Interleukin 15 as Selectable Marker for Gene Transfer in Lymphocytes

摘要

The present invention relates to the use of interleukin-15 (IL-15) as selectable marker for gene transfer, preferably of at least one gene of therapeutic interest, into a mammalian cell or cell line, in particular a human cell or cell line. The present invention furthermore relates to transgenic mammalian cells or cell lines expressing IL-15 as selectable marker and co-expressing at least one protein of interest encoded by at least one gene of interest, which is preferably a protein of therapeutic interest. The present invention is in particular suitable for chimeric antigen receptors (CARs) as the gene or protein of interest and their expression in lymphocytes. The transgenic mammalian cells and cell lines are furthermore suitable for use as a medicament, in particular in the treatment of cancer and in immunotherapy, such as adoptive, target-cell specific immunotherapy.

主权项

1. A method for ex vivo or in vitro gene transfer into a mammalian cell, wherein said method comprises the use of interleukin-15 (IL-15) as a selectable marker,
wherein the mammalian cell is a T lymphocyte (T cell) or natural killer (NK) cell, wherein the method comprises the transfer, into the mammalian cell, of one or more expression constructs comprising a first nucleotide sequence encoding IL-15 and a second nucleotide sequence encoding a protein of interest, which is a protein other than IL-15, and wherein the protein of interest is a protein of therapeutic interest wherein the expression of the IL-15 as a selectable marker results in survival or growth of the mammalian T cell or NK cell in the absence of exogenously added IL-15, and wherein the IL-15 has the amino acid sequence of SEQ ID NO. 1 or an amino acid sequence that has at least 95% sequence identity to SEQ ID NO. 1, or the IL-15 is encoded by the nucleotide sequence of SEQ ID NO. 2 or a nucleotide sequence with at least 95% sequence identity to SEQ ID NO. 2, and wherein the expression of the IL-15 does not result in the secretion of IL-15 into the culture supernatant in amounts sufficient to support survival and growth of cells that are not transformed or transduced with the IL-15 and/or are not expressing the IL-15 themselves.