Optimizing the Procedure to Manufacture Clinical-Grade NK Cells for Adoptive Immunotherapy

Fernández, Adrián; Navarro-Zapata, Alfonso; Escudero, Adela; Matamala, Nerea; Ruz-Caracuel, Beatriz; Mirones, Isabel; Pernas, Alicia; Cobo, Marta; Casado, Gema; Lanzarot, Diego; Rodriguez-Antolín, Carlos; Vela, María; Ferreras, Cristina; Mestre, Carmen; Viejo, Aurora; Leivas, Alejandra; Martı́nez, Javier; Fernández, Lucía; Pérez‐Martínez, Antonio

doi:10.3390/cancers13030577

Cited by 19 publications

(22 citation statements)

References 50 publications

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“…Incubation of PB-derived NK cells with modified K562 cells expressing 4-1BBL and mbIL-15 (K562-mbIL-15.4-1BBL cells) resulted in 1000-fold expansion over a three-week period with no co-stimulation of T lymphocytes [ 45 ]. Similar results were reported using K562-mbIL-21.4-1BBL cells [ 46 ]. Yet another approach is to genetically modify HLA-A- and HLA-B-negative K562 cells to express CD48, 4-1BBL and mbIL-21.…”

Section: Production Process Of Car-nk Cellssupporting

confidence: 90%

CAR-NK Cells in the Treatment of Solid Tumors

Wrona

Borowiec

Potemski

2021

IJMS

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CAR-T (chimeric antigen receptor T) cells have emerged as a milestone in the treatment of patients with refractory B-cell neoplasms. However, despite having unprecedented efficacy against hematological malignancies, the treatment is far from flawless. Its greatest drawbacks arise from a challenging and expensive production process, strict patient eligibility criteria and serious toxicity profile. One possible solution, supported by robust research, is the replacement of T lymphocytes with NK cells for CAR expression. NK cells seem to be an attractive vehicle for CAR expression as they can be derived from multiple sources and safely infused regardless of donor–patient matching, which greatly reduces the cost of the treatment. CAR-NK cells are known to be effective against hematological malignancies, and a growing number of preclinical findings indicate that they have activity against non-hematological neoplasms. Here, we present a thorough overview of the current state of knowledge regarding the use of CAR-NK cells in treating various solid tumors.

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Section: Production Process Of Car-nk Cellssupporting

confidence: 90%

CAR-NK Cells in the Treatment of Solid Tumors

Wrona

Borowiec

Potemski

2021

IJMS

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“…The median content of NK cells in seeded PBMCs was 11.5% (range 4.2-29.4%), with a median absolute number of NK cells of 12.0 × 10 6 (range 5.8-44.1 × 10 6 ). The median duration of expansion was 18 days (range [14][15][16][17][18][19][20][21][22][23][24][25]. At the end of culturing, the total number of yielded live cells was 2.26 × 10 9 (0.89-5.5 × 10 9 ).…”

Section: Growth Kinetics Of Pbmc Subsetsmentioning

confidence: 99%

Phenotypic and functional characterisation of locally produced natural killer cells ex vivo expanded with the K562-41BBL-mbIL21 cell line

Шман

Vashkevich

Migas

et al. 2022

Preprint

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We characterise the expansion, phenotype and functional activity of natural killer (NK) cells obtained for a linical trial. Nineteen expansion procedures were performed to obtain NK cell products for 16 patients. NK cells were ex vivo expanded from haploidentical donor peripheral blood mononuclear cells in the presence of the locally generated feeder cell line K-562 with ectopic expression of 4-1BBL and mbIL-21.The median duration of expansion was 18 days (range 14–25). The median number of live cells yielded was 2.26 × 109 (range 0.89–5.5 × 109) with an NK content of 96.6% (range 89.0%–98.8%). The median NK cell fold expansion was 224.7 (range 42–647). The majority of expanded NK cells had the phenotype of immature activated cells (NKG2A+, double bright CD56++CD16++, CD57-) expressing NKp30, NKp44, NKp46, NKG2D, CD69, HLA-DR and CD96. Despite the expression of exhaustion markers, expanded NK cells exhibited high cytolytic activity against leukaemia cell lines, high degranulation activity and production of cytokines. There was noted decreased functional activity of NK cells in tests against the patient’s blasts.NK cells obtained by ex vivo expansion with locally generated K562-41BBL-mbIL21 cells have both a relatively undifferentiated phenotype and enhanced cytolytic activity against cancer cell lines. Expansion of NK cells with the feeder cells allows obtaining a sufficient quantity of the NK cell product to reach high cell doses or increase the frequency of cell infusions for adoptive immunotherapy. Registered at clinicaltrials.gov as NCT04327037.

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“…Although large-scale generation of GMP-grade products is possible using serum-free commercial media such as AIMV media (Life Technologies, USA), X-VIVO media (BioWhittaker, Belgium) and SCGM media (CellGenix, Germany) [ 91 ], there are no standard manufacturing techniques because of the variation of culture conditions throughout clinical trials. In an optimization study of GMP-compliant manufacturing methods in CliniMACS Prodigy, NK MACS and TexMACs media achieved the highest NK cell purity; the highest fold of expansion was achieved by K562mbIL21-41BBL cells [ 92 ].…”

Section: Challenges In Manufacturing Of Clinical-grade Nk and Car-nk Cell Therapies And Strategies To Overcome Themmentioning

confidence: 99%

New Orders to an Old Soldier: Optimizing NK Cells for Adoptive Immunotherapy in Hematology

Gündüz

Atilla

2021

Biomedicines

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NK (Natural Killer) cell-mediated adoptive immunotherapy has gained attention in hematology due to the progressing knowledge of NK cell receptor structure, biology and function. Today, challenges related to NK cell expansion and persistence in vivo as well as low cytotoxicity have been mostly overcome by pioneering trials that focused on harnessing NK cell functions. Recent technological advancements in gene delivery, gene editing and chimeric antigen receptors (CARs) have made it possible to generate genetically modified NK cells that enhance the anti-tumor efficacy and represent suitable “off-the-shelf” products with fewer side effects. In this review, we highlight recent advances in NK cell biology along with current approaches for potentiating NK cell proliferation and activity, redirecting NK cells using CARs and optimizing the procedure to manufacture clinical-grade NK and CAR NK cells for adoptive immunotherapy.

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Optimizing the Procedure to Manufacture Clinical-Grade NK Cells for Adoptive Immunotherapy

Cited by 19 publications

References 50 publications

CAR-NK Cells in the Treatment of Solid Tumors

CAR-NK Cells in the Treatment of Solid Tumors

Phenotypic and functional characterisation of locally produced natural killer cells ex vivo expanded with the K562-41BBL-mbIL21 cell line

New Orders to an Old Soldier: Optimizing NK Cells for Adoptive Immunotherapy in Hematology

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