Methods for enhancing natural killer cell proliferation and activity

A natural killer and cell technology, applied in cell culture active agents, biochemical equipment and methods, animal cells, etc., can solve problems such as restricted NK cells

Inactive Publication Date: 2012-11-14
GAMIDA CELL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, none of the protocols have significantly achieved an expanded NK cell population capable of surviving and expanding in suitable host target organs after transplantation (steady-state proliferation), and NK cell immunotherapy using ex vivo expansion remains limited. Limited availability of adequate numbers of highly purified functionally appropriate NK cells for use in clinical protocols (Bachanova et al., Canc Immunol. Immunother. 2010; 59:739-44; Guven, Karolinska Institute, 2005; Schuster et al., E.J. Immunology 2009; 34:2981-90; Bernardini et al. Blood 2008; 111:3626-34)

Method used

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  • Methods for enhancing natural killer cell proliferation and activity
  • Methods for enhancing natural killer cell proliferation and activity
  • Methods for enhancing natural killer cell proliferation and activity

Examples

Experimental program
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Embodiment

[0237] Reference is now made to the following examples, which, together with the foregoing description, illustrate some embodiments of the invention in a non-limiting manner.

[0238]Generally, the nomenclature used herein and the laboratory methods and procedures utilized in the present invention include molecular, biochemical, microbiological and recombinant DNA techniques. These techniques are explained in depth in the literature. See, for example, "Molecular Cloning: A laboratory Manual" Sambrook et al., (1989); "Current Protocols in Molecular Biology" Volumes I-III Ausubel, R.M., ed. (1994); Ausubel et al., "Current Protocols in Molecular Biology", John Wiley and Sons, Baltimore, Maryland (1989); Perbal, "A Practical Guide to Molecular Cloning", John Wiley & Sons, New York (1988); Watson et al., "Recombinant DNA", Scientific American Books, New York; Birren et al. (eds) "Genome Analysis: A Laboratory Manual Series", Vols.1-4, Cold Spring Harbor Laboratory Press, New York...

Embodiment I

[0278] Example 1: Nicotinamide enhances the ex vivo proliferation of NK cells

[0279] To evaluate the effect of added nicotinamide on NK cell growth in vitro, cord blood or bone marrow cells were cultured with growth factors (cytokines) and increasing concentrations of nicotinamide without the use of feeder cells or feeder layers, whereas The fraction of NK and non-NK (eg, CD3+) cells was determined at different time points.

[0280] CD56+ cells derived from umbilical cord blood were found to be enriched in the CD56+CD3- NK cell population and contained considerably fewer CD56+CD3+ NKT cells. When purified cord blood NK cells (CD56+) were cultured with nicotinamide in the presence of IL-2 and IL-15, a substantial increase in NK cell proliferation was evident as early as day 14 in culture and was tested at all concentrations. Figure 1A showed that the proliferation of NK cells treated with 2.5mM nicotinamide at 14 days was 4 times greater than that of cells cultured with on...

Embodiment II

[0291] Example II - ex vivo exposure to nicotinamide enhances NK cell function

[0292] NK cells are characterized by their ability to respond to both inhibitory and activating stimuli, and the production of functional NK cells with potent and specific cytotoxicity is critical to any consideration of ex vivo NK cell culture. The effect of nicotinamide on NK cell function was assessed by its effect on cell markers and analyzed using chemotaxis "Tranwell" migration and target cell "killing" assays. To detect changes in the prevalence of inhibitory and activated NK cell fractions, growth factors [10 ng / mL Flt-3, 20 ng / mL interleukin-15 (IL-15) and 5 ng / mL interleukin-2 (IL -2)], with or without 1, 2.5 and 5 mM nicotinamide, purified cord blood-derived CD56+ cells were cultured in the wells of the well. After 3 weeks of culture, cord blood-derived NK cells ( Figure 7 ), the prevalence of the inhibitory CD56+ NKG2A cell fraction in NK cells was significantly and dose-dependent...

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Abstract

Methods of ex-vivo culture of natural killer (NK) cells are provided and, more particularly, methods for enhancing propagation and / or functionality of NK cells by treating the cells with a nicotinamide or other nicotinamide moiety in combination with cytokines driving NK cell proliferation. Also envisioned are compositions comprising cultured NK cells and therapeutic uses thereof.

Description

technical field [0001] The present invention, in certain embodiments thereof, relates to the ex vivo (ex-vivo) culture of natural killer (NK) cells, and more particularly, but not exclusively, to the treatment of said cells with nicotinamide and binding Compositions and methods for enhancing NK cell proliferation and / or functionality through cytokine-promoted NK cell proliferation. Background technique [0002] Natural killer (hereinafter also referred to simply as "NK") cells are lymphoid cells that participate in the immune response. These cells have multiple functions, especially killing tumor cells, cells undergoing oncogenic transformation and other abnormal cells in vivo, and are an important part of the innate immune surveillance mechanism. Clinical experience with immunotherapy with NK cells highlights the need for better methods to potently and efficiently expand NK cell populations while maintaining or even enhancing their in vivo functionality (killing capacity, ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/455A61K35/17
CPCA61K35/17A61K38/2086C12N5/0646A61K31/455A61K38/2013A61K45/06C12N2501/2302C12N2501/2315C12N2510/00C12N2500/38A61P31/12A61P35/00A61P35/02A61P37/02A61P37/06A61P43/00A61K2300/00
Inventor 托尼·佩勒德加比·M·弗雷
Owner GAMIDA CELL
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