臺灣博碩士論文加值系統

樟芝 (Taiwanofungus camphoratus) 為台灣傳統極具發展潛力之藥用真菌，並具有免疫調節及抗腫瘤之活性。樟芝免疫調節蛋白 ACA1 為醣蛋白，其 cDNA已成功選殖並構築於 pET-32a 表現載體，以大腸桿菌為宿主進行表現，可得重組樟芝免疫調節蛋白 rACA1。本論文之研究目的為探討 rACA1 活化巨噬細胞的分子機制。首先我們發現 rACA1 可誘導巨噬細胞株 RAW 264.7 和小鼠腹腔巨噬細胞產生一氧化氮和腫瘤壞死因子 (TNF-alpha)。為了進一步探討細胞膜上 receptor 是否參與 rACA1 活化巨噬細胞產生腫瘤壞死因子，我們利用正常小鼠 (C57BL/6) 和TLR4 免疫缺陷鼠 (C57BL/10ScN) 之腹腔巨噬細胞進行試驗，結果顯示 rACA1 可活化正常小鼠 (TLR4+/+) 及免疫缺陷鼠 (TLR4-/-) 之腹腔巨噬細胞。另一方面，利用小鼠 TLR2 單株抗體進行中和試驗，則發現 rACA1 誘導巨噬細胞產生腫瘤壞死因子之作用，會顯著受到 anti-TLR2 抗體中和而下降。此外，我們轉染 dominant-negative MyD88載體至 RAW 264.7 中表現，亦發現 rACA1 活化巨噬細胞之訊息傳導路徑，會受到 dominant-negative MyD88 之阻礙，無法活化巨噬細胞產生腫瘤壞死因子。綜合上述，我們推測 rACA1 活化巨噬細胞至少會經由 TLR2/MyD88 訊息傳導路徑。

Taiwanofungus camphoratus, a traditional and potential medicinal mushroom in Taiwan, is known to have immunomodulatory and antitumor activities. ACA1, which is a glycoprotein obtained from T. camphoratus and is capable to activate mouse macrophages and splenocytes, has been cloned into pET32 vector and successfully expressed in E. coli to obtain normal protein, rACA1. Here, we investigate the bioactivity of rACA1 on murine macrophages and the membrane receptor and the intracellular signaling pathway responsible for the activation of macrophages by rACA1. rACA1 is demonstrated to the production of nitric oxide (NO) and TNF-alpha both in RAW 264.7 cells and peritoneal macrophages from normal mice. To investigate the membrane receptor involved in the activation of TNF-alpha production, we further examine the effects of rACA1 on the production of TNF-alpha in mouse peritoneal macrophages isolated from wild type C57BL/6 and from functional Toll-like receptor 4 (TLR4)-deficient C57BL/10ScN mice. The results show that rACA1 induces TNF-alpha production by macrophages isolated from both C57BL/6 and C57BL/10ScN mice. However, anti-TLR2 mAb is demonstrated to block the activation of rACA1 toward peritoneal cells, which indicates that TLR2 is highly involved in rACA1 signaling. Further we examine the effect of rACA1 on the activation of an adapter protein, MyD88, which is a downstream mediator of TLR2. rACA1 of macrophages activation was obstructed by dominant-negative MyD88 to produce TNF-alpha. Taken together, these results suggest that rACA1-mediated induction of TNF-alpha production in macrophages is mediated, at least in part, by TLR2/MyD88 signaling pathway.

中文摘要 1
Abstract 2
Chapter 1 Preface 3
Chapter 2 Introduction 5
2.1 Introduction of Taiwanofungus camphoratus 5
2.1.1 Morphology, component and bioactivity of Taiwanofungus camphoratus 5
2.1.2 Immunomodulatory protein ACAs and its activity 13
2.2 Mechanism of pathogen activating innate immunity 17
2.2.1 Physiological function of macrophages 17
2.2.2 The relationship between Toll-like receptor family and macrophage activation 19
2.2.3 Innate recognition of lipopolysaccharide by Toll-like receptor 4-MD-2 22
2.2.4 MyD88 dependent and independent signaling pathway 24
2.3 Recognition of fungal pathogens by Toll-like receptors 32
2.4 Objectives and strategy of this study 34
Chapter 3 Materials and Methods 37
3.1 Materials 37
3.1.1 Reagents 37
3.1.2 Systems and Kits 38
3.1.3 Equipments 39
3.1.4 Animals 41
3.2 Methods 41
3.2.1 Sample preparation 41
3.2.1.1 Preparation of recombinant ACA1 (rACA1) 41
3.2.1.2 Bioactivity of rACA1 41
3.2.1.3 LPS contamination of ACA1 45
3.2.2 Detection of TLRs-dependent signaling pathway by TNF-alpha secretion 48
3.2.2.1 Interaction between ACA1 and macrophages 48
3.2.2.2 Determination of TLR4-dependent pathway 51
3.2.2.3 Neutralization test of alphaTLR2 monoclonal antibody 53
3.2.3 Establishment of dominant negative MyD88 transient transfection system 55
3.2.3.1 Cell culture 55
3.2.3.2 Preparation of murine macrophage cDNA 55
3.2.3.3 Construction of dominant negative MyD88 57
3.2.3.4 Transient transfection 61
3.2.3.5 Bioassay 63
3.2.3.6 Reporter assay 63
3.2.4 Statistical analysis 63
Chapter 4 Results 65
4.1 Sample preparation 65
4.1.1 Preparation of recombinant ACA1 (rACA1) 66
4.1.1.1 Bioactivity of rACA1 67
(1) Effect of rACA1 on macrophage-like RAW 264.7 65
(2) Effect of rACA1 on Balb/C peritoneal cells 66
4.1.1.2 Exclusion of LPS contamination of rACA1 68
(1) LAL assay 68
(2) Autocalved rACA1 activity 69
4.2 Involvement of rACA1 on TLRs-dependent signaling pathway 71
4.2.1 Interaction between ACA1 and macrophages 71
4.2.2 Investigation of TLR4-dependent pathway 72
4.2.3 Involvement of TLR2 in rACA1 activating macrophages 74
4.3 Involvement of MyD88 in rACA1 activating macrophages 76
4.3.1 Construction of dominant negative MyD88 76
4.3.2 Transient transfection of RAW 264.7 macrophages 77
Chapter 5 Discussion and Conclusion 80
5.1 Discussion 80
5.2 Conclusion 86
References 87
Figures and Appendix 97

Adachi O, K. T., Takeda K, Matsumoto M, Tsutsui H, Sakagami M, Nakanishi K, Akira S. 1998. Targeted disruption of the MyD88 gene results in loss of IL-1- and IL-18-mediated function. Immunity 9:143.

Akira S, Y. M., Takeda K. 2003. Role of adapters in Toll-like receptor signalling. Biochem Soc Trans. 31:637.

Alexopoulou L, H. A., Medzhitov R, Flavell RA. 2001. Recognition of double-stranded RNA and activation of NF-kappaB by Toll-like receptor 3. Nature 413:732.

Aliprantis AO, Y. R., Mark MR, Suggett S, Devaux B, Radolf JD, Klimpel GR, Godowski P, Zychlinsky A. 1999. Cell activation and apoptosis by bacterial lipoproteins through toll-like receptor-2. Science 285:736.

Anderson KV, J. G., Nusslein-Volhard C. 1985. Establishment of dorsal-ventral polarity in the Drosophila embryo: genetic studies on the role of the Toll gene product. Cell 42:779.

B, B. 2000. Tlr4: central component of the sole mammalian LPS sensor. Curr Opin Immunol. 12:20.

B, B. 2004. Inferences, questions and possibilities in Toll-like receptor signalling. Nature 430:257.

Bellocchio S, M. C., Bozza S, Gaziano R, Rossi G, Mambula SS, Vecchi A, Mantovani A, Levitz SM, Romani L. 2004. The contribution of the Toll-like/IL-1 receptor superfamily to innate and adaptive immunity to fungal pathogens in vivo. J Immunol. 172:3059.

Brightbill HD, L. D., Krutzik SR, Yang RB, Belisle JT, Bleharski JR, Maitland M, Norgard MV, Plevy SE, Smale ST, Brennan PJ, Bloom BR, Godowski PJ, Modlin RL. 1999. Host defense mechanisms triggered by microbial lipoproteins through toll-like receptors. Science 285:732.

Brown GD, H. J., Williams DL, Willment JA, Marshall AS, Gordon S. 2003. Dectin-1 mediates the biological effects of beta-glucans. J Exp Med. 197:1119.

Campos MA, A. I., Takeuchi O, Akira S, Valente EP, Procopio DO, Travassos LR, Smith JA, Golenbock DT, Gazzinelli RT. 2001. Activation of Toll-like receptor-2 by glycosylphosphatidylinositol anchors from a protozoan parasite. J Immunol. 167:416.

Cao Z, H. W., Gao X. 1996a. IRAK: a kinase associated with the interleukin-1 receptor. Science 271:1128.

Cao Z, X. J., Takeuchi M, Kurama T, Goeddel DV. 1996b. TRAF6 is a signal transducer for interleukin-1. Nature 383:443.

Chaing HC, W. D., Cherng IW, Ueng CH. 1995. A sesquiterpene lactone, phenyl and biphenyl compounds from Antrodia cinnamomea. Phytochemistry:613.

Chang TT, C. W. 1995. Antrodia cinnamomea sp. nov. on Cinnamomum kane hirai in Taiwan. Mycol. Res.:756.

Chang TT, C. W., Hua J. 1997. The cultural atlas of wood-inhabiting Aphyllophorales in Taiwan. Mycol. Monog. 2:1.

Chen PC. High throughput screening of anti-liver-cancer drug candidate from Antrodia champhorata mycelia. 2004. Graduate Institute of Chemistry and Biochemistry National Chung Cheng University. Master Thesis.

Cherng, I., Chiang, HC. 1995. Three new triterpenoids from Antrodia cinnamomea. J. Nat. Prod.:365.

Cherng, I., Wu DP, Chiang HC. 1996. Triterpenoids from Antrodia cinnamomea. Phytochemistry:263.

Chin KL. Molecular cloning and gene expression of the immunomodulatory protein Aca1 from Taiwanofungus camphoratus. 2004. Graduate Institute of Horticulture National Taiwan University. Master Thesis.

Coutinho A, M. T. 1978. Genetic basis for unresponsiveness to lipopolysaccharide in C57BL/10Cr mice. Immunogenetics:17.

Damais C, G. A., Parant M. 1977. Non-specific responses of C3H/He low responder mice to LPS and to TCA-extracted endotoxin. Ann Immunol 128:67.

Duerksen-Hughes PJ, D. D., Laster SM, Zachariades NA, Aquino L, Gooding LR. 1992. Both tumor necrosis factor and nitric oxide participate in lysis of simian virus 40-transformed cells by activated macrophages. J Immunol. 149:2114.

Fan CC. Study on biological activities of Antrodia camphorata mycelial fractions. 2004. Graduate insititute of chemical and material engineering Sounthern Taiwan University of Technology. Master Thesis.

Farias-Eisner R, S. M., Aeberhard E, Chaudhuri G. 1994. Nitric oxide is an important mediator for tumoricidal activity in vivo. Proc Natl Acad Sci 91:9407.

Fitzgerald KA, P.-M. E., Bowie AG, Jefferies CA, Mansell AS, Brady G, Brint E, Dunne A, Gray P, Harte MT, McMurray D, Smith DE, Sims JE, Bird TA, O''Neill LA. 2001. Mal (MyD88-adapter-like) is required for Toll-like receptor-4 signal transduction. Nature 413:78.

Fitzgerald KA, R. D., Barnes BJ, Caffrey DR, Visintin A, Latz E, Monks B, Pitha PM, Golenbock DT. 2003. LPS-TLR4 signaling to IRF-3/7 and NF-kappaB involves the toll adapters TRAM and TRIF. J Exp Med. 198:1043.

Gantner BN, S. R., Canavera SJ, Akira S, Underhill DM. 2003. Collaborative induction of inflammatory responses by dectin-1 and Toll-like receptor 2. J Exp Med. 197:1107.

Gorelik L, B.-D. Y., Mokyr MB. 1996. Insight into the mechanism(s) through which TNF promotes the generation of T cell-mediated antitumor cytotoxicity by tumor bearer splenic cells. J Immunol. 156:4298.

Hacker H, V. R., Takeuchi O, Hoshino K, Akira S, Wagner H. 2000. Immune cell activation by bacterial CpG-DNA through myeloid differentiation marker 88 and tumor necrosis factor receptor-associated factor (TRAF)6. J Exp Med. 192:595.

Hayashi F, S. K., Ozinsky A, Hawn TR, Yi EC, Goodlett DR, Eng JK, Akira S, Underhill DM, Aderem A. 2001. The innate immune response to bacterial flagellin is mediated by Toll-like receptor 5. Nature 410:1099.

Hemmi H, K. T., Takeuchi O, Sato S, Sanjo H, Hoshino K, Horiuchi T, Tomizawa H, Takeda K, Akira S. 2002. Small anti-viral compounds activate immune cells via the TLR7 MyD88-dependent signaling pathway. Nat Immunol. 3:196.

Hirschfeld M, K. C., Schwandner R, Wesche H, Weis JH, Wooten RM, Weis JJ. 1999. Cutting edge: inflammatory signaling by Borrelia burgdorferi lipoproteins is mediated by toll-like receptor 2. J Immunol. 163:2382.

Horng T, B. G., Medzhitov R. 2001. TIRAP: an adapter molecule in the Toll signaling pathway. Nat Immunol. 2:935.

Horng T, B. G., Flavell RA, Medzhitov R. 2002. The adaptor molecule TIRAP provides signalling specificity for Toll-like receptors. Nature 420:329.

Hoshino K, T. O., Kawai T, Sanjo H, Ogawa T, Takeda Y, Takeda K, Akira S. 1999. Cutting edge: Toll-like receptor 4 (TLR4)-deficient mice are hyporesponsive to lipopolysaccharide: evidence for TLR4 as the Lps gene product. J Immunol. 162:3749.

Hseu YC, C. W., Hseu YT, Lee CY, Yech YJ, Chen PC, Chen JY, Yang HL. 2002. Protection of oxidative damage by aqueous extract from Antrodia camphorata mycelia in normal human erythrocytes. Life Sci. 71:469.

Hsiao G, S. M., Lin KH, Lan MH, Wu LY, Chou DS, Lin CH, Su CH, Sheu JR. 2003. Antioxidative and hepatoprotective effects of Antrodia camphorata extract. J Agric Food Chem. 51:3302.

Huang LC. Antioxidant properties and polysaccharide composition analysis of Antrodia camphorata and Agaricus blazei. National Chung-Hsing University, Taichung, Taiwan. 63-76.

Jiang Z, M. T., Sen G, Li X. 2004. Toll-like receptor 3-mediated activation of NF-kappaB and IRF3 diverges at Toll-IL-1 receptor domain-containing adapter inducing IFN-beta. Proc Natl Acad Sci 101:3533.

Jiang Z, Z.-D. M., Nie H, Silva AM, Williams BR, Li X. 2003. Poly(I-C)-induced Toll-like receptor 3 (TLR3)-mediated activation of NFkappa B and MAP kinase is through an interleukin-1 receptor-associated kinase (IRAK)-independent pathway employing the signaling components TLR3-TRAF6-TAK1-TAB2-PKR. J Biol Chem. 278:16713.

K, M. 2004. Innate recognition of lipopolysaccharide by Toll-like receptor 4-MD-2. Trends Microbiol. 12:186.

Kawai T, A. S. 2005. Pathogen recognition with Toll-like receptors. Curr Opin Immunol.

Kitchens RL, W. P., Munford RS. 1998. Bacterial lipopolysaccharide can enter monocytes via two CD14-dependent pathways. J Immunol. 161:5534.

Klis FM, d. G. P., Hellingwerf K. 2001. Molecular organization of the cell wall of Candida albicans. Med Mycol.:1.

Kobayashi K, H. L., Galan JE, Janeway CA Jr, Medzhitov R, Flavell RA. 2002. IRAK-M is a negative regulator of Toll-like receptor signaling. Cell 110:191.

L, R. 2004. Immunity to fungal infections. Nat Rev Immunol. 4:1.

Lee IH, H. R., Chen CT, Chen HC, Hsu WC, Lu MK. 2002. Antrodia camphorata polysaccharides exhibit anti-hepatitis B virus effects. FEMS Microbiol Lett. 209:63.

Lemaitre B, N. E., Michaut L, Reichhart JM, Hoffmann JA. 1996. The dorsoventral regulatory gene cassette spatzle/Toll/cactus controls the potent antifungal response in Drosophila adults. Cell 86:973.

Li S, S. A., Fontana EJ, Wesche H. 2002. IRAK-4: a novel member of the IRAK family with the properties of an IRAK-kinase. Proc Natl Acad Sci. 99:5567.

Lin WH, H. C., Hsu CI, Lin JY. 1997. Dimerization of the N-terminal amphipathic alpha-helix domain of the fungal immunomodulatory protein from Ganoderma tsugae (Fip-gts) defined by a yeast two-hybrid system and site-directed mutagenesis. J Biol Chem. 272:20044.

Liu JJ, H. T., Hsu ML, Chen CC, Lin WS, Lu FJ, Chang WH. 2004. Antitumor effects of the partially purified polysaccharides from Antrodia camphorata and the mechanism of its action. Toxicol Appl Pharmacol. 201:186.

LM, W. 2003. The role of Toll-like receptor 2 in microbial disease and immunity. Vaccine 21:55.

Lord KA, H.-L. B., Liebermann DA. 1990. Nucleotide sequence and expression of a cDNA encoding MyD88, a novel myeloid differentiation primary response gene induced by IL-6. Oncogene 5:1095.

Marr KA, B. S., Hawn TR, Ozinsky A, Pham U, Akira S, Aderem A, Liles WC. 2003. Differential role of MyD88 in macrophage-mediated responses to opportunistic fungal pathogens. Infect Immun. 71:5280.

McAdam KP, R. J. 1978. C57BL/10/CR mice: nonresponders to activation by the lipid a moiety of bacterial lipopolysaccharide. J Immunol. 120:249.

Muzio M, N. J., Feng P, Dixit VM. 1997. IRAK (Pelle) family member IRAK-2 and MyD88 as proximal mediators of IL-1 signaling. Science 278:1612.

Netea MG, V. D. G. C., Vonk AG, Verschueren I, Van Der Meer JW, Kullberg BJ. 2002. The role of toll-like receptor (TLR) 2 and TLR4 in the host defense against disseminated candidiasis. J Infect Dis. 185:1483.

Netea MG, v. d. G. C., Van der Meer JW, Kullberg BJ. 2004a. Toll-like receptors and the host defense against microbial pathogens: bringing specificity to the innate-immune system. J Leukoc Biol. 75:749.

Netea MG, V. d. G. C., Van der Meer JW, Kullberg BJ. 2004b. Recognition of fungal pathogens by Toll-like receptors. Eur J Clin Microbiol Infect Dis. 23:672.

Ninomiya-Tsuji J, K. K., Hiyama A, Inoue J, Cao Z, Matsumoto K. 1999. The kinase TAK1 can activate the NIK-I kappaB as well as the MAP kinase cascade in the IL-1 signalling pathway. Nature 398:252.

O''Neill LA, F. K., Bowie AG. 2003. The Toll-IL-1 receptor adaptor family grows to five members. Trends Immunol. 24:286.

Oshiumi H, M. M., Funami K, Akazawa T, Seya T. 2003a. TICAM-1, an adaptor molecule that participates in Toll-like receptor 3-mediated interferon-beta induction. Nat Immunol. 4:161.

Oshiumi H, S. M., Shida K, Fujita T, Matsumoto M, Seya T. 2003b. TIR-containing adapter molecule (TICAM)-2, a bridging adapter recruiting to toll-like receptor 4 TICAM-1 that induces interferon-beta. J Biol Chem. 278:49751.

Ozinsky A, U. D., Fontenot JD, Hajjar AM, Smith KD, Wilson CB, Schroeder L, Aderem A. 2000. The repertoire for pattern recognition of pathogens by the innate immune system is defined by cooperation between toll-like receptors. Proc Natl Acad Sci. 97:13766.

Palmer RM, A. D., Moncada S. 1988. Vascular endothelial cells synthesize nitric oxide from L-arginine. Nature 333:664.

Palsson-McDermott EM, O. N. L. 2004. Signal transduction by the lipopolysaccharide receptor, Toll-like receptor-4. Immunology 2:153.

Poltorak A, H. X., Smirnova I, Liu MY, Van Huffel C, Du X, Birdwell D, Alejos E, Silva M, Galanos C, Freudenberg M, Ricciardi-Castagnoli P, Layton B, Beutler B. 1998. Defective LPS signaling in C3H/HeJ and C57BL/10ScCr mice: mutations in Tlr4 gene. Science 282:2085.

Qureshi ST, L. L., Leveque G, Clermont S, Moore KJ, Gros P, Malo D. 1999. Endotoxin-tolerant mice have mutations in Toll-like receptor 4 (Tlr4). J Exp Med. 189:615.

Rock FL, H. G., Timans JC, Kastelein RA, Bazan JF. 1998. A family of human receptors structurally related to Drosophila Toll. Proc Natl Acad Sci 95:588.

Roeder A, K. C., Rupec RA, Schaller M, Korting HC. 2004. Toll-like receptors and innate antifungal responses. Trends Microbiol. 12:44.

Sato S, S. M., Yamamoto M, Watanabe Y, Kawai T, Takeda K, Akira S. 2003. Toll/IL-1 receptor domain-containing adaptor inducing IFN-beta (TRIF) associates with TNF receptor-associated factor 6 and TANK-binding kinase 1, and activates two distinct transcription factors, NF-kappa B and IFN-regulatory factor-3, in the Toll-like receptor signaling. J Immunol. 171:4304.

Schnare M, H. A., Takeda K, Akira S, Medzhitov R. 2000. Recognition of CpG DNA is mediated by signaling pathways dependent on the adaptor protein MyD88. Curr Biol. 10:1139.

Schumann RR, L. S., Flaggs GW, Gray PW, Wright SD, Mathison JC, Tobias PS, Ulevitch RJ. 1990. Structure and function of lipopolysaccharide binding protein. Science 249:1429.

Shen YC, Y. S., Lin CS, Chen CH, Kuo YH, Chen CF. 1997. Zhankuic acid F: a new metabolic from Formosan fungus Antrodia cinnamomea. Planta medica.:86.

Shimazu R, A. S., Ogata H, Nagai Y, Fukudome K, Miyake K, Kimoto M. 1999. MD-2, a molecule that confers lipopolysaccharide responsiveness on Toll-like receptor 4. J Exp Med. 189:1777.

Shirakabe K, Y. K., Shibuya H, Irie K, Matsuda S, Moriguchi T, Gotoh Y, Matsumoto K, Nishida E. 1997. TAK1 mediates the ceramide signaling to stress-activated protein kinase/c-Jun N-terminal kinase. J Biol Chem. 272:8141.

Shoham S, H. C., Chen JM, Golenbock DT, Levitz SM. 2001. Toll-like receptor 4 mediates intracellular signaling without TNF-alpha release in response to Cryptococcus neoformans polysaccharide capsule. J Immunol. 166:4620.

SM, L. 2004. Interactions of Toll-like receptors with fungi. Microbes Infect. 6:1351.

Song TY, Y. G. 2002. Antioxidant properties of Antrodia camphorata in submerged culture. J Agric Food Chem. 50:3322.

Song TY, Y. G. 2003. Protective effects of fermented filtrate from Antrodia camphorata in submerged culture against CCl4-induced hepatic toxicity in rats. J Agric Food Chem. 51:1571.

Takeda K, A. S. 2004. TLR signaling pathways. Semin Immunol. 16:3.

Takeuchi O, A. S. 2001. Toll-like receptors; their physiological role and signal transduction system. Int Immunopharmacol. 1:625.

Takeuchi O, K. A., Grote K, Kawai T, Hoshino K, Morr M, Muhlradt PF, Akira S. 2000. Cutting edge: preferentially the R-stereoisomer of the mycoplasmal lipopeptide macrophage-activating lipopeptide-2 activates immune cells through a toll-like receptor 2- and MyD88-dependent signaling pathway. J Immunol. 164:554.

Takeuchi O, K. T., Sanjo H, Copeland NG, Gilbert DJ, Jenkins NA, Takeda K, Akira S. 1999. TLR6: A novel member of an expanding toll-like receptor family. Gene 231:59.

Tsao CY. Studies on the purification and bioactivity of immunomodulatory proteins from Taiwanofugus camphoratus. 2003. Graduate Institute of Horticulture National Taiwan University. Master Thesis.

Ulevitch RJ, T. P. 1995. Receptor-dependent mechanisms of cell stimulation by bacterial endotoxin. Annu Rev Immunol.:437.

Underhill DM, O. A., Hajjar AM, Stevens A, Wilson CB, Bassetti M, Aderem A. 1999. The Toll-like receptor 2 is recruited to macrophage phagosomes and discriminates between pathogens. Nature 401:811.

Vogel SN, H. C., Rosenstreich DL. 1979. Characterization of a congenitally LPS-resistant, athymic mouse strain. J Immunol. 122:619.

Vogel SN, J. D., Perera PY, Medvedev A, Lariviere L, Qureshi ST, Malo D. 1999. Cutting edge: functional characterization of the effect of the C3H/HeJ defect in mice that lack an Lpsn gene: in vivo evidence for a dominant negative mutation. J Immunol. 162:5666.

Wesche H, G. X., Li X, Kirschning CJ, Stark GR, Cao Z. 1999. IRAK-M is a novel member of the Pelle/interleukin-1 receptor-associated kinase (IRAK) family. J Biol Chem. 274:19403.

Wesche H, H. W., Shillinglaw W, Li S, Cao Z. 1997. MyD88: an adapter that recruits IRAK to the IL-1 receptor complex. Immunity 7:837.

Wesche H, N. D., Resch K, Martin MU. 1996. Co-expression of mRNA for type I and type II interleukin-1 receptors and the IL-1 receptor accessory protein correlates to IL-1 responsiveness. FEBS Lett. 391:104.

Wright SD, R. R., Tobias PS, Ulevitch RJ, Mathison JC. 1990. CD14, a receptor for complexes of lipopolysaccharide (LPS) and LPS binding protein. Science 249:1431.

Wu SH, R. L., Chang TT. 1997. Antrodia champhorata (“niu-chang-chig”), new combination of a medicinal fungus in Taiwan. Bot. Bull. Acad. Sin.:273.

Wu SH, Y. Z., Dai YC, Chen CT, Su CH, Chen LC, Hsu WC, Hwang GY. 2004. Taiwanofungus, a polypore new genus. Fung. Sci. 19:109.

Yamaguchi K, S. K., Shibuya H, Irie K, Oishi I, Ueno N, Taniguchi T, Nishida E, Matsumoto K. 1995. Identification of a member of the MAPKKK family as a potential mediator of TGF-beta signal transduction. Science 270:2008.

Yamamoto M, S. S., Hemmi H, Hoshino K, Kaisho T, Sanjo H, Takeuchi O, Sugiyama M, Okabe M, Takeda K, Akira S. 2003a. Role of adaptor TRIF in the MyD88-independent toll-like receptor signaling pathway. Science 301:640.

Yamamoto M, S. S., Hemmi H, Uematsu S, Hoshino K, Kaisho T, Takeuchi O, Takeda K, Akira S. 2003b. TRAM is specifically involved in the Toll-like receptor 4-mediated MyD88-independent signaling pathway. Nat Immunol. 4:1144.

Yang SW, S. Y., Chen Ch. 1996. Streoids and triterpenoids of Antrodia cinnamomea-fungus parasitic on cinnamomum micranthum. Phytochemistry:1389.

Yoon YD, H. S., Kang JS, Lee CW, Park SK, Lee HS, Kang JS, Kim HM. 2003. Toll-like receptor 4-dependent activation of macrophages by polysaccharide isolated from the radix of Platycodon grandiflorum. Int Immunopharmacol. 3:1873.

Zang M, S., Q. 1990. Ganoderma comphoratum, a new taxon in genus Ganoderma from Taiwan, China. Acta Bot. Yuunnanica:395.