IP Library › Granted Patent US 12,473,336
Granted Patent B2
US 12,473,336 · App. 16/970,937 · Granted Nov 18, 2025

Methods for activation and expansion of natural killer cells and uses thereof

Inventors: Lucila Nassif Kerbauy (Houston, TX); Elizabeth Shpall (Houston, TX); Katy Rezvani (Houston, TX)
Assignee: Board of Regents, The University of Texas System
C07K14/5434A61K40/15A61K40/31A61K40/35A61K40/4211A61K40/4217C07K14/5443C07K14/55C07K14/70592C07K14/70596C12N5/0646
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Quick Facts
Patent No.
US 12,473,336
App. No.
16/970,937
Granted
Nov 18, 2025
Kind
B2
Abstract

Provided herein are methods for pre-activating and expanding an isolated population of NK cells. Further provided herein are methods for the treatment of cancer by administering the pre-activated and expanded NK cells.

Claims (19)

1 . An in vitro method for expanding natural killer (NK) cells comprising:

(a) pre-activating a population of cord blood (CB) NK cells in a pre-activation culture comprising an effective concentration of IL-12, IL-15, and IL-18 to obtain pre-activated NK cells; and then

(b) expanding the pre-activated NK cells in an expansion culture comprising artificial antigen presenting cells (aAPCs) expressing CD137 ligand, thereby producing expanded NK cells,

wherein the expanded NK cells display enhanced antibody-dependent cellular cytotoxicity (ADCC) when compared with a control.

2 . The method of claim 1 , wherein the CB is pooled from 2 or more individual cord blood units.

3 . The method of claim 1 , wherein the population of NK cells are further defined as CD56 + NK cells.

4 . The method of claim 1 , wherein the aAPCs further express a membrane-bound cytokine.

5 . The method of claim 4 , wherein the membrane-bound cytokine is membrane-bound IL-21 (mIL-21) or membrane-bound IL-15 (mIL-15).

6 . The method of claim 1 , wherein the aAPCs have no expression of endogenous HLA class I, II, or CD1d molecules.

7 . The method of claim 1 , wherein the aAPCs express ICAM-1 (CD54) and LFA-3 (CD58).

8 . The method of claim 1 , wherein the aAPCs are further defined as leukemia cell-derived aAPCs.

9 . The method of claim 8 , wherein the leukemia-cell derived aAPCs are K562 cells engineered to express CD137 ligand, mIL-21, or both CD137 and mIL-21.

10 . The method of claim 9 , wherein the K562 cells are engineered to express CD137 ligand and mIL-21.

11 . The method of claim 9 , wherein the aAPCs have been engineered by retroviral transduction.

12 . The method of claim 1 , wherein the aAPCs are irradiated.

13 . The method of claim 1 , wherein the pre-activating step is for 10-20 hours.

14 . The method of claim 1 , wherein the pre-activated NK cells and aAPCs are present in the expansion culture at a ratio of 3:1 to 1:3.

15 . The method of claim 1 , wherein the expansion culture further comprises IL-2.

16 . The method of claim 1 , wherein the NK cells are engineered to express a chimeric antigen receptor (CAR).

Assignments (1)
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Aug 18, 2020
From: KERBAUY, LUCILA NASSIF; SHPALL, ELIZABETH; REZVANI, KATY
To: BOARD OF REGENTS, THE UNIVERSITY OF TEXAS SYSTEM
Reel/Frame 053531/0722 →
Continuity (2)
Provisional Application 62633592 · Feb 21, 2018
Related Publication 20200390816A1 · Dec 17, 2020
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