Replicative potential of human natural killer cells

The replicative potential of human CD56+ CD3− natural killer (NK) cells is unknown. We found that by exposing NK cells to the leukaemic cell line K562 genetically modified to express 4‐1BB ligand and interleukin 15 (K562‐mb15‐41BBL), they expanded for up to 30 population doublings, achieving numbers that ranged from 1·6 × 105 to 1·2 × 1011% (median, 5·9 × 106%; n = 7) of those originally seeded. However, NK cells eventually became unresponsive to stimulation and died. Their demise could be suppressed by enforcing the expression of the human telomerase reverse transcriptase gene (TERT). TERT‐overexpressing NK cells continued to proliferate in response to K562‐mb15‐41BBL stimulation for more than 1 year of culture, while maintaining a normal karyotype and genotype. Long‐lived NK cells had high cytotoxicity against myeloid and T‐lineage leukaemic cells. They remained susceptible to genetic manipulation, becoming highly cytotoxic to B‐lineage leukaemic cells after expression of anti‐CD19 signaling receptors. Thus, human NK cells have a replicative potential similar to that of T lymphocytes and their lifespan can be significantly prolonged by an increase in TERT activity. We suggest that the methods described here should have many applications in studies of NK cell biology and NK cell‐based therapies.

[1]  Eric Vivier,et al.  Functions of natural killer cells , 2008, Nature Immunology.

[2]  L. Zitvogel,et al.  Natural killer cell–directed therapies: moving from unexpected results to successful strategies , 2008, Nature Immunology.

[3]  C. Demanet,et al.  NK cell receptors and their ligands in leukemia , 2008, Leukemia.

[4]  J. Chewning,et al.  Hierarchy of the human natural killer cell response is determined by class and quantity of inhibitory receptors for self-HLA-B and HLA-C ligands , 2007, The Journal of Immunology.

[5]  J. Chewning,et al.  Hierarchy of the Human Natural Killer Cell Response Is Determined by Class and Quantity of Inhibitory Receptors for Self-HLA-B and HLA-C Ligands1 , 2007, The Journal of Immunology.

[6]  Christopher B. Miller,et al.  Genome-wide analysis of genetic alterations in acute lymphoblastic leukaemia , 2007, Nature.

[7]  D. Campana,et al.  Genetic modification of primary natural killer cells overcomes inhibitory signals and induces specific killing of leukemic cells. , 2005, Blood.

[8]  Peter M Lansdorp,et al.  Telomere loss, senescence, and genetic instability in CD4+ T lymphocytes overexpressing hTERT. , 2005, Blood.

[9]  C. Le,et al.  Successful adoptive transfer and in vivo expansion of human haploidentical NK cells in patients with cancer. , 2005, Blood.

[10]  A. Gratwohl,et al.  Purified donor NK-lymphocyte infusion to consolidate engraftment after haploidentical stem cell transplantation , 2004, Leukemia.

[11]  M. Wigler,et al.  Circular binary segmentation for the analysis of array-based DNA copy number data. , 2004, Biostatistics.

[12]  D. Campana,et al.  Chimeric receptors with 4-1BB signaling capacity provoke potent cytotoxicity against acute lymphoblastic leukemia , 2004, Leukemia.

[13]  L. Moretta,et al.  Unravelling natural killer cell function: triggering and inhibitory human NK receptors , 2004, The EMBO journal.

[14]  E. Kimby,et al.  Expansion of natural killer (NK) and natural killer-like T (NKT)-cell populations derived from patients with B-chronic lymphocytic leukemia (B-CLL): a potential source for cellular immunotherapy , 2003, Leukemia.

[15]  H. Spits,et al.  Ectopic hTERT expression extends the life span of human CD4+ helper and regulatory T-cell clones and confers resistance to oxidative stress-induced apoptosis. , 2003, Blood.

[16]  Todd A Fehniger,et al.  Natural killer cell receptors: new biology and insights into the graft-versus-leukemia effect. , 2002, Blood.

[17]  H. Harada,et al.  Selective Expansion of Human Natural Killer Cells from Peripheral Blood Mononuclear Cells by the Cell Line, HFWT , 2002, Japanese journal of cancer research : Gann.

[18]  N. Rufer,et al.  Transfer of the human telomerase reverse transcriptase (TERT) gene into T lymphocytes results in extension of replicative potential. , 2001, Blood.

[19]  Erik Hooijberg,et al.  Immortalization of Human CD8+ T Cell Clones by Ectopic Expression of Telomerase Reverse Transcriptase1 , 2000, The Journal of Immunology.

[20]  G. Pawelec,et al.  Replicative senescence of T cells: does the Hayflick Limit lead to immune exhaustion? , 1997, Immunology today.

[21]  C. Croce,et al.  A potential role for interleukin-15 in the regulation of human natural killer cell survival. , 1997, The Journal of clinical investigation.

[22]  Jeffrey S. Miller,et al.  Role of monocytes in the expansion of human activated natural killer cells. , 1992, Blood.

[23]  B. Naume,et al.  A comparative study of IL-12 (cytotoxic lymphocyte maturation factor)-, IL-2-, and IL-7-induced effects on immunomagnetically purified CD56+ NK cells. , 1992, Journal of immunology.

[24]  L. Lanier,et al.  A model for the differentiation of human natural killer cells. Studies on the in vitro activation of Leu-11+ granular lymphocytes with a natural killer-sensitive tumor cell, K562 , 1985, The Journal of experimental medicine.

[25]  G. Trinchieri,et al.  Response of resting human peripheral blood natural killer cells to interleukin 2 , 1984, The Journal of experimental medicine.

[26]  G. Todaro,et al.  Growth factors from murine sarcoma virus-transformed cells. , 1978, Proceedings of the National Academy of Sciences of the United States of America.

[27]  R. Kiessling,et al.  „Natural”︁ killer cells in the mouse. I. Cytotoxic cells with specificity for mouse Moloney leukemia cells. Specificity and distribution according to genotype , 1975, European journal of immunology.

[28]  J. D. Di Santo,et al.  Natural killer cells: diversity in search of a niche , 2008, Nature Immunology.

[29]  Lewis L Lanier,et al.  NK cell recognition. , 2005, Annual review of immunology.

[30]  J. Ritz,et al.  Costimulation of human natural killer cell proliferation: role of accessory cytokines and cell contact-dependent signals. , 1996, Natural immunity.

[31]  G. Trinchieri,et al.  Preferential proliferation of natural killer cells among peripheral blood mononuclear cells cocultured with B lymphoblastoid cell lines. , 1987, Natural immunity and cell growth regulation.