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Intrinsic Functional Potential of NK-Cell Subsets Constrains Retargeting Driven by Chimeric Antigen Receptors
- Source :
- Cancer Immunology Research
- Publication Year :
- 2018
- Publisher :
- American Association for Cancer Research (AACR), 2018.
-
Abstract
- Natural killer (NK) cells hold potential as a source of allogeneic cytotoxic effector cells for chimeric antigen receptor (CAR)-mediated therapies. Here, we explored the feasibility of transfecting CAR-encoding mRNA into primary NK cells and investigated how the intrinsic potential of discrete NK-cell subsets affects retargeting efficiency. After screening five second- and third-generation anti-CD19 CAR constructs with different signaling domains and spacer regions, a third-generation CAR with the CH2-domain removed was selected based on its expression and functional profiles. Kinetics experiments revealed that CAR expression was optimal after 3 days of IL15 stimulation prior to transfection, consistently achieving over 80% expression. CAR-engineered NK cells acquired increased degranulation toward CD19+ targets, and maintained their intrinsic degranulation response toward CD19− K562 cells. The response of redirected NK-cell subsets against CD19+ targets was dependent on their intrinsic thresholds for activation determined through both differentiation and education by killer cell immunoglobulin-like receptors (KIR) and/or CD94/NKG2A binding to self HLA class I and HLA-E, respectively. Redirected primary NK cells were insensitive to inhibition through NKG2A/HLA-E interactions but remained sensitive to inhibition through KIR depending on the amount of HLA class I expressed on target cells. Adaptive NK cells, expressing NKG2C, CD57, and self-HLA–specific KIR(s), displayed superior ability to kill CD19+, HLA low, or mismatched tumor cells. These findings support the feasibility of primary allogeneic NK cells for CAR engineering and highlight a need to consider NK-cell diversity when optimizing efficacy of cancer immunotherapies based on CAR-expressing NK cells. Cancer Immunol Res; 6(4); 467–80. ©2018 AACR.
- Subjects :
- Cytotoxicity, Immunologic
0301 basic medicine
Cancer Research
Immunology
Receptors, Antigen, T-Cell
Gene Expression
Human leukocyte antigen
Lymphocyte Activation
CD19
Mice
03 medical and health sciences
Receptors, KIR
Antigen
Antigens, Neoplasm
HLA Antigens
Cell Line, Tumor
Animals
Humans
Cytotoxic T cell
Receptors, Chimeric Antigen
biology
Chemistry
Degranulation
Transfection
Lymphocyte Subsets
Chimeric antigen receptor
Cell biology
Killer Cells, Natural
Electroporation
030104 developmental biology
biology.protein
NK Cell Lectin-Like Receptor Subfamily C
Subjects
Details
- ISSN :
- 23266074 and 23266066
- Volume :
- 6
- Database :
- OpenAIRE
- Journal :
- Cancer Immunology Research
- Accession number :
- edsair.doi.dedup.....6f597f15119efb89acff003de8dd2fcd