Expression level of BTN3A1 on the surface of CD14+ monocytes is a potential predictor of γδ T cell expansion efficiency

Tomogane, Mako; Omura, Maho; Sano, Yusuke; Shimizu, Daiki; Toda, Yuki; Hosogi, Shigekuni; Kimura, Shinya; Ashihara, Eishi

doi:10.1016/j.bbrc.2021.12.060

Cited by 5 publications

(3 citation statements)

References 27 publications

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“…Peripheral blood mononuclear cells (PBMCs) were isolated using the Ficoll-Paque (GE Healthcare, Little Chalfont, Bucks, UK) density gradient centrifugation. The γδ T cells were expanded from cultured PBMCs using ZOL and rhIL-2, as described previously (15). We ensured that the proportion of γδ T cells within the expanded PBMC preparations exceeded 90% prior to use in experiments.…”

Section: Methodsmentioning

confidence: 99%

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Hypoxia-adapted Multiple Myeloma Stem Cells Resist γδ-T-Cell-mediated Killing by Modulating the Mevalonate Pathway

et al. 2023

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Background/Aim: The prognosis of patients with multiple myeloma (MM) has recently improved due to the emergence of new molecular targeting agents. However, MM remains incurable because MM stem cells are resistant to these agents. Therefore, it is essential to develop strategies to eradicate MM stem cells. We have previously demonstrated that MM cells cultured under prolonged hypoxic conditions (1% O 2 ) (i. e., hypoxia-adapted MM cells; MM-HA cells) exhibited stem-celllike characteristics. γδ T cells attack tumor cells by recognizing butyrophilin (BTN) 3A1 and BTN2A1, which are activated by the intracellular accumulation of isopentenyl pyrophosphate (IPP), an intermediate in the mevalonate pathway. In the present study, we investigated the cytotoxicity of γδ T cells against MM-HA stem-like cells. Materials and Methods: We used a combination of flow cytometry, liquid chromatography-tandem mass spectrometry, and western blotting methods to investigate the cytotoxicity of γδ T cells against MM-HA cells and measured the amounts of IPP in MM-HA cells and their supernatants.Results: The cytotoxicity of γδ T cells against MM-HA cells was significantly lower than that against MM cells cultured under normoxic conditions (20% O 2 ; MM-Normo). Furthermore, the concentration of IPP in MM-HA cells was lower than that in MM-Normo cells. The expression of mevalonate decarboxylase and farnesyl diphosphate synthase proteins were decreased in MM-HA-cells. Conclusion: The cytotoxicity of γδ T cells against MM-HA cells was suppressed by the reduced IPP accumulation by modulating the mevalonate pathway in MM-HA cells.

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Section: Methodsmentioning

confidence: 99%

“…Western blotting. Preparation of MM cells lysates was performed as described previously (15). Samples were blotted with primary antibodies against the following proteins: mevalonate decarboxylase (MVD; #sc-376975, Santa Cruz Biotechnology, Dallas, TX, USA), FDPS/FPS (#ab189874, Abcam, Cambridge, UK), and β-actin (#A5441, Sigma-Aldrich).…”

Section: Methodsmentioning

confidence: 99%

Hypoxia-adapted Multiple Myeloma Stem Cells Resist γδ-T-Cell-mediated Killing by Modulating the Mevalonate Pathway

et al. 2023

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“…In the PBMCs, there are monocytes that can function as the phosphoantigen-containing cells ( Roelofs et al., 2009 ; Tomogane et al., 2022 ). In the co-cultures, most human cells lines that we have evaluated can function in this assay.…”

Section: Before You Beginmentioning

confidence: 99%

Generation of effector Vγ9Vδ2 T cells and evaluation of their response to phosphoantigen-loaded cells

Hsiao

Wiemer

2022

STAR Protocols

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Glycolysis inhibition affects proliferation and cytotoxicity of Vγ9Vδ2 T cells expanded for adoptive cell therapy

Aehnlich,

Santiago,

Dam

et al. 2024

Cytotherapy

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Expression level of BTN3A1 on the surface of CD14+ monocytes is a potential predictor of γδ T cell expansion efficiency

Cited by 5 publications

References 27 publications

Hypoxia-adapted Multiple Myeloma Stem Cells Resist γδ-T-Cell-mediated Killing by Modulating the Mevalonate Pathway

Hypoxia-adapted Multiple Myeloma Stem Cells Resist γδ-T-Cell-mediated Killing by Modulating the Mevalonate Pathway

Generation of effector Vγ9Vδ2 T cells and evaluation of their response to phosphoantigen-loaded cells

Glycolysis inhibition affects proliferation and cytotoxicity of Vγ9Vδ2 T cells expanded for adoptive cell therapy

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