deficient viral vector comprises a polynucleotide encoding:
- the p30Gag-vRaf-T2A-vMyc fusion protein of SEQ ID NO:
- 1;
- the Thy1.1 protein of SEQ ID NO:
- 3 and the p30Gag-vRaf-T2A-vMyc fusion protein of SEQ ID NO:
- 1;
- or
- the Thy1.1-P2A-p30Gag-vRaf-T2A-vMyc fusion protein of SEQ ID NO:
- 2.
Dependencies are inferred from the claim text.