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Discovery of a novel natural killer cell line with distinct immunostimulatory and proliferative potential as an alternative platform for cancer immunotherapy
- Source :
- Journal for ImmunoTherapy of Cancer, Vol 7, Iss 1, Pp 1-17 (2019), Journal for Immunotherapy of Cancer
- Publication Year :
- 2019
- Publisher :
- BMJ Publishing Group, 2019.
-
Abstract
- Background Human natural killer (NK) cell lines serve as an attractive source for adoptive immunotherapy, but NK-92 remains the only cell line being assessed in the clinic. Here, we established a novel NK cell line, NK101, from a patient with extra-nodal natural killer/T-cell lymphoma and examined its phenotypic, genomic and functional characteristics. Methods Single cell suspensions from lymphoma tissue were expanded with anti-NKp46/anti-CD2-coated beads in the presence of IL-2. A continuously growing CD56+ cell clone was selected and designated as NK101. Flow cytometry and RNA sequencing were used to characterize phenotypic and genomic features of NK101. In vitro cytotoxicity and IFN-γ/TNF-α secretion were measured by flow cytometry-based cytotoxicity assay and enzyme-linked immunosorbent assay, respectively, after direct co-culture with tumor cells. Immunomodulatory potential of NK101 was assessed in an indirect co-culture system using conditioned medium. Finally, in vivo antitumor efficacy was evaluated in an immunocompetent, syngeneic 4T1 mammary tumor model. Results NK101 displayed features of CD56dimCD62L+ intermediate stage NK subset with the potential to simultaneously act as a cytokine producer and a cytotoxic effector. Comparative analysis of NK101 and NK-92 revealed that NK101 expressed lower levels of perforin and granzyme B that correlated with weaker cytotoxicity, but produced higher levels of pro-inflammatory cytokines including IFN-γ and TNF-α. Contrarily, NK-92 produced greater amounts of anti-inflammatory cytokines, IL-1 receptor antagonist and IL-10. Genome-wide analysis revealed that genes associated with positive regulation of leukocyte proliferation were overexpressed in NK101, while those with opposite function were highly enriched in NK-92. The consequence of such expressional and functional discrepancies was well-represented in (i) indirect co-culture system where conditioned medium derived from NK101 induced greater proliferation of human peripheral blood mononuclear cells and (ii) immunocompetent 4T1 tumor model where peritumoral injections of NK101 displayed stronger anti-tumor activities by inducing higher tumor-specific immune responses. In a manufacturing context, NK101 not only required shorter recovery time after thawing, but also exhibited faster growth profile than NK-92, yielding more than 200-fold higher cell numbers after 20-day culture. Conclusion NK101 is a unique NK cell line bearing strong immunostimulatory potential and substantial scalability, providing an attractive source for adoptive cancer immunotherapy. Electronic supplementary material The online version of this article (10.1186/s40425-019-0612-2) contains supplementary material, which is available to authorized users.
- Subjects :
- 0301 basic medicine
Cytotoxicity, Immunologic
Male
Cancer Research
medicine.medical_treatment
Immunology
NK cell line
Biology
lcsh:RC254-282
Natural killer cell
Flow cytometry
03 medical and health sciences
Mice
0302 clinical medicine
Immune system
Cancer immunotherapy
Cell Line, Tumor
Neoplasms
medicine
Immunology and Allergy
Cytotoxic T cell
Leukocyte proliferation
Animals
Humans
Cell Proliferation
Pharmacology
Mice, Inbred BALB C
medicine.diagnostic_test
Immunostimulatory potential
Middle Aged
lcsh:Neoplasms. Tumors. Oncology. Including cancer and carcinogens
Adoptive Transfer
Granzyme B
030104 developmental biology
medicine.anatomical_structure
Oncology
Cell culture
030220 oncology & carcinogenesis
Adoptive cancer immunotherapy
Cancer research
Molecular Medicine
Female
Immunotherapy
Off-the-shelf platform
Research Article
Subjects
Details
- Language :
- English
- ISSN :
- 20511426
- Volume :
- 7
- Issue :
- 1
- Database :
- OpenAIRE
- Journal :
- Journal for ImmunoTherapy of Cancer
- Accession number :
- edsair.doi.dedup.....6ddbf35cd31666ad3899a96a62ae17cf