银屑病T细胞活化和信号转导的进展

银屑病发病与辅助性T细胞亚群分化失衡，活化T细胞信号转导失控及特定基因表达异常密切相关。T细胞可经抗原或非抗原刺激而活化，活化T细胞的信号转导途径有：Ca^2＋离子依赖的蛋白激酶C途径、Ras丝裂原活化的蛋白激酶途径和詹纳斯激酶一信号转导及转录活化因子途径等。其中詹纳斯激酶一信号转导及转录活化因子途径是细胞因子信号转导的主要途径。有效调控T细胞活化和信号转导途径对治疗银屑病有重要意义。     

原发性皮肤T细胞淋巴瘤的T细胞受体基因重排分析

目的：评价Ｓｏｕｔｈｅｒｎ印迹分析（ＳＢＡ）和聚合酶链反应（ＰＣＲ）检测原发性皮肤Ｔ细胞淋巴瘤（ＰＣＴＣＬ）Ｔ细胞受体（ＴＣＲ）基因重排（ＧＲ）的意义。方法：以ＰＣＲ扩增ＴＣＲγ的结合Ⅴ（可变区）－Ｊ（结合区）序列（ＴＣＲγＰＣＲ）和ＳＢＡ分析ＴＣＲβ链基因（ＴＣＲβＳＢＡ）检测克隆性ＧＲ。结果：蕈样肉芽肿（ＭＦ）：ＴＣＲγＰＣＲ和ＴＣＲβＳＢＡ检测６例ⅡＡ期和７例ⅡＢ期皮损标本的ＧＲ分别为５例和４例以及６例和５例，外周血分别有４例、２例和５例、３例示ＧＲ；而７例ⅠＡ期和１０例ⅠＢ期的ＴＣＲγＧＲ和ＴＣＲβＧＲ皮肤组织为４例、１例和７例、１例，外周血为３例、阴性和４例、１例。１例ＭＦⅡＡ表现为皮病性淋巴结病患者的淋巴结中证实有ＧＲ。疑诊ＭＦ：１１例患者的皮损和外周血标本经ＴＣＲγＰＣＲ检测５例皮肤和３例外周血见ＧＲ。非蕈样肉芽肿、Ｓéｚａｒｙ综合征的ＰＣＴＣＬ：ＰＣＲ和ＳＢＡ显示ＴＣＲＧＲ分别为皮肤组织占９例／１０例和６例／８例，外周血占９例／１０例和６例／１１例。Ｓéｚａｒｙ综合征和淋巴瘤样丘疹病：２例Ｓéｚａｒｙ综合征外周血和其中１例皮肤标本同时见ＴＣＲγＧＲ和ＴＣＲβＧＲ；２例淋巴瘤样丘疹病的皮肤标本     

寻常性银屑病患者外周血T细胞受体β可变区优势表达分析

目的 分析寻常性银屑病患者外周血中T细胞受体β可变区(TRBV)的优势表达状况,探讨其在银屑病发病中的作用.方法 以人功能性TRBV家族设计33个上游引物,共同的T细胞受体β恒定区(TRBC)基因设计下游引物,在T细胞受体α恒定区(TRAC)设计引物作为内参,分析寻常性银屑病患者、正常人对照外周血样本各10例,检测各PCR反应管荧光强度,以大于正常人外周血T细胞相应TRBV基因相对表达量(-x+3s)筛选TRBV优势表达基因家族.结果 TRAC扩增产物阈值循环数(Ct)值集中于21～24,银屑病组TRBV2扩增产物Ct与TRAC产物Ct差值平均数银屑病患者为2.98,TRBV5-7为3.24,TRBV6-6/6-9为2.52,TRBV12为2.04,TRBV24为3.56,TRBV29为4.12,其表达与正常人比较均明显增高(P值均＜0.05),其中TRBV6-6/6-9、TRBV12、TRBV29在银屑病患者呈优势表达.结论 银屑病患者外周血TRBV基因家族存在优势表达,可能在银屑病T细胞异常免疫反应中起重要作用.

Abstract：

Objective To assess the preferential expressions of peripheral blood T cell receptor beta chain variable region (TRBV) subfamilies in patients with psoriasis vulgaris(PV), and to estimate their role in the pathogenesis of psoriasis. Methods Thirty-three upstream primers were designed to target the human functional TRBV genes, downstream primers to target the common T cell receptor beta constant (TRBC) gene,with T cell receptor alpha constant (TRAC) gene as the internal reference. Total RNA was extracted from the peripheral blood T cells of 10 health human controls and 10 patients with PV, and transcribed into cDNA.Then, TRBV genes were amplified by real-time fluorescence quantitative PCR (RFQ-PCR) and the fluorescence intensity of each samples was detected. The expression levels of TRBV genes in the control group were used to calculate the cut-off values (mean expression levels of TRBV subfamilies in the 10 normal controls + 3 standard deviations). When the expression level of a TRBV subfamily from patients with PV was equal to or higher than the cut-off value, it was considered as the preferentially expressed TRBV subfamily. Results The threshold cycle (Ct) value varied from 21 to 24 for TRAC gene. The difference in the Ct value between TRBV subfamily genes and TRAC gene in patients with PV was 2.98 for TRBV2 gene, 3.24 for TRBV5-7 gene, 2.52 for TRBV6-6/6-9 gene, 2.04 for TRBV 12 gene, 3.56 for TRBV 24 gene, and 4.12 for TRBV 29 gene, and the expression levels of these subfamily genes were significantly higher than those in the normal controls (all P ＜ 0.05). According to the above standard, TRBV6-6/6-9, TRBV12 and TRBV29 were considered to be preferentially expressed subfamilies. Conclusions There is a preferential expression of TRBV gene subfamilies in peripheral blood of patients with psoriasis vulgaris, which may play a vital role in the abnormal T cell-mediated immune responses in psoriasis.     

组织常驻记忆性T细胞与银屑病

银屑病为免疫介导的慢性炎症性皮肤病,复发是其特点之一,且复发一般位于原皮损部位［1?3］。白细胞介素23（IL?23）/IL?17轴在银屑病的发病中具有重要作用［4］。γδT细胞为银屑病产生关键致病性细胞因子IL?17最主要的细胞［5］,应用咪喹莫特诱导的银屑病样鼠模型中,γδT细胞Vγ4+T细胞亚群具有记忆功能并长期存在于小鼠皮肤,再次经咪喹莫特刺激后能够产生比初次反应更强更快的反应［6］。Vγ4+ T细胞的特征与组织常驻记忆性T细胞（TRM）的特征相似,可能也是银屑病在相同部位复发的关键性细胞。我们综述银屑病与TRM之间的联系……     

原发性皮肤T细胞淋巴瘤外周血克隆性T细胞的检测与研究

为了解原发性皮肤Ｔ细胞淋巴瘤（ＰＣＴＣＬ）外周血中克隆性Ｔ细胞的存在及其与临床疗铲的相关性,分别对２５例ＰＣＴＣＬ患者经重组α干扰素治疗前后的外周血标本,应用聚合酶链反应（ＰＣＲ）方法,以通用引物扩增Ｔ细胞受体（ＣＲ）γ链编码基因中Ⅴ（可变区）－Ｊ（连接区）的结合序列,检测代表克隆性Ｔ细胞分子标志的ＴＣＲ基因重排（ＧＲ）。结果发现,２５例ＰＣＴＣＬ外周血样品,１７例见克隆性Ｔ细胞（示ＴＣＲＧＲ）,     

Efalizumab in the treatment of psoriasis

Efalizumab is a humanized, monoclonal antibody, which targets CD11a, one of the subunits of leukocyte function-associated antigen-1. Administered subcutaneously once weekly, it decreases the activation of T lymphocytes as a primary or secondary process and interferes with the trafficking of T cells into sites of inflammation. Clinically, improvement in psoriasis can be observed as early as two to four weeks. The percent of patients experiencing a 75% reduction in PASI (PASI-75) was 27%, 44%, 47% at 12 weeks, 24 weeks, and 24 months, respectively. During the trials, the safety profile was highly favorable, with minor headaches and myalgias occurring after the initial doses. Rebound on abrupt discontinuation can be problematic for some patients and avoided by transition to an alternative therapy. Efalizumab appears to be a valuable option for patients requiring long-term control of their psoriasis.     

Hyporesponsiveness of peripheral blood lymphocytes to streptococcal superantigens in patients with guttate psoriasis: evidence for systemic stimulation of T cells with superantigens released from focally infecting Streptococcus pyogenes

Abstract Throat infection with Streptococcus pyogenes is the most important trigger for acute guttate psoriasis. We examined the in vitro responses of peripheral blood mononuclear cells (PBMC) to streptococcal superantigens, SPEA and SPEC, and staphylococcal superantigens, SEB and TSST-1, in patients with guttate psoriasis, in patients with chronic plaque psoriasis, and in healthy subjects. PBMC from patients with guttate psoriasis responded poorly to SPEA and SPEC at concentrations of 0.1 and 1 ng/ml as compared with those from patients with plaque psoriasis, but showed high responses to SEB and TSST-1. The hyporesponsiveness recovered after improvement of the skin eruption. There was no significant difference between guttate and chronic types of psoriasis in the percentage of circulating T-cell receptor BV2 or BV8-bearing T cells, responsive to streptococcal superantigens, indicating that T-cell clonal anergy was a mechanism underlying the hyporesponsiveness. Our results suggest that superantigens released from focally infecting S. pyogenes induce a transient activation of relevant T cells, leading to the development of skin eruption and, subsequently, temporary T-cell anergy to these toxins. Received: 4 September 1998 / Received after revision: 28 December 1998 / Accepted: 21 January 1999     

Adenosine deaminase in adult T cell leukemia

Increased adenosine deaminase activity was observed in peripheral lymphocytes from a patient with adult T cell leukemia. Gel filtration pattern on a Sephadex G-150 revealed that the enzyme from normal lymphocytes existed in two forms with different molecular weights (large and small), whereas the enzyme from the patient existed solely as the small molecular weight form. Comparison of this small form adenosine deaminase from the patient with that from normal controls revealed that both enzymes were quite similar in relative substrate specificity, Km values for adenosine, pH optima, heat stability pattern, and sensitivity to inhibition by coformycin, a tight binding inhibitor of adenosine deaminase. Increased adenosine deaminase activity and the lack of the large form of the enzyme might be significant features of adult T cell leukemia lymphocytes.     

银屑病发病机制中T细胞作用的进展

银屑病是一种Th1细胞介导的自身免疫性皮肤病,近来的研究表明,其他细胞也参与其发病过程,尤其是树突细胞、内皮细胞、Th17细胞及调节性T细胞等,其中T细胞的活化、增殖及分化是发病的主要环节,银屑病特征性的角质形成细胞增殖和异常分化及炎性细胞浸润是继发于T细胞活化后释放的细胞因子.一些黏附分子(E/P选择素等)及皮肤淋巴细胞相关抗原-P选择素糖蛋白配体-1复合体和皮肤淋巴细胞相关抗原-CD43复合体在银屑病发病中也起着一定的作用.     

异位性皮炎的记忆性T细胞浸润及ICAM－1表达

为了解粘连分子在异位性皮炎（ＡＤ）炎症及免疫反应过程中的作用，对ＡＤ皮损部位细胞间粘连分子－１（ＩＣＡＭ－１）的表达作了研究。结果虽然正常皮肤表皮不表达ＩＣＡＭ－１，但ＡＤ皮损处角朊细胞则局灶性表达ＩＣＡＭ－１，尤其在有严重单个核细胞浸润及表皮内淋巴细胞移入的部位。免疫表型研究表明，ＡＤ真皮浸润中ＣＤ４＋／ＣＤｗ２９＋／ＣＤ４５ＲＡ－记忆性Ｔ细胞占主导，推测它们可能通过分泌某些细胞因子而诱导角朊细胞表达ＩＣＡＭ－１。ＩＣＡＭ－１与淋巴细胞表面的淋巴细胞功能相关抗原－１（ＬＦＡ－１）之间相互作用可能对淋巴细胞在皮肤内的运行起调控作用。     

银屑病患者皮肤中原位记忆T淋巴细胞的检测

目的 研究皮肤组织原位记忆T淋巴细胞在银屑病发病过程中的作用。 方法 收集32例进行期斑块状银屑病患者的临床资料,采集每例患者的皮损和非皮损组织、9例患者皮损消退后皮肤标本;10例健康人皮肤为对照。采用免疫组化法检测组织原位记忆T淋巴细胞的特征性表面标志CD69和CD103,分析组织原位记忆T淋巴细胞在银屑病发病不同时期的情况。两组免疫组化结果进行t检验。 结果 32例进行期银屑病患者皮损和非皮损每高倍视野中CD69+CD103+ T淋巴细胞数量分别为11.34 ± 7.60和2.72 ± 4.20,皮损区明显高于非皮损区（t = 8.46,P ＜ 0.01）;其中9例患者皮损消退前后每高倍视野中CD69+CD103+ T淋巴细胞数量分别为14.33 ± 2.21和12.00 ± 4.58,皮损消退前后比较差异无统计学意义（t = 1.98,P = 0.08）;健康对照组为1.70 ± 2.98,与银屑病患者非皮损区比较,差异无统计学意义（t = 0.71,P > 0.05）。 结论 银屑病患者皮肤中原位记忆T淋巴细胞可能在皮损的形成和复发过程中起作用。     

体外研究银屑病患者T淋巴细胞对表皮通过时间的影响

目的　揭示T细胞在银屑病表皮动力学紊乱中的作用。方法　应用T细胞与皮肤组织体外共培养的方法检测银屑病T细胞对表皮通过时间产生的影响 ;采用3 H胸腺嘧啶核苷 ( 3 H TdR)标记体外培养的皮肤组织基底层有丝分裂S期细胞 ,放射自显影法追踪标记细胞从基底层到颗粒层的表皮通过时间。结果　未受T细胞作用的银屑病皮损表皮通过时间为 ( 4 .5± 2 .1)天 ,较正常人皮肤 ( 11.5± 3 .8)天显著缩短 (P <0 .0 1) ;银屑病外观正常皮肤 ( 8.7± 3 .2 )天与正常人皮肤比较差异无显著性 (P >0 .0 1)。银屑病外观正常皮肤及正常人皮肤分别与银屑病T细胞共培养后 ,表皮通过时间均显著缩短。结论　表皮通过时间缩短 ,表皮周转速率加快是银屑病皮损重要的表皮动力学改变 ,与银屑病皮损形态学改变密切相关 ;银屑病T细胞可以影响正常皮肤的表皮通过时间 ,表明T细胞在银屑病表皮动力学紊乱中发挥重要作用。     

Dohi memorial lecture. Cutaneous T cell lymphoma

Cutaneous T cell lymphoma (CTCL) is a generic classification of clonally-derived malignancies of phenotypic helper/inducer T cells with a propensity to infiltrate the skin, migrate into the epidermis, localize in T cell zones of lymphoid structures and spare the bone marrow. One clinical presentation has a tendency to evolve into another as subclones of progressively less mature and less epidermotropic neoplastic cells arise and overgrow the relatively more mature other subclones. In this manner, localized, scaling plaques of typical parapsoriasis develop into more infiltrated plaques of classical mycosis fungoides which are themselves predecessors of tumor or erythroderma stage CTCL. With loss of epidermotropism, systemic dissemination occurs: first microscopically with blood involvement and seeding of internal organs and finally with visceral tumor formation. The relationship between CTCL and adult T cell lymphoma/leukemia is unclear because of tremendous overlap in the clinical and immunologic findings and because a fraction of patients with classical CTCL have low titers of anti-HTLV-1 antibodies. It is important to distinguish the skin-limited and systemic phases of CTCL in order to select appropriate treatments. New diagnostic techniques which are quite helpful include immunotyping with monoclonal antibodies, karyotype analysis and T cell receptor studies. Photopheresis, a very promising new therapeutic approach for the management of patients with systemically disseminated disease, and a suggested pathogenetic scheme are discussed.     

寻常性银屑病患者外周血Th17／Treg平衡偏移的研究

目的通过检测寻常性银屑病患者血清中卟17、Treg细胞的水平,探讨其在银屑病发病机制中的作用。方法选择46例寻常性银屑病患者和20例健康人为研究对象,采用流式细胞技术检测外周血Thl7细胞和CD4＋CD25＊Treg细胞亚群。结果银屑病患者Thl7表达水平明显高于正常对照组,而Treg与正常对照组比较差异无统计学意义;患者Thl7／Treg表达水平与患者PASI评分呈正相关。结论银屑病患者外周血中Thl7／Treg细胞表达失衡可能拳与了其发病的过程。     

Stimulation of T cells by autologous mononuclear leukocytes and epidermal cells in psoriasis

Summary Based on reports suggesting aberrant cell-mediated immunity and altered infiltration of immunocompetent cells into the skin in psoriasis, we studied the stimulation of T cells by autologous non-T mononuclear leukocytes (autologous mixed lymphocyte reaction, AMLR) and by epidermal cells isolated from lesional and clinically uninvolved skin in psoriasis (autologous mixed epidermal cell lymphocyte reaction, AMECLR). Age- and sex-matched individuals served as controls. We found that the AMLR in psoriasis (n=11) was similar to that in healthy controls (n=16); furthermore, cell proliferation was alike in the presence of either 5% AB-serum or autologous serum. By contrast, while the AMECLR in healthy controls (n=9) resembled that in psoriatics employing epidermal cells from univolved skin, epidermal cells from lesional sites (n=10) induced a significantly higher proliferation of autologous T cells in the AMECLR (P<0.01). We conclude that the in vitro stimulation of T cells by non-T mononuclear leukocytes is normal in psoriasis and is not regulated by autologous serum. Lesional psoriatic epidermal cells, however, are more active in stimulating autologous T cell proliferation than cells from univolved psoriatic or normal epidermis.     

156例银屑病患者外周血T淋巴细胞亚群的表现

笔者对 15 6例银屑病患者的外周血进行了T淋巴细胞亚群的检查 ,结果显示 ,CD3 明显低于正常 ,而CD8高于正常 ,检验分析有显著性意义。