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Antibody-dependent cellular cytotoxicity toward neuroblastoma enhanced by activated invariant natural killer T cells.

Mise N, Takami M, Suzuki A, Kamata T, Harada K, Hishiki T, Saito T, Terui K, Mitsunaga T, Nakata M, Ikeuchi T, Nakayama T, Yoshida H, Motohashi S - Cancer Sci. (2016)

Bottom Line: For further improvement in patient outcomes, ways to maximize the cytotoxic effects of anti-GD2 therapies with minimal toxicity are required.Although some of the expanded iNKT cells expressed natural killer (NK) cell markers, including FcγR, iNKT cells were not directly associated with ADCC.In conclusion, iNKT cell-based immunotherapy could be an appropriate candidate for anti-GD2 antibody therapy for neuroblastoma.

View Article: PubMed Central - PubMed

Affiliation: Department of Medical Immunology, Graduate School of Medicine, Chiba University, Chiba, Japan.

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Related in: MedlinePlus

Blocking antibodies against IFNγ, IL‐2, TNFα and GM‐CSF did not abrogate the ADCC of activated NK cells. Using Transwell plates, NK cells were seeded in the lower wells, and invariant natural killer T (iNKT) cells and moDC were seeded in the upper wells. Neutralizing mAbs against IFNγ (a), IL‐2 (b), TNFα (c) and GM‐CSF (d) or isotype controls were also added. Two days later, NK cells were collected from the lower wells and cytotoxicity assays were performed using NMB NB cells with anti‐GD2 Abs. (e) NK cells were precultured with iNKT cells and αGalCer‐pulsed moDC for two days, and CD3−CD56+ NK cells were then purified by MACS and mixed together with NMB NB cells. The cytotoxicity assays were performed under anti‐GD2 Abs with or without anti‐IFNγ Abs.
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cas12882-fig-0007: Blocking antibodies against IFNγ, IL‐2, TNFα and GM‐CSF did not abrogate the ADCC of activated NK cells. Using Transwell plates, NK cells were seeded in the lower wells, and invariant natural killer T (iNKT) cells and moDC were seeded in the upper wells. Neutralizing mAbs against IFNγ (a), IL‐2 (b), TNFα (c) and GM‐CSF (d) or isotype controls were also added. Two days later, NK cells were collected from the lower wells and cytotoxicity assays were performed using NMB NB cells with anti‐GD2 Abs. (e) NK cells were precultured with iNKT cells and αGalCer‐pulsed moDC for two days, and CD3−CD56+ NK cells were then purified by MACS and mixed together with NMB NB cells. The cytotoxicity assays were performed under anti‐GD2 Abs with or without anti‐IFNγ Abs.

Mentions: To assess the responsible cytokines for ADCC enhancement, neutralization of IFNγ, IL‐2, TNFα and GM‐CSF was performed. Using Transwell plates, NK cells were seeded in the lower well, iNKT cells and moDC were seeded in the upper well, and neutralizing mAbs or isotype controls were also added. NK cells were collected after 48 h, and cytotoxicity assays were performed using NMB NB cells with anti‐GD2 Abs. None of the neutralizing mAbs abrogated the cytotoxicity of NK cells activated by iNKT cells and αGalCer‐pulsed moDC (Fig. 7a–d).


Antibody-dependent cellular cytotoxicity toward neuroblastoma enhanced by activated invariant natural killer T cells.

Mise N, Takami M, Suzuki A, Kamata T, Harada K, Hishiki T, Saito T, Terui K, Mitsunaga T, Nakata M, Ikeuchi T, Nakayama T, Yoshida H, Motohashi S - Cancer Sci. (2016)

Blocking antibodies against IFNγ, IL‐2, TNFα and GM‐CSF did not abrogate the ADCC of activated NK cells. Using Transwell plates, NK cells were seeded in the lower wells, and invariant natural killer T (iNKT) cells and moDC were seeded in the upper wells. Neutralizing mAbs against IFNγ (a), IL‐2 (b), TNFα (c) and GM‐CSF (d) or isotype controls were also added. Two days later, NK cells were collected from the lower wells and cytotoxicity assays were performed using NMB NB cells with anti‐GD2 Abs. (e) NK cells were precultured with iNKT cells and αGalCer‐pulsed moDC for two days, and CD3−CD56+ NK cells were then purified by MACS and mixed together with NMB NB cells. The cytotoxicity assays were performed under anti‐GD2 Abs with or without anti‐IFNγ Abs.
© Copyright Policy - creativeCommonsBy-nc-nd
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4814252&req=5

cas12882-fig-0007: Blocking antibodies against IFNγ, IL‐2, TNFα and GM‐CSF did not abrogate the ADCC of activated NK cells. Using Transwell plates, NK cells were seeded in the lower wells, and invariant natural killer T (iNKT) cells and moDC were seeded in the upper wells. Neutralizing mAbs against IFNγ (a), IL‐2 (b), TNFα (c) and GM‐CSF (d) or isotype controls were also added. Two days later, NK cells were collected from the lower wells and cytotoxicity assays were performed using NMB NB cells with anti‐GD2 Abs. (e) NK cells were precultured with iNKT cells and αGalCer‐pulsed moDC for two days, and CD3−CD56+ NK cells were then purified by MACS and mixed together with NMB NB cells. The cytotoxicity assays were performed under anti‐GD2 Abs with or without anti‐IFNγ Abs.
Mentions: To assess the responsible cytokines for ADCC enhancement, neutralization of IFNγ, IL‐2, TNFα and GM‐CSF was performed. Using Transwell plates, NK cells were seeded in the lower well, iNKT cells and moDC were seeded in the upper well, and neutralizing mAbs or isotype controls were also added. NK cells were collected after 48 h, and cytotoxicity assays were performed using NMB NB cells with anti‐GD2 Abs. None of the neutralizing mAbs abrogated the cytotoxicity of NK cells activated by iNKT cells and αGalCer‐pulsed moDC (Fig. 7a–d).

Bottom Line: For further improvement in patient outcomes, ways to maximize the cytotoxic effects of anti-GD2 therapies with minimal toxicity are required.Although some of the expanded iNKT cells expressed natural killer (NK) cell markers, including FcγR, iNKT cells were not directly associated with ADCC.In conclusion, iNKT cell-based immunotherapy could be an appropriate candidate for anti-GD2 antibody therapy for neuroblastoma.

View Article: PubMed Central - PubMed

Affiliation: Department of Medical Immunology, Graduate School of Medicine, Chiba University, Chiba, Japan.

Show MeSH
Related in: MedlinePlus