NK Cell Amplification via T-Cell Depletion and High-Dose IL-2

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Solution Overview

Problem

Current methods for amplifying NK cells for adoptive immunotherapy are limited by low cytotoxic activity and the need for frequent transplants due to insufficient cell numbers, and they often require feeder cells that pose contamination risks.

Innovation Solution

A method involving the removal of T cells and hematopoietic progenitor cells from a cell population, followed by cultivation in a medium supplemented with IL-2, to selectively amplify NK cells without using feeder cells, thereby achieving high purity and cytotoxic activity.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Quantity of substance

If conventional NK cell amplification methods are used, then NK cells can be expanded in vitro, but the cytotoxic activity remains low and insufficient for effective therapy

Engineering Contradiction:
ImproveNK cell numberVSAvoidcytotoxic activity
Core Design Contradiction:
Quantity of substanceVSReliability

Solution Approach 1:

The patent extracts and removes T cells (CD3-positive cells) from the cell population before NK cell amplification. This selective removal eliminates a major source of contamination and allows for pure NK cell expansion with high cytotoxic activity, resolving the contradiction between cell quantity and therapeutic reliability

Inventive Principle:
Principle #2Taking out (Extraction)

Solution Approach 2:

The patent changes the cytokine concentration parameter by using high-dose IL-2 (2,500-2,813 IU/mL) during the amplification phase, which selectively promotes NK cell proliferation while maintaining their cytotoxic potential, thereby achieving both high cell numbers and high therapeutic activity

Inventive Principle:
Principle #35Parameter changes

2Quantity of substance

If feeder cells are used to amplify NK cells, then cell numbers can be increased, but the risk of contamination increases

Engineering Contradiction:
ImproveNK cell numberVSAvoidcontamination risk
Core Design Contradiction:
Quantity of substanceVSObject-affected harmful factors

Solution Approach 1:

The patent completely removes the feeder cell component from the amplification system by using T cell-depleted mononuclear cells as the starting material. This eliminates the source of contamination while still achieving sufficient NK cell expansion through high-dose IL-2 stimulation alone

Inventive Principle:
Principle #2Taking out (Extraction)

Solution Approach 2:

The patent enables NK cells to amplify themselves without external feeder cell support. By removing T cells and using high-dose IL-2, the system allows NK cells to proliferate autonomously, eliminating contamination risk while maintaining amplification efficiency

Inventive Principle:
Principle #25Self-service

3Quantity of substance

If frequent transplants are performed to maintain adequate NK cell numbers, then therapeutic coverage can be maintained, but treatment complexity and burden increase

Engineering Contradiction:
ImproveNK cell numberVSAvoidtreatment frequency
Core Design Contradiction:
Quantity of substanceVSDevice complexity

Solution Approach 1:

The patent performs preliminary amplification of NK cells in vitro to achieve very high cell numbers (sufficient for 1:1 E:T ratio) before transplantation. This pre-expansion allows a single transplant to provide adequate therapeutic coverage for an extended period, eliminating the need for frequent repeat transplants

Inventive Principle:
Principle #10Preliminary action

Applied Scientific Principles

This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.

Function Achieved in This Case

This method results in a significant amplification of NK cells with high cytotoxic activity and purity, reducing the need for frequent transplants and eliminating the risk of feeder cell contamination, effectively enhancing the therapeutic efficacy of NK cells for cancer and infectious disease treatment.

Implementation Method 1

cultivating the remaining cells in a medium supplemented with 2500 to 2813 IU/mL of IL-2

Methodology Applied
Scientific EffectCytokine stimulation:

Implementation Method 2

removing T cells from the cell population which is comprised of NK cells

Methodology Applied
Scientific EffectCell separation:

Data Source

PatentEP2725100B1Method for amplifying NK cells
Publication Date: 2019.09.04 GAIA BIOMEDICINE INC
  • EP2725100B1 patent drawingFigure 1A~1B
  • EP2725100B1 patent drawingFigure 2A~3A
  • EP2725100B1 patent drawingFigure 3B~4B

AI summary

A technique is needed which can amplify NK cells in vitro and prepare optimum number of NK cells for the adoptive immunotherapy. A method for amplifying NK cells is provided which comprises steps of: preparing cell population which is comprised of NK cells, removing T cells from the cell population which is comprised of NK cells, and, after removal of T cells, cultivating the remaining cells in a medium supplemented with 2500 to 2831 IU/mL of IL-2. The method for amplifying NK cells of the present invention may comprise a step of removing hematopoietic progenitor cells from the cell population. The present invention provides a pharmaceutical composition for adoptive immunotherapy, comprising NK cells which are prepared by the amplifying method of the present invention.