8 Fungal immunomodulatory proteins allergies and in 30 min after the final injection of BSA were IFN-Y with dose dependent manner in spleen cells(Ko et al., dead.However,the experimental group of mice injected with 1995). FIP-fve for 6-7 times did not appear allergies,which suggests Compared with Ganoderma polysaccharides,an effective that the FIP-fve has anti-allergy effects(Rincon et al.,1998). components for immunomodulatory (Zhou et al.,2007),LZ- In addition,FIP-fve can improve the concentration of cancer 8 could activate murine macrophages and T lymphocytes. tumor-specific antibodies in mice serum by feeding FIP-fve, Further experimental results showed that both the purified the concentration of tumor specific antibodies is increased by polysaccrides and reLZ-8 were effective in the induction 386.6 of the control group (Kong et al.,2006).Therefore, toward murine primary macrophage and T lymphocytic cells. FIP-fve was able to inhibit tumor formation.FIP-fve could But their mechanism of action for inducing expression activate the specific and non-specific immunocyte and cytokine production is different.In TLR4-deletion and wild- associated immune response.Therefore,FIP-fve could extend type mice in primary macrophages,the inference is dpPS-G the life of mice with cancer(Rincon et al.,1998). TLR4 receptor,mediates cytokine TNF-a,IL-1B,IL-12p70 Oral administration of Fip-fve during allergen sensitization and CD86,and major histocompatibility complex II (MHC could induce a Th,-predominant allergen-specific immune II)secretion.The reLZ-8 by other means induces cytokine response in mice and protect the mice from the systemic IL-1B,IL-12p70,CD86,and MHC II expression. anaphylaxis-like symptoms after subsequent oral challenge FIP can be mass-produced during the fermentation industry with same allergen.It is worth noting that FIP-fve could be through the heterologous expression.It is an obvious administrated orally,while most protein drugs cannot(Hsieh advantage in health product and drug development.If we etal.,2003) have a better understanding of its cell surface receptors or protein structure and function,designing inhibitor for the Activation of Immune Effector Cells Th,inflammatory response or Th,allergic reactions will The immune effector cells include T lymphocytes,B- become possible.In addition,drugs can be developed using lymphocytes,natural killer (NK)cells,and macrophages.In these fungal active substances.We can combine cellular-mediated immunity,the reLZ-8 in p.pastoris KM71 immunotherapy with existing treatments to solve many clinical has a significant regulatory role for human monocyte-derived problems. dendritic cell(DC)cells. Treating of DC cells with reLZ-8 could result in the ACKNOWLEDGEMENTS enhanced expression level of cell surface CD80,CD86,CD83 This research is financially supported by the National and Human leukocyte antigen DR (HLA-DR)and also Natural Science Foundation of China,Shanghai Science and improve the cytokine's expression level of IL-12 p40,IL-10 Technology Committee and Shanghai Leading Academic and IL-23,and the capacity for endocytosis is suppressed in Discipline Project(Project Number:B209). DCs.At the same time,reLZ-8 enhances the regulation of T cells,thereby reducing the cytokine IFN-y and IL-10 REFERENCES secretion.Target lesion revascularization (TLR4)antibodies An,M.,Gao,E.G.,Qi,J.X.,Li,F.and Liu,X.Z.(2010). of TLR4's neutralization inhibit the secretion of IL-12p40, Expression and Crystallographic Studies of A Fungal IL-10 in the DC cells.ReLZ-8 can stimulate TLR4 or Immunomodulatory Protein LZ-8 from A Medicinal Fungus HEK293 cells transfected by the TLR4/MD2 to produce IL- Ganoderma lucidum.Chinese Journal of Biotechnology 8(Lin et al.,2009b).These results suggest an important role 26:1563-1568.(In Chinese with English abstract). for TLR4 in signaling DCs upon incubation with reLZ-8. Further studies showed that reLZ-8 could increase the IKK, Bai,J.Y.,Zeng,L.,Liu,Y.,Li Y.F.,Lin Z.P.and Hu,Y.L. NF-KB's activity,MAPK and IKBo's phosphorylation level that (2006).Expression of LZ-8 from Garnodum lucidum in is of great significance for the signal transduction process. Transgenic Tobacco and Primary Study on Characteristic of Microphylla streptozotocin suppresses NF-KB and prevents Recombinant LZ-8.Molecular Plant Breeding 4:645-649.(In reLZ-8-induced CD80,CD86,CD83 and HLA-DR Chinese with English abstract). expression and IL-12 p40 and IL-10 secretion. To immune Bal b/c mice with ovalbumin (OVA)and reLZ- Berovic,M.,Habijanic,J.,Zore,I.,Wraber,B.,Hodzar,D., 8,the anti-OVA IgG2a,IFN-Y and IL-2 expression levels has Boh,B.and Pohleven,F.(2003).Submerged Cultivation of significantly improved,compared with only OVA immune Ganoderma Lucidum Biomass and Immunostimulatory mice in the control group.The results showed that reLZ-8 Effects of Fungal Polysaccharides.Journal of Biotechnology could significantly induce Th,cell response and promote the 103:77-86. maturation of immature DC process(Lin et al.,2009b).FIP- fve could selectively activate Th,cells response and turn on Haak-Frendscho,M.,Kino,K.,Sone,T.and Jardieu,B(1993). the expression of cytokines thus inhibit Th,cells development Ling Zhi-8:A Novel T Cell Mitogen Induces Cytokine and then indirectly inhibit the secretion of IgE.Experiments Production and Upregulation of ICAM-1 Expression.Cellular showed that FIP-fve could increase mRNA levels of IL-2 and Immunology 105:101-113.8 Fungal immunomodulatory proteins allergies and in 30 min after the final injection of BSA were FIP-fve for 6-7 times did not appear allergies, which suggests that the FIP-fve has anti-allergy effects (Rincón et al., 1998). In addition, FIP-fve can improve the concentration of cancer tumor-specific antibodies in mice serum by feeding FIP-fve, the concentration of tumor specific antibodies is increased by 386.6 % of the control group (Kong et al., 2006). Therefore, FIP-fve was able to inhibit tumor formation. FIP-fve could activate the specific and non-specific immunocyte and associated immune response. Therefore, FIP-fve could extend the life of mice with cancer (Rincón et al., 1998). Oral administration of Fip-fve during allergen sensitization could induce a Th1 -predominant allergen-specific immune response in mice and protect the mice from the systemic anaphylaxis-like symptoms after subsequent oral challenge with same allergen. It is worth noting that FIP-fve could be administrated orally, while most protein drugs cannot (Hsieh et al., 2003). Activation of Immune Effector Cells The immune effector cells include T lymphocytes, B￾lymphocytes, natural killer (NK) cells, and macrophages. In cellular-mediated immunity, the reLZ-8 in p. pastoris KM71 has a significant regulatory role for human monocyte-derived dendritic cell (DC) cells. Treating of DC cells with reLZ-8 could result in the enhanced expression level of cell surface CD80, CD86, CD83 and Human leukocyte antigen DR (HLA-DR) and also improve the cytokine’s expression level of IL-12 p40, IL-10 and IL-23, and the capacity for endocytosis is suppressed in DCs. At the same time, reLZ-8 enhances the regulation of T cells, thereby reducing the cytokine IFN-γ and IL-10 secretion. Target lesion revascularization (TLR4) antibodies of TLR4’s neutralization inhibit the secretion of IL-12p40, IL-10 in the DC cells. ReLZ-8 can stimulate TLR4 or HEK293 cells transfected by the TLR4/MD2 to produce IL- 8 (Lin et al., 2009b). These results suggest an important role for TLR4 in signaling DCs upon incubation with reLZ-8. Further studies showed that reLZ-8 could increase the IKK, NF-κB’s activity, MAPK and IκBα’s phosphorylation level that is of great significance for the signal transduction process. Microphylla streptozotocin suppresses NF-κB and prevents reLZ-8-induced CD80, CD86, CD83 and HLA-DR expression and IL-12 p40 and IL-10 secretion. To immune Bal b/c mice with ovalbumin (OVA) and reLZ- 8, the anti-OVA IgG2a, IFN-γ and IL-2 expression levels has significantly improved, compared with only OVA immune mice in the control group. The results showed that reLZ-8 could significantly induce Th1 cell response and promote the maturation of immature DC process (Lin et al., 2009b). FIP￾fve could selectively activate Th1 cells response and turn on the expression of cytokines thus inhibit Th2 cells development and then indirectly inhibit the secretion of IgE. Experiments showed that FIP-fve could increase mRNA levels of IL-2 and IFN-γ with dose dependent manner in spleen cells (Ko et al., 1995). Compared with Ganoderma polysaccharides, an effective components for immunomodulatory (Zhou et al., 2007), LZ- 8 could activate murine macrophages and T lymphocytes. Further experimental results showed that both the purified polysaccrides and reLZ-8 were effective in the induction toward murine primary macrophage and T lymphocytic cells. But their mechanism of action for inducing expression cytokine production is different. In TLR4-deletion and wild￾type mice in primary macrophages, the inference is dpPS-G TLR4 receptor, mediates cytokine TNF-α, IL-1β, IL-12p70 and CD86, and major histocompatibility complex II (MHC II) secretion. The reLZ-8 by other means induces cytokine IL-1β, IL-12p70, CD86, and MHC II expression. FIP can be mass-produced during the fermentation industry through the heterologous expression. It is an obvious advantage in health product and drug development. If we have a better understanding of its cell surface receptors or protein structure and function, designing inhibitor for the Th1 inflammatory response or Th2 allergic reactions will become possible. In addition, drugs can be developed using these fungal active substances. We can combine immunotherapy with existing treatments to solve many clinical problems. ACKNOWLEDGEMENTS This research is financially supported by the National Natural Science Foundation of China, Shanghai Science and Technology Committee and Shanghai Leading Academic Discipline Project (Project Number: B209). REFERENCES An, M., Gao, F.G., Qi, J.X., Li, F. and Liu, X.Z. (2010). Expression and Crystallographic Studies of A Fungal Immunomodulatory Protein LZ-8 from A Medicinal Fungus Ganoderma lucidum. Chinese Journal of Biotechnology 26:1563-1568. (In Chinese with English abstract). Bai, J.Y., Zeng, L., Liu, Y., Li Y.F., Lin Z.P. and Hu, Y.L. (2006). Expression of LZ-8 from Garnodum lucidum in Transgenic Tobacco and Primary Study on Characteristic of Recombinant LZ-8. Molecular Plant Breeding 4:645-649. (In Chinese with English abstract). Berovic, M., Habijanic, J., Zore, I., Wraber, B., Hodzar, D., Boh, B. and Pohleven, F. (2003). Submerged Cultivation of Ganoderma Lucidum Biomass and Immunostimulatory Effects of Fungal Polysaccharides. Journal of Biotechnology 103:77-86. Haak-Frendscho, M., Kino, K., Sone, T. and Jardieu, P. (1993). Ling Zhi-8: A Novel T Cell Mitogen Induces Cytokine Production and Upregulation of ICAM-1 Expression. Cellular Immunology 105:101-113. dead. However, the experimental group of mice injected with