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Natural Killer Cells Generated From Human Induced Pluripotent Stem Cells Mature to CD56 bright CD16 + NKp80 +/- In-Vitro and Express KIR2DL2/DL3 and KIR3DL1.
- Source :
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Frontiers in immunology [Front Immunol] 2021 May 04; Vol. 12, pp. 640672. Date of Electronic Publication: 2021 May 04 (Print Publication: 2021). - Publication Year :
- 2021
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Abstract
- The differentiation of human induced pluripotent stem cells (hiPSCs) into T and natural killer (NK) lymphocytes opens novel possibilities for developmental studies of immune cells and in-vitro generation of cell therapy products. In particular, iPSC-derived NK cells gained interest in adoptive anti-cancer immunotherapies, since they enable generation of homogenous populations of NK cells with and without genetic engineering that can be grown at clinical scale. However, the phenotype of in-vitro generated NK cells is not well characterized. NK cells derive in the bone marrow and mature in secondary lymphoid tissues through distinct stages from CD56 <superscript>bright</superscript> CD16 <superscript>-</superscript> to CD56 <superscript>dim</superscript> CD16 <superscript>+</superscript> NK cells that represents the most abandoned population in peripheral blood. In this study, we efficiently generated CD56 <superscript>+</superscript> CD16 <superscript>+</superscript> CD3 <superscript>-</superscript> NK lymphocytes from hiPSC and characterized NK-cell development by surface expression of NK-lineage markers. Hematopoietic priming of hiPSC resulted in 31.9% to 57.4% CD34 <superscript>+</superscript> CD45 <superscript>+</superscript> hematopoietic progenitor cells (HPC) that did not require enrichment for NK lymphocyte propagation. HPC were further differentiated into NK cells on OP9-DL1 feeder cells resulting in high purity of CD56 <superscript>bright</superscript> CD16 <superscript>-</superscript> and CD56 <superscript>bright</superscript> CD16 <superscript>+</superscript> NK cells. The output of generated NK cells increased up to 40% when OP9-DL1 feeder cells were inactivated with mitomycine C. CD7 expression could be detected from the first week of differentiation indicating priming towards the lymphoid lineage. CD56 <superscript>bright</superscript> CD16 <superscript>-/+</superscript> NK cells expressed high levels of DNAM-1, CD69, natural killer cell receptors NKG2A and NKG2D, and natural cytotoxicity receptors NKp46, NKp44, NKp30. Expression of NKp80 on 40% of NK cells, and a perforin <superscript>+</superscript> and granzyme B <superscript>+</superscript> phenotype confirmed differentiation up to stage 4b. Killer cell immunoglobulin-like receptor KIR2DL2/DL3 and KIR3DL1 were found on up to 3 and 10% of mature NK cells, respectively. NK cells were functional in terms of cytotoxicity, degranulation and antibody-dependent cell-mediated cytotoxicity.<br />Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.<br /> (Copyright © 2021 Euchner, Sprissler, Cathomen, Fürst, Schrezenmeier, Debatin, Schwarz and Felgentreff.)
- Subjects :
- CD56 Antigen immunology
Cell Culture Techniques methods
Cell Degranulation immunology
GPI-Linked Proteins immunology
Humans
Induced Pluripotent Stem Cells cytology
Induced Pluripotent Stem Cells immunology
Killer Cells, Natural cytology
Lymphocyte Activation immunology
Lymphocyte Subsets cytology
Receptors, IgG immunology
Receptors, KIR2DL2 immunology
Receptors, KIR2DL3 immunology
Receptors, KIR3DL1 immunology
Cell Differentiation immunology
Killer Cells, Natural immunology
Lymphocyte Subsets metabolism
Subjects
Details
- Language :
- English
- ISSN :
- 1664-3224
- Volume :
- 12
- Database :
- MEDLINE
- Journal :
- Frontiers in immunology
- Publication Type :
- Academic Journal
- Accession number :
- 34017328
- Full Text :
- https://doi.org/10.3389/fimmu.2021.640672