人类白细胞抗原-G和人类白细胞抗原-E在妊娠免疫耐受中的作用

人类白细胞抗原（HLA）-G、-E同属非经典主要组织相容性复合物（MHC）I类分子,于母胎界面滋养层细胞高表达,在妊娠免疫耐受中发挥重要作用。HLA—G和HLA—E主要通过和自然杀伤（NK）细胞、T淋巴细胞表面抑制性CD94／NKG2C受体或激活性CD94／NKG2A受体结合,抑制或激活免疫细胞效应,维持正常妊娠免疫耐受。HLA-G和HLA—E基因结构及表达异常可能与妊娠并发症相关,可作为判定正常妊娠和妊娠相关疾病指标。本文综述HLA-G和HLA—E结构和分布、与免疫细胞可能的作用机制以及相关临床应用,讨论其在维持母胎免疫耐受中的作用。     

人类白细胞抗原-G在临床医学中的研究进展

人类白细胞抗原(HLA)-G是一类非经典HLAⅠb类分子, 早期的研究主要集中在妊娠免疫, HLA-G可保护胎儿免受母体免疫系统的攻击。近几年越来越多的研究发现HLA-G在诱导器官移植物免疫耐受、协助肿瘤细胞以及病毒的免疫逃逸中发挥着重要作用。因此将HLA-G作为器官移植、恶性肿瘤和病毒感染等疾病诊断和治疗的靶点已逐渐成为学者关注的热点。本文就HLA-G的基因多态性与分子结构、HLA-G在临床医学中应用的最新进展做一总结。     

黑色素瘤抗原基因在肝癌细胞系中的表达

目的：研究黑色素瘤抗原（MAGE）基因在肝癌（HCC）细胞系中的表达，为应用MAGE抗原肽作为疫苗治疗肝癌的体外实验筛选靶细胞。方法：分别用逆转录-聚合酶链反应（RT-PCR）和免疫组织化学方法对MAGE-1和MAGE-3mRNA及MAGE-1蛋白在8种HCC细胞系中的表达进行检测，用微量淋巴细胞毒法测定HCC细胞系的主要组织相容性复合物（MHC)I类分子的表达。结果：6种HCC细胞系MAGE-1基因mRNA阳性表达，2种MAGE-3阳性；有5种细胞系MAGE-1基因蛋白阳性。BEL7405HLA-A24阳性，HLE和HpG2HLA-A2阳性。结论：MAGE基因在HCC细胞系中有较高的表达率，HLE和BEL7405可作为阳性靶细胞，其他细胞系可作为相应的对照细胞，应用于用MAGE抗原肽疫苗治疗HCC的体外实验研究中。     

人类白细胞抗原-G蛋白在星形细胞肿瘤组织的表达

胶质瘤存在免疫耐受。而人类白细胞抗原-G(HLA-G)是一种非经典人类白细胞抗原,有研究表明HLA-G作为免疫耐受分子,在肿瘤免疫逃逸中可能具有重要作用。我们采用免疫组织化学、蛋白印迹实验观察HLA-G蛋白在星形细胞性肿瘤表达情况。     

肝癌HLA-E基因的表达及对判断肝癌肝移植疗效的意义

目的 观察肝癌患者人类白细胞抗原-E(HLA-E)及相关抗原的表达,以及其对判断肝癌肝移植预后的可行性和价值.方法 回顾性研究40例肝癌肝移植患者,检测HLA-ABC、HLA-E、抗原提呈辅助分子(β2-M、TAP)抗原在其原发灶和癌旁硬化组织中的表达,逆转录PCR法检测石蜡标本中HLA-E mRNA的表达,并将各抗原表达情况与肝移植的预后进行统计学分析.结果 与癌旁组织相比,癌巢HLA-ABC表达增强12例(30%),一致21例(52.5%),相对下调7例(17.5%).HLA-E抗原在癌旁组织中表达不明显,癌巢阳性表达(++/+)为29例(72.5%),P＜0.01,肝癌标本可检测出HLA-E mRNA的表达.β2-M和TAP在癌巢和癌旁组织中均有相同程度的表达,无统计学意义.随访期内HLA-E(++/+)患者移植后无瘤生存期较短(P＜0.05).结论 肝癌HLA-E异常表达影响肝癌肝移植的疗效,提示其在肝癌免疫逃避中的重要作用.     

RNA干扰白细胞抗原-G增强自然杀伤细胞对肝癌的免疫监视

目的 构建表达人类白细胞抗原-G(HLA-G)基因shRNA载体,探讨其对自然杀伤细胞(NK)细胞杀伤效应的影响.方法 构建4个HLA-G-shRNA真核表达质粒,转染至Huh7细胞.检测HLA-G mRNA及蛋白质水平的表达.将NK-92MI细胞与靶细胞共同培养,以LDH释放法观察不同效靶比时NK-92MI细胞对靶细胞的杀伤效应.结果 经酶切及测序鉴定证实,插入序列与设计的序列相符.Real Time-PCR和Western-blot结果均表明重组载体抑制了Huh7细胞HLA-G基因的表达.NK-92MI细胞对转染HLA-G的shRNA的Huh7细胞杀伤作用明显升高(P<0.01).封闭NK-92MI细胞ILT2受体后,杀伤作用降低(P<0.01).结论 HLA-G shRNA可以增强NK细胞对肝癌细胞的杀伤效应.     

人类白细胞抗原-G与免疫耐受的研究进展

人类白细胞抗原(HLA)-G是位于人6号染色体短臂上的一群紧密连锁基因群,属于人类一种非经典的主要组织相容性复合体(MHC)的Ⅰ类分子,选择性高表达于侵入子宫蜕膜的绒毛外滋养细胞。母胎免疫耐受是天然的同种异体免疫耐受现象,虽然胎儿含有一半与母体基因无关的父源性基     

人类白细胞抗原-GRNA干扰载体的构建及效应检测

目的构建表达人类白细胞抗原-G(HLA-G)基因shRNA载体,探讨其对NK细胞杀伤效应的影响。方法构建4个HLA-G-shRNA真核表达质粒,瞬时转染至人肝癌Bel-7402细胞,检测HLA-G mRNA和蛋白水平的表达。将NK-92MI细胞(效应细胞)与转染后的Bel-7402细胞(靶细胞)共同培养,以LDH释放法观察不同效靶比时NK-92MI细胞对靶细胞的杀伤效应。结果经酶切及测序鉴定证实,插入序列与设计的序列相符。RT-PCR和Western blot结果均表明重组载体抑制了Bel-7402细胞HLA-G基因的表达。NK-92MI细胞对转染HLA-GshRNA的Bel-7402细胞杀伤作用明显增强(P<0.01)。封闭NK-92MI细胞KIR2DL4受体后,杀伤作用减弱(P<0.01)。结论成功构建了HLA-G shRNA载体,下调HLA-G可以增强NK细胞对肝癌细胞的杀伤效应,HLA-G与NK细胞表面KIR2DL4受体结合后,向NK细胞传递抑制效应。     

PCNA表达与原发性肝癌生物学行为及预后关系

目的:探讨增殖细胞核抗原(proliferating cell nuclear antigen,PCNA)表达与肝细胞性肝癌(hepatocellular carcinoma,HCC)生物学行为及临床预后的关系。方法:用免疫组织化学方法检测59例HCC癌组织和40例癌旁肝组织中PCNA的表达情况。结果:59例癌组织的PCNA标记指数(Labelling index,LI)(41.9％±21.8％)显著高于40例癌旁肝组织(1.9％±1.0％)(P<0.001)。分化程度Ⅰ、Ⅱ、Ⅲ、Ⅳ级的癌组织其平均PCNA LI分别为14.6％±3.5％、26.7％±6.9％、50.4％±16.7％和72.7％±14.7％,经统计学分析,PCNA LI与肝癌组织的组织学分级有显著相关性(P值均小于0.005)。高PCNALI组(≥42％)与低PCNA LI组(<42％)相比,前组的肿瘤包膜浸润率(91.7％)和静脉癌栓形成率(83.3％)显著高于后组(分别为45.0％和42.9％)(P值均小于0.05)。术后存活期<5年组的PCNA LI(59.3％±15.6％)显著高于术后存活期≥15年组(32.7％±9.8％)(P<0.001)。结论:测定PCNA LI对于判定HCC组织学分化程度、评估其生物学行为、判断HCC患者预后具有一定意义。     

人类白细胞抗原-G与早期胚胎生长发育和着床

人类白细胞抗原-G(HLA-G)是人类组织相容性抗原复合物,在母胎免疫耐受中起作用。近年的研究发现,HLA-G在着床前胚胎表达。我们和国际上其他的临床研究显示,分泌可溶性HLA-G的胚胎在体外受精和胚胎移植中的临床妊娠率和种植率较高。进一步的研究发现,HLA-G与胚胎的免疫耐受、生长、着床相关的粘附和浸润相关,并通过MEK1/2信号传导通路促进人绒毛膜促性腺激素的分泌。我们还发现,子宫内膜细胞与可溶性HLA-G结合,并表达其受体;可溶性HLA-G还促进子宫内膜细胞与滋养细胞的粘附;白血病抑制因子和miR-152参与HLA-G在滋养细胞表达的调控。HLA-G表达和功能相关的分子机制研究显示,HLA-G在胚胎生长和着床上起重要作用,可能是胚胎发育潜能的标志分子。     

Expression of Human Leukocyte Antigen G (HLA-G) Correlates with Poor Prognosis in Gastric Carcinoma

Objective We had previously demonstrated that human leukocyte antigen G (HLA-G) was expressed in a majority of primary colorectal carcinomas and that the detection of HLA-G expression had a strong and independent prognostic value for that cancer. Currently, we investigate whether or not HLA-G is also expressed in patients with gastric carcinoma and whether the expression has any clinical application value. Methods The expression of HLA-G was investigated immunohistochemically in 160 patients with gastric carcinoma. The correlation between HLA-G status and various clinicopathological parameters was analyzed with the levels of HLA-G expression used to compare the survival length amongst patients. Results HLA-G protein expression was observed in 71% (113 of 160) of the primary site of gastric carcinomas, but not in the normal stomach tissues. HLA-G expression in the tumors was significantly correlated with the tumor location, histological grade, depth of invasion, lymph nodal metastasis, clinical stages of the disease, and host immune response (P = .012, .008, .001, .038, .030, and .016, respectively). Patients with HLA-G positive tumors had a significantly shorter survival time than those patients with tumors that were HLA-G negative (P = .001). As well, in multivariate analysis, HLA-G demonstrated an independent prognostic factor (P = .0001, relative risk 9.08; 95% confidence interval, 3.44–24.0). Conclusions Overall, our results indicated that the expression of HLA-G is a characteristic feature of gastric carcinoma and that immunostaining by anti-HLA-G antibody may be a potentially useful prognostic indicator.     

挽救性肝移植治疗肝癌切除术后复发的价值研究

目的 探讨挽救性肝移植(SLT)的手术安全性及对患者预后的影响.方法 回顾性分析复旦大学附属中山医院肝外科2001年6月至2008年12月期间连续289例肝癌肝移植(符合UCSF标准)患者的临床资料,其中242例患者行初始肝移植(PLT),即PLT组,47例患者行SLT,即SLT组,比较2组患者围手术期及长期生存情况的差异.结果 2组患者的平均年龄、性别构成及肿瘤情况差异均无统计学意义(P>0.05).SLT组的手术时间要长于PLT组[(7.1±1.8)h比(6.4±1.4)h,P=0.004],但2组患者的术中出血量[(2 560.5±2 683.6)ml比(2 042.9±2 006.2)ml,P=0.173]及术中输血量[(13.8±12.9)U比(9.9±12.6)U,P=0.087]比较差异均无统计学意义,SLT组患者从第1次手术切除至行肝移植的间隔时间为(32.8±32.4)个月.截至2009年12月,2组患者中位随访时间为38.7个月,SLT组与PLT组患者的3年生存率(82.3%比75.5%,P=0.312)和3年无瘤生存率(78.8%比70.1%,P＝0.755)之间比较差异均无统计学意义.但按意向性治疗分析,SLT组患者的3年生存率明显优于PLT组(88.4%比76.2%,P＝0.047).结论 SLT并不增加移植手术的风险,也不影响患者的长期预后,对部分病例,先行手术切除再行肝移植可作为肝癌治疗的一种有效策略.     

Prognostic Impact of Sarcomatous Change of Hepatocellular Carcinoma in Patients Undergoing Liver Resection and Liver Transplantation

Sarcomatous change has been rarely observed in hepatocellular carcinoma (HCC), but it is usually associated with very aggressive tumor behavior and widespread metastasis. To assess the impact of sarcomatous changes, we analyzed the outcomes of 15 patients with sarcomatous HCC after resection (n = 11) or liver transplantation (LT) (n = 4). No imaging findings characteristic of sarcomatous changes were observed. According to modified pathological tumor-node metastasis staging, the HCC lesions were classified as stage II in five patients, stage III in six, stage IVa2 in two, and stage IVb in one. The Milan criteria were met in 7 of 15 patients, including 3 of 4 in the LT group. R0 resection was achieved in 9 of 11 resected patients, and their 3-year overall and disease-free survival rates were both 18.2%. In the LT group, 3-year overall and disease-free survival rates were 37.5 and 25%, respectively. In patients within the Milan criteria, 2-year overall survival rate was 25% after resection and 33% after LT, showing no prognostic difference. Extrahepatic metastasis as initial recurrence was detected in 80% after resection and 66.7% after LT. In conclusion, we found that the prognosis of patients with sarcomatous HCC was very unfavorable after either resection or LT and that, except for liver biopsy, no diagnostic method could distinguish between sarcomatous and ordinary HCC. Vigorous postoperative systemic surveillance may be helpful for timely detection and treatment of localized metastases.     

射频消融在肝细胞肝癌根治性治疗中的临床价值

目的探索射频消融术（RFA）在肝细胞肝癌（简称肝癌）根治性治疗中的临床应用价值。方法检索近年来有关RFA在肝癌治疗中应用的文献并进行综述。结果肝脏移植、肝切除术和RFA是目前被认为具有治愈性治疗效果的3种方法，其中RFA由于具有较好的局部肿瘤控制效果，近年来较多地用于肝脏移植术前的减瘤治疗，在延长患者的等待期的同时不增加病例脱落及死亡的危险。虽然RFA与肝脏切除术对小肝癌的疗效目前尚有争论，但是RFA联合肝脏切除术，扩大了肝癌患者的手术指征，提高了疗效。结论由于RFA技术良好的局部肿瘤控制能力和微创特点，使其在肝癌的各种治疗策略中发挥越来越重要的作用，并与肝移植及肝部分切除术的优点相互补充，使更多的肝癌患者受益。无论是哪一种治疗方法，术中最大程度地减少残癌的发生率，术后密切随访，复发后积极地治疗才是提高疗效的根本。     

Two Rare Metachronous Metastases of Hepatocellular Carcinoma After Liver Transplantation

A 59-year-old male with hepatocellular carcinoma (HCC) due to liver cirrhosis caused by the hepatitis C virus underwent cadaveric whole liver transplantation. Two years later, he had a metastatic HCC in the superior mediastinum. Over the following postoperative year, he underwent transcatheter arterial chemoembolization (TACE) for 4 tumors in the implanted liver. In the third post-TACE month, he was emergently hospitalized due to intracerebral hematoma with a tumor invading the bone in the medial frontal segment. He underwent emergency intracranial tumorectomy and hemorrhage removal. The histopathologic diagnosis was metastatic HCC. He regained consciousness as well as the ability to speak and to feed himself, resulting in an improved quality of life. The incidence of HCC recurrence after liver transplantation is observed in approximately 8% to 11% of selected cases, with frequent relapses observed in the implanted liver, bones, adrenal glands, and lungs. Mediastinal and intracranial metastases from HCC post-liver transplantation are very rare.     

Applicability of the Milan Criteria for Determining Liver Transplantation as a First-Line Treatment for Hepatocellular Carcinoma

Background To determine whether or not the Milan criteria (MC) should be used to determine the applicability of liver transplantation (LT) as a first-line treatment for patients with cirrhosis with hepatocellular carcinoma (HCC) who are able to endure hepatectomy.Methods Retrospective analysis of 82 patients with cirrhosis with HCC who were treated by hepatectomy without LT at our institution between 1990 and 2003.Results Of these 82 patients, 48 met the MC. Proportional hazard regression analyses to determine the independent prognostic factors for postoperative cumulative patient and disease-free survival showed that meeting the MC is the strongest prognostic factor for both patient and disease-free survival. The cumulative patient and disease-free survival rates were 76.7% and 28.9%, respectively, at 5 years in patients who met the MC. The cumulative disease-free survival was markedly inferior to those in previously reported series of LT for HCC who met the MC, but the cumulative patient survival was comparable to those in the previously reported series. A comparison of cumulative postoperative survival between patients who met the MC and fulfilled all five factors listed below and patients who met the MC but did not fulfill any of the five factors demonstrated that the latter patients showed statistically significantly worse postoperative patient survival than the former. The five factors included: Model for End-Stage Liver Disease score <10, indocyanine green retention rate at 15 minutes <20%, absence of microscopic fibrous capsular invasion and microscopic intrahepatic metastases, and earlier grade (T1 or T2) of American Joint Committee on Cancer tumor classification.Conclusions The MC should not be used to determine the applicability of LT as a first-line treatment for patients with HCC considered able to endure hepatectomy. However, modifying MC with some clinicopathological factors could satisfy the appropriate criteria for applying LT as a first-line treatment for these patients.     

Bridging Locoregional Therapy for Hepatocellular Carcinoma Prior to Liver Transplantation

Introduction  The impact of locoregional therapy prior to liver transplantation for hepatocellular carcinoma utilizing either transcatheter arterial chemoembolization (TACE), yttrium-90 (⁹⁰Y), radiofrequency ablation (RFA), or resection prior to orthotopic liver transplantation (OLT) is largely unknown. We sought to examine locoregional therapies and their effect on survival compared with transplantation alone. Methods  A retrospective review of a prospectively collected database. Results  123 patients were included. Patients were analyzed in two groups. Group I consisted of 50 patients that received therapy (20 TACE; 16 ⁹⁰Y; 13 RFA, 3 resections). Group II consisted of 73 patients transplanted without therapy. Median list time was 28 days (range 2–260 days ) in group I, and 24 days (range 1–380 days) in group II. Median time from therapy to OLT was 3.8 months (range 9 days to 68 months). Twelve patients (24%) were successfully downstaged (8 TACE, 2 ⁹⁰Y, 2 RFA/resection). Overall 1-, 3-, and 5-year survival were 81%, 74%, and 74%, respectively. Survival was not statistically significantly different between the two groups (P = 0.53). The 12 patients downstaged did not have a significant difference in survival as compared with the patients who received therapy but did not respond or the patients who were transplanted without therapy (P = 0.76). Conclusion  Our report addresses locoregional therapy for hepatocellular carcinoma as a bridge to transplant. There was no statistical difference in overall survival between patients treated and those not treated prior to transplant. We provide further evidence that locoregional therapy is a safe tool for patients on the transplant list, does not impact survival, and can downstage selected patients to allow life-saving liver transplantation.     

肝癌肝移植术后复发的危险因素分析

目的探讨原发性肝癌(HCC)肝移植术后肿瘤复发或转移的危险因素。方法回顾性我院2003年4月至2007年11月期间76例HCC患者行肝移植的临床资料,根据随访期间是否有复发分为复发组(n=23)和未复发组(n=53),总结肿瘤复发的特点。结果 76例患者中23例(30.3%)术后复发。单因素分析显示患者性别(P=0.449)、年龄(P=0.091)、术前是否治疗(P=0.958)、肿瘤数目(P=0.212)和是否伴有HBV/HCV感染(P=0.220)与肿瘤的复发无关,而肿瘤包膜完整性(P=0.009)、肿瘤分期(P=0.002)、肿瘤直径(P<0.001)、血管侵犯(P<0.001)以及术前AFP水平(P=0.044)与肿瘤的复发有关,其中肿瘤直径<5.0 cm(P=0.001)和术后2个月AFP水平恢复正常者(P<0.001)1年复发率更低。多因素分析显示肿瘤直径(P=0.001,OR=6.456,95%CI为2.356～17.680)、血管侵犯(P=0.030,OR=10.653,95%CI为1.248～90.910)以及术前AFP水平(P=0.017,OR=2.601,95%CI为2.196～5.658)是肝移植术后肿瘤复发的独立危险因素。结论对于肿瘤直径>5.0 cm、伴有血管侵犯以及术前AFP水平≥400μg/L尤其术后2个月AFP水平仍高于正常者术后需加强监测,必要时尽早给予抗肿瘤治疗。