系统性红斑狼疮外周血单一核细胞Fas和FasL mRNA的表达

目的：探讨系统性红斑狼疮（SLE）患者外周血淋巴细胞Fas及其相应配体FasL的表达水平及在SLE发病中的作用。方法：应用逆转录-聚合酶链反应（RT-PCR）检测34例活动期SLE患者和30例正常人外周血单一核细胞（PBMC）中Fas和FasL mRNA的表达水平。结果：活动期SLE患者PBMC中FasL的阳性检出率为91.18%,明显高于正常对照组（63.30%),差异非常显著（P＜0.01);Fas的阳性检出率与对照组相比无明显差异（P＞0.05)。活动期SLE患者PBMC中Fas和FasL mRNA的平均表达水平均高于正常对照组,差异显著（P＜0.05)。结论：Fas和FasL的高水平表达可能与SLE患者外周血T淋巴细胞活化、凋亡增加有关,Fas介导的淋巴细胞凋亡可能参与了SLE的发病过程。平表达可能与SLE患者外周血T淋巴细胞活化、凋亡增加有关,Fas介导的淋巴细胞凋亡可能参与了SLE的发病过程。     

Fas FasL TRAIL分子在尖锐湿疣皮损中的分布

目的　探讨Fas、FasL和TRAIL介导的细胞凋亡在尖锐湿疣发病中的可能作用及意义。方法　用免疫组化方法检测了 30例尖锐湿疣患者皮损中Fas、FasL和TRAIL的表达。结果　所有尖锐湿疣标本 ( 10 0 % )Fas均阳性 ,2 3例 ( 76 .7% )FasL阳性 ,2 1例 ( 70 .0 % )TRAIL阳性 ,三者分布基本一致 ,正常表皮基本不表达。且FasL和TRAIL分子与真皮单一核细胞浸润呈显著负相关 (γ =-0 .5 0 6和γ =-0 .45 0 ,P均 <0 .0 5 )。结论　尖锐湿疣组织中细胞凋亡相关分子Fas、FasL和TRAIL表达异常 ,且Fas、TRAIL分子表达影响局部免疫反应 ,考虑凋亡失控可能参与尖锐湿疣发病 ,引起复发。     

银屑病皮损中基质金属蛋白酶2、Fas和FasL表达

目的 通过检测银屑病患者皮损中基质金属蛋白酶2(MMP-2)、Fas、FasL的表达，探讨MMP-2对银屑病细胞凋亡的影响。方法 采用免疫组化ABC法对银屑病患者皮损中MMP-2、Fas、FasL表达进行检测。结果 银屑病皮损中既可表达MMP-2，又可表达Fas、FasL而存正常皮肤不表达。结论 MMP-2、Fas、FasL参与了银屑病的发病过程。     

芪加真武汤对系统性红斑狼疮患者外周血淋巴细胞凋亡及Fas和FasL表达的影响

目的探讨中药芪加真武汤对系统性红斑狼疮(SLE)患者外周血淋巴细胞(PBL)凋亡及Fas和FasL表达的影响。方法采用血清药理学的原理制取兔芪加真武汤含药血清。选取活动期SLE患者19例和正常人10名,分离PBL,将植物血凝素刺激后的PBL分别加入RPMI1640(含10%小牛血清)、含药血清(5%,10%,20%)培养72h,应用流式细胞仪检测其凋亡及Fas和FasL的表达。结果①SLE患者PBL凋亡率明显高于正常人(P<0.01),Fas表达明显升高(P<0.01)。②10%,20%含药血清组PBL的凋亡均较SLE显著减少(P<0.01),Fas表达显著降低(P<0.01),FasL的阳性表达率升高(P<0.05)。结论SLE患者PBL存在凋亡及Fas和FasL表达的紊乱。中药芪加真武汤可降低SLE患者外周血淋巴细胞Fas的表达和增加FasL的细胞阳性表达率,进而影响其凋亡。     

芪加真武汤联合糖皮质激素体外对SLE患者外周血淋巴细胞凋亡及Fas、FasL表达的影响

目的 探讨中药芪加真武汤联合糖皮质激素对系统性红斑狼疮(SLE)患者外周血淋巴细胞(PBL)凋亡及Fas、FasL表达的影响。方法 采用血清药理学方法 ,制取芪加真武汤含药兔血清。运用流式细胞仪检测19例SLE患者外周血淋巴细胞在不同浓度的芪加真武汤的含药血清(20%、10%、5%)、地塞米松(0.001mol/L)及两者联合刺激下细胞凋亡和Fas、FasL表达的变化。结果 ①SLE患者PBL凋亡率明显高于正常组(P<0.001),Fas表达明显多于正常组(P<0.01),FasL表达与正常组差异不明显(P>0.05)。②10%、20%含药血清显著减少SLE患者PBL的凋亡率(P<0.01),显著降低Fas表达(P<0.01),升高FasL的阳性细胞率(P<0.05);③地塞米松可增高SLE患者PBL的凋亡率(P<0.05),但对Fas、FasL的表达无影响(P>0.05);④含药血清联合地塞米松显著减少PBL凋亡率(P<0.01),降低Fas表达(P<0.05),明显升高FasL的阳性细胞率(P<0.01)。结论 SLE患者PBL存在凋亡及凋亡相关蛋白表达的紊乱,中药芪加真武汤在体外可纠正其紊乱,缓解由地塞米松引起的PBL过度凋亡。     

局部中晚期宫颈癌新辅助化疗对凋亡细胞因子Fas/FasL表达水平影响分析

目的:考察局部中晚期宫颈癌新辅助化疗对凋亡细胞因子Fas/FasL表达水平影响。方法:将50例宫颈癌患者随机分为观察组和对照组,每组25例。比较两组患者手术病理情况、治疗前后Fas/Fas L表达水平和生存期。结果:观察组患者淋巴结转移、脉管受累和宫颈深层间质浸润发生率显著低于对照组(χ~2=3.947,χ~2=5.094,χ~2=7.219,P0.05,P0.01)。两组患者治疗前Fas/FasL表达水平差异无统计学意义(P0.05)。治疗后观察组患者Fas表达水平显著升高、FasL表达水平显著降低(P0.05)。观察组患者5年生存率显著高于对照组(χ~2=3.947,P0.05)。结论:局部中晚期宫颈癌新辅助化疗对凋亡细胞因子Fas/FasL表达水平具有显著影响可能是影响患者预后的原因之一。     

银屑病中Fas、FasL和穿孔素的检测

目的：通过对银屑病中Fas、FasL和穿孔素的检测,探讨细胞凋亡和银屑病的关系。方法：采用免疫组化ABC法对银屑病皮损中Fas、FasL和穿孔素进行检测。结果：在银屑病组织中Fas、FasL和穿孔素均阳性表达。结论：在银屑病中,细胞毒性T淋巴细胞可能通过FasL和穿孔素两条途径引起细胞凋亡,细胞凋亡参与了银屑病的发病。     

白癜风患者外周血淋巴细胞Fas及Fas配体和可溶性Fas的研究

目的探讨Fas抗原与Fas配体(FasL)及可溶性Fas(sFas)在白癜风发病中的作用。方法应用流式细胞计数仪对白癜风患者外周血淋巴细胞(PBLC)上Fas和FasL的表达情况进行了检测,用双抗体夹心酶联免疫吸附试验检测患者血清中sFas的含量。结果寻常型白癜风患者PBLC上Fas的表达(43.45%)较正常人(58.30%)显著降低(P<0.01),寻常型白癜风患者PBLC上FasL的表达(58.40%)亦较正常人(64.65%)显著降低(P<0.05);寻常型白癜风患者血清中sFas水平(3.95±3.23μg/L)较正常人(1.51±0.71μg/L)明显增高(P<0.01),散发型白癜风患者进展期血清sFas水平(5.55±4.82μg/L)明显高于稳定期(2.94±2.29μg/L)(P<0.05),治疗后血清sFas水平较治疗前明显下降(P<0.001)。结论白癜风患者PBLC上Fas/FasL及循环sFas的表达均有异常,淋巴细胞Fas/FasL异常表达及Fas介导的细胞凋亡机制可能与白癜风的免疫异常及发病有关。     

银屑病皮损细胞凋亡的研究

目的：通过对银屑病皮损细胞凋亡以及凋亡相关基因bcl-2、Fas、bax和Fas-L、穿孔素的检测，探讨细胞凋亡与银屑病的关系。方法：采用原位末端标记及免疫组化ABC法、SABC法。结果：银屑病中细胞凋亡较正常人增多。抑凋亡基因bcl．2几乎不表达：而促凋亡基因Fas、bax阳性表达。细胞毒性T淋巴细胞可能通过Fas-L和穿孔素两条途径起作用。结论：细胞凋亡在银屑病发病机理中有十分重要的意义。     

Fas/FasL在基底细胞癌和鳞状细胞癌中的表达

目的:检测Fas/FasL在基底细胞癌(BCC)和鳞状细胞癌(SCC)中的表达.方法:采用SABC技术分别检测Fas/FasL在BCC、SCC及正常人对照皮肤中的表达.结果:Fas/FasL在BCC组不表达或弱表达,较正常人组无显著差异(P＞0.05);SCC组Fas和FasL的表达均高于对照皮肤(P＜0.05和＜0.01).Fasl染色主要集中于肿瘤细胞,弥漫染色,在肿瘤团块边缘表达更强;Fas在角珠处表达较强.结论:FasL在基底细胞癌不表达或弱表达,表明BCC侵袭扩散能力低,而在鳞状细胞癌中高表达可能与肿瘤的侵袭扩散能力高有关.     

系统性红斑狼疮患者bcl-2及Fas、Fas配体表达与淋巴细胞凋亡的关系

目的：对SLE患者外周血淋巴细胞凋亡率以及凋亡凋控基因表达蛋白bcl-2、Fas抗原和Fas配体进行分析,以进一步探讨其在SLE发病中的作用及相互关系。方法：应用流式细胞仪检测。结果：SLE患者外周血淋巴细胞凋亡率明显视于正常人（P＜0.01）和其他疾病患者（P＜0.05）;bcl-2抗原表达显著低于正常人和其他疾病患者（P＜0.01）,而Fas抗原、Fas配体表达三组间无明显差异。结论：SLE患者体内淋巴细胞凋亡的明显增加可能与其发病有关,而bcl-2对淋巴细胞凋亡抑制作用的减弱可能较Fas／Fas配体诱导促进淋巴细胞的凋亡更为重要。     

系统性红斑狼疮外周血单个核细胞Fas FasL及bcl-2抗原的表达

目的探讨系统性红斑狼疮（ＳＬＥ）免疫功能失调与细胞凋亡的关系。方法应用免疫细胞化学技术对ＳＬＥ患者外周血单个核细胞（ＰＢＭＣ）上某些凋亡调节分子的表达进行了检测。结果ＳＬＥ患者ＰＢＭＣ上Ｆａｓ及ＦａｓＬ抗原均高表达,ｂｃｌ ２蛋白正常表达。结论ＳＬＥ患者体内存在较多活化的淋巴细胞,Ｆａｓ和ＦａｓＬ抗原表达异常促进了ＳＬＥ的ＰＢＭＣ凋亡。     

Membranous and Soluble Forms of Fas Antigen in Cutaneous Lupus Erythematosus

The role of Fas-mediated apoptosis in cutaneous lupus erythematosus (LE) is still unclear, although the Fas/FasL system has been investigated in autoimmune diseases in relation to impaired apoptosis. In order to elucidate the connections between acute cutaneous LE (ACLE) and chronic cutaneous LE (CCLE), we determined the expression of membranous Fas antigen (mFas) on peripheral blood mononuclear cells (PBMC) by flow cytometry and the levels of the soluble form of the Fas antigen (sFas) in sera. The ratio and the mean fluorescence intensity of mFas were much higher in ACLE patients than in others, including patients with CCLE and atopic dermatitis and normal healthy controls. The levels of sFas in ACLE and CCLE patients were also elevated, and there was a significant increase in sFas levels in ACLE patients over that in CCLE patients. Immunohistochemical studies revealed that Fas antigen was predominantly expressed on infiltrating cells around blood vessels and appendages in ACLE and CCLE patients. Based on these findings, it is suggested that the expression of Fas antigen is closely associated with the activation of circulating lymphocytes, especially in ACLE patients, but is not directly associated with keratinocyte damage.     

Proapoptotic and antiapoptotic markers in cutaneous T-cell lymphoma skin infiltrates and lymphomatoid papulosis

BACKGROUND: In cutaneous T-cell lymphoma (CTCL) lesions, both reactive T cells and malignant T cells intermingle. The disease progression is mostly slow. Recent evidence suggests that even if clinical remission is reached, malignant cells persist and a relapse follows sooner or later. To wha extent tumour cell apoptosis occurs in the skin lesions either due to the reactive T cells or t therapeutic efforts is not known. OBJECTIVES: To determine the extent of tumour cell apoptosis and the expression of proapoptotic an antiapoptotic markers in serial skin lesion samples from patients with CTCL, and to compare th findings with those in patients with lymphomatoid papulosis (LyP). METHODS: Thirty-four skin samples were obtained from 12 patients with CTCL at the time o diagnosis and at a mean of 1.6, 3 and 6 years later. The patients received psoralen plus ultraviolet (PUVA), electron beam or cytostatic treatments. In addition, fresh post-treatment samples fro three patients with CTCL undergoing PUVA therapy were obtained. For comparison, skin biopsies o five patients with LyP were studied. Immunohistochemical demonstration of the expression of th following markers was performed on formalin-fixed skin sections: Fas (CD95), Fas ligand (FasL) bcl-2, granzyme B, the tumour-suppressor protein PTEN and the effector caspase, caspase-3. Th malignant cells were identified morphologically, and apoptotic cells were identified with th terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end labelling method on parallel sections. RESULTS: In untreated CTCL lesions, apoptotic lymphocytes were extremely rare, and no increase in the number of apoptotic cells was observed after any of the treatments used. In LyP, apoptotic cell were more frequent, comprising on average 5% of the infiltrate. The apoptosis-associated marker Fas, FasL, caspase-3 and granzyme B were expressed by morphologically neoplastic cells in CTCL and by large atypical cells in LyP, with no significant differences. However, only a few reactive cell in CTCL infiltrates expressed granzyme B while about 10% of the corresponding cells were positive in LyP. The expression of antiapoptotic bcl-2 was more frequent in CTCL than in LyP, while PTE expression was high in both instances. The number of bcl-2 + cells tended to decrease after therapy When comparing the findings between the first and the last samples, a decrease in the number of bcl-2+ cells and an increase in Fas+ cells was associated with disease progression, despite therapy, while the opposite was true for remissions. CONCLUSIONS: Apoptosis was found to be a rare event in CTCL lesions irrespective of precedin therapy During patient follow-up, no significant differences in the expression of apoptotic marker was observed while in most cases a lower level of antiapoptotic bcl-2 expression was observed after all types of therapies and in association with disease progression when compared with high expression in the untreated lesions. The absence of apoptosis and high expression of bcl-2 together with a low expression of apoptosis-inducing granzyme B in the reactive lymphocytes in CTC could explain the chronic nature of the disease and the poor response to therapy, while th more frequent occurrence of granzyme B and apoptosis together with a lower level of expressio of bcl-2 by the large atypical cells in LyP could contribute to the favourable outcome of the latter.     

Fas and Fas ligand: expression and soluble circulating levels in cutaneous malignant melanoma

BACKGROUND: The Fas/Fas ligand (FasL) system plays a key part in maintaining tissue homeostasis via the induction of apoptosis. Functional impairment of the Fas/FasL system is associated with the development and progression of malignancies. Malignant melanoma cells and tissues have been shown to express Fas and FasL to variable extents. OBJECTIVES: To demonstrate the expression and the presence of soluble circulating levels of Fas and FasL in cutaneous malignant melanoma. METHODS: Biopsy specimens of 42 patients with primary melanoma and nine patients with cutaneous metastatic melanoma were obtained for immunohistochemistry studies. All patients were followed for at least 5 years. In another 46 patients with melanoma (15 stage I and II; 11 stage III; and 20 stage IV) and in 10 healthy volunteer control subjects circulating levels of Fas and FasL were analysed with commercial ELISA tests. RESULTS: FasL was strongly positive in 38 (90%) of 42 primary melanomas; two of nine metastases did not express FasL. In the primary melanomas Fas was strongly or intensely positive in 17 (40%), moderately or weakly positive in 10 (24%) and negative in 15 (36%) of 42 melanomas. Soluble Fas plasma levels in patients with metastatic malignant melanoma were significantly elevated over those in the control group (P = 0.01). CONCLUSIONS: The absence of Fas in most of the thick melanomas that did not metastasize, and in insitu melanomas, might be taken as a theoretical factor for a good prognosis. Soluble Fas is increased in patients with metastatic melanoma and might be associated with poor prognosis.     

Resistance to activation-induced cell death and bystander cytotoxicity via the Fas/Fas ligand pathway are implicated in the pathogenesis of cutaneous T cell lymphomas

By engaging Fas, Fas ligand (FasL) on activated T lymphocytes induces activation-induced cell death (AICD), and also triggers apoptosis of target cells during immune downregulation. We previously showed that within cutaneous T cell lymphoma (CTCL) lesions, malignant CD4(+) T cells expressing FasL accumulated, and were inversely distributed with CD8(+) T cells. We thus determined the responses of human CTCL cells to AICD and their cytotoxic to Fas(+) target T cells in vitro. CTCL cells expressing Fas were resistant to AICD following activation by CD3 monoclonal antibody (mAb) whereas still undergoing apoptosis if Fas was ligated to Fas mAb. CTCL cell lines, as well as Sezary Syndrome patients' peripheral blood lymphocytes, exhibited ratio-dependent cytotoxicity to Fas(+) Jurkat cells. The kinetic study showed that FasL surface expression was absent before activation, and its expression was low and/or delayed after activation. We therefore hypothesize that CTCL cells express functional FasL possibly contributing to bystander cytotoxicity within tumor infiltrates. In addition, decreased and/or delayed FasL surface expression following activation may in part contribute to their resistance to AICD. Both bystander cytotoxicity and resistance to AICD are likely to contribute to the loss of cytotoxic anti-tumor CD8(+) T cells as well as the accumulation of malignant T cells in CTCL.     

Increased expression of Fas on human epidermal cells after in vivo exposure to single-dose ultraviolet (UV) B or long-wave UVA radiation

BACKGROUND: Apoptosis has been proposed to act as an important mechanism for eliminating keratinocytes that have been irreversibly damaged by ultraviolet (UV) irradiation. One way to induce apoptosis in keratinocytes is through activation of the cell surface receptor Fas (CD95), either with the ligand (FasL) or directly with UV radiation. OBJECTIVES: To investigate the regulation of Fas and FasL expression in human skin and the formation of apoptotic cells after in vivo exposure to UVB or long-wave UVA radiation. METHODS: Volunteers were irradiated with either 3 minimal erythema doses (MED) of UVB (n = 6) or 3 MED of long-wave UVA (n = 6) on buttock skin 12, 24 and 72 h before skin punch biopsies were taken. Expression of Fas and FasL was demonstrated by immunohistochemistry on cryostat sections. Apoptosis was assessed by the terminal deoxynucleotidyl transferase-mediated fluorescein-deoxyuridine triphosphate nick-end labelling reaction. RESULTS: In five of six subjects, exposure to UVB radiation resulted in increased homogeneous expression of Fas on epidermal cells, with greatest expression at 24 and 72 h after irradiation. In all subjects, exposure to long-wave UVA resulted in increased homogeneous expression of Fas on epidermal cells, with greatest expression at 12 h after irradiation. In five of six subjects, exposure to UVB radiation resulted in temporarily decreased expression of FasL, but after 72 h the expression of FasL had returned to the preirradiation level. The expression of FasL on epidermal cells after exposure to long-wave UVA showed considerable variation. UVB irradiation was a stronger inducer of epidermal apoptosis than was UVA irradiation. The number of apoptotic epidermal cells did not correlate with expression of Fas or FasL. CONCLUSIONS: In human skin the expression of Fas on epidermal cells increases after in vivo exposure to UVB or long-wave UVA. Exposure to UVB causes a temporary decrease in the expression of FasL on epidermal cells.