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Transforming growth factor β (TGF-β), interleukin-10 (IL-10), and vascular endothelial growth factor (VEGF) are three of the commonly studied cytokines playing an important role in tumor initiation and progression. Besides their promotional effects on tumor progression, the three cytokines have immunosuppressive effects that facilitate tumor initiation and progression as well. Ganoderma lucidum polysaccharides (Gl-PS) with multiple bioactivities may have the effect on B16F10 melanoma cells to induce stronger antitumor immune response that has been demonstrated. Gl-PS may have the suppressive effects on the production of these three cytokines, which has yet to be demonstrated. In this study, we tested these effects of Gl-PS by incubating Gl-PS with malignant tumor cells such as B16F10 cells, a melanoma cell line, and LA795 cells, a lung carcinoma cell line. RT-qPCR and enzyme-linked immunosorbent assay showed that the production of TGF-β1, IL-10, and VEGF in B16F10 melanoma cells and LA795 lung carcinoma cells was suppressed by Gl-PS at both mRNA and protein levels, suggesting that the suppression on production of TGF-β, IL-10, and VEGF in B16F10 melanoma cells and LA795 lung carcinoma cells by Gl-PS may contribute to the therapy on melanoma and lung carcinoma along with the induction of stronger antitumor immune response.

It is well-documented that macrophages have the functions to regulate antitumor immune response. Antitumor response can be launched by a series of events, starting with inflammation mediated by monocyte/macrophages, which stimulates natural killer and dendritic cells and finally activates the cytotoxic lymphoid system. Monocytes/macrophages may be the first line of defense in tumors. However, specific and nonspecific immunotherapy for human cancer has shown no success or limited success in clinical trials. Part of the reasons attribute to tumor-derived soluble factors that suppress functions of immune cells or induce apoptosis of these cells, including macrophages. Therefore, antagonism of the suppression on the macrophages is an important goal for tumor immunotherapy. To achieve this purpose, Ganoderma lucidum polysaccharides (Gl-PS) with multiple bioactivities were used on mouse peritoneal macrophages incubating with culture supernatants of B16F10 melanoma cells (B16F10-CS). It was shown that the viability, phagocytic activity, NO production, TNF-α production and activity in peritoneal macrophages after activation by lipopolysaccharide were suppressed by B16F10-CS, while the suppressions were fully or partially antagonized by Gl-PS. In conclusion, B16F10-CS is suppressive to the viability, phagocytic activity, NO production, TNF-α production and activity in peritoneal macrophages while Gl-PS had the antagonistic effects against this suppression, suggesting this potential of Gl-PS to facilitate cancer immunotherapy., (Copyright © 2013 John Wiley & Sons, Ltd.)

Purpose: It is obvious that malignant cells evade from immune system in patients with manifest malignancy. Deficient major histocompatibility complex (MHC) class I and costimulatory molecules on malignant cells partially consist of evasion strategy since antigen bond MHC and costimulatory molecules provide two signals necessary for T cell activation. Therefore, enhancement of MHC-I and costimulatory molecules may favor restraint of the evasion. For this purpose, Ganoderma lucidum Polysaccharides (Gl-PS) was used on B16F10 melanoma cells in this study., Methods: Immunocytochemistry and flowcytometry were used to determine the H-2K(b) and H-2D(b) (two prominent MHC class I molecules in C57BL mouse) as well as B7-1 and B7-2 (two prominent costimulatory molecules) expression on B16F10 cells after incubation with Gl-PS, while messenger ribonucleic acid (mRNA) of these molecules was detected by reverse transcription polymerase chain reaction (RT-PCR)., Results: The H-2K(b) and H-2D(b), and B7-1 and B7-2 on B16F10 cells and mRNAs of these molecules were enhanced by Gl-PS, and more efficient antitumor cytotoxicity was induced by the Gl-PS treated cells., Conclusions: The MHC class I molecules and costimulatory molecules may be enhanced by Gl-PS, and more efficient immune cell mediated cytotoxicity against these B16F10 cells may be induced, which may favor cancer therapy.

Objectives: Restoration of epithelial continuity in the intestinal surface after extensive destruction is important since intestinal epithelial cells stand as a boundary between the body's internal and external environment. Polysaccharides from Ganoderma lucidum (Gl-PS) may benefit intestinal epithelial wound healing in different aspects, which awaits clarification. To identify potential effects, a non-transformed small-intestinal epithelial cell line, IEC-6 cells, was used., Methods: Effects on epithelial cell proliferation, migration, morphology of differentiation and transforming growth factor beta (TGF-β) protein expression, as well as the cellular ornithine decarboxylase (ODC) mRNA and c-Myc mRNA expression, were assessed, respectively, by MTT assay, wound model in vitro, observation under a microscope after hematoxylin and eosin staining, enzyme-linked immunosorbent assay and reverse transcription-polymerase chain reaction assays., Key Findings: It was shown that Gl-PS stimulated IEC-6 cell proliferation and migration significantly in a dose-dependent manner; 10 µg/ml Gl-PS improved the morphology of differentiation in IEC-6 cells. Inefficacy in expression of TGF-β in IEC-6 cells indicated a possible TGF-β independent action of Gl-PS. However, Gl-PS increased ODC mRNA and c-Myc mRNA expression in a dose-dependent manner, indicating, at least partially possible involvement of ODC and c-Myc gene expression in improvement of intestinal wound healing., Conclusions: These results suggest the potential usefulness of Gl-PS to cure intestinal disorders characterized by injury and ineffective repair of the intestinal mucosa., (© 2011 The Authors. JPP © 2011 Royal Pharmaceutical Society.)

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., (© 2011 The Authors. JPP © 2011 Royal Pharmaceutical Society.)

The immune system in patients with cancer often fails to control tumour growth because of deficient immunogenicity of tumour cells. Ganoderma lucidum polysaccharides (Gl-PS) are believed to have anti-tumour effects by boosting host immune function. Additionally, Gl-PS may have some direct effects on tumour cells in the activation of lymphocytes, thus enhancing the immunogenicity of tumour cells. We tested the effects of Gl-PS in lymphocyte activation by incubating Gl-PS with a tumour cell line deficient in antigen presentation. Our study showed that Gl-PS can promote B16F10 melanoma cells to induce lymphocyte proliferation, CD69 and FasL expression and IFN-γ production, indicating that Gl-PS can improve the nature of B16F10 cells to activate lymphocytes. Furthermore, H-2D(b) [a major histocompatibility (MHC) class I molecule], and B7-1 and B7-2 (two prominent co-stimulatory molecules expressed on B16F10 cells) were enhanced by Gl-PS, suggesting that these molecules may at least partially be involved in the process of Gl-PS on B16F10 cells to activate lymphocytes., (© 2010 The Authors. Basic & Clinical Pharmacology & Toxicology © 2010 Nordic Pharmacological Society.)

Objective: To establish precolumn derivation HPLC method to determine the monosaccharides in Heterosmilax japonica polysaccharide., Methods: Heterosmilax japonica sample was hydrolyzed with trifluoroacetic (TFA)and derivated by 1-phenyl-3-methyl-5-pyrazolone (PMP). The monosaccharide composition of Heterosmilax japonica polysaccharide was carried out by reversed-phase technique on a Kromasil C18 column with a mobile phase composed of 100 mmol/L ammonium acetate buffer (pH5.5) and acetonitrile in the ratio of 77: 23. The detection was carried out at 245 nm., Results: The Heterosmilax japonica polysaccharide composed of mannose, rhamnose, glucose, galactose and xylose with amolar ratio of 66: 1:7590: 452: 528., Conclusion: The HPLC method with precolumn derivatization is appropriate for the analysis of monosaccharide composition of Heterosmilax japonica polysaccharide and the method is simple, quick and accurate.