WO2020143042A1 - Procédé de restauration de fonction cellulaire - Google Patents

Procédé de restauration de fonction cellulaire Download PDF

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Publication number
WO2020143042A1
WO2020143042A1 PCT/CN2019/071416 CN2019071416W WO2020143042A1 WO 2020143042 A1 WO2020143042 A1 WO 2020143042A1 CN 2019071416 W CN2019071416 W CN 2019071416W WO 2020143042 A1 WO2020143042 A1 WO 2020143042A1
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WO
WIPO (PCT)
Prior art keywords
cells
ctl
nkg2d
ctl cells
restoring
Prior art date
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Ceased
Application number
PCT/CN2019/071416
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English (en)
Chinese (zh)
Inventor
周双念
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Shenzhen Shuangke Biotechnology Co Ltd
Original Assignee
Shenzhen Shuangke Biotechnology Co Ltd
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Shenzhen Shuangke Biotechnology Co Ltd filed Critical Shenzhen Shuangke Biotechnology Co Ltd
Priority to PCT/CN2019/071416 priority Critical patent/WO2020143042A1/fr
Publication of WO2020143042A1 publication Critical patent/WO2020143042A1/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
    • C12N5/06—Animal cells or tissues; Human cells or tissues
    • C12N5/0602—Vertebrate cells
    • C12N5/0634—Cells from the blood or the immune system
    • C12N5/0636—T lymphocytes
    • C12N5/0638—Cytotoxic T lymphocytes [CTL] or lymphokine activated killer cells [LAK]
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2502/00—Coculture with; Conditioned medium produced by
    • C12N2502/11—Coculture with; Conditioned medium produced by blood or immune system cells
    • C12N2502/1164—NK cells
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2510/00—Genetically modified cells

Definitions

  • the invention relates to the field of cell separation and combination, in particular to a method for restoring cell performance.
  • CTL cells cytotoxic T lymphocytes
  • the object of the present invention is to provide a method for restoring the performance of cells to restore the performance of CTL cells.
  • a method to restore cell performance including:
  • NKg2d is isolated from the NK cells
  • chimerizing the NKg2d with the CTL cell includes:
  • the NKg2d was chimeric on the surface of the CTL cells.
  • the method further includes:
  • NKg2d in CTL cells of the sample is lower than a threshold
  • NKg2d in the CTL cells is lower than the threshold, and the expression value of the MHC-1 is within a preset range, then NK cells and CTL cells are extracted.
  • the method further includes:
  • the NK cells and the CTL cells are separated, cultivated and expanded.
  • NKg2d is isolated from NK cells (natural killer cells), and NKg2d is chimeric with CTL cells, thereby restoring the ability of CTL cells to recognize and kill abnormal cells.
  • FIG. 1 is a flowchart of a method for restoring cell performance provided by an embodiment of the present invention.
  • the method for restoring cell performance includes:
  • Step S101 Extract NK cells and CTL cells in the same body.
  • NK cells and CTL cells are from the same body to ensure the success rate of NKg2d and CTL cell chimerism.
  • the method further includes: detecting whether NKg2d in the CTL cells of the sample is lower than a threshold; detecting whether the MHC-1 expression value of the CTL cells in the sample is within a preset range; if the CTL cells NKg2d in is lower than the threshold or NKg2d falls off, and the expression value of the MHC-1 is within a preset range, then NK cells and CTL cells are extracted, and then NK cells and CTL cells are extracted.
  • the sample is generally a sample containing tumor and virus cells.
  • NK cells are in the state of normally expressing NKg2d or less expressing NKg2d. NK cells can be isolated from NKg2d. If the NKg2d on the surface of CTL cells falls off to a preset value, it will affect the CTL cells. Identification and killing of tumor and virus cells. First check whether the NKg2d in the CTL cells in the sample is below the threshold. If it is below the threshold or NKg2d falls off, the CTL cells need to be repaired to extract the NK cells and CTL cells in the sample.
  • Step S102 Isolate NKg2d from the NK cells.
  • the cells need to be cultured, and the NK cells and CTL cells are allowed to stand for a preset time to separate, cultivate, and expand the cells, so as to extract the desired NKg2d.
  • Step S103 chimeric the NKg2d with the CTL cells.
  • Chimerizing NKg2d with CTL cells includes: chimerizing NKg2d on the surface of the CTL cells so that NKg2d does not miss the target on the surface of the CTL cells, so as to restore the performance of the CTL cells and restore the identification and lethality of abnormal cells.
  • the method for restoring cell performance isolates NKg2d from NK cells (natural killer cells), and chimerizes NKg2d with CTL cells to restore the performance of CTL cells.

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  • Health & Medical Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Immunology (AREA)
  • Biomedical Technology (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Zoology (AREA)
  • Organic Chemistry (AREA)
  • Biotechnology (AREA)
  • Chemical & Material Sciences (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Genetics & Genomics (AREA)
  • Wood Science & Technology (AREA)
  • Microbiology (AREA)
  • Cell Biology (AREA)
  • Biochemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Hematology (AREA)
  • Medicines Containing Material From Animals Or Micro-Organisms (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)

Abstract

L'invention concerne un procédé de restauration de fonction cellulaire consistant à collecter des cellules NK et des cellules CTL du même organisme, à séparer NKg2d des cellules NK, et à réaliser une chimérisation de NKg2d et des cellules CTL, ce qui permet de restaurer la capacité des cellules CTL à reconnaître et à tuer des cellules anormales.
PCT/CN2019/071416 2019-01-11 2019-01-11 Procédé de restauration de fonction cellulaire Ceased WO2020143042A1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
PCT/CN2019/071416 WO2020143042A1 (fr) 2019-01-11 2019-01-11 Procédé de restauration de fonction cellulaire

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
PCT/CN2019/071416 WO2020143042A1 (fr) 2019-01-11 2019-01-11 Procédé de restauration de fonction cellulaire

Publications (1)

Publication Number Publication Date
WO2020143042A1 true WO2020143042A1 (fr) 2020-07-16

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Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/CN2019/071416 Ceased WO2020143042A1 (fr) 2019-01-11 2019-01-11 Procédé de restauration de fonction cellulaire

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WO (1) WO2020143042A1 (fr)

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2010141093A2 (fr) * 2009-06-04 2010-12-09 The University Of Maryland, Baltimore Méthodes de co-signalement permettant de traiter les cancers
CN104293815A (zh) * 2014-09-18 2015-01-21 扬州大学 一种纳米基因疫苗及其制备方法和应用
US20150299252A1 (en) * 2010-11-01 2015-10-22 Susavion Biosciences, Inc. Compositions and methods for modulating innate and adaptive immune systems
CN108048404A (zh) * 2017-12-08 2018-05-18 新乡医学院 一种新型抗肿瘤nk细胞及其制备方法和应用

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2010141093A2 (fr) * 2009-06-04 2010-12-09 The University Of Maryland, Baltimore Méthodes de co-signalement permettant de traiter les cancers
US20150299252A1 (en) * 2010-11-01 2015-10-22 Susavion Biosciences, Inc. Compositions and methods for modulating innate and adaptive immune systems
CN104293815A (zh) * 2014-09-18 2015-01-21 扬州大学 一种纳米基因疫苗及其制备方法和应用
CN108048404A (zh) * 2017-12-08 2018-05-18 新乡医学院 一种新型抗肿瘤nk细胞及其制备方法和应用

Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
ARTHUR I. ROBERTS, LEANNE LEE, ELIEZER SCHWARZ, VERONIKA GROH, THOMAS SPIES, ELLEN C. EBERT AND BANA JABRI: "Cutting Edge: NKG2D Receptors Induced by IL-15 Costimulate CD28-negative Effector CTL in the Tissue Microenvironment.", JOURNAL OF IMMUNOLOGY, vol. 167, no. 10, 2001115, pages 5527 - 5530, XP055683855, ISSN: 0022-1767, DOI: 10.4049/jimmunol.167.10.5527 *
MARINE CHAMPSAUR , LEWIS L. LANIER: "Effect of NKG2D Ligand Expression on Host Immune Responses.", IMMUNOLOGICAL REVIEWS, vol. 235, 31 December 2010 (2010-12-31), XP055618469, ISSN: 0105-2896, DOI: 10.1111/j.0105-2896.2010.00893.x *
MARY A. MARKIEWICZ, LEONIDAS N. CARAYANNOPOULOS, OLGA V. NAIDENKO, KEN MATSUI, W. RICHARD BURACK, ERICA L. WISE, DAVED H. FREMONT,: "Costimulation through NKG2D Enhances Murine CD8+ CTL Function: Similarities and Differences between NKG2D and CD28 Costimulation.", JOURNAL OF IMMUNOLOGY, vol. 175, no. 5, 1 September 2005 (2005-09-01), XP055718717, ISSN: 0022-1767, DOI: 10.4049/jimmunol.175.5.2825 *
MERESSE BERTRAND , CHEN ZHANGGUO, CISZEWSKI CEZARY, TRETIAKOVA MARIA, BHAGAT GOVIND, KRAUSZ THOMAS N, RAULET DAVID H, LANIER LEWI: "Coordinated Induction by IL15 of a TCR-independent NKG2D Signaling Pathway Converts CTL into Lymphokine-activated Killer Cells in Celiac Disease.", IMMUNITY, vol. 21, no. 3, 30 September 2004 (2004-09-30), pages 357 - 366, XP055718714, DOI: 10.1016/j.immuni.2004.06.020 *

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