银屑病皮损TLRs的表达研究
摘要
银屑病是一种在遗传易感背景下受多种环境因素影响的慢性炎症性皮肤病,为多因素复杂性疾病。近年来,在众多的危险因素中,感染引起国内外学者的广泛重视。研究显示,细菌和病毒感染与银屑病的发生发展密切相关。目前研究发现TLR家族中TLR2、TLR4和TLR9与病原微生物的感染关系密切,Toll样受体是一种模式识别受体(PRR),通过识别微生物病原体相关模式(PAMPs)来诱导非特异性免疫应答,介导相关疾病的发生发展。表皮是机体抵抗病原微生物感染的第一道屏障,主要由角质形成细胞构成,角质形成细胞不但维持表皮结构的完整性,而且能分泌多种具有生物活性的细胞因子,参与并反馈性调节皮肤免疫反应,在银屑病的发生发展中发挥了关键性的作用。角质形成细胞在病理上发生的改变是引起和维持银屑病大多数临床症状的主要原因。因此研究角质形成细胞是否表达TLRs以及TLRs是否介导了银屑病发病中细菌病毒感染诱发的免疫反应,是阐明银屑病的发生与感染相关的核心所在。本实验通过研究正常皮肤组织和银屑病患者皮损处角质形成细胞上的TLRs的表达,推论和探讨TLRs介导的感染诱发银屑病的机制。
本研究共分为两部分。第一部分:采用免疫组织化学方法检测正常皮肤组织和银屑病患者皮损处角质形成细胞上的TLR2、TLR4和TLR9的表达。第二部分:采用流式细胞仪检测角质形成细胞上TLR2、TLR4和TLR9的表达。结果显示,正常皮肤组织角质形成细胞检测到TLR2、TLR4和TLR9的表达,其中TLR2主要表达在细胞膜上,表达呈弱阳性;TLR4表达更为微弱,几乎可以忽略不计;TLR9表达阳性,阳性细胞主要位于基底细胞层和棘细胞层下部。在银屑病患者皮损处角质形成细胞,TLR2、TLR4和TLR9均呈强阳性表达,其中TLR2阳性细胞主要位于棘细胞层上部;TLR4呈强阳性表达,棘细胞层上部表达最强,逐渐向基底层细胞减弱;TLR9阳性细胞主要位于基底细胞层和棘细胞层下部。另外,在银屑病患者皮损处真皮层内,单核细胞上可见TLR2、TLR4和TLR9的阳性表达,TLR2和TLR4阳性细胞散在分布;TLR9阳性细胞则聚集分布;血管内皮细胞上还可见到TLR9的阳性表达。流式细胞仪检测到了皮肤角质形成细胞上TLR2、TLR4和TLR9的表达。
实验结论:在正常皮肤组织角质形成细胞TLR2、TLR4弱阳性表达,TLR4的表达更微弱,可以忽略不计。而角质形成细胞TLR9的表达阳性,主要在基底细胞层和棘细胞层下部。提示了角质形成细胞具有非特异性免疫功能;在银屑病皮损处角质形成细胞上TLR2、TLR4和TLR9的表达增强,其中TLR2、TLR4主要表达在表皮棘细胞层上部,TLR9表达在表皮基底细胞层和棘细胞层下部。提示了银屑病和细菌病毒感染密切相关,感染是诱发银屑病的重要因素之一,TLR2、TLR4和TLR9参与了银屑病的发生发展,在介导感染因素诱发银屑病的信号转导通路的过程中,TLRs发挥了关键性的作用;在银屑病皮损真皮内单核细胞上有TLR2、TLR4和TLR9的表达,血管内皮细胞上有TLR9的表达。提示这些细胞参与了银屑病的发生发展,非特异性免疫反应紊乱是银屑病的主要发病机制。
Psoriasis is a chronic, inflammatory skin disease, which is easily affected by hereditary and environmental factors, and the cause of this disease is complicated and indefinite.Now studies indicate that TLR2,TLR4 and TLR9 from TLRs family closely relate with the infection by pathogenic microorganism. Toll-like receptor, a kind of pattern recognition recetor, induces nonspecific immune response and mediates the nosogenesis and development of the correlated diseases by recognizing pathogen associated molecular patterns from bacterial and virus. Epidermis, which is mainly constituted by keratinocytes,is the first barrier of bodies to resist the infection by pathogen microorganism. Keratinocytes not only keep the integrality of epidermis,but also can secrete many kinds of bioactive cytokine, participate and regulate the immune reaction of skin. This kind of cell plays a critical role in the nosogenesis ang development of psoriasis. The pathological change of keratinocytes is the cause of producing and retaining the major clinical symptom of psoriasis. Our experiments invested the expression of TLRs on the keratinocyte from normal skin organism and psoriatic lession, deducted and approached the mechanism of psoriasis by infection.
Our research can be divided into two parts. Part one: detect the expression of TLR2,TLR4 and TLR9 on the keratinocytes from normal skin organism and psoriasis lession by immunohistochemistral methods. Part two: detect the expression of TLR2,TLR4 and TLR9 on the keratinocytes from normal skin organism and psoriasis lession by flow cytometer.Results indicate that keratinocytes from normal skin organism can express TLR2TLR4 and TLR9. Among of the TLRs, TLR2 mainly expresses on the cellucar membrance, and the expression is weak; The expression of TLR4 is enough weak to be ignored;TLR9 expresses strongly and masculine cells chiefly lie in the basal cell layer and the bottom of prickle cell layer. On the kertinocytes from psoriatic lession, TLR2,TLR4 and TLR9 express strongest. Cells expressing TLR2 mainly lie in the suprior part of prickle cell layer;The expression of TLR4 in the in the suprior part of prickle cell layer is strongest,and becomes weaker and weaker from the prickle cell layer to the basal cell layer. Cells expressing TLR9 mainly lie in the basal cell layer and the bottom of prickle cell layer, Besides, in the dermis layer of psoriatic lession,there are TLR2,TLR4 and TLR9 on the monocytes. Monocytes expressing TLR2 and TLR4 are rare,but Monocytes expressing TLR9 are concentrated. There are TLR9 on the vascular endothelial cells. With flow cytometry, we have detected the TLR2,TLR4 and TLR9 on the keratinocytes from normal skin organism.
Conclusion: There are fewer TLR2 and TLR4 but more TLR9 on the keratinocytes from normal skin organism. Cells expressing TLR9 mainly in the basal cell layer and the bottom of prickle cell layer. These demonstrate that keratinocytes can mediate nonspecific immumnologic reaction. The expression of TLR2,TLR4 and TLR9 up-regulates on the kerationcytes from psoriatic lession. Cells expressing TLR2 and TLR4 mainly lie in the superior part of the prickle cell layer. Cells with TLR9 chiefly lie in the basal cell layer and the inferior part of the prickle cell layer. These testify that psoriasis correlates with infection closely. Infection is an important factors inducing psoriasis. TLR2,TLR4 and TLR9 take part in the nosogenesis and development of psoriasis, and play a critical role in the progress of mediating the signal transduction of psoriasis by infections. TLR2 and TLR4 on the monocytes and TLR9 on the vascular endothelial cell in the dermis of psoriatic lession indicate that these TLRs participate and mediate the nosogenesis and development of psoriasis ,and that the disord of nonspecific immune reaction is the main pathogenesy of psoriasis.
本研究共分为两部分。第一部分:采用免疫组织化学方法检测正常皮肤组织和银屑病患者皮损处角质形成细胞上的TLR2、TLR4和TLR9的表达。第二部分:采用流式细胞仪检测角质形成细胞上TLR2、TLR4和TLR9的表达。结果显示,正常皮肤组织角质形成细胞检测到TLR2、TLR4和TLR9的表达,其中TLR2主要表达在细胞膜上,表达呈弱阳性;TLR4表达更为微弱,几乎可以忽略不计;TLR9表达阳性,阳性细胞主要位于基底细胞层和棘细胞层下部。在银屑病患者皮损处角质形成细胞,TLR2、TLR4和TLR9均呈强阳性表达,其中TLR2阳性细胞主要位于棘细胞层上部;TLR4呈强阳性表达,棘细胞层上部表达最强,逐渐向基底层细胞减弱;TLR9阳性细胞主要位于基底细胞层和棘细胞层下部。另外,在银屑病患者皮损处真皮层内,单核细胞上可见TLR2、TLR4和TLR9的阳性表达,TLR2和TLR4阳性细胞散在分布;TLR9阳性细胞则聚集分布;血管内皮细胞上还可见到TLR9的阳性表达。流式细胞仪检测到了皮肤角质形成细胞上TLR2、TLR4和TLR9的表达。
实验结论:在正常皮肤组织角质形成细胞TLR2、TLR4弱阳性表达,TLR4的表达更微弱,可以忽略不计。而角质形成细胞TLR9的表达阳性,主要在基底细胞层和棘细胞层下部。提示了角质形成细胞具有非特异性免疫功能;在银屑病皮损处角质形成细胞上TLR2、TLR4和TLR9的表达增强,其中TLR2、TLR4主要表达在表皮棘细胞层上部,TLR9表达在表皮基底细胞层和棘细胞层下部。提示了银屑病和细菌病毒感染密切相关,感染是诱发银屑病的重要因素之一,TLR2、TLR4和TLR9参与了银屑病的发生发展,在介导感染因素诱发银屑病的信号转导通路的过程中,TLRs发挥了关键性的作用;在银屑病皮损真皮内单核细胞上有TLR2、TLR4和TLR9的表达,血管内皮细胞上有TLR9的表达。提示这些细胞参与了银屑病的发生发展,非特异性免疫反应紊乱是银屑病的主要发病机制。
Psoriasis is a chronic, inflammatory skin disease, which is easily affected by hereditary and environmental factors, and the cause of this disease is complicated and indefinite.Now studies indicate that TLR2,TLR4 and TLR9 from TLRs family closely relate with the infection by pathogenic microorganism. Toll-like receptor, a kind of pattern recognition recetor, induces nonspecific immune response and mediates the nosogenesis and development of the correlated diseases by recognizing pathogen associated molecular patterns from bacterial and virus. Epidermis, which is mainly constituted by keratinocytes,is the first barrier of bodies to resist the infection by pathogen microorganism. Keratinocytes not only keep the integrality of epidermis,but also can secrete many kinds of bioactive cytokine, participate and regulate the immune reaction of skin. This kind of cell plays a critical role in the nosogenesis ang development of psoriasis. The pathological change of keratinocytes is the cause of producing and retaining the major clinical symptom of psoriasis. Our experiments invested the expression of TLRs on the keratinocyte from normal skin organism and psoriatic lession, deducted and approached the mechanism of psoriasis by infection.
Our research can be divided into two parts. Part one: detect the expression of TLR2,TLR4 and TLR9 on the keratinocytes from normal skin organism and psoriasis lession by immunohistochemistral methods. Part two: detect the expression of TLR2,TLR4 and TLR9 on the keratinocytes from normal skin organism and psoriasis lession by flow cytometer.Results indicate that keratinocytes from normal skin organism can express TLR2TLR4 and TLR9. Among of the TLRs, TLR2 mainly expresses on the cellucar membrance, and the expression is weak; The expression of TLR4 is enough weak to be ignored;TLR9 expresses strongly and masculine cells chiefly lie in the basal cell layer and the bottom of prickle cell layer. On the kertinocytes from psoriatic lession, TLR2,TLR4 and TLR9 express strongest. Cells expressing TLR2 mainly lie in the suprior part of prickle cell layer;The expression of TLR4 in the in the suprior part of prickle cell layer is strongest,and becomes weaker and weaker from the prickle cell layer to the basal cell layer. Cells expressing TLR9 mainly lie in the basal cell layer and the bottom of prickle cell layer, Besides, in the dermis layer of psoriatic lession,there are TLR2,TLR4 and TLR9 on the monocytes. Monocytes expressing TLR2 and TLR4 are rare,but Monocytes expressing TLR9 are concentrated. There are TLR9 on the vascular endothelial cells. With flow cytometry, we have detected the TLR2,TLR4 and TLR9 on the keratinocytes from normal skin organism.
Conclusion: There are fewer TLR2 and TLR4 but more TLR9 on the keratinocytes from normal skin organism. Cells expressing TLR9 mainly in the basal cell layer and the bottom of prickle cell layer. These demonstrate that keratinocytes can mediate nonspecific immumnologic reaction. The expression of TLR2,TLR4 and TLR9 up-regulates on the kerationcytes from psoriatic lession. Cells expressing TLR2 and TLR4 mainly lie in the superior part of the prickle cell layer. Cells with TLR9 chiefly lie in the basal cell layer and the inferior part of the prickle cell layer. These testify that psoriasis correlates with infection closely. Infection is an important factors inducing psoriasis. TLR2,TLR4 and TLR9 take part in the nosogenesis and development of psoriasis, and play a critical role in the progress of mediating the signal transduction of psoriasis by infections. TLR2 and TLR4 on the monocytes and TLR9 on the vascular endothelial cell in the dermis of psoriatic lession indicate that these TLRs participate and mediate the nosogenesis and development of psoriasis ,and that the disord of nonspecific immune reaction is the main pathogenesy of psoriasis.
引文
[1]Bos JD,De Rie MA.The pathogenesis of psoriasis:immunological facts and speculations.[J].Immunol Today,1999,20(1):40-46
[2]解士海,马鹏程。银屑病的生物治疗进展。国外医学皮肤性病学分册[J].2003,29(2):78-81.
[3]Komai-Koma M,Jones L,Ogg GS,et al.TLR2 is expressed on activated T cells as a costimulatory receptor.[J].Proc Natl Acad Sci USA.2004,101(9):3029-3034.
[4]Chuang Y H,Ulevitch R J.Cloning and charcterization of a subfamily of human Toll-like receptors:hTLR7,hTLR8 and hTLR9.[J].Eur Cytokine Network,2000,11(3):372-378
[5]Michael G.Interleukin-4 reduces toll-like receptor expression.[J].Biotech,2003,7(23):327.
[6]临床皮肤病学 江苏省科学技术出版社 第五版 赵辩
[7]Kreutz M,Ackermann U,Hauschildt S,et al.A comparative analysis of cytokine production and tolerance induction by bacterial lipopeptides,lipopolysacchaides and Staphyloccous aureus in human monocytes.[J].Immunology,1997,92(7):396-401
[8]Kollisch G,Kalali BN,Voelcker V,et al.Various members of the Toll-like receptor family contribute to the innate immune response of human epidermal keratinocytes.[J].Immunology.2005,114(4):531-541
[9]Baker BS,et al.Normal keratinocytes express Toll-like receptors(TLRs)1,2 and 5:modulation of TLR expression in chronic plaque psoriasis.[J].Br J Dermatol.2003,148(4):670-679
[10]Heumann D,Roger T。 Initial responses to endotoxins and Gram-negative bacteria.[J].Clin Chim Acta,2002,323(1-2):59-72.
[11]Hashimoto M,Asai Y,Ogawa T.Separation and structural analysis of lipoprotein in a lipopolysaccharide preparation from porphyromonas gingivalis.[J].Int Immunol 2004,16(10):1431-1437.
[12]Mirlashari MR,Lyberg T.Expression and involvement of Toll-like receptors.TLR2,TLR4 and CD14 in m onocyte TNF-alpha production induced by lipopolysaccharides from Neisseria meningitidis.[J].Med Sci Monit,2003,9(8):316-324.
[13]沙国华 孙琮琮.982例银屑病病因调查及防治.中国社区医师,2008,22(302)
[14]王红艳 张学军等银屑病危险因素研究。中华流行病学杂志 2001,22(3)
[15]Kirby B.Griffiths CE.Novel immune-based therapies for psoriasis.[J].Br J DermatoI.2002.146(24):546-551
[16]Zepter K,Haffner A,Soohoo LF,et al.Induction of biologieally active IL-1beta-converting enzyme and mature IL-lbeta in human keratinocytes by inflammatory and immunologic stimuli.[J].Immunol 1997;159(13):6203-6208.
[17]Delaporte E,Bieber T,Viac J,et al.Cytokines epi- -dermique at inflammation cutanee.[J].Ann Dennatol Venereo1 1994:121(4):836-843.
[18]Muzio M,Bosiso D,Polenamtfi N,et al.Diferential expression and regulation of Toll-like receptors(TLRs)in human leukcocytes:selective expression of TLRs in dendritic cells.[J].Immunol,2000,164(8):5998-6004.
[19]Pivarcsi A,Bodai L,Rethi B,et al.Expression and function of Toll-like receptors 2 and 4 in human keratinocytes.[J].Int Immunol.2003,15(6):721-730.
[20]Song PI,Park YM,Abraham T,et al.Human keratinocytes express functional CD14 and toll-like receptor 4.[J].Invest Dermatol.2003,121(1):217.
[21]Kawai K,Shimura H,Minagawa M.Expression of functional Toll-like receptor 2on human epidermal keratinocytes.[J].Dermatol Sci.2002,30(3):185-194.
[22]Fieflbeck G.Rasser G.Muller C.Psoriasis induced at the injection site of recmbenant interferon gamma.[J].Arcb Dermatol Res 1990;126(9):351-355.
[23]Takeshita F,Bernasconi NL,Onai N,et al.A role of toll-like receptor 9 in CpG DNA-induced activation of human cell。[J].Immunol,2001,167(4):3555-3558
[24]Lund J.Innate immunity is regulated by CD38,an eto-enzyme withADP-ribosy 1 cyclaseactivity[J].Microbes-Infect,2003,5(1):49-58
[25]Michel G,Mirmohammadsadegh A,Olasz E,et al.Dem- -onstration and functional analysis of IL-10 rec- -eptors in human epidermal cells:decreased expression in psoriation by an antipsoriatc gluccocorticoid in normal cultured keratinocytes.[J]. Immunol 1997; 159(12): 6291-6297.
[26] Liu Y J. IPC: professional type 1 interferon-producing cells and plasmacytoid dendritic cell precursors. [J]. Annu Rev Immunol. 2005, 23(3): 275-306.
[27] Nestle FO, Conrad C, Tun-Kyi A, et al. Plasmacytoid predendritic cells initiate psoriasis through interferon-alpha production. [J]. Exp Med. 2005, 202(1): 135-143
[28]Bos JD, De Rie MA. The pathogenesis of psoriasis: immunological facts and speculations. [J].Immunol Today, 1999, 20(1): 40-46
[1] Medzhitov R, Preston-Hurlburt P, Janeway CA Jr. A human homoelogue of the Drosophila Toll protein signals activation of adaptive immunity [J] . Nature.l997,388(6640): 394-397.
[2] Seitz M. Toll-like receptors: sensors of the innate immune system [J]. Allergy, 2003 ,58 (12):1247-1249.
[3] Bell JK, Mullen GE, Leifer CA, et al . Leucine-rich repeats a- -nd pathogen recognition in Toll-like receptors [J] .Trends Im- -munol,2003,24(10):528-533.
[4] Akira S , Hemmi H. Recognition of pathogen-associated molecular patterns by TLR family. [J]. Immunol Lett, 2003, 85(2):85-95.
[5] Kriey AM et al. CpG motifs in bacterial DNA trigger direct B-cell activation. [J].Nature, 1995; 374(6522):546-549.
[6] Hemmi H et al. A Toll-like receptor recognizes bacterial DNA. [J].Nature, 2000; 408(6813):740-745.
[7] Chuang TH et al. Toll-like receptor 9 mediates CpG-DNA signaling. [J]. Leukoc Biol, 2002; 71(3): 538-544.
[8] Chuang T H, Ulevitch R J. Cloning and charcterization of a subfamily of human Toll-like receptors: hTLR7, hTLR8 and hTLR9. [J].Eur Cytokine Network, 2000,11(3):372-378
[9] Du X, Poltorak A,Wei Y,et al .Three novel mammalian toll- like receptors: gene structure, expresion, and evolution.[J]. Eur Cytokine Network,2000,11 (3):362-371
[10] Martin M U ,Wesche H. Summary and comparison of the signaling mechanisms of the Toll/interlenkin-1 receptor family [J] .Biochim Biophy Acta , 2002,1592 (3): 265-280.
[11] Shimosto T, Kitazawa T, Katoh H, et al. Swine Toll-like receptor 9 recognizcs CpG motifs of human cell stimulant.[J].Bi- -ochim Biophys Acta, 2003,1627(1): 56-60
[12] Mambula SS , Sau K, Henneke P , et al. Toll-like receptor (TLR)signaling in response to Aspergillus fumigatus. [J]. Biol Chem, 2002 ,277 (1): 39320-39326
[13] Bourkc E, Bosisio D, Golay J, et al. The toll like receptor repertoire of human B lymphocytes: inducible and selective ex- -pression of TLR9 and TLR10 in normal and transformed cells [J].Blood,2003,102(3): 956-963
[14]Yeo SJ, Yoon JG, Hong SC, et al. CpG DNA induces self and crosshypo-rcsponsivenes of RAW264.7 cells in response to CpG DNA and lipopolysccharide: alteration in IL-1 receptor-associated kinase expression [J]. Immunol, 2003,170(2): 1032—1061.
[15] Takeshita F,Bernasconi NL,Onai N,et al. A role of toll—like receptor 9 in CpG DNA-induced activation of human cell [J].Immunol,2001,167(7):3555-3558.
[16] An HZ,Xu HM, Cao XT, et al. Up-regulation of TLR9 gene expresion by LPS in mouse macrophages via activation of NF- kB , ERK and p38 MAPK signal pathways[J].Immunology Lett. 2002,81(3): 165-169.
[17]Mempel M, Voelcker V, Plank C, et al. Toll-like receptor expression in human keratiocytes: nuclear factor kappa B cont- -rolled gene activation by Staphylococcus aureus is toll-like re- -ceptor 2 but not toll-like receptor 4 or platelet activating factor receptor dependent[J]. J Invest Dermatol,2003,121(6):1389—1396.
[18] Michael G.Interleukin-4 reduces toll-like receptor expression.[J]. Biotech, 2003, 7(23):327.
[19] Bottcher T,Von Mering M,Ebert S,et al .Differential regula- -tion of Toll-like receptor mRNAs in experimental murine centr- -al nervous system infections [J]. Neuroscience Lettes,2003, 344 (1): 17-20.
[20] Liu L,ZhouX,Shi J,et al.Toll-like receptor 9 induced by physical trauma medi--ates release of cytokines following exposure to CpGmotif in mouse skin [J] . Immunology, 2003, 110(3): 341-347.
[21] Pisetsky DS. Mechanisms of immune stimulation by bacterial DNA. [J]. Springer Semin Immunopathol, 2000, 22(1-2): 21-33.
[22] Messina JP, Gilkesen GS, Pisetsky DS, et al. Stimulation of in vitro murine lymphocyte proliferation by bacterial DNA[J]. J Immunol, 1991, 147(6): 1759-1764.
[23] Stacey KJ, Sweet MJ, Hume DA, et al. Macrophages ingest and are activated by bacterial DNA[J]. Immunol, 1996, 157(5): 2116-212.
[24] Sparwasser T, Koch ES, Vabulas RM , et al. Bacterial DNA and immuno-stimulatory CpG oligenudeotides trigger mamratien and activation of muline dendritic ceus[J]. Eur J Immunol, 1998, 28(6): 2045—2054.
[25] Verthelyi D, Ishii KJ, Gursel M, etal. Human peripheral blood cells diferentially recognize and respond to two distinct CPG motifs[J]. Immunol, 2001, 166(4): 2372-2377.
[26] Bauer S, Kirschning CJ, Hacker H, et al .Human TLR9 confers responsiveness to bacterial DNA via species-specific CpG motif recognition[J]. Prac Nail Acad Sci USA, 2001, 98(16): 9237-9242.
[27] Verthelyi D, Zenner RA. Diferential signaling by CpG DNA in DC and B cells: not just TLR9[J]. Trends Immunol, 2003, 24(10): 519—522.
[28] Takeshita F, LeiferCA, Gursel I, etal. Role of Toll—like receptor 9 in CpG DNA-induced activation of human cells [J]. Immunol, 2001, 167(7): 3555—3558.
[29] Kerkmann M, Rothelafusser S, Homung V. et al. Activation with CpG—A and CpG—B oligonudeotides reveals two distinct regulatory pathways of type I IFN synthesis in human plasmacytoid dendritic cells [J]. Immunol, 2003, 170(9): 4465
[30] Takauji R. Iho S. Takatsuka H. et al. CpC—DNA—induced IFN-alpha production involves p38 MAPK—dependent STAT1 phosphorylation in human plasmacytoid dendritic cell precursors[J]. Leukoc Biol,2002 . 72(5): 1011
[31] Underhill DM. Toll-like receptors: networking for success[J]. Eur J Immunol, 2003, 33(7): 176
[32] Kawai T , Adachi O , Ogawa T , et al. Unresponsiveness of MyD88-deficient mice to endotoxin[J]. Immunity, 1999, 11(6): 1152-1221
[33] Yamamoto M , Sato S , Hemmi H , et al. Essential role of TIRAP/Mal for activation of the signaling cascade shared by TLR2 and TLR4[J]. Nature , 2002 , 420(8) :3242-3291
[34] Yamamoto M, Sato S , Hemmi H, et al. Role of adaptor TRIF inthe MyD88--independent toll-like receptor signaling pathway[J]. Science, 2003, 301(2):640-643.
[35] Sharma S , tenOever BR , Grandvaux N , et al. Triggering the interferon antiviral response through an IKK-related pathway[J] Science,2003,300 (6):11482-11511
[36] Horng T , Barton GM , Flavell RA , et al. The adaptor molecule TIRAP provides signalling specificity for Toll-like receptors [J]. Nature , 2002 , 420(7) :3292-3331
[37] Yamaoto, et al. Peripheral blood mononuclear cell proliferative response against staphylococcal superantigens in patients with psoriasis arthropathy[J]. Eur J Dermatol, 1999, 9(1): 17-21
[38] Nielsen, et al. Staphylococcal enterotoxin-A directly stimu- -lates signal transduction and interferon-gamma production in psoriatic T-cell lines[J]. Tissue Antigens. 1998, 52(6):530-8.
[39] Weissenborn , et al. High prevalence of a variety of epidermodysplasia verruciformis-associated human papill- -omaviruses in psoriatic skin of patients treated or not treated with PUVA[J]. Invest Dermatol, 1999, 113(1): 122-6
[40] Favre M, et al. Psoriasis: A possible reservoir for human papillomavirus type 5, the virus associated with skin carcinomas of epidermodysplasia verruciformis [J]. Invest Dermatol. 1998 , 110(4):311-7
[41] Tokura,et al .Hyporesponsiveness of peripheral blood lymphocytes to streptoc--occal superantigens in patients with guttate psoriasis: evidence for systemic stimulation of T cells with superantigens released from focally infecting Streptococcus pyogenes[J]. Arch Dermatol Res,1999,291(7-8):382-389.
[42]Valdimarsson H , et al. Is psoriasis induced by streptococcal superantigens and maintained by M-protein-specific T cells that cross-react with keratin? [J]. Clin Exp Immunol. 1997, 107 (Supp 1) 1:21-4
[43] Valdimarsson H, et al. Psoriasis: a T-cell-mediated autoimmune disease induced by streptococcal superantigens? [J]. Immunol Today. 1995 16(3):145-149
[44] Villeda, et al. Clinical trial of ototopical ofloxacin for treatment of chronic suppurative otitis media [J].ArchMed Res, 1998, 29(2): 143-146
[45] Bonifati C, et al. Correlated increases of tumour necrosis factor-alpha, interleukin-6 and granulocyte monocyte-colony stimulating factor levels in suction blister fluids and sera of psoriatic patients-relationships with disease severity [J]. Clin ExpDermatol, 1994, 19(5): 383-7
[46] Kristensen M , et al. Localization of tumour necrosis factor-alpha (TNF-alpha) and its receptors in normal and pso -riatic skin: epidermal cells express the 55-kD but not the 75-kD TNF receptor[J]. Clin Exp Immunol, 1993, 94(2): 354-62
[47] Bonifati C, et al. Correlated increases of tumour necrosis factor-alpha, interleukin-6 and granulocyte monocyte-colony stimulating factor levels in suction blister fluids and sera of pso -riatic patients-relationships with disease severity[J]. Clin Exp Der-matol, 1994, 19(5): 383-387
[48] Nickolof, et al. Superantigens, autoantigens, and pathogenic T cells in psoriasis [J]. Invest Dermatol. 1998 Apr;110(4):459-460.
[49] Weitzul, Duvic M. HIV-related psoriasis and Reiter's syndrome[J]. Semin Cutan MedSurg, 1997,16(3):213-218
[50] Bunse T. Mahrle G.Soluble Pityrosporum-derived chemoattractant for polym--orphonuclear leukocytes of psoriatic patients[J]. Arch Dermatol Venereol.1996, 76(1):10-15
[2]解士海,马鹏程。银屑病的生物治疗进展。国外医学皮肤性病学分册[J].2003,29(2):78-81.
[3]Komai-Koma M,Jones L,Ogg GS,et al.TLR2 is expressed on activated T cells as a costimulatory receptor.[J].Proc Natl Acad Sci USA.2004,101(9):3029-3034.
[4]Chuang Y H,Ulevitch R J.Cloning and charcterization of a subfamily of human Toll-like receptors:hTLR7,hTLR8 and hTLR9.[J].Eur Cytokine Network,2000,11(3):372-378
[5]Michael G.Interleukin-4 reduces toll-like receptor expression.[J].Biotech,2003,7(23):327.
[6]临床皮肤病学 江苏省科学技术出版社 第五版 赵辩
[7]Kreutz M,Ackermann U,Hauschildt S,et al.A comparative analysis of cytokine production and tolerance induction by bacterial lipopeptides,lipopolysacchaides and Staphyloccous aureus in human monocytes.[J].Immunology,1997,92(7):396-401
[8]Kollisch G,Kalali BN,Voelcker V,et al.Various members of the Toll-like receptor family contribute to the innate immune response of human epidermal keratinocytes.[J].Immunology.2005,114(4):531-541
[9]Baker BS,et al.Normal keratinocytes express Toll-like receptors(TLRs)1,2 and 5:modulation of TLR expression in chronic plaque psoriasis.[J].Br J Dermatol.2003,148(4):670-679
[10]Heumann D,Roger T。 Initial responses to endotoxins and Gram-negative bacteria.[J].Clin Chim Acta,2002,323(1-2):59-72.
[11]Hashimoto M,Asai Y,Ogawa T.Separation and structural analysis of lipoprotein in a lipopolysaccharide preparation from porphyromonas gingivalis.[J].Int Immunol 2004,16(10):1431-1437.
[12]Mirlashari MR,Lyberg T.Expression and involvement of Toll-like receptors.TLR2,TLR4 and CD14 in m onocyte TNF-alpha production induced by lipopolysaccharides from Neisseria meningitidis.[J].Med Sci Monit,2003,9(8):316-324.
[13]沙国华 孙琮琮.982例银屑病病因调查及防治.中国社区医师,2008,22(302)
[14]王红艳 张学军等银屑病危险因素研究。中华流行病学杂志 2001,22(3)
[15]Kirby B.Griffiths CE.Novel immune-based therapies for psoriasis.[J].Br J DermatoI.2002.146(24):546-551
[16]Zepter K,Haffner A,Soohoo LF,et al.Induction of biologieally active IL-1beta-converting enzyme and mature IL-lbeta in human keratinocytes by inflammatory and immunologic stimuli.[J].Immunol 1997;159(13):6203-6208.
[17]Delaporte E,Bieber T,Viac J,et al.Cytokines epi- -dermique at inflammation cutanee.[J].Ann Dennatol Venereo1 1994:121(4):836-843.
[18]Muzio M,Bosiso D,Polenamtfi N,et al.Diferential expression and regulation of Toll-like receptors(TLRs)in human leukcocytes:selective expression of TLRs in dendritic cells.[J].Immunol,2000,164(8):5998-6004.
[19]Pivarcsi A,Bodai L,Rethi B,et al.Expression and function of Toll-like receptors 2 and 4 in human keratinocytes.[J].Int Immunol.2003,15(6):721-730.
[20]Song PI,Park YM,Abraham T,et al.Human keratinocytes express functional CD14 and toll-like receptor 4.[J].Invest Dermatol.2003,121(1):217.
[21]Kawai K,Shimura H,Minagawa M.Expression of functional Toll-like receptor 2on human epidermal keratinocytes.[J].Dermatol Sci.2002,30(3):185-194.
[22]Fieflbeck G.Rasser G.Muller C.Psoriasis induced at the injection site of recmbenant interferon gamma.[J].Arcb Dermatol Res 1990;126(9):351-355.
[23]Takeshita F,Bernasconi NL,Onai N,et al.A role of toll-like receptor 9 in CpG DNA-induced activation of human cell。[J].Immunol,2001,167(4):3555-3558
[24]Lund J.Innate immunity is regulated by CD38,an eto-enzyme withADP-ribosy 1 cyclaseactivity[J].Microbes-Infect,2003,5(1):49-58
[25]Michel G,Mirmohammadsadegh A,Olasz E,et al.Dem- -onstration and functional analysis of IL-10 rec- -eptors in human epidermal cells:decreased expression in psoriation by an antipsoriatc gluccocorticoid in normal cultured keratinocytes.[J]. Immunol 1997; 159(12): 6291-6297.
[26] Liu Y J. IPC: professional type 1 interferon-producing cells and plasmacytoid dendritic cell precursors. [J]. Annu Rev Immunol. 2005, 23(3): 275-306.
[27] Nestle FO, Conrad C, Tun-Kyi A, et al. Plasmacytoid predendritic cells initiate psoriasis through interferon-alpha production. [J]. Exp Med. 2005, 202(1): 135-143
[28]Bos JD, De Rie MA. The pathogenesis of psoriasis: immunological facts and speculations. [J].Immunol Today, 1999, 20(1): 40-46
[1] Medzhitov R, Preston-Hurlburt P, Janeway CA Jr. A human homoelogue of the Drosophila Toll protein signals activation of adaptive immunity [J] . Nature.l997,388(6640): 394-397.
[2] Seitz M. Toll-like receptors: sensors of the innate immune system [J]. Allergy, 2003 ,58 (12):1247-1249.
[3] Bell JK, Mullen GE, Leifer CA, et al . Leucine-rich repeats a- -nd pathogen recognition in Toll-like receptors [J] .Trends Im- -munol,2003,24(10):528-533.
[4] Akira S , Hemmi H. Recognition of pathogen-associated molecular patterns by TLR family. [J]. Immunol Lett, 2003, 85(2):85-95.
[5] Kriey AM et al. CpG motifs in bacterial DNA trigger direct B-cell activation. [J].Nature, 1995; 374(6522):546-549.
[6] Hemmi H et al. A Toll-like receptor recognizes bacterial DNA. [J].Nature, 2000; 408(6813):740-745.
[7] Chuang TH et al. Toll-like receptor 9 mediates CpG-DNA signaling. [J]. Leukoc Biol, 2002; 71(3): 538-544.
[8] Chuang T H, Ulevitch R J. Cloning and charcterization of a subfamily of human Toll-like receptors: hTLR7, hTLR8 and hTLR9. [J].Eur Cytokine Network, 2000,11(3):372-378
[9] Du X, Poltorak A,Wei Y,et al .Three novel mammalian toll- like receptors: gene structure, expresion, and evolution.[J]. Eur Cytokine Network,2000,11 (3):362-371
[10] Martin M U ,Wesche H. Summary and comparison of the signaling mechanisms of the Toll/interlenkin-1 receptor family [J] .Biochim Biophy Acta , 2002,1592 (3): 265-280.
[11] Shimosto T, Kitazawa T, Katoh H, et al. Swine Toll-like receptor 9 recognizcs CpG motifs of human cell stimulant.[J].Bi- -ochim Biophys Acta, 2003,1627(1): 56-60
[12] Mambula SS , Sau K, Henneke P , et al. Toll-like receptor (TLR)signaling in response to Aspergillus fumigatus. [J]. Biol Chem, 2002 ,277 (1): 39320-39326
[13] Bourkc E, Bosisio D, Golay J, et al. The toll like receptor repertoire of human B lymphocytes: inducible and selective ex- -pression of TLR9 and TLR10 in normal and transformed cells [J].Blood,2003,102(3): 956-963
[14]Yeo SJ, Yoon JG, Hong SC, et al. CpG DNA induces self and crosshypo-rcsponsivenes of RAW264.7 cells in response to CpG DNA and lipopolysccharide: alteration in IL-1 receptor-associated kinase expression [J]. Immunol, 2003,170(2): 1032—1061.
[15] Takeshita F,Bernasconi NL,Onai N,et al. A role of toll—like receptor 9 in CpG DNA-induced activation of human cell [J].Immunol,2001,167(7):3555-3558.
[16] An HZ,Xu HM, Cao XT, et al. Up-regulation of TLR9 gene expresion by LPS in mouse macrophages via activation of NF- kB , ERK and p38 MAPK signal pathways[J].Immunology Lett. 2002,81(3): 165-169.
[17]Mempel M, Voelcker V, Plank C, et al. Toll-like receptor expression in human keratiocytes: nuclear factor kappa B cont- -rolled gene activation by Staphylococcus aureus is toll-like re- -ceptor 2 but not toll-like receptor 4 or platelet activating factor receptor dependent[J]. J Invest Dermatol,2003,121(6):1389—1396.
[18] Michael G.Interleukin-4 reduces toll-like receptor expression.[J]. Biotech, 2003, 7(23):327.
[19] Bottcher T,Von Mering M,Ebert S,et al .Differential regula- -tion of Toll-like receptor mRNAs in experimental murine centr- -al nervous system infections [J]. Neuroscience Lettes,2003, 344 (1): 17-20.
[20] Liu L,ZhouX,Shi J,et al.Toll-like receptor 9 induced by physical trauma medi--ates release of cytokines following exposure to CpGmotif in mouse skin [J] . Immunology, 2003, 110(3): 341-347.
[21] Pisetsky DS. Mechanisms of immune stimulation by bacterial DNA. [J]. Springer Semin Immunopathol, 2000, 22(1-2): 21-33.
[22] Messina JP, Gilkesen GS, Pisetsky DS, et al. Stimulation of in vitro murine lymphocyte proliferation by bacterial DNA[J]. J Immunol, 1991, 147(6): 1759-1764.
[23] Stacey KJ, Sweet MJ, Hume DA, et al. Macrophages ingest and are activated by bacterial DNA[J]. Immunol, 1996, 157(5): 2116-212.
[24] Sparwasser T, Koch ES, Vabulas RM , et al. Bacterial DNA and immuno-stimulatory CpG oligenudeotides trigger mamratien and activation of muline dendritic ceus[J]. Eur J Immunol, 1998, 28(6): 2045—2054.
[25] Verthelyi D, Ishii KJ, Gursel M, etal. Human peripheral blood cells diferentially recognize and respond to two distinct CPG motifs[J]. Immunol, 2001, 166(4): 2372-2377.
[26] Bauer S, Kirschning CJ, Hacker H, et al .Human TLR9 confers responsiveness to bacterial DNA via species-specific CpG motif recognition[J]. Prac Nail Acad Sci USA, 2001, 98(16): 9237-9242.
[27] Verthelyi D, Zenner RA. Diferential signaling by CpG DNA in DC and B cells: not just TLR9[J]. Trends Immunol, 2003, 24(10): 519—522.
[28] Takeshita F, LeiferCA, Gursel I, etal. Role of Toll—like receptor 9 in CpG DNA-induced activation of human cells [J]. Immunol, 2001, 167(7): 3555—3558.
[29] Kerkmann M, Rothelafusser S, Homung V. et al. Activation with CpG—A and CpG—B oligonudeotides reveals two distinct regulatory pathways of type I IFN synthesis in human plasmacytoid dendritic cells [J]. Immunol, 2003, 170(9): 4465
[30] Takauji R. Iho S. Takatsuka H. et al. CpC—DNA—induced IFN-alpha production involves p38 MAPK—dependent STAT1 phosphorylation in human plasmacytoid dendritic cell precursors[J]. Leukoc Biol,2002 . 72(5): 1011
[31] Underhill DM. Toll-like receptors: networking for success[J]. Eur J Immunol, 2003, 33(7): 176
[32] Kawai T , Adachi O , Ogawa T , et al. Unresponsiveness of MyD88-deficient mice to endotoxin[J]. Immunity, 1999, 11(6): 1152-1221
[33] Yamamoto M , Sato S , Hemmi H , et al. Essential role of TIRAP/Mal for activation of the signaling cascade shared by TLR2 and TLR4[J]. Nature , 2002 , 420(8) :3242-3291
[34] Yamamoto M, Sato S , Hemmi H, et al. Role of adaptor TRIF inthe MyD88--independent toll-like receptor signaling pathway[J]. Science, 2003, 301(2):640-643.
[35] Sharma S , tenOever BR , Grandvaux N , et al. Triggering the interferon antiviral response through an IKK-related pathway[J] Science,2003,300 (6):11482-11511
[36] Horng T , Barton GM , Flavell RA , et al. The adaptor molecule TIRAP provides signalling specificity for Toll-like receptors [J]. Nature , 2002 , 420(7) :3292-3331
[37] Yamaoto, et al. Peripheral blood mononuclear cell proliferative response against staphylococcal superantigens in patients with psoriasis arthropathy[J]. Eur J Dermatol, 1999, 9(1): 17-21
[38] Nielsen, et al. Staphylococcal enterotoxin-A directly stimu- -lates signal transduction and interferon-gamma production in psoriatic T-cell lines[J]. Tissue Antigens. 1998, 52(6):530-8.
[39] Weissenborn , et al. High prevalence of a variety of epidermodysplasia verruciformis-associated human papill- -omaviruses in psoriatic skin of patients treated or not treated with PUVA[J]. Invest Dermatol, 1999, 113(1): 122-6
[40] Favre M, et al. Psoriasis: A possible reservoir for human papillomavirus type 5, the virus associated with skin carcinomas of epidermodysplasia verruciformis [J]. Invest Dermatol. 1998 , 110(4):311-7
[41] Tokura,et al .Hyporesponsiveness of peripheral blood lymphocytes to streptoc--occal superantigens in patients with guttate psoriasis: evidence for systemic stimulation of T cells with superantigens released from focally infecting Streptococcus pyogenes[J]. Arch Dermatol Res,1999,291(7-8):382-389.
[42]Valdimarsson H , et al. Is psoriasis induced by streptococcal superantigens and maintained by M-protein-specific T cells that cross-react with keratin? [J]. Clin Exp Immunol. 1997, 107 (Supp 1) 1:21-4
[43] Valdimarsson H, et al. Psoriasis: a T-cell-mediated autoimmune disease induced by streptococcal superantigens? [J]. Immunol Today. 1995 16(3):145-149
[44] Villeda, et al. Clinical trial of ototopical ofloxacin for treatment of chronic suppurative otitis media [J].ArchMed Res, 1998, 29(2): 143-146
[45] Bonifati C, et al. Correlated increases of tumour necrosis factor-alpha, interleukin-6 and granulocyte monocyte-colony stimulating factor levels in suction blister fluids and sera of psoriatic patients-relationships with disease severity [J]. Clin ExpDermatol, 1994, 19(5): 383-7
[46] Kristensen M , et al. Localization of tumour necrosis factor-alpha (TNF-alpha) and its receptors in normal and pso -riatic skin: epidermal cells express the 55-kD but not the 75-kD TNF receptor[J]. Clin Exp Immunol, 1993, 94(2): 354-62
[47] Bonifati C, et al. Correlated increases of tumour necrosis factor-alpha, interleukin-6 and granulocyte monocyte-colony stimulating factor levels in suction blister fluids and sera of pso -riatic patients-relationships with disease severity[J]. Clin Exp Der-matol, 1994, 19(5): 383-387
[48] Nickolof, et al. Superantigens, autoantigens, and pathogenic T cells in psoriasis [J]. Invest Dermatol. 1998 Apr;110(4):459-460.
[49] Weitzul, Duvic M. HIV-related psoriasis and Reiter's syndrome[J]. Semin Cutan MedSurg, 1997,16(3):213-218
[50] Bunse T. Mahrle G.Soluble Pityrosporum-derived chemoattractant for polym--orphonuclear leukocytes of psoriatic patients[J]. Arch Dermatol Venereol.1996, 76(1):10-15