Fas-L ( CD95-Ligand ) 是屬於腫瘤壞死因子家族的一員。Fas-L 作用在帶有 Fas 的細胞上而導致細胞進行細胞凋亡。Fas/FasL 的系統在免疫反應中扮演一個重要的角色。Fas-L 不僅表現在活化的 T 細胞以及自然殺手細胞上，也表現在許多不同的腫瘤細胞上。若使不表現 Fas-L 的腫瘤細胞表現 Fas-L 後有時會誘導大量的嗜中性球浸潤，引發強烈的發炎反應，使腫瘤細胞受到排斥與破壞。Fas-L 誘導嗜中性球浸潤的機制，目前還不清楚。我們推測 Fas-L 誘發嗜中性球浸潤的機制，可能與刺激內皮細胞或是上皮細胞表現趨化因子有關。我們使用 in vitro 細胞培養系統研究 Fas-L 如何影響內皮細胞或上皮細胞趨化因子的表現。人類微血管內皮細胞 HMEC-1 以及人類呼吸道上皮細胞 A549 上都能表現 Fas。以抗體活化 Fas 後，能引起 HMEC-1 細胞凋亡，但不能誘發 HMEC-1 表現 IL-8 及GRO-a。但是，A549 細胞上的 Fas 受到刺激後，不僅引起細胞凋亡，同時也能表現 IL-8 及 GRO-a。雖然性荷爾蒙可能具有調控免疫反應與基因表現的功能，但是在本研究中，性荷爾蒙對於 A549 細胞表現 IL-8 及 GRO-a 的程度並無影響。另外，當 A549 細胞在CH-11 刺激時合併處理 caspase 抑制劑：Z-VAD-fmk、NF-kB 抑制劑：SN-50、JNK 抑制劑：SP600125、MAPK/ERK 抑制劑：PD98059、p38 MAPK 抑制劑：SB202190、PKA 抑制劑：KT5720以及HSP70抑制劑後，由 CH-11 所誘發的 IL-8 及 GRO-a 表現會受到抑制。綜合以上的研究結果證明，Fas 訊號能誘發 IL-8 及 GRO-a 表現。是否它是導至免疫細胞浸潤的主因則有待進一步的研究來驗證。

Fas-L ( CD95-Ligand ) is a member of the TNF family. Engagement of Fas-L with Fas induces apoptosis in Fas-bearing cells. Fas-L is not only expressed on activated CTL and NK cells but also on tumors of diverse cellular organs. Recent reports suggested that forced expression of Fas-L in tumors unexpectedly induced migration of neutrophils into the tumors and the FasL-expressing tumors were rejected due to the inflammatory reaction. The mechanism of the neutrophilic response to Fas-L expression is unknown. We speculate that Fas-L can induce the chemokines expression on endothelial or epithelial cells which in turn leading to cell infiltration. Fas were detected on the surface of human microvascular endothelial cell line-1 (HMEC-1). When treated with anti-Fas antibody, CH-11, HMEC-1 cells became apoptotic, but did not express IL-8 and GRO-a at transcription and translation level. Besides, A549 cells, a human airway epithelial cell line expressing Fas on the surface, were apoptotic and expressed IL-8 and GRO-a upon CH-11 treatment. In addition, sex hormones had no effects on CH-11-induced IL-8 and GRO-a productions in A549 cells. Application of caspase inhibitor：Z-VAD-fmk, NF-kB inhibitor：SN-50, JNK inhibitor：SP600125, MAPK/ERK inhibitor：PD98059, p38 MAPK inhibitor：SB202190, PKA inhibitor：KT5720 or HSP70 inhibitor could decrease the CH-11 induced productions of IL-8 and GRO-a in A549. These data suggest that Fas signal mediates induce IL-8 and GRO-a expression in A549 cells.

Ades EW, Candal FJ, Swerlick RA, George VG, Summers S, Bosse DC, Lawley TJ. HMEC-1: establishment of an immortalized human microvascular endothelial cell line. J Invest Dermatol. 99(6): 683-90,1992. (52)

Ahuja SK, Murphy PM. The CXC chemokines growth-regulated oncogene (GRO) alpha, GRObeta, GROgamma, neutrophil-activating peptide-2, and epithelial cell-derivedneutrophil-activating peptide-78 are potent agonists for the type B, but not the type A, human interleukin-8 receptor. J Biol Chem. 271(34): 20545-50,1996. (32)

Akoum A, Lemay A, Paradis I, Rheault N, Maheux R. Secretion of interleukin-6 by human endometriotic cells and regulation by proinflammatory cytokines and sex steroids. Hum Reprod. 11(10): 2269-75,1996. (39)

Arai H, Gordon D, Nabel EG, Nabel GJ. Gene transfer of Fas ligand induces tumor regression in vivo. Proc Natl Acad Sci U S A. 94(25): 13862-7,1997. (18)

Aragane Y, Kulms D, Metze D, Wilkes G, Poppelmann B, Luger TA, Schwarz T. Ultraviolet light induces apoptosis via direct activation of CD95 (Fas/APO-1) independently of its ligand CD95L. J Cell Biol. 140(1): 171-82,1998. (51)

Arici A, Senturk LM, Seli E, Bahtiyar MO, Kim G. Regulation of monocyte chemotactic protein-1 expression in human endometrial stromal cells by estrogen and progesterone. Biol Reprod. 61(1): 85-90,1999. (40)

Aubry F, Habasque C, Satie AP, Jegou B, Samson M. Expression and regulation of the CXC-chemokines, GRO/KC and IP-10/mob-1 in rat seminiferous tubules.
Eur Cytokine Netw. 11(4): 690-8,2000. (31)

Aust G, Steinert M, Boltze C, Kiessling S, Simchen C. GRO-alpha in normal and pathological thyroid tissues and its regulation in thyroid-derived cells. J Endocrinol. 170(3): 513-20,2001. (30)

Berry KK, Varney ML, Dave BJ, Bucana CD, Fidler IJ, Singh RK. Expression of interleukin-8 in human metastatic endometrial carcinoma cells and its regulation by inflammatory cytokines. Int J Gynecol Cancer. 11(1): 54-60,2001. (22)
Bhattacharyya A, Pathak S, Kundu M, Basu J. Mitogen-activated protein kinases regulate Mycobacterium avium-induced tumor necrosis factor-alpha release from macrophages. FEMS Immunol Med Microbiol. 34(1): 73-80,2002. (43)

Brutkiewicz RR, Sriram V. Natural killer T (NKT) cells and their role in antitumor immunity. Crit Rev Oncol Hematol. 41(3): 287-98,2002. (45)

Budd RC. Death receptors couple to both cell proliferation and apoptosis.
J Clin Invest. 109(4): 437-41,2002. (57)

Choi C, Xu X, Oh JW, Lee SJ, Gillespie GY, Park H, Jo H, Benveniste EN. Fas-induced expression of chemokines in human glioma cells: involvement of extracellular signal-regulated kinase 1/2 and p38 mitogen-activated protein kinase. Cancer Res. 61(7): 3084-91,2001. (59)

Chang YH, Hsieh SL, Chen MC, Lin WW. Lymphotoxin beta receptor induces interleukin 8 gene expression via NF-kappaB and AP-1 activation. Exp Cell Res. 278(2): 166-74,2002. (42)

Chen YL, Wang JY, Chen SH, Yang BC. Granulocytes mediates the Fas-L-associated apoptosis during lung metastasis of melanoma that determines the metastatic behaviour. Br J Cancer. 87(3): 359-65,2002. (14)

Chen JJ, Yao PL, Yuan A, Hong TM, Shun CT, Kuo ML, Lee YC, Yang PC. Up-regulation of tumor interleukin-8 expression by infiltrating macrophages: its correlation with tumor angiogenesis and patient survival in non-small cell lung cancer. Clin Cancer Res. 9(2): 729-37,2003. (27)

Fujiwara K, Matsukawa A, Ohkawara S, Takagi K, Yoshinaga M. Functional distinction between CXC chemokines, interleukin-8 (IL-8), and growth related oncogene (GRO)alpha in neutrophil infiltration. Lab Invest. 82(1): 15-23,2002. (47)

Flynn G, Maru S, Loughlin J, Romero IA, Male D. Regulation of chemokine receptor expression in human microglia and astrocytes. J Neuroimmunol. 136(1-2):84-93, 2003. (25)

Griffith TS, Yu X, Herndon JM, Green DR, Ferguson TA. CD95-induced apoptosis of lymphocytes in an immune privileged site induces immunological tolerance. Immunity. 5(1): 7-16,1996. (10)

Gabai VL, Meriin AB, Mosser DD, Caron AW, Rits S, Shifrin VI, Sherman MY. Hsp70 prevents activation of stress kinases. A novel pathway of cellular thermotolerance. J Biol Chem. 272(29): 18033-7,1997. (63)

Gabai VL, Meriin AB, Yaglom JA, Volloch VZ, Sherman MY. Role of Hsp70 in regulation of stress-kinase JNK: implications in apoptosis and aging. FEBS Lett. 438(1-2): 1-4,1998. (64)

Hsu PN, Lin HH, Tu CF, Chen NJ, Wu KM, Tsai HF, Hsieh SL. Expression of human Fas ligand on mouse beta islet cells does not induce insulitis but is insufficient to confer immune privilege for islet grafts. J Biomed Sci. 8(3): 262-9,2001. (17)

Hohlbaum AM, Saff RR, Marshak-Rothstein A. Fas-ligand--iron fist or Achilles' heel? Clin Immunol. 103(1): 1-6,2002. (62)

Heidemann J, Ogawa H, Dwinell MB, Rafiee P, Maaser C, Gockel HR, Otterson MF, Ota DM, Lugering N, Domschke W, Binion DG. Angiogenic effects of interleukin 8 (CXCL8) in human intestinal microvascular endothelial cells are mediated by CXCR2. J Biol Chem. 278(10): 8508-15,2003. (28)

Hor WS, Huang WL, Lin YS, Yang BC. Cross-talk between tumor cells and neutrophils through the Fas (APO-1, CD95)/FasL system: human glioma cells enhance cell viability and stimulatecytokine production in neutrophils. J Leukoc Biol. 73(3): 363-8,2003. (61)

Itoh N, Yonehara S, Ishii A, Yonehara M, Mizushima S, Sameshima M, Hase A,Seto Y, Nagata S. The polypeptide encoded by the cDNA for human cell surface antigen Fas can mediate apoptosis. Cell. 66(2): 233-43,1991. (1)

Ju ST, Matsui K, Ozdemirli M. Molecular and cellular mechanisms regulating T and B cell apoptosis through Fas/FasL interaction. Int Rev Immunol. 18(5-6): 485-513,
1999. (4)

Koga M, Hiromatsu Y, Jimi A, Inoue Y, Nonaka K. Possible involvement of Fas-mediated apoptosis in eye muscle tissue from patients with thyroid-associated ophthalmopathy. Thyroid. 8(4): 311-8,1998. (5)
Kayagaki N, Yamaguchi N, Nakayama M, Kawasaki A, Akiba H, Okumura K, Yagita H. Involvement of TNF-related apoptosis-inducing ligand in human CD4+ T cell-mediated cytotoxicity. J Immunol. 162(5): 2639-47,1999. (49)

Kang SM, Braat D, Schneider DB, O'Rourke RW, Lin Z, Ascher NL, Dichek DA,Baekkeskov S, Stock PG. A non-cleavable mutant of Fas ligand does not prevent neutrophilic destructionof islet transplants. Transplantation. 69(9): 1813-7,2000. (2)

Kataoka T, Budd RC, Holler N, Thome M, Martinon F, Irmler M, Burns K, Hahne M, Kennedy N, Kovacsovics M, Tschopp J. The caspase-8 inhibitor FLIP promotes activation of NF-kappaB and Erk signaling pathways. Curr Biol. 10(11): 640-8,2000. (56)

Liu J, Kuszynski CA, Baxter BT. Doxycycline induces Fas/Fas ligand-mediated apoptosis in Jurkat T lymphocytes. Biochem Biophys Res Commun. 260(2): 562-7,
1999. (15)

Le X, Shi Q, Wang B, Xiong Q, Qian C, Peng Z, Li XC, Tang H, Abbruzzese JL, Xie K. Molecular regulation of constitutive expression of interleukin-8 in human pancreatic adenocarcinoma. J Interferon Cytokine Res. 20(11): 935-46,2000. (23)

Luscinskas FW, Gerszten RE, Garcia-Zepeda EA, Lim YC, Yoshida M, Ding HA,
Gimbrone MA Jr, Luster AD, Rosenzweig A. C-C and C-X-C chemokines trigger firm adhesion of monocytes to vascular endothelium under flow conditions. Ann N Y Acad Sci. 902: 288-93,2000. (21)

Li J, Kartha S, Iasvovskaia S, Tan A, Bhat RK, Manaligod JM, Page K, Brasier AR, Hershenson MB. Regulation of human airway epithelial cell IL-8 expression by MAP kinases. Am J Physiol Lung Cell Mol Physiol. 283(4): L690-9,2002. (41)

Li JH, Kluger MS, Madge LA, Zheng L, Bothwell AL, Pober JS. Interferon-gamma augments CD95(APO-1/Fas) and pro-caspase-8 expression and sensitizes human vascular endothelial cells to CD95-mediated apoptosis. Am J Pathol. 161(4): 1485-95,2002. (50)

Li A, Dubey S, Varney ML, Dave BJ, Singh RK. IL-8 directly enhanced endothelial cell survival, proliferation, and matrix metalloproteinases production and regulated angiogenesis. J Immunol. 170(6): 3369-76,2003. (26)
Modur V, Zimmerman GA, Prescott SM, McIntyre TM. Endothelial cell inflammatory responses to tumor necrosis factor alpha. Ceramide-dependent and -independent mitogen-activated protein kinase cascades. J Biol Chem. 271(22): 13094-102,1996. (53)

May AE, Neumann FJ, Schomig A, Preissner KT. VLA-4 (alpha(4)beta(1)) engagement defines a novel activation pathway for beta(2) integrin-dependent leukocyte adhesion involving the urokinase receptor. Blood. 96(2): 506-13,2000. (55)

Maaser C, Schoeppner S, Kucharzik T, Kraft M, Schoenherr E, Domschke W, Luegering N. Colonic epithelial cells induce endothelial cell expression of ICAM-1 and VCAM-1 by a NF-kappaB-dependent mechanism. Clin Exp Immunol. 124(2): 208-13,2001. (35)

Mulayim N, Palter SF, Kayisli UA, Senturk L, Arici A. Chemokine receptor expression in human endometrium. Biol Reprod. 68(5): 1491-5,2003. (24)

O'Connell J, O'Sullivan GC, Collins JK, Shanahan F. The Fas counterattack: Fas-mediated T cell killing by colon cancer cells expressing Fas ligand. J Exp Med. 184(3): 1075-82,1996. (9)

O'Flaherty E, Ali S, Pettit SJ, Kirby JA. Examination of the sensitivity of T cells to Fas ligation: induction of allospecific apoptosis. Transplantation. 66(8): 1067-73,1998. (20)

Petrofsky M, Bermudez LE. Neutrophils from Mycobacterium avium-infected mice produce TNF-alpha, IL-12, and IL-1 beta and have a putative role in early host response. Clin Immunol. 91(3): 354-8,1999. (33)

Park SH, Kyin T, Bendelac A, Carnaud C. The contribution of NKT cells, NK cells, and other gamma-chain-dependent non-T non-B cells to IL-12-mediated rejection of tumors. J Immunol. 170(3): 1197-201,2003. (44)

Rabinowich H, Reichert TE, Kashii Y, Gastman BR, Bell MC, Whiteside TL. Lymphocyte apoptosis induced by Fas ligand- expressing ovarian carcinoma cells.Implications for altered expression of T cell receptor in tumor-associatedlymphocytes. J Clin Invest. 101(11): 2579-88,1998. (7)

Ronald JA, Ionescu CV, Rogers KA, Sandig M. Differential regulation of transendothelial migration of THP-1 cells by ICAM-1/LFA-1 and VCAM-1/VLA-4.
J Leukoc Biol. 70(4): 601-9,2001. (54)

Strand S, Hofmann WJ, Hug H, Muller M, Otto G, Strand D, Mariani SM,Stremmel W, Krammer PH, Galle PR. Lymphocyte apoptosis induced by CD95 (APO-1/Fas) ligand-expressing tumor cells--a mechanism of immune evasion? Nat Med. 2(12): 1361-6,1996. (8)

Seino K, Kayagaki N, Tsukada N, Fukao K, Yagita H, Okumura K. Transplantation of CD95 ligand-expressing grafts: influence of transplantation site and difficulty in protecting allo- and xenografts. Transplantation. 64(7): 1050-4,1997. (19)

Shimizu M, Fontana A, Takeda Y, Yagita H, Yoshimoto T, Matsuzawa A. Induction of antitumor immunity with Fas/APO-1 ligand (CD95L)-transfected neuroblastoma neuro-2a cells. J Immunol. 162(12): 7350-7,1999. (12)

Schafer C, Williams JA. Stress kinases and heat shock proteins in the pancreas: possible roles innormal function and disease. J Gastroenterol. 35(1): 1-9,2000. (65)

Seghaye MC, Qing M, von Bernuth G. Systemic inflammatory response to cardiac surgery: does female sex really protect? Crit Care. 5(6): 280-2,2001. (37)

Suhara T, Mano T, Oliveira BE, Walsh K. Phosphatidylinositol 3-kinase/Akt signaling controls endothelial cell sensitivity to Fas-mediated apoptosis via regulation of FLICE-inhibitory protein(FLIP). Circ Res. 89(1): 13-9,2001. (48)

Sano Y, Sotozono C. Role of Fas ligand in ocular tissue. Cornea. 21(2 Suppl 1): S30-2,2002. (16)

Scheller C, Sopper S, Ehrhardt C, Flory E, Chen P, Koutsilieri E, Ludwig S, Meulen V, Jassoy C. Caspase inhibitors induce a switch from apoptotic to proinflammatory signaling inCD95-stimulated T lymphocytes. Eur J Immunol. 32(9): 2471-80,2002. (58)

Spandau UH, Toksoy A, Verhaart S, Gillitzer R, Kruse FE. High expression of chemokines Gro-alpha (CXCL-1), IL-8 (CXCL-8), and MCP-1 (CCL-2) in inflamed human corneas in vivo. Arch Ophthalmol. 121(6): 825-31,2003. (29)
Takeuchi T, Ueki T, Nishimatsu H, Kajiwara T, Ishida T, Jishage K, Ueda O,Suzuki H, Li B, Moriyama N, Kitamura T. Accelerated rejection of Fas ligand-expressing heart grafts. J Immunol. 162(1): 518-22,1999. (3)

Turvey SE, Gonzalez-Nicolini V, Kingsley CI, Larregina AT, Morris PJ, CastroMG, Lowenstein PR, Wood KJ. Fas ligand-transfected myoblasts and islet cell transplantation. Transplantation. 69(9): 1972-6,2000. (11)

Tada Y, O-Wang J, Seimiya M, Takiguchi Y, Tatsumi K, Kuriyama T, Tagawa M. Antitumor effects are produced by forced expression of membrane-bound but not soluble Fas ligand in murine lung carcinoma cells. Anticancer Res. 22(2A): 831-6,
2002. (13)

Verthelyi D, Klinman DM. Sex hormone levels correlate with the activity of cytokine-secreting cells in vivo. Immunology. 100(3): 384-90,2000. (38)

Vermijlen D, Froelich CJ, Luo D, Suarez-Huerta N, Robaye B, Wisse E. Perforin and granzyme B induce apoptosis in FasL-resistant colon carcinoma cells. Cancer Immunol Immunother. 50(4): 212-7,2001. (46)

Verthelyi D. Sex hormones as immunomodulators in health and disease.
Int Immunopharmacol. 1(6): 983-93,2001. (36)

Walker PR, Saas P, Dietrich PY. Role of Fas ligand (CD95L) in immune escape: the tumor cell strikes back. J Immunol. 158(10): 4521-4,1997. (6)

Yamaoka-Tojo M, Yamaguchi S, Nitobe J, Abe S, Inoue S, Nozaki N, Okuyama M, Sata M, Kubota I, Nakamura H, Tomoike H. Dual response to Fas ligation in human endothelial cells: apoptosis and induction of chemokines, interleukin-8 and monocyte chemoattractant protein-1. Coron Artery Dis. 14(1): 89-94,2003. (60)

Zanone MM, Favaro E, Conaldi PG, Greening J, Bottelli A, Perin PC, Klein NJ, Peakman M, Camussi G. Persistent infection of human microvascular endothelial cells by Coxsackie Bviruses induces increased expression of adhesion molecules.
J Immunol. 171(1): 438-46,2003. (34)