Toll样受体7与皮肤病的研究进展

Toll样受体是一类模式识别受体,通过识别病原相关分子模式参与先天性免疫和获得性免疫.Toll样受体有10余种家族成员,Toll样受体7是其中一种,分布于细胞内噬体膜上,主要识别内噬体中的单链核酸,在病毒识别及病毒感染的传递中起重要作用.近期研究发现,Toll样受体7在一些皮肤病中起重要作用,Toll样受体7既可诱发和加重白癜风、红斑狼疮、银屑病,也可在部分皮肤病中发挥积极的作用,如Toll样受体7可激发宿主抗真菌感染的免疫应答;在特应性皮炎中促进向Th1型免疫应答转化;Toll样受体7还可通过其特效激动剂咪喹莫特辅助治疗皮肤肿瘤及尖锐湿疣.     

Intercellular adhesion molecule-1 suppression in skin by topical delivery of anti-sense oligonucleotides

We topically applied 20 nucleotide phosphorothioate intercellular adhesion molecule-1 anti-sense oligodeoxynucleotide in a cream formulation. It effectively inhibited tumor necrosis factor-alpha-induced expression of intercellular adhesion molecule-1 in human skin transplanted on severe compromised immunodeficient mice. The effects were concentration dependent, sequence specific, and resulted from reduction of intercellular adhesion molecule-1 mRNA levels in the skin. Intravenous administration of the drug did not show pharmacologic effects, probably due to insufficient drug concentrations in skin. Topical delivery, however, produced a rapid and a significantly higher accumulation of oligodeoxynucleotide in the epidermis and dermis. The results strongly suggest that topically applied anti-sense oligonucleotides can be delivered to target sites in the skin and may be of considerable value in the treatment of psoriasis and other inflammatory skin disorders.     

孢子丝菌病患者皮损中Toll样受体2、4及髓样分化因子88的表达

【摘要】 目的 通过观察孢子丝菌病患者皮损中Toll样受体2、4（TLR2、TLR4）以及髓样分化因子88（MyD88）的表达情况,探讨其在孢子丝菌病免疫识别及免疫介导中的作用。 方法 选择孢子丝菌病患者19例及健康人对照12例,应用免疫组化法分别检测患者组皮损及对照组皮肤组织中TLR4及MyD88的表达情况,同时应用实时荧光定量PCR技术检测TLR2及MyD88 mRNA的表达情况。所有结果数据以■ ± s表示,采用SPSS17.0统计软件进行数据分析,两组间比较采用独立样本t检验,P ＜ 0.05认为差异有统计学意义。 结果 免疫组化分析：孢子丝菌病患者皮损中TLR4、MyD88的表达部位主要是除角质层外的表皮全层和真皮及皮下组织中的浆细胞和淋巴细胞。对照组皮肤几乎不表达TLR4,MyD88于真皮及皮下组织呈弱表达。孢子丝菌病组TLR4表达水平（63.767 ± 3.829）高于对照组（5.167 ± 3.246）,差异有统计学意义（t = 4.82,P ＜ 0.05）;MyD88表达水平（57.236 ± 4.744）亦高于对照组（10.588 ± 1.640）,差异有统计学意义（t = 3.30,P ＜ 0.05）。实时荧光定量PCR分析：孢子丝菌病患者皮损TLR2 mRNA和MyD88 mRNA的相对表达水平分别为1.974 ± 1.452和2.028 ± 2.061,均显著高于对照组（分别为1.430 ± 1.073和0.688 ± 0.422）,均P ＜ 0.05。 结论 孢子丝菌病发病可能与真菌通过Toll样受体途径作用于机体免疫系统有关。     

The epidermal growth factor receptor system in skin repair and inflammation

The epidermal growth factor (EGF) family comprises multiple mediators such as transforming growth factor-alpha, amphiregulin, heparin binding-EGF, and epiregulin, which are crucially involved in the tissue-specific proliferation/differentiation homeostasis. Typically, they act in an autocrine and paracrine manner on their specific cell membrane receptor and mount an effective reparative response to any attack to biophysical integrity. In addition, the EGFR can be activated by transactivation from a variety of G-protein-coupled receptors, integrins, and cytokine receptors, so that it acts as the major transducer of disparate cell functions, including changes in proliferation rate, cellular shape, attachment and motility, and regulation of proinflammatory activation. However, numerous experimental observations indicate that the different EGFR ligands are not redundant, but may rather provide distinct and specific contributions to keratinocyte functions. Importantly, increasing evidence now suggests that the EGFR pathway has a major impact on the inflammatory/immune reactions of the skin, in the apparent effort of enhancing innate immune defense while opposing overactivation of keratinocyte pro-inflammatory functions. This review covers the molecular mechanisms and functions activated by this major growth factor system in the regulation of keratinocyte biology and focuses on the complex contribution of EGFR signaling to the inflammatory processes in the skin.     

Human keratinocytes express functional Toll-like receptor 3, 4, 5, and 9

Keratinocytes are continuously in contact with external stimuli and have the capacity to produce several soluble mediators. Pathogen-associated molecular patterns (PAMPs) are recognized, among others, by Toll-like receptors (TLRs). The functional responses of keratinocytes to different PAMPs have not yet been fully established. Here we show that keratinocytes constitutively express TLR1, 2, 3, 4, 5, 6, 9, and 10 mRNA, but not TLR7 and 8. Stimulation of keratinocytes with TLR3, 4, 5, and 9 ligands resulted in differential immune-associated responses. Tumor necrosis factor-alpha, CXC chemokine ligand 8 (CXCL8), CCL2, and C chemokine ligand 20 (CCL20) release was enhanced in response to all PAMPs tested, in a time- and dose-dependent manner. Only TLR9 ligand CpG-oligodeoxynucleotides (ODNs) and TLR3 ligand poly-I:C could additionally induce type I IFNs. CCL27 production was selectively induced by poly-I:C and flagellin, whereas CXCL9 and CXCL10 were exclusively induced by CpG-ODNs and/or poly-I:C. Upregulation of ICAM-1, HLA-DR, HLA-ABC, FasR, and CD40 was mainly observed in response to poly-I:C, flagellin, and lipopolysaccharide. Furthermore, PAMP triggering resulted in the phosphorylation of phosphorylated-IkappaB alpha and in the nucleus translocation of NF-kappaB p65. Altogether, these findings stress an unexpectedly multifaceted role of keratinocytes in innate immunity as evident by their differential, TLR-mediated responses to PAMPs associated with different classes of pathogens.     

Toll样受体9基因多态性与尖锐湿疣的研究

目的探讨Toll样受体9的单核苷酸多态性与尖锐湿疣(CA)发病的相关性.方法抽取63例CA患者和23例正常人对照组的外周静脉血,采用TLR9基因直接测序的方法分析TLR9受体的单核苷酸多态性.结果 TLR9基因自翻译起始点第1 174、1635、1269、1724位均存在单核苷酸多态性位点,分别称为SNP1、SNP2、SNP3和SNP4,其中SNP3、SNP4为新发现位点,SNP1、SNP2为NCBI数据库中公布的位点,登录号分别为rs352139、rs352140.CA患者与正常人对照组的SNP1位点的等位基因A的频率分别为0.690、0.609,等位基因G的频率0.309、0.391,两组间比较差异均无统计学意义(P>0.05).SNP2位点的等位基因A的频率分别为0.302、0.369,等位基因G的频率0.698、0.630,两组间比较差异无统计学意义(P>0.05).SNP1、SNP2两个多态性位点存在4种单倍体型,分别为AA、AG、GA、GG.每种单倍体型在CA患者组和正常人对照组间比较差异均无统计学意义(P>0.05).结论 TLR9基因在广东汉族人群中存在4个单核苷酸多态性位点,分别为SNP1、SNP2、SNP3、SNP4.SNP1、SNP2两位点与CA的发病易感性可能无相关性.     

Toll样受体9基因多态性与尖锐湿疣的研究

目的探讨Toll样受体9的单核苷酸多态性与尖锐湿疣(CA)发病的相关性.方法抽取63例CA患者和23例正常人对照组的外周静脉血,采用TLR9基因直接测序的方法分析TLR9受体的单核苷酸多态性.结果 TLR9基因自翻译起始点第1 174、1635、1269、1724位均存在单核苷酸多态性位点,分别称为SNP1、SNP2、SNP3和SNP4,其中SNP3、SNP4为新发现位点,SNP1、SNP2为NCBI数据库中公布的位点,登录号分别为rs352139、rs352140.CA患者与正常人对照组的SNP1位点的等位基因A的频率分别为0.690、0.609,等位基因G的频率0.309、0.391,两组间比较差异均无统计学意义(P>0.05).SNP2位点的等位基因A的频率分别为0.302、0.369,等位基因G的频率0.698、0.630,两组间比较差异无统计学意义(P>0.05).SNP1、SNP2两个多态性位点存在4种单倍体型,分别为AA、AG、GA、GG.每种单倍体型在CA患者组和正常人对照组间比较差异均无统计学意义(P>0.05).结论 TLR9基因在广东汉族人群中存在4个单核苷酸多态性位点,分别为SNP1、SNP2、SNP3、SNP4.SNP1、SNP2两位点与CA的发病易感性可能无相关性.     

Toll样受体9基因多态性与尖锐湿疣的研究

目的 探讨Toll样受体9的单核苷酸多态性与尖锐湿疣（CA）发病的相关性。方法 抽取63例CA患者和23例正常人对照组的外周静脉血,采用TLR9基因直接测序的方法分析TLR9受体的单核苷酸多态性。结果 TLR9基因自翻译起始点第1174、1635、1269、1724位均存在单核苷酸多态性位点,分别称为SNP1、SNP2、SNP3和SNP4,其中SNP3、SNP4为新发现位点,SNP1、SNP2为NCBI数据库中公布的位点,登录号分别为rs352139、rs352140。CA患者与正常人对照组的SNP1位点的等位基因A的频率分别为0.690、0.609,等位基因G的频率0.309、0.391,两组间比较差异均无统计学意义（P ＞ 0.05）。SNP2位点的等位基因A的频率分别为0.302、0.369,等位基因G的频率0.698、0.630,两组间比较差异无统计学意义（P ＞ 0.05）。SNP1、SNP2两个多态性位点存在4种单倍体型,分别为AA、AG、GA、GG。每种单倍体型在CA患者组和正常人对照组间比较差异均无统计学意义（P ＞ 0.05）。结论 TLR9基因在广东汉族人群中存在4个单核苷酸多态性位点,分别为SNP1、SNP2、SNP3、SNP4。SNP1、SNP2两位点与CA的发病易感性可能无相关性。     

Toll样受体9基因多态性与尖锐湿疣的研究

目的探讨Toll样受体9的单核苷酸多态性与尖锐湿疣(CA)发病的相关性.方法抽取63例CA患者和23例正常人对照组的外周静脉血,采用TLR9基因直接测序的方法分析TLR9受体的单核苷酸多态性.结果 TLR9基因自翻译起始点第1 174、1635、1269、1724位均存在单核苷酸多态性位点,分别称为SNP1、SNP2、SNP3和SNP4,其中SNP3、SNP4为新发现位点,SNP1、SNP2为NCBI数据库中公布的位点,登录号分别为rs352139、rs352140.CA患者与正常人对照组的SNP1位点的等位基因A的频率分别为0.690、0.609,等位基因G的频率0.309、0.391,两组间比较差异均无统计学意义(P>0.05).SNP2位点的等位基因A的频率分别为0.302、0.369,等位基因G的频率0.698、0.630,两组间比较差异无统计学意义(P>0.05).SNP1、SNP2两个多态性位点存在4种单倍体型,分别为AA、AG、GA、GG.每种单倍体型在CA患者组和正常人对照组间比较差异均无统计学意义(P>0.05).结论 TLR9基因在广东汉族人群中存在4个单核苷酸多态性位点,分别为SNP1、SNP2、SNP3、SNP4.SNP1、SNP2两位点与CA的发病易感性可能无相关性.     

带状疱疹患者单一核细胞Toll样受体3和9的表达

目的　探讨Toll样受体TLR3和TLR9在带状疱疹发病机制中的作用。方法　采用流式细胞仪检测40例带状疱疹患者外周血单一核细胞（PBMC）上TLR3和TLR9的表达水平。16例正常人群为对照组。结果 带状疱疹患者PBMC上TLR3和TLR9的表达水平均明显高于正常人对照组（ P < 0. 01）。带状疱疹患者PBMC上TLR3和TLR9的表达水平与发病时间存在负相关关系（ r值分别为- 6.23,- 5.88, P值均 < 0. 01） ,但与性别、年龄无相关性。带状疱疹患者PBMC上TLR3和TLR9表达在各年龄组间（< 40岁, 40 ~ 50岁, > 50岁）,差异具有统计学意义（F = 3.410,P < 0. 01）,且< 40岁年龄组TLR3和TLR9的表达明显高于其他两组。结论　TLR3和TLR9可能参与带状疱疹发病。     

Toll-like receptor 3 ligand polyinosinic:polycytidylic acid promotes wound healing in human and murine skin

Toll-like receptors (TLRs) are pattern-recognition receptors and have a critical role in both innate and adaptive responses to tissue injury. Our previous study showed that wound healing was impaired in TLR3-deficient mice. In this study, we investigated the capacity of the TLR3 agonist polyriboinosinic-polyribocytidylic acid (poly(I:C)) to promote the healing of skin wounds in humans and mice. We found that topical application with poly(I:C) accelerated the closure of wounds in patients with laser plastic surgery. In a mouse model, topical application of poly(I:C) markedly enhanced re-epithelialization, granulation, and neovascularization required for wound closure. Further studies revealed that poly(I:C) treatment resulted in enhanced recruitment of neutrophils and macrophages in association with upregulation of a chemokine, macrophage inflammatory protein-2 (MIP-2/CXCL2), in the wounds. The effect of poly(I:C) was abolished in TLR3-deficient mice or by treatment with MIP-2/CXCL2-neutralizing antibodies. These results suggest a potential therapeutic value of the TLR3 activator poly(I:C) for wound healing.     

白癜风患者外周血单一核细胞Toll样受体7、9 mRNA表达的初步研究

目的 探讨Toll样受体7、9 （TLR7、TLR9）mRNA在白癜风患者外周血单一核细胞（PBMC）中的表达及其意义。方法 应用实时荧光定量逆转录聚合酶链反应（RT-PCR）技术检测50例白癜风患者及25例正常人对照PBMC中TLR7、TLR9 mRNA的表达。采用SPSS11.5软件进行统计学分析,两样本均数比较采用t检验。结果 白癜风患者PBMC中TLR7 mRNA（目的基因绝对定量校正拷贝数为0.85 ± 1.90）、TLR9 mRNA（0.94 ± 2.25）表达均明显高于对照组（分别为0.44 ± 1.18和0.11 ± 0.31,P ＜ 0.05和 ＜ 0.01）;稳定期与进展期、局限型与泛发型白癜风患者TLR7、TLR9 mRNA表达差异均无统计学意义（P值均 ＞ 0.05）。结论 白癜风患者PBMC中TLR7、TLR9 mRNA的表达均上调,TLR7、TLR9的异常表达可能参与白癜风的发病机制。     

Type I IL-1 receptor mediates IL-1 and intracellular IL-1 receptor antagonist effects in skin inflammation

The IL-1 system plays a key role in skin physiology and pathology. In this study, we used mutant mice lacking the type I IL-1 receptor (IL-1RI), lacking IL-1 receptor antagonist (IL-1Ra), or overexpressing the human intracellular (ic) IL-1Ra1 isoform, as well as combinations thereof, to dissect the role of the IL-1 system in phorbol 13-myristate 12-acetate (PMA)-induced skin inflammation. In wild-type (WT) mice, PMA application induced epidermal thickening and dermal inflammation. Skin IL-1alpha production and circulating levels of the acute-phase protein serum amyloid A (SAA) were elevated. In mice lacking IL-1RI or overexpressing icIL-1Ra1, PMA induced similar epidermal thickening as in WT mice, but dermal inflammation was partially prevented. Skin IL-1alpha mRNA expression was similar in PMA-treated IL-1RI-/- and WT mice, whereas the increase in serum SAA was suppressed in IL-1RI-/- mice. Interestingly, PMA-induced IL-1alpha mRNA expression was further enhanced by icIL-1Ra1 overexpression in an IL-1RI-dependent manner. Finally, IL-1Ra-/- mice spontaneously displayed skin lesions characterized by high IL-1beta, but not IL-1alpha, expression. In conclusion, PMA-induced epidermal thickening and skin IL-1alpha expression were independent of IL-1 signaling, in contrast to dermal inflammation and systemic inflammatory response.     

广西壮族、汉族系统性红斑狼疮与Toll样受体9基因多态性研究

目的 探讨TLR9单核苷酸多态性与广西系统性红斑狼疮（SLE）患者发病的相关性,以及其在壮族、汉两个民族间的差异。 方法 聚合酶链反应-限制性酶切技术、聚合酶链反应-限制性片段长度多态性分析技术（PCR-RFLP）对97例广西SLE患者和202例广西健康对照者的TLR9基因多态性进行检测,分析其基因型和等位基因频率与SLE部分临床实验室指标的关联性,并比较两民族间的差异。 结果 TLR9 rs352140CC、CT、TT基因型频率在汉族SLE组与汉族对照组中分别为42.9%、41.1%、16.1%和38.3%、55.8%、5.8%,差异有统计学意义（P < 0.05）,其C、T等位基因频率差异无统计学意义（P > 0.05）。TLR9rs352140 C/T基因型频率和等位基因频率在壮族SLE组与壮族健康对照组间、壮族SLE组与汉族SLE组间差异无统计学意义（均P > 0.05）。TLR9 rs352140TT基因型频率在ds-DNA阳性组比阴性组高（P < 0.05）,而T等位基因频率在两组间差异无统计学意义（P > 0.05）。TLR9 rs352140TT基因型频率和T等位基因的频率在SLEDAI ≥ 9组比SLEDAI < 9组高,差异有统计学意义（均P < 0.05）。TLR9 rs352140TT基因型频率和T等位基因的频率在ANA阳性组与阴性组间差异无统计学意义（均P > 0.05）。 结论 TLR9 rs352140基因多态性可能与广西汉族人SLE的易感性有关, TLR9基因多态性与部分SLE指标可能具有相关性。     

Leukotrienes as mediators of skin inflammation

Leukotrienes (LTs) derived from the 5-lipoxygenase pathways of arachidonic acid metabolism, are a new group of biologically active mediators. LTC⁴, LTD⁴ and LTE⁴ are considered collectively to account for the activity of slow reacting substance of anaphylaxis (SRS-A). They have potent smooth muscle contracting activity (Drazen et al ., 1980; Holme et al ., 1980) and cause vascular permeability changes in guinea-pig skin (Peck, Piper & Williams, 1981). LTB⁴ has been shown to be a potent chemotactic and chemokinetic agent for polymorphonuclear leucocytes (PMNs) (Ford-Hutchinson et al ., 1980; Smith, Ford-Hutchinson & Bray, 1980) and to cause vascular permeability changes in rabbit, rat and guinea-pig skin (Bray et al ., 1981a).
In human skin LTB⁴ has been shown to be a chemotactic agent for neutrophils (Bray, Ford-Hutchinson & Smith, 1981b; Camp et al ., 1982). LTC⁴ and LTD⁴ have pronounced inflammatory actions in human skin causing weal and flare responses and increases in blood flow at low concentrations (Camp et al ., 1982; Bisgard, Kristensen & Sondergaard, 1982). The present studies were designed to investigate a variety of leukotrienes on permeability responses in rabbit and guinea-pig skin.     

Neurovascular aspects of skin neurogenic inflammation

Neurogenic inflammation is involved in skin inflammation. It is hypothesized that it is involved in the pathogenesis of the common chronic cutaneous vascular disorder rosacea, but the exact mechanism of action is currently unknown. Transient receptor potential vanilloid 1 (TRPV1) and ankyrin 1 (TRPA1) are widely expressed on primary sensory neuron endings and non-neuronal cells such as keratinocytes. Here we describe the potential for TRPV1 and TRPA1 receptors to be involved in the pathophysiology of rosacea due to their polymodal activation, including cold and hot temperature, pungent products from vegetable and spices, reactive oxygen species, and mechanical stimuli. We discuss the role of both receptors and the sensory neuropeptides that they release in inflammation and pain sensation and evidence suggesting that both TRPV1 and TRPA1 receptors may be promising therapeutic targets for the treatment of the inflammatory symptoms of rosacea.