Ilmmune Enhancing Effect On Mice Of EPS From Trichoderma Pseudokoning Ii SDTP1

Biologically active polysaccharide is a kind of biological response modifier (BRM) which is widely researched now. As one of the most important part of the polysaccharides, the biologically active functions of fungi polysaccharide have also been studied. Studies have shown the good immune regulation and anti-tumor functions of the fungi polysaccharides, with bolstering immunity, resistance to viruses and so on. Some polysaccharides extracted from the fungus have been developed into immune enhancement and anti-tumor agents on clinical application.Trichoderma pseudokoningi is a Trichoderma species and was isolated in our laboratory. Two kinds of extracellular polysaccharides, TPS1-1 and TPS1-2 are extracted from its fermentation liquid. Preliminary tests confirmed the strong antagonism to a variety of pathogens of the TPS. Meanwhile they show the ability of improving the system resistance of the plants and inducing the generation of enzymes to prevent infection caused by certain bacteria. But the research of the TPS is focused on the plant resistance. As a fungi polysaccharide, rare research is about the immune regulation of animals. We isolated a new Trichoderma extracellular polysaccharide named TPS1-1 from the previous polysaccharide. After testing,we determined it a single component. The molecular weight of TPS1-1 was 31,930Da, and its monosaccharides consist of rhamnose, xylose, fructose, mannose, glucan and galactose in the relative proportions 16.2:14.4:1:25.8:23.6:48.1. The research interest of the paper is focused on the immune modulation on animals of TPS1-1. We explored how the macrophages and the lymphocytes react to the TPS1-1, the effects on the immune function of normal mice and cyclophosphamide (CTX)-induced immunosuppressive mice. The phagocytosis, NO production and acid phosphatase activities in macrophage, the level of serum cytokine, lymphocyte proliferation were tested to investigate the cellular and humoral immunity in animal models.1.In vitro experiments, the impact of TPS1-1, directly and co ConA or LPS on lymphocyte proliferation and secretion of IL-2 was rescarchd,using MTTmethod and the ELISA method respectively; Respectively, neutral red assay, pNPP method and Griess reagent method were carried out to study the impact of the TPS1-1 on peritoneal macrophage phagocytosis, acid phosphatase content and NO content. The results show that, TPS1-1 alone, in the concentration range 25-400μg/mLcan significantly stimulated lymphocyte proliferation in a dose dependent manner. In 6.25-400μg/mL range, TPS1-1 synergistically stimulated spleen lymphocyte proliferation together with ConA or LPS, with highly significant differences; TPS1-1 can increase the secretion of mouse spleen lymphocyte IL-2, the secretion of polysaccharide is 2.8 times compared with the control group, at 100μg/mL; TPS1-1 in 200μg/ml can significantly promote the phagocytosis of macrophages;In the dose 25-800μg/ml, a significant amount of NO production was improved,In the 6.25-400μg/mL range, TPS1-1 can significantly increase the activity of acid phosphatase in macrophage.2. In vivo, we studied the effects of TPS1-1 in normal mice and immunosuppressive mice. The results show that, after treated with TPS1-1 orally in dose of 50,100,200 mg/kg/day for 7 days. Normal rats significantly increased serum IL-2 and TNF-a, compared with the control rats, with an average of 16.0% and 17.0%, respectively; TPS1-1 can improve the serum IL-2 and TNF-a levels of the immunosuppressive mice, compared with the negative control group,10.0% and 32.5% in maximum respectively; TPS1-1 cansignificantly increase delayed type hypersensitivity of normal mice and immunosuppressive mice by the thickness of the toe, compared with the controlthe highest increase rate reached 97.0% and 350%, respectively. TPS1-1 could extend the swimming time of immunosuppressive mice induced by cyclophosphamide, and improved its lymphocyte proliferation and organ index.The results demonstrated that the EPS from Trichoderma TPS1-1 was a good immunomodulator. It had performed well in enhancing humoral immunity, cellular immunity, or specific, non-specific immunity. It is likely to have a good anti-tumor effect. Our study lay the foundation of the further research on its anti-tumor effect.