PRDM1 decreases sensitivity of human NK cells to IL2-induced cell expansion by directly repressing CD25 (IL2RA)

Akman, Burcu; Hu, Xiaozhou; Liu, Xuxiang; Hatipoğlu, Tevfik; You, Hua; Chan, Wing C.; Küçük, Can

doi:10.1002/jlb.2a0520-321rr

Cited by 11 publications

(8 citation statements)

References 70 publications

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“…In the current study, we noticed that IL-15 was superior to IL-2 in enhancing the NK cell cytotoxicity in vitro with all used protocols, except the continuous IL-2/15 expanded NK cells. The low CD25 expression, as seen here with the IL-2/15 expanded NK cells, has an impact on the NK cells' sensitivity to IL-2 (47,48). This is possibly related to the continuous presence of soluble IL-2, which inhibits the expression of the high affinity IL-2 receptor and its alpha chain (CD25) (47,48).…”

Section: Discussionmentioning

confidence: 66%

Cytokines impact natural killer cell phenotype and functionality against glioblastoma in vitro

Sivonen,

Sirviö,

Wojciechowski

et al. 2023

Front. Immunol.

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ObjectiveNatural killer (NK) cells are a part of the innate immune system and first-line defense against cancer. Since they possess natural mechanisms to recognize and kill tumor cells, NK cells are considered as a potential option for an off-the-shelf allogeneic cell-based immunotherapy. Here, our objective was to identify the optimal cytokine-based, feeder-free, activation and expansion protocol for cytotoxic NK cells against glioblastoma in vitro.MethodsNK cells were enriched from human peripheral blood and expanded for 16 days with different activation and cytokine combinations. The expansion conditions were evaluated based on NK cell viability, functionality, expansion rate and purity. The cytotoxicity and degranulation of the expanded NK cells were measured in vitro from co‑cultures with the glioma cell lines U‑87 MG, U‑87 MG EGFR vIII, LN-229, U-118 and DK-MG. The best expansion protocols were selected from ultimately 39 different conditions: three magnetic cell‑selection steps (Depletion of CD3+ cells, enrichment of CD56+ cells, and depletion of CD3+ cells followed by enrichment of CD56+ cells); four activation protocols (continuous, pre-activation, re-activation, and boost); and four cytokine combinations (IL-2/15, IL‑21/15, IL‑27/18/15 and IL-12/18/15).ResultsThe expansion rates varied between 2-50-fold, depending on the donor and the expansion conditions. The best expansion rate and purity were gained with sequential selection (Depletion of CD3+ cells and enrichment of CD56+ cells) from the starting material and pre-activation with IL‑12/18/15 cytokines, which are known to produce cytokine-induced memory-like NK cells. The cytotoxicity of these memory-like NK cells was enhanced with re-activation, diminishing the donor variation. The most cytotoxic NK cells were produced when cells were boosted at the end of the expansion with IL-12/18/15 or IL-21/15.ConclusionAccording to our findings the ex vivo proliferation capacity and functionality of NK cells is affected by multiple factors, such as the donor, composition of starting material, cytokine combination and the activation protocol. The cytokines modified the NK cells' phenotype and functionality, which was evident in their reactivity against the glioma cell lines. To our knowledge, this is the first comprehensive comparative study performed to this extent, and these findings could be used for upscaling clinical NK cell manufacturing.

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Section: Discussionmentioning

confidence: 66%

Cytokines impact natural killer cell phenotype and functionality against glioblastoma in vitro

Sivonen,

Sirviö,

Wojciechowski

et al. 2023

Front. Immunol.

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“…This analysis revealed that interactions between monocytes and NK cells in severe COVID-19 were predicted to upregulate genes involved in NK cell cytotoxicity [ GZMB ( 48 ), PRF1 ( 49 ), LGALS1 ( 50 ), TNFSF10 ( 51 )], proliferation [ MCM6 ( 52 ), KNTC1 ( 53 )], apoptosis [ ANXA2 ( 54 ), ANXA5 ( 54 ), BAX ( 55 ), FAS ( 56 ), CASP1 ( 57 ), PML ( 58 )], cellular adhesion [ STMN1 ( 59 ), VCAN ( 35 )], and migration [ CX3CR1 ( 60 )]. Other predicted downstream targets in severe COVID-19 included several genes associated with decreased NK cell functionality [ EZH2 ( 61 ), PRDM1 ( 62 ), SERPINB1 ( 63 )], two STAT family transcription factors [ STAT1 , STAT2 ( 64 )], and signaling adaptor MYD88 ( 65 ) ( Fig. 2C ).…”

Section: Resultsmentioning

confidence: 99%

NK Cell–Monocyte Cross-talk Underlies NK Cell Activation in Severe COVID-19

Lee,

de los Rios Kobara,

Barnard

et al. 2024

The Journal of Immunology

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NK cells in the peripheral blood of severe COVID-19 patients exhibit a unique profile characterized by activation and dysfunction. Previous studies have identified soluble factors, including type I IFN and TGF-β, that underlie this dysregulation. However, the role of cell–cell interactions in modulating NK cell function during COVID-19 remains unclear. To address this question, we combined cell–cell communication analysis on existing single-cell RNA sequencing data with in vitro primary cell coculture experiments to dissect the mechanisms underlying NK cell dysfunction in COVID-19. We found that NK cells are predicted to interact most strongly with monocytes and that this occurs via both soluble factors and direct interactions. To validate these findings, we performed in vitro cocultures in which NK cells from healthy human donors were incubated with monocytes from COVID-19+ or healthy donors. Coculture of healthy NK cells with monocytes from COVID-19 patients recapitulated aspects of the NK cell phenotype observed in severe COVID-19, including decreased expression of NKG2D, increased expression of activation markers, and increased proliferation. When these experiments were performed in a Transwell setting, we found that only CD56bright CD16− NK cells were activated in the presence of severe COVID-19 patient monocytes. O-link analysis of supernatants from Transwell cocultures revealed that cultures containing severe COVID-19 patient monocytes had significantly elevated levels of proinflammatory cytokines and chemokines, as well as TGF-β. Collectively, these results demonstrate that interactions between NK cells and monocytes in the peripheral blood of COVID-19 patients contribute to NK cell activation and dysfunction in severe COVID-19.

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“… 14 , 28 , 29 Studies indicated IL-2Rα essential for the sensitivity of human NK cells. 30 Therefore, we speculated that simulating high-affinity IL-2Rαβγc trimeric receptor complexes would benefit NK cell activation and persistence. In parallel, to limit the systemic toxicity of IL-2, we constructed a cell-restricted mbIL-2 by fusion of human IL-2 and human IL-2Rα with a (G 4 S) 3 linker.…”

Section: Discussionmentioning

confidence: 99%

“… 14 , 28 , 29 However, the low-affinity receptor IL-2Rα is essential for the sensitivity of human NK cells. 30 To simulate the biological process of high-affinity trimeric receptor complex in NK cells and avoid the side effects caused by IL-2, we constructed a cell-restricted artificial IL-2, named membrane-bound IL-2 (mbIL-2), by fusion of human IL-2 and human IL-2Rα with a (G 4 S) 3 linker. Here, we demonstrated that mbIL-2 can vigorously improve NK-92 cell survival and proliferation, as well as enhance NK-92 cell-mediated antitumor activity by tuning IL-2 receptor downstream signals and NK cell receptor repertoire expression.…”

Section: Introductionmentioning

confidence: 99%

A novel membrane-bound interleukin-2 promotes NK-92 cell persistence and anti-tumor activity

Xiong

Zhang

et al. 2022

OncoImmunology

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A major challenge in natural killer (NK) cell immunotherapy is the limited persistence of NK cells in vivo . However, the proliferation of NK cells is dependent on cytokines such as interleukin-2 (IL-2). Although IL-2 is a critical cytokine for NK cell activation and survival, IL-2 administration in adoptive NK cell therapy can induce adverse toxicities. To improve the persistence of NK cells and attenuate the systemic toxicity of IL-2, we constructed a cell-restricted artificial IL-2, named membrane-bound IL-2 (mbIL-2), comprising human IL-2 and human IL-2Rα joined by a classic linker. We found that mbIL-2-activated NK-92 cells can survive and proliferate in vitro and in vivo , independent of exogenous IL-2, while mbIL-2-expressing NK-92 cells do not support bystander cell survival or proliferation. Additionally, mbIL-2 enhanced NK-92 cell-mediated antitumor activity by tuning the IL-2 receptor downstream signals and NK cell receptor repertoire expression. To conclude, our novel mbIL-2 improves NK-92 cell persistence and enhances NK-92 cell-mediated antitumor activity. NK-92 cells genetically modified to express the novel mbIL-2 with potential significance for clinical development.

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PRDM1 decreases sensitivity of human NK cells to IL2-induced cell expansion by directly repressing CD25 (IL2RA)

Cited by 11 publications

References 70 publications

Cytokines impact natural killer cell phenotype and functionality against glioblastoma in vitro

Cytokines impact natural killer cell phenotype and functionality against glioblastoma in vitro

NK Cell–Monocyte Cross-talk Underlies NK Cell Activation in Severe COVID-19

A novel membrane-bound interleukin-2 promotes NK-92 cell persistence and anti-tumor activity

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