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Methods and compositions of natural killer cells

A composition and cell technology, applied in the field of natural killer cells and compositions, can solve problems such as the worsening outcome of GVHD

Active Publication Date: 2021-02-12
UNIV OF CENT FLORIDA RES FOUND INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, clinical benefit is limited and observed only in a minority of patients with a small tumor burden, and T cell-mediated GVHD often further worsens outcome

Method used

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  • Methods and compositions of natural killer cells
  • Methods and compositions of natural killer cells
  • Methods and compositions of natural killer cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0135] Example 1: Expansion of NK cells with crude exosome preparations

[0136] result

[0137] As demonstrated herein, expansion of NK cells can occur by using exosomes derived from K562-mb21-41BBL stimulating cell cultures. Cultures of K562-mb21-41BBL were selected for exosome isolation because these cells are reported to expand NK cells very robustly and do not require isolation of NK cells from peripheral blood mononuclear cell (PBMC) mixtures prior to culture initiation . Furthermore, the presence of the stimulatory ligands 41BBL and mbIL21 could be easily followed by antibody staining to confirm the expression of these molecules on feeder cells and their presence in isolated exosomes.

[0138] This experiment was performed to test whether exosomes derived from stimulator cell cultures support the expansion of NK cells in a similar manner as stimulator cells. When PBMC were exposed in culture to 50 U / mL of IL-2 over a period of 24 days and reduced the concentratio...

Embodiment 2

[0142] Example 2: Characterization of exosomes

[0143] NK cell-stimulating exosomes were isolated from cultures of K562-mb21-41bbl cells. Cells were grown to approximately 1 x 10 6cells / mL, washed, resuspended in serum-free RPMI and treated with 2 μM monensin. Cells were removed by centrifugation at 1000 xg for 10 minutes and then filtered through a 0.22 μm PES membrane. The filtered medium was then concentrated by ultrafiltration using a 100,000 KDa MWCO membrane.

[0144] Figure 2A to Figure 2D Characterization of exosomes isolated from K562-mb21-41bbl cell cultures is shown. Exosomes were suspended in PBS and characterized by nanoparticle tracking analysis (NTA) with a NanoSight NS300 (Malvern) video microscope. NTA determines particle size based on the analysis of light scattering intensity and diffusion kinetics. A single frame of light scattering image is shown ( Figure 2A ) and the grouped histogram of particle size distribution ( Figure 2B ). The presenc...

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Abstract

The present invention discloses novel compositions and methods for stimulating and generating or expanding natural killer (NK) cells. The number of NK cells can be increased following contact with exosomes modified with one or more stimulatory peptides. The present invention also describes methods and compositions for producing exosomes, wherein the exosomes include stimulating peptides. The present invention also describes methods of treating cancer using the disclosed NK stimulating exosomes or NK cells stimulated by the disclosed methods.

Description

[0001] Cross references to related patent applications [0002] This application claims the benefit of US Provisional Application 62 / 069,057, filed October 27, 2014, which is hereby incorporated by reference in its entirety. technical field [0003] The present application relates generally to compositions and methods comprising natural killer (NK) cells. More specifically, the present application relates to the in vivo, ex vivo or in vitro stimulation and expansion of endogenous natural killer (NK) cells capable of attacking and killing cancer cells, virus-infected cells and certain immune cells. Background technique [0004] Hematopoietic stem cell transplantation (HSCT) from genetically HLA-matched siblings has improved long-term survival in patients with hematologic malignancies and bone marrow failure syndromes. Each year, more than 10,000 Americans develop life-threatening diseases for which the only hope of cure is a bone marrow transplant from an unrelated donor or ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K47/64A61K48/00C12N15/88
CPCC07K14/54C12N5/0646C12N2500/84C07K14/5434C07K14/55C07K14/70525C07K14/70596A61K38/177A61K38/20A61K38/2013A61P31/12A61P35/00A61P37/02A61K35/17C12N2501/2321A61K35/13C12N2501/599A61K2035/124C12N2501/2302
Inventor A·J·科皮克J·L·奥耶R·Y·五十岚D·阿尔托马尔
Owner UNIV OF CENT FLORIDA RES FOUND INC