Department of Microbiology, The School of Preclinical Medicine, Guangxi Medical University, Nanning, Guangxi, China.
Department of Pharmaceutical and Medical Equipment, Trading Center of Guangxi Public Resources, Nanning, Guangxi, China.
Anat Rec (Hoboken). 2019 Oct;302(10):1718-1725. doi: 10.1002/ar.24177. Epub 2019 Jun 12.
The objective of this article is to evaluate whether the tumoricidal activity of mouse IFN R nature killer (NK) cells is induced by Newcastle disease virus hemagglutinin-neuraminidase (NDV-HN) stimulation, and to investigate what is the mechanism of the HN-stimulated NK cells to kill mouse hepatoma cell line in vitro. The mouse IFN R NK cells were stimulated for 16 hr with 500 ng/mL NDV-HN in 1640 medium. Quantify the cytotoxic activities of NK cells against mouse hepatoma cells (Hepa1-6) by flow cytometry. Granzymes B (GrB) and Fas/FasL concentrations in the supernatants of IFN R NK cells medium were determined by specific ELISA assay. The expression of cell surface GrB and Fas was determined by Western blot. NDV-HN stimulation enhanced tumoricidal activity of IFN R NK cells toward Hepa1-6 in vitro. Treating with anti-HN neutralizing mAb induced significant decline in the cytotoxicity of IFN R NK cells toward Hepa1-6 cell line (P < 0.05). After treating with anti-HN protein (1 μL/mL), Syk-specific inhibitor Herbimycin A(250 ng/mL) and NF-κB inhibitor PDTC (500 ng/mL) downregulated the tumoricidal activity of HN-stimulated IFN R NK cells (P < 0.05). Moreover, significant suppressions in the production of GrB and Fas/FasL were observed in HN-stimulated IFN R NK cells (P < 0.05). Thus, we concluded that killer activation receptors pathway is involved in the IFN-γ-independent GrB and Fas/FasL expression of NDV-HN-stimulated IFN R NK cells, and these are activated by Syk and NF-κB. Anat Rec, 302:1718-1725, 2019. © 2019 The Authors. The Anatomical Record published by Wiley Periodicals, Inc. on behalf of American Association for Anatomy.
本文旨在评估新城疫病毒血凝素神经氨酸酶（NDV-HN）刺激是否能诱导鼠 IFNγR 自然杀伤（NK）细胞的细胞毒性作用，并探讨 HN 刺激的 NK 细胞在体外杀伤鼠肝癌细胞的机制。用 500ng/mL 的 NDV-HN 在 1640 培养基中刺激 IFNγR NK 细胞 16 小时。通过流式细胞术定量 NK 细胞对鼠肝癌细胞（Hepa1-6）的细胞毒性作用。通过特异性 ELISA 测定 IFNγR NK 细胞培养基上清液中颗粒酶 B（GrB）和 Fas/FasL 的浓度。通过 Western blot 测定细胞表面 GrB 和 Fas 的表达。NDV-HN 刺激增强了 IFNγR NK 细胞在体外对 Hepa1-6 的杀伤活性。用抗 HN 中和 mAb 处理可显著降低 IFNγR NK 细胞对 Hepa1-6 细胞系的细胞毒性（P<0.05）。用抗 HN 蛋白（1μL/mL）、Syk 特异性抑制剂 Herbimycin A（250ng/mL）和 NF-κB 抑制剂 PDTC（500ng/mL）处理后，下调了 HN 刺激的 IFNγR NK 细胞的杀伤活性（P<0.05）。此外，在 HN 刺激的 IFNγR NK 细胞中观察到 GrB 和 Fas/FasL 的产生明显受到抑制（P<0.05）。因此，我们得出结论，杀伤激活受体途径参与了 IFNγ 非依赖性的 NDV-HN 刺激的 IFNγR NK 细胞中 GrB 和 Fas/FasL 的表达，这些表达由 Syk 和 NF-κB 激活。解剖记录，302:1718-1725,2019. © 2019 作者。解剖记录由 Wiley 期刊出版公司代表美国解剖学会出版。
