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灵芝多糖拮抗 B16F10 黑色素瘤细胞培养上清液对淋巴细胞的抑制作用。

Ganoderma lucidum polysaccharides antagonize the suppression on lymphocytes induced by culture supernatants of B16F10 melanoma cells.

机构信息

Department of Pharmacology, Peking University Health Science Center, School of Basic Medical Sciences, Beijing, China.

出版信息

J Pharm Pharmacol. 2011 May;63(5):725-35. doi: 10.1111/j.2042-7158.2011.01266.x.

Abstract

OBJECTIVES

Tumour cells produce factors such as interleukin 10 (IL-10), transforming growth factor β1 (TGF-β1) and vascular endothelial growth factor (VEGF) that suppress the function of immune cells or induce apoptosis of immune cells. One of the most important goals of tumour immunotherapy is to antagonize this suppression on immune cells. Ganoderma lucidum polysaccharides (Gl-PS) may have this potential. The purpose of this study was to determine the antagonistic effects of Gl-PS on the suppression induced by B16F10 melanoma cell culture supernatant (B16F10-CS) on lymphocytes.

METHODS

Gl-PS was used on lymphocytes incubated with B16F10-CS. Enzyme-linked immunosorbent assay was used to determine the levels of IL-10, TGF-β1 and VEGF in B16F10-CS. The MTT assay was used to determine the proliferation of lymphocytes. Immunocytochemistry and Western blot assay were used to determine perforin and granzyme B production in lymphocytes.

KEY FINDINGS

There were elevated levels of IL-10, TGF-β1 and VEGF in B16F10-CS. The lymphocyte proliferation, and perforin and granzyme B production in lymphocytes after induction with phytohemagglutinin, as well as lymphocyte proliferation in the mixed lymphocyte reaction, were suppressed by B16F10-CS. This suppression was fully or partially antagonized by Gl-PS.

CONCLUSIONS

B16F10-CS suppressed lymphocyte proliferation and perforin and granzyme B production in lymphocytes after induction with phytohemagglutinin, as well as lymphocyte proliferation in the mixed lymphocyte reaction. This suppression may be associated with elevated levels of immunosuppressive IL-10, TGF-β1 and VEGF in B16F10-CS. Gl-PS had antagonistic effects on the immunosuppression induced by B16F10-CS, suggesting the potential for Gl-PS in cancer immunotherapy.

摘要

目的

肿瘤细胞产生白细胞介素 10(IL-10)、转化生长因子 β1(TGF-β1)和血管内皮生长因子(VEGF)等因子,抑制免疫细胞的功能或诱导免疫细胞凋亡。肿瘤免疫治疗的最重要目标之一是拮抗免疫细胞的这种抑制作用。灵芝多糖(Gl-PS)可能具有这种潜力。本研究旨在确定 Gl-PS 对 B16F10 黑色素瘤细胞培养上清液(B16F10-CS)诱导的淋巴细胞抑制作用的拮抗作用。

方法

用 Gl-PS 孵育与 B16F10-CS 共培养的淋巴细胞。酶联免疫吸附试验测定 B16F10-CS 中 IL-10、TGF-β1 和 VEGF 的水平。MTT 法测定淋巴细胞增殖。免疫细胞化学和 Western blot 法测定淋巴细胞中穿孔素和颗粒酶 B 的产生。

主要发现

B16F10-CS 中 IL-10、TGF-β1 和 VEGF 水平升高。植物血凝素诱导的淋巴细胞增殖以及穿孔素和颗粒酶 B 的产生,以及混合淋巴细胞反应中的淋巴细胞增殖均受 B16F10-CS 抑制。Gl-PS 可完全或部分拮抗这种抑制作用。

结论

B16F10-CS 抑制植物血凝素诱导的淋巴细胞增殖以及穿孔素和颗粒酶 B 的产生,以及混合淋巴细胞反应中的淋巴细胞增殖。这种抑制作用可能与 B16F10-CS 中免疫抑制性 IL-10、TGF-β1 和 VEGF 水平升高有关。Gl-PS 对 B16F10-CS 诱导的免疫抑制具有拮抗作用,提示 Gl-PS 在癌症免疫治疗中的潜力。

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