IL-4–induced apoptosis in peripheral blood eosinophils

Background: The regulation of eosinophil survival and apoptosis may play a major role in diseases demonstrating increased numbers of circulating and tissue eosinophils such as allergic reactions. Because few promoters of eosinophil apoptosis have been described so far, the objective of this study was to elucidate the role of endogenous factors on eosinophil survival and apoptosis. Methods and Results: Highly purified peripheral blood eosinophils were analyzed in the time course from 24 up to 144 hours in culture. Eosinophil survival was assessed with trypan blue dye exclusion and apoptosis was determined by DNA fragmentation gel analysis and ELISA technique with anti-histone antibodies. We confirmed previous results demonstrating prolonged eosinophil survival and inhibited apoptosis by IL-3, IL-5, and GM-CSF. In contrast, eosinophil apoptosis was significantly enhanced by corticosteroids, particularly dexamethasone and hydrocortisone. However, IL-1β, IL-8, IL-12, platelet-activating factor, TNF-α, and eotaxin had no effect on eosinophil survival or apoptosis when compared with culture medium alone. In contrast, IL-4 at concentrations of 100 U/mL or more inhibited eosinophil survival and induced apoptosis. This effect was time dependent and abrogated by preincubation with neutralizing anti–IL-4 antibodies. However, after 96 hours in coincubation, IL-4 did overcome the survival-prolonging effect of IL-3, IL-5, and GM-CSF. IL-4 did not enhance eosinophil surface expression of APO-1/Fas antigen (CD95). In assessing IL-4–mediated effects on eosinophil function, we found no response by means of the release of eosinophil cationic protein or reactive oxygen species. Conclusions: Taken together, our data present direct evidence for the presence of functional IL-4 receptors on human eosinophils and indicate that IL-4 may lead to resolution of chronic inflammation by induction of eosinophil apoptosis. (J Allergy Clin Immunol 1998;102:1013-20)     

IL—18对狼疮性肾炎患者外周血淋巴细胞凋亡及相关基因表达的研究

目的：研究LN患者外周血淋巴细胞（PBL）在IL－18刺激下细胞凋亡及凋亡相关基因表达情况。方法：Anniexin V联合PI染色定量法及免疫荧光染色法,分析了12例活动期和26例非活动期狼疮性肾炎患者PBL在IL－18刺激培养后凋亡发生率,凋亡相关基因bcl-2和Fas蛋白的表达以及疾病活动性与淋巴细胞凋亡发生率的相关性。结果：在IL－18刺激培养作用下,活动期LN淋巴细胞凋亡发生率明显增高（P＜0．01）,而非活动期则无明显区别（P＞0．05）,疾病活动性与体外淋巴细胞凋亡率呈正相关（P＜0．01）。bcl-2表达活动期显著性升高（P＜0．05）,非活动期无显著性差异（P＞0．05）。Fas表达活动期显著升高（P＜0．001）,非活动期明显升高（P＜0．05）。结论：IL－18可引起LN患者外周血淋巴细胞凋亡率的增高,表明IL－18在体内凋亡或凋亡相关性免疫机制中起着一定的作用。     

FasL在膀胱移行细胞癌中的表达及意义

目的：探讨膀胱移行细胞癌中Fas配体（FasL）的表达情况和临床意义。方法：对42例膀胱移行细胞癌组织及10例正常膀胱粘膜组织石蜡包埋标本应用免疫组化SP法检测FasL。结果：42例膀胱移行细胞癌细胞中FasL阳性32例（76．2％）,而正常膀胱粘膜FasL呈阴性表达（P＜0．01）,FasL阳性率与膀胱移行细胞癌的分级分期有关（P＜0．05）。结论：FasL的异常表达可能参与 膀胱肿瘤的免疫逃逸机制,并在膀胱肿瘤的发生发展过程中起重要作用。     

脊髓全横断损伤后凋亡因子Fas和FasL的表达变化

本研究通过制备SD大鼠脊髓全横断性损伤模型,应用免疫组织化学ABC法和灰度值测定揭示促凋亡因子Fas和FasL在脊髓完全性损伤后,不同时间点之间的相互变化趋势,以探讨Fas和FasL在脊髓损伤中的作用。结果显示:正常和脊髓全横断损伤后的不同时间组SD大鼠脊髓灰质的前角神经元中均可观察到Fas和FasL的免疫阳性产物。与正常组比较,Fas、FasL阳性神经元细胞数在术后1、3、7、14d组均升高(P<0.05),21d组则降低(P<0.05)。前角Fas、FasL阳性神经元的灰度值在术后1、3、7d组较正常组降低(P<0.05),14、21d组无显著性差异(P>0.05)。结果提示大鼠脊髓全横断损伤后,在损伤节段尾侧脊髓前角神经元Fas与FasL的表达经历了由增高到降低的过程,Fas和FasL表达增加有诱导神经细胞凋亡的趋势,可能参与诱导了脊髓继发性损伤。     

氧化苦参碱对小鼠急性肝损伤早期肝细胞凋亡的作用

以往研究表明[1 ] ,小鼠腹腔注射脂多糖 (L PS) +D-氨基半乳糖 (Gal N )后肝组织出现严重出血、坏死 ,氧化苦参碱 (oxy-matrine,OM) 5 0 mg/ kg腹腔注射预保护能明显减轻上述病理损害。为了探讨 OM对肝损伤早期肝细胞凋亡的影响 ,本实验从血清学、形态学和免疫组化等方面进行如下研究。1　材料和方法1.1　药品和试剂　 L PS由本校基础医学部微生物学教研室馈赠 ;Gal N购自重庆医科大学化学教研室 ;氧化苦参碱 ,为宁夏制药厂产品 ;纯化抗鼠 TNFα- Ig G EL ISA试剂盒购自深圳晶美生物公司 ;Fas及其配体 (Fas L )免疫组化试剂盒购…     

小儿幽门螺杆菌感染与胃粘膜上皮细胞凋亡

目的　探讨幽门螺杆菌 (H .pylori)相关性小儿慢性胃炎胃粘膜上皮细胞凋亡和相关基因 Fas的变化 .方法　采用 TUNEL技术和 Envision免疫组化方法检测 19例 H .py-lori阴性慢性胃炎和 17例 H.pylori阳性慢性胃炎患儿 H.pylori清除前后胃粘膜上皮细胞凋亡和相关基因 Fas的变化 .结果　 19例 H.pylori阴性小儿胃炎胃粘膜上皮细胞凋亡指数和 Fas指数分别为 (2 .7± 0 .8) %和 (11.7± 9.0 ) % ;17例 H.pylori阳性小儿慢性胃炎胃粘膜凋亡指数和 Fas指数分别为 (8.5± 1.8) %和 (31.3± 1.9) % ;H.pylori阳性组凋亡指数和 Fas指数明显高于 H .pylori阴性组 P<0 .0 1. H .py-lori清除后胃粘膜上皮凋亡指数和 Fas指数均明显减少(P<0 .0 1) ,治疗前凋亡指数和 Fas指数分别为 (8.6± 2 .3) %和 (32 .6± 12 .7) % ;清除 H.pylori后 ,凋亡和 Fas指数分别为 (3.6± 1.8) %和 (13.3± 6 .4) % .结论　 H .pylori感染促进胃粘膜上皮细胞的凋亡 ;Fas基因过度表达可能是 H.py-lori诱导胃粘膜上皮细胞凋亡增加的机制之一 .     

慢性乙型肝炎患者PBMC Fas表达和凋亡的检测及意义

目的探讨活化诱导细胞死亡（Activation-inducedcelldealth,AICD）在乙型肝炎发病机制中的意义。方法分离30例乙型肝炎患者和12例健康献血员外周血单个核细胞(PBMC),利用间接免疫荧光染色法,通过流式细胞仪检测PBMCFas的表达;分离20例乙型肝炎病人和10例健康献血员PBMC,在PHA-P的刺激下短期培养,采用PI染色法,经流式细胞仪检测PBMC的凋亡。结果慢性乙型肝炎组PBMCFas表达率[(53.31±14.53)%]高于正常对照组PBMCFas表达率[(39.32±11.28)%,P<0.05]和慢性重型乙型肝炎组PBMCFas表达率[(37.93±15.89)%,P<0.01]。慢性乙型肝炎组PBMC凋亡率[(30.57±13.43)%]高于正常对照组PBMC凋亡率[(11.45±5.27)%,P<0.01]和慢性重型乙型肝炎组PBMC凋亡率[(13.59±6.44)%,P<0.01];HBeAg(+)组PBMC凋亡率高于正常对照组PBMC凋亡率[(29.50±12.25)%vs(11.45±5.27)%,P<0.01]。结论HBV感染能诱导慢性乙型肝炎外周血淋巴细胞Fas表达增加,通过Fas/FasL途径使Fas+淋巴细胞发生AICD,这可能是形成HBV慢性感染免疫耐受的一个重要机制。     

胃粘膜Hp感染与细胞凋亡和Fas抗原表达的关系

目的　通过观察幽门螺杆菌 (Hp)感染诱导胃粘膜上皮细胞凋亡与Fas抗原表达的关系 ,探讨Hp诱导胃粘膜上皮细胞凋亡的机制。方法　采用脱氧核糖核酸末端转移酶介导的缺口末端标记 (TUNEL)技术原位观察和比较Hp根除前后胃粘膜上皮细胞凋亡的变化 ,采用免疫组织化学染色检测Fas抗原表达变化。结果　Hp阳性患者胃粘膜上皮细胞凋亡指数为 12 .78% ,明显高于Hp阴性者 (3 .63 % ,P <0 .0 1) ;Hp根除后胃粘膜上皮细胞凋亡指数 (4.3 7% )较治疗前明显降低 (12 .46% ,P <0 .0 1) ,而持续阳性者凋亡指数无明显降低。Hp阳性患者胃粘膜上皮细胞Fas抗原表达率为 67.92 % ,高于Hp阴性者 (45 .0 0 % ) ;Fas抗原表达阳性的Hp阳性患者在Hp根除后Fas抗原细胞密度显著减少 (P <0 .0 1) ,而Hp未被根除者Fas抗原表达阳性细胞密度无明显变化。结论　Hp感染可诱导胃粘膜上皮细胞凋亡 ,这可能是Hp参与胃癌发生的重要机制之一 ;Hp感染可促进Fas抗原表达增加 ,这可能是Hp感染诱导胃粘膜上皮细胞凋亡的机制之一。     

乙型肝炎患者血清可溶性Fas和可溶性Fas配体水平的研究

目的 研究可溶性Ｆａｓ（ｓＦａｓ）和可溶怀Ｆａｓ配体（ｓＦａｓＬ）在乙型肝炎发病中的意义。方法 用ＥＬＩＳＡ双抗体夹心法检测３９例各种类型地乙型肝炎患者血清ｓＦａｓ和ｓＦａｓＬ含量及其与病变的关系。并以８例健康献血员血清作对照。结果 慢性肝炎（中～重型）血清中ｓＦａｓ含量较高,与其它各组比较有显著性差异（Ｐ〈０．０１）;重型肝炎血清中ｓＦａｓＬ含量较高,与其它各组比较亦有显著性差异（Ｐ〈０．０１）     

Participation of the Fas-Signaling System in the Initiation of Germ Cell Apoptosis in Young Rat Testes after Exposure to Mono-(2-Ethylhexyl) Phthalate

The Fas-signaling system is composed of the interacting proteins Fas (CD95/APO-1) and Fas ligand (FasL, CD95L, APO-1L) and is proposed to act in the testis as a paracrine signaling mechanism by which FasL-expressing Sertoli cells initiate apoptosis of Fas-bearing germ cells. Here we describe alterations in the expression of Fas and FasL in the testis after the intimate physical association between Sertoli cells and germ cells is disrupted by exposure to the Sertoli cell toxicant mono-2-(ethylhexyl) phthalate (MEHP). Young, 28-day-old Fisher rats were treated with MEHP (2 g/kg po) and killed 0, 3, 6, and 12 h after exposure. Immunohistochemical analyses revealed a significant increase in the numbers of Fas-positive germ cells as well as increases in the expression of Sertoli cell FasL. Western blot analysis demonstrated a time-dependent increase in the production of the soluble form of FasL after MEHP exposure and suggests that it may participate in triggering apoptosis in germ cells that have lost their intimate association with the Sertoli cells. Measurement of Fas in cytosolic and membrane fractions of testis homogenates by Western blot analysis revealed a significant shift of Fas expression into the membrane fraction after MEHP exposure. Taken together, these observations indicate that the Fas-mediated paracrine signaling mechanism participates in triggering apoptosis of germ cells despite the loss of their close physical association with Sertoli cells. A working model is presented to explain the involvement of the Fas-system in stimulating germ cell apoptosis after MEHP exposure.