移植物浸润T淋巴细胞肿瘤坏死因子相关凋亡诱导配体及受体DR4和DR5表达与大鼠小肠移植急性排斥反应的相关性

目的 探讨浸润T淋巴细胞七的DR4、DR5表达同小肠移植急性排斥反应的关系.方法 将2种近交系大鼠(SD、Wistar)54只按随机配对法分为A、B、C3组.A组大鼠为对照组(18只)行虚拟手术;B组(18只)大鼠行同系小肠移植;C组(18只)大鼠行不同品系小肠移植.各组大鼠于术后5 d取移植肠样本分别做HE染色和免疫荧光双标记染色.采用免疫荧光染色、激光共聚焦技术测定各组标本浸润T淋巴细胞肿瘤坏死因相关凋亡诱导配体及DR4、DR5的表达情况.结果 A组大鼠小肠黏膜正常,B组大鼠小肠表现为免疫耐受,C组大鼠表现为急性排斥反应.C组大鼠高T淋巴细胞表达肿瘤坏死因子相关凋亡诱导配体与A、B组大鼠比较差异有统计学意义(P < 0.01);A、B组大鼠浸润T淋巴细胞DR4、DR5均呈高表达,C组大鼠呈低表达,C组大鼠与A、B组比较差异有统计学意义(P < 0.01);A、B组大鼠比较差异无统计学意义(P > 0.05).结论 急性排斥反应的发生可能与浸润T淋巴细胞DR的低表达有关.减少浸润淋巴细胞DR表达的下调,或者上调DR将有助于控制急性排斥反应,诱导免疫耐受.     

移植物浸润T淋巴细胞肿瘤坏死因子相关凋亡诱导配体及受体DR4和DR5表达与大鼠小肠移植急性排斥反应的相关性

目的 探讨浸润T淋巴细胞七的DR4、DR5表达同小肠移植急性排斥反应的关系.方法 将2种近交系大鼠(SD、Wistar)54只按随机配对法分为A、B、C3组.A组大鼠为对照组(18只)行虚拟手术;B组(18只)大鼠行同系小肠移植;C组(18只)大鼠行不同品系小肠移植.各组大鼠于术后5 d取移植肠样本分别做HE染色和免疫荧光双标记染色.采用免疫荧光染色、激光共聚焦技术测定各组标本浸润T淋巴细胞肿瘤坏死因相关凋亡诱导配体及DR4、DR5的表达情况.结果 A组大鼠小肠黏膜正常,B组大鼠小肠表现为免疫耐受,C组大鼠表现为急性排斥反应.C组大鼠高T淋巴细胞表达肿瘤坏死因子相关凋亡诱导配体与A、B组大鼠比较差异有统计学意义(P < 0.01);A、B组大鼠浸润T淋巴细胞DR4、DR5均呈高表达,C组大鼠呈低表达,C组大鼠与A、B组比较差异有统计学意义(P < 0.01);A、B组大鼠比较差异无统计学意义(P > 0.05).结论 急性排斥反应的发生可能与浸润T淋巴细胞DR的低表达有关.减少浸润淋巴细胞DR表达的下调,或者上调DR将有助于控制急性排斥反应,诱导免疫耐受.     

PD142893诱导活化T细胞凋亡减轻大鼠肺移植急性排斥反应

T淋巴细胞在器官移植急性排斥反应中起核心作用，目前许多免疫抑制剂都是通过阻止T淋巴细胞的活化而发挥抗急性排斥效应的。本研究旨在探讨PD142893参与大鼠肺移植后对急性移植排斥反应的影响及对活化T细胞的作用。     

肿瘤坏死因子相关凋亡诱导配体与胰腺癌及相关药物研究

自肿瘤坏死因子家族的新成员肿瘤坏死因子相关凋亡诱导配体被发现以来,它一直是近些年来在肿瘤研究方面的热点.本文综述了近些年肿瘤坏死因子相关凋亡诱导配体在胰腺癌方面的相关研究进展,从肿瘤坏死因子相关凋亡诱导配体诱导胰腺癌肿瘤细胞凋亡机制人手,简要介绍胰腺癌对肿瘤坏死因子相关凋亡诱导配体的耐药机制.并根据目前已认同的耐药机制...     

肿瘤坏死因子相关凋亡诱导配体治疗肿瘤存在的问题及策略

肿瘤坏死因子相关凋亡诱导配体(TRAIL)是近几年发现的TNF超家族成员 ,为Ⅱ型跨膜蛋白 ,能与其受体(TRAILR)结合 ,启动细胞内的信号转导 ,激活Caspase级联反应诱导细胞凋亡。这种新发现的凋亡诱导途径有重要的生物学意义 ,尤其是在抗肿瘤方面具有选择性杀伤肿瘤细胞同时赦免正常细胞的作用 ,因而引起了人们的高度重视 ,认为有望找到一个全新的肿瘤治疗方法。然而 ,随着对TRAIL研究的深入 ,TRAIL治疗肿瘤可能存在的障碍不断出现 ,人们对TRAIL能否成为一个理想的抗癌新药提出质疑 ,现从TRAIL治疗肿瘤可能存在的问题作一综述 ,并…     

肿瘤坏死因子相关诱导配体受体在前列腺癌组织中的表达

目的：研究肿瘤坏死因子相关诱导配体受体在前列腺癌组织中的表达情况。方法：应用RT—PCR检测20例良性前列腺增生和20例前列腺癌组织中的死亡受体DR4、DR5、假受体DcRl、DcR2的mRNA表达。结果：良性前列腺增生组织中DR4、DR5、DcR1均为85％(17／20)；DcR2为75％(15／20)。前列腺癌组织中死亡受体DR41、DR5为80％(16／20)；DcR1阳性表达率为15％(3／20)；假受体DcR2于前列腺癌组织中未见表达。结论：前列腺癌组织中的DcR1、DcR2受体缺乏表达，DcR1、DcR2在前列腺癌的凋亡调控途径中可能有重要作用。     

大鼠肝移植急性排斥反应中肿瘤坏死因子受体配体的表达及其意义

目的 观察肝移植排斥反应中移植肝脏内糖皮质激素诱导的肿瘤坏死因子受体配体(GITRL)的表达.方法 采用Kamada's二袖套法建立从Lewis到Brown Norway(BN)大鼠的肝移植排斥模型为排斥组(n=5),从BN到BN的肝移植模型为耐受组(n=5).术后24h,抽取血液,取肝脏及分离库普弗(Kupffer)细胞,检测肝脏上GITRL及肿瘤坏死因子(TNF)-α的表达,Kupffer细胞上GITRL的表达,检测血清及细胞上清液中TNF-α的表达.免疫组织化学染色强度采用Image-Pro Plus 6.0图像分析软件分析.结果 免疫耐受组和排斥组肝脏内的CITRL的平均染色强度分别为0.113±0.007和0.270±0.018(P＜0.05),TNF-α平均染色强度分别为0.114±0.004和0.141±0.005(P＜0.05),耐受组和排斥组的Kupffer细胞GITRL平均染色强度分别为0.206±0.017和0.337±0.018(P＜0.05),Kupffer细胞的培养上清液中,耐受组和排斥组TNF-α的值分别为(68.66±21.12)、(178.33±29.39)ng/L(P＜0.05).结论 在排斥的早期阶段肝脏及Kupffer细胞的GITRL表达增高,监测和干扰GITRL可能有益于肝移植急性排斥反应的早期诊断和处理.

Abstract：

Objective To investigate te changes of glucocorticoid induced tumor necrosis factor receptor ligand (GITRL) in hepatic allograft rejection. Methods Liver transplantation from Lewis rats (n = 5 ) to Brown Norway (BN) rats was performed by Kamada' s two-cuff technique as acute rejection group. Liver transplantation from BN to BN rats ( n = 5 ) was performed as tolerance group. Recipients were sacrificed at 24th h postoperation. Blood samples were collected and grafts were harvested, then Kupffer cells were isolated. GITRL and tumor necrosis factor (TNF)-α protein expression in the hver was tested by immunohistochemistry, and the GITRL expression in Kupffer cells by immunocytochemistry. Enzyme linked immunosorbent assay (ELISA) was employed to detect the changes of TNF-α protein in the serum and supernatant. The staining intensity was analyzed by Image-Pro Plus 6. 0 image analysis software. Results At 24th h postoperation, the liver GITRL expression levels in tolerance and rejection groups were 0. 113 ± 0. 007 and 0. 270 ±0. 018, respectively (P ＜0. 05). The TNF-α expression levels in the liver in tolerance and rejection groups were 0. 114 ± 0. 004 and 0. 141 ± 0. 005 respectively ( P ＜ 0.05 ). The GITRL expression levels in Kupffer cells in tolerance and rejection groups were 0. 206 ±0. 017 and 0. 337 ±0. 018 respectively (P ＜0. 05 ). As compared with tolerance group (68. 66 ±21.12) ng/L, TNF-α protein expression levels were up-regulated in the supernatant of rejection group ( 178.33 ± 29. 39 ) ng/L ( P ＜ 0. 05 ).Conclusion The expression of GITRL in the liver and Kupffer cells was increased in the early stage of rejection, and monitoring and interfering GITRL may be useful for the early diagnosis and management of an acute rejection in liver transplantation.     

转染肿瘤坏死因子相关凋亡诱导配体对膀胱癌细胞的抑制作用

目的探讨肿瘤坏死因子相关凋亡诱导配体（TRAIL）对膀胱癌细胞的抑制作用。方法构建TRAIL融合增强绿色荧光蛋白（EGFP）真核表达载体，脂质体法转染膀胱癌细胞株玎细胞。荧光显微镜下计算转染率。分别采用逆转录-聚合酶链反应（RT-PCR）、Western blot检测TRAIL的表达。采用流式细胞仪检测细胞凋亡，噻唑蓝（MTT）法、细胞倍增时间和集落形成率观察TRAIL的抑制作用。结果48h观察重组载体转染率为38．4％，空载体转染率为35．3％（P〉0．05）．RT-PCR和Western blot证实转染后EJ细胞中TRAIL的表达明显增加；凋亡率明显高于转染空白载体和未转染组（P〈0．01）；EJ细胞增殖受到明显抑制（P〈0．01）。结论TRAIL具有诱导膀胱癌细胞凋亡、抑制增殖的作用，有望成为治疗膀胱癌的新方法。     

肿瘤坏死因子相关凋亡诱导配体及其受体与抗肿瘤治疗

肿瘤坏死因子相关凋亡诱导配体(ｔｕｍｏｒｎｅｃｒｏｓｉｓｆａｃｔｏｒｒｅｌａｔｅｄａｐｏｐｔｏｓｉｓ ｉｎｄｕｃｉｎｇｌｉｇａｎｄ ,ＴＲＡＩＬ)是近几年发现的肿瘤坏死因子 (ｔｕｍｏｒｎｅｃｒｏｓｉｓｆａｃｔｏｒ,ＴＮＦ)超家族成员 ,为Ⅱ型跨膜蛋白 ,能与其受体 (ｔｕｍｏｒｎｅｃｒｏｓｉｓｆａｃｔｏｒｒｅｌａｔｅｄａｐｏｐｔｏｓｉｓ ｉｎｄｕｃｉｎｇｌｉｇａｎｄｒｅｃｅｐｔｏｒ,ＴＲＡＩＬＲ)结合 ,启动细胞内的信号转导 ,激活半胱—天冬氨酸蛋白酶 (ｃｙｓｔｅｉｎｅｃｏｎｔａｉｎｉｎｇａｓｐａｒａｔｅｓｐｅｃｉ…     

肿瘤坏死因子相关凋亡诱导配体介导脓毒症免疫抑制的研究进展

背景 肿瘤坏死因子相关凋亡诱导配体(tumor necrosis factor related apoptosis induced ligand,TRAIL)是肿瘤坏死因子超家族成员之一,其与受体DR4/DR5结合可以诱导肿瘤细胞的凋亡,而备受关注. 目的 总结TRAIL通路在脓毒症中的可能机制,探讨其在脓毒症疾病过程中的调控作用. 内容 脓毒症动物模型的研究发现,TRAIL可以加快活化的中性粒细胞发生凋亡并参与脓毒症免疫耐受的形成. 趋势 TRAIL受体在肿瘤、自身免疫性疾病、感染、心血管疾病、器官移植等领域的研究越来越多,作用机制也在不断地阐明.TRAIL及其受体在脓毒症疾病过程中发挥重要调节作用.     

Migration of host and donor T cells in small bowel transplantation

In this study migration of host and donor CD4⁺ and CD8⁺ T cells in a fully allogeneic model was described and compared with the migration pattern in a graft-versushost reaction (GVHR) model, where the T-cell traffic in the graft served as a physiological control. Heterotopic small bowel transplantations were performed in a rat model, with animals being sacrificed on postoperative days (POD) 2, 3, 4, 5, and 7. Graft and host mesenteric lymph nodes were harvested, homogenized, and stained with monoclonal antibodies against MHC class 1, CD4⁺, and CD8⁺ antigens. The host and donor T cell migration patterns were studied using a doublestaining flow cytometric technique. We found that during the development of rejection, the normal physiological circulation of graft and host T cells was disrupted. In the graft of the allogeneic model, a shift from host cell to graft cell dominance occurred on POD 3–4. This change in migration pattern coincided in the host with a 6 % peak in graft cell infiltration, which disappeared on POD 7. These patterns of T-cell migration may be further explored for diagnostic purposes.     

Migration of host and donor T cells in small bowel transplantation

Abstract In this study migration of host and donor CD4⁺ andCD8⁺ T cells in a fully allogeneic model was described and compared with the migration pattern in a graft-versus-host reaction (GVHR) model, where the T-cell traffic in the graft served as a physiological control. Heterotopic small bowel transplantations were performed in a rat model, with animals being sacrificed on postoperative days (POD) 2, 3, 4, 5, and 7. Graft and host mesenteric lymph nodes were harvested, homogenized, and stained with monoclonal antibodies against MHC class I, CD4 ⁺, and CD8 ⁺ antigens. The host and donor T cell migration patterns were studied using a double-staining flow cytometric technique. We found that during the development of rejection, the normal physiological circulation of graft and host T cells was disrupted. In the graft of the allogeneic model, a shift from host cell to graft cell dominance occurred on POD 3–4. This change in migration pattern coincided in the host with a 6 % peak in graft cell infiltration, which disappeared on POD 7. These patterns of T-cell migration may be further explored for diagnostic purposes.     

小肠移植后亚临床排斥反应的临床病理研究一例

目的 探讨小肠移植后亚临床型细胞性排斥反应(SCR)的临床表现、肠镜下改变和病理特点.方法 小肠移植1例,受者为女性,34岁,供者为男性.应用抗CD52单克隆抗体行诱导治疗,术后单用他克莫司(Tac)、无皮质激素的维持治疗方案.怀疑发生可疑排斥反应(IND)级至I级排斥反应时,提高血Tac浓度,行短程小剂量皮质激素治疗,排斥反应控制不佳时,则行甲泼尼龙冲击治疗;发生中度排斥反应时提高血Tac浓度,给予甲泼尼龙2 g,随后应用皮质激素递减方案.术后头2个月内,每周行2次肠镜和病理学检查,之后频次减为1次/周.结果 至随访结束患者已存活19个月(611 d),期间共发生有临床症状的急性排斥反应6次,亚临床排斥反应3次.3次亚临床排斥反应时,2次肠镜下未见明显改变,1次表现为斑点状的充血、水肿,放大肠镜下绒毛数量轻度减少.3次亚临床排斥反应的病理检查中,1次以黏膜上皮剥脱为主,隐窝上皮损伤较轻,组织学改变符合轻度急性排斥反应;而另2次以隐窝上皮的损伤为主,为可疑急性排斥反应.结论 SCR可能是临床排斥反应的早期阶段,无明显临床症状,肠镜下改变不明显,诊断主要依靠病理学检查,但必须排除移植肠血管病变、肠梗阻、各类炎症等术后并发症.

Abstract：

Objective To investigate the clinical presentation, endoscopy and pathological features of subclinical cellular rejection (SCR) of small bowel allotransplantation. Methods Three times of SCR in a patient after isolated small bowel transplantation were studied by endoscopy and microscopy, and the clinical data and literature were reviewed. Results SCR was an unusual type of acute rejection after small bowel transplantation. SCR showed low-grade morphological changes of acute rejection, and may be relived after low-dose steroid or bolus steroid was given. Conclusion The causes of SCR are not clear now. SCR may be the early stage of clinical acute rejections, and may be correlated with unexpected high grade acute rejection, and chronic loss function of graft. The biopsy through ileoscopy is a "golden standard" of diagnosis of SCR in small bowel transplantation.However, the vessel lesions of graft, ileus, and inflammation should be excluded before diagnosis.     

Altered distribution of MHC class II antigens on enterocytes during acute small bowel allograft rejection in rats

Class II major histocompatibility complex (MHC) antigen induction was investigated on enterocytes of heterotopic rat small bowel allografts in the Lewis-Brown Norway strain combination and on isografts in the Lewis-Lewis strain combination. I a antigens were detected with monoclonal antibodies using an immunoperoxidase technique. Generally, MHC class II antigens were not exhibited in the isografted group, with the exception of two long-term isografts that presented the same pattern as normal small bowel. In these cases, I a was expressed in a patchy distribution predominantly in the villi, and only very few enterocytes stained positive in Lieberkühn's crypts. Allografted rats showed a typical pattern of I a expression on the enterocytes during the rejection course. The initial expression was confined to the crypts, indicating a very early stage of rejection when compared to histological findings. More advanced stages of rejection were accompanied by increasing I a biosynthesis in the crypts and I a expression by the epithelium lining the villi. Cyclosporin (CyA) was not able to fully inhibit MHC class II antigen expression; however, the appearance of I a was delayed.     

辅助性同种异体肝肠联合移植术后急性排斥反应的监测

目的评价猪同种异体辅助性肝肠联合移植术后排斥反应的监测方法。方法将50头杂交长白猪分为3组．A、B组各20头各完成10次猪辅助性带胰头及十二指肠的同种异体肝肠联合移植术．其中B组术后予以免疫抑制治疗：C组10头完成交互的同种异体节段性小肠移植术10例。术后1、3、5、7、14、21及30d经移植肠远端造口取小肠黏膜经常规处理后。分别在光镜和电镜下观察并进行排斥反应评分。结果术后A组出现排斥反应的中位时间为8（7～12）d．迟于c组的5（3～5）d（P〈0．05）。术后1周,A组的排斥反应评分为1．11±0．20。低于C组（2．56±0．18,P〈0．05）;但比B组高（O．20±0．13,P〈O．05）。A组移植术后中位存活时问为9（7～25）d,C组为12（7～20）d．而B组术后全部成活超过30d．与以上两组比较,P〈0.05．差异有统计学意义。结论移植术后排斥反应通过肠造口取材进行监测方便有效。     

Relevance of tumor necrosis factor to graft-versus-host disease after small bowel transplantation

The small bowel (SB), an organ replete with lymphocytes, may provoke graft-versus-host disease (GVHD) after transplantation (Tx). Since tumor necrosis factor (TNF) has been suspected of mediating the tissue lesions of GVHD, we sought to determine whether TNF could be detected in the serum of rats undergoing GVHD after SBTx or lymphocyte transfer. For this purpose, post-operative serum TNF activity was determined in Lewis x Brown after undergoing transplantation of an entire (group 1; n=8) or a segmental (group 2; n=4) Lew SB, or after i. p. injection with lethal doses (500×10⁶) of Lew lymphocytes (group 3; n=3). Control LBNF1 received i.p. small doses (50×10⁶) of Lew lymphocytes (group 4; n=4). Serum TNF activity was assessed using the WEHI bioassay. In rats with acute and lethal GVHD after entire SBTx (group 1) or injection with large doses of lymphocytes (group 3), TNF activity gradually increased and reached high levels by the time the rats were agonal. In segmental SBTx rats (group 2), GVHD was less severe than in entire SBTx rats. Similarly, the increase in TNF activity was less intense and only transient since it had returned to control levels by the time the rats had completely recovered from GVHD. In control rats primed with small doses of lymphocytes (group 4), GVHD did not occur and no increase in TNF activity was detected. We conclude that: (1) GVHD after SBTx or lymphocyte transfer is associated with the appearance of TNF in the serum and (2) the intensity and the reversibility of this phenomenon correlate with clinical severity and lethality of GVHD. These data strongly suggest that TNF is involved in the pathogenesis of GVHD.     

Value of mucosal biopsies in the monitoring of acute small bowel rejection

The value of mucosal biopsies in evaluating small bowel rejection is controversial. In this study, the value of mucosal biopsies was estimated in unmodified porcine small bowel rejection. Ten animals received the distal half of the small bowel as a heterotopic loop (Thiry-Vella loop). The allografts were followed by proximally and distally harvested full-thickness and mucosal biopsies every other day, starting from the 3rd day and continuing until the grafts became necrotic. The histological parameters in both types of biopsies were semiquantitatively scored from 0 to 3 and compared with each other. The difference in mean values on subsequent days was not remarkable, the results favoring slightly higher values in full-thickness than in mucosal biopsies. Our results suggest that multiple mucosal biopsies are adequate in monitoring morphological changes of small bowel grafts during rejection and that the proximal and distal ileum are similarly affected by acute rejection. Received: 17 June 1996 Received after revision: 20 November 1996 Accepted: 6 December 1996     

小肠移植术后内镜引导下移植肠黏膜活检的时机及诊断价值

目的 总结小肠移植术后内镜引导下移植肠黏膜活检的时机及该技术对急性排斥反应和感染的诊断价值.方法 根据免疫抑制方案的不同,将15例小肠移植受者分为3个阶段.1994-1995年为第1阶段(3例),2003-2006年为第2阶段(7例),2007年以后为第3阶段(5例).第3阶段进行计划性内镜引导下移植肠黏膜活检的监测,既术后第3天进行首次内镜引导下移植肠黏膜活检,此后活检的频次在术后第1个月为2次/周,术后第2～3个月为1次/周,术后第4～6个月为1次/2周,术后7个月以后为1次/月,在受者出现排斥反应的临床症状和抗排斥反应治疗期间,也进行内镜引导下移植肠黏膜活检.结果 15例共进行内镜引导下移植肠黏膜活检255次,移植肠腹壁造口肉眼直视下取材活检21次.以上276份样本中,诊断排斥反应共51份(18.5%),其中诊断不确定急性排斥反应至轻度排斥反应32份(11.6%)、中度排斥反应9份(3.3%)、重度排斥反应10份(3.6%),巨细胞病毒(CMV)感染2份(0.7%),细菌感染2份(0.7%).15例共发生病理证实并需l临床治疗的排斥反应20次,其中不确定急性排斥反应至轻度排斥反应11次、中度5次、重度4次,发生细菌性和CMV肠炎各1次.结论 内镜引导下移植肠黏膜活检及其病理学检查是小肠移植术后诊断排斥反应和感染的重要手段,有计划的进行该检查对排斥反应有术后监测、早期诊断、鉴别诊断和指导治疗的价值.