丙型肝炎病毒包膜E2蛋白DNA疫苗诱导小鼠产生中和抗体研究

目的 探讨HCV包膜E2蛋白DNA疫苗诱导小鼠产生中和抗体的可行性.方法 分别构建HCV E2蛋白全长表达质粒pcDNA3.1-1a746、截除疏水性羧基末端的表达质粒pcDNA3.1-1a661以及同时截除疏水性羧基末端和高变区1(HVR1)的表达质粒pcDNA3.1-1a661Δ,转染293T细胞,以Western blot和ELISA法检测细胞内和培养上清中的HCV E2蛋白,将3种表达质粒以及空载体分别肌注免疫 BALB/c小鼠,检测小鼠血清的E2及HVR1抗体,以HCV假病毒模型分析小鼠血清的中和活性.结果 3种E2表达质粒均能有效表达HCV E2蛋白,其中pcDNA3.1-1a746质粒的表达产物不能分泌,而pcDNA3.1-1a661和pcDNA3.1-1a661Δ均能分泌表达E2蛋白.分泌表达E2蛋白可显著增强小鼠的抗体应答,pcDNA3.1-1a661免疫血清对HCV假病毒颗粒(HCVpp)的中和活性明显高于pcDNA3.1-1a661Δ免疫血清.pcDNA3.1-1a661免疫血清中的HVR1抗体量仅占总E2抗体的一小部分,却是中和抗体的重要成分.结论 表达E2蛋白的DNA疫苗能有效诱导HCV中和抗体的产生,HVR1不仅是重要的中和抗体表位,而且能增强E2蛋白其他抗原表位的抗体应答.     

h5n1禽流感假病毒体外中和试验技术平台的建立与应用

目的 建立h5n1禽流感假病毒的体外中和试验技术平台,并系统评价2份h5n1人禽流感患者恢复期血清的中和效价.方法 通过磷酸钙沉淀法将转移载体phr-luc、包装载体pcmv△8.2和包膜载体cmv/r-sz(或cmv/r-th)共转染人胚肾细胞(293t),72 h后收集假病毒上清液,免疫印迹(western blot)鉴定假病毒颗粒ha、p24蛋白的表达,并取200μl假病毒上清液测定感染活性.收集制备假病毒过程中整合ha蛋白的293t细胞,通过流式细胞术测定2份恢复期血清中的ha抗体.利用2份恢复期血清分别与深圳株和泰国株假病毒进行中和试验,通过计算ic5o判断不同血清对2株假病毒中和效价的差异.结果 构建的2株假病毒颗粒不仅含有hiv基质蛋白p24,还可以整合ha蛋白,并且能够由前体蛋白ha0裂解为具有生物活性的ha1和ha2.200μl假病毒上清液感染靶细胞后rla值约为2×104,具有较高的感染效价.facs检测结果显示2份血清中均含有ha抗体,而且对不同分枝的假病毒具有交叉反应性.中和试验表明2份血清对深圳株和泰国株假病毒均具有中和能力,而且对不同分枝的假病毒具有交叉中和活性,但对深圳株的中和效价均高于泰国株.结论 本研究成功构建具有感染活性的深圳株和泰国株h5n1禽流感假病毒,并可以快速、安全、高效的评价血清的中和效价,具有广阔的应用前景. abstract： objective to establish neutralization assay based on h5n1 avian influenza pseudotyped virus in vitro and to evaluate neutralizing titer of convalescent serum from 2 patients with h5n1 avian influenza.methods phr-luc,pcmv△8.2 and cmv/r-sh or cmv/r-th were cotransfected into 293t cell by co-precipitation with calcium phosphate.pseudotyped virus supernatant was harvested 72 h posttranofection and identified the expression of ha and p24 by western blot,and then we analyzed infective activity of 200 μl supernatant of pseudotyped virus.293t cell integrated ha was prepared and anti-ha antibodies in convalescent serum were measured with facs assay.neutralizing titers of convalescent serums against shenzhen and thailand pseudotyped virus were determined based on calculating ic50 with neutralizing assay.results pseudotyped virus involved p24 and ha,and precursor protein ha0 could cleavage into ha1 and ha2 with biological activity.pseudotyped virus possessed better infective activity,and rla value was about 2 × 104 with 200 μl supernatant.both convalescent serums contained anti-ha antibodies and had cross-reactivity against different virus clades with facs assay.both convalescent serums had neutralizingactivity and could cross-neutralize different virus clades.however,both serums'neutralizing titers against shenzhen virus were higher than thailand.conclusion we successfully constructed infectious pseudotyped virus which integrated ha of shenzhen or thailand virus,and it could be used for evaluation of serum neutralizing activity fast,efficiently and safely with broadly application prospect.     

蝙蝠来源的重症急性呼吸综合征样冠状病毒WIV1刺突蛋白利用浣熊狗血管紧张素转换酶Ⅱ受体侵入细胞能力的研究

目的研究蝙蝠来源的重症急性呼吸综合征（SARS）样冠状病毒WIV1刺突蛋白利用浣熊狗血管紧张素转换酶Ⅱ（ACE2）受体侵入细胞的能力,并探究SARS样冠状病毒WIV1潜在的跨宿主传播风险。 方法构建来自浣熊狗、果子狸、中华菊头蝠和人等不同动物来源的ACE2表达质粒,转染至293T细胞并利用免疫印迹方法检测其在293T细胞中的表达水平;建立SARS冠状病毒（Tor2株系）及蝙蝠SARS样冠状病毒（WIV1株系）假病毒感染系统,并进行假病毒感染实验;利用假病毒感染系统及荧光素酶报告基因检测WIV1刺突蛋白对浣熊狗等不同动物来源ACE2受体利用能力;构建跨膜丝氨酸蛋白酶2（TMPRSS2）质粒,并转染至T Rex 293细胞,借助假病毒感染系统检测TMPRSS2对WIV1侵入能力的影响。 结果本研究所获赠及构建的不同动物来源的ACE2质粒可经瞬时转染至细胞进行表达;与pcDNA3.1载体相比,浣熊狗、中华菊头蝠、果子狸和人的ACE2均可使蝙蝠SARS样冠状病毒WIV1侵入细胞的能力增加上万倍,上述ACE2组与载体组的荧光素酶活性差异均具有统计学意义（t = 27.744、P < 0.001,t = 18.740、P < 0.001,t = 32.297、P < 0.001,t = 15.902、P < 0.001）;与TMPRSS2阴性组相比,TMPRSS2在靶细胞的表达可使WIV1假病毒的感染能力增加10倍以上,两组荧光素酶活性差异具有统计学意义（t = 29.460、P < 0.001）。 结论蝙蝠SARS样冠状病毒WIV1的刺突蛋白除了利用人类、果子狸及中华菊头蝠的ACE2受体感染细胞,还可利用浣熊狗的ACE2受体侵入细胞,且TMPRSS2可显著促进其侵入能力,提示WIV1可能存在多种跨宿主传播的风险。     

HIV-1假病毒包装方法的探讨

目的：通过不同方案包装HIV-1假病毒,确定获得高重复性、高稳定性和高滴度假病毒的方法。方法从转染效率影响因素出发,通过不同的转染方案,利用pNL4-3 LucR-E-质粒与VSV-G质粒共转染293细胞包装病毒,分别于48 h和72 h收取假病毒并重新感染293细胞,48 h后测荧光素酶报告基因。结果成功包装出HIV-1假病毒,不同包装方案的病毒滴度不同,测得报告基因最高接近107 RLUs。结论应用PEI转染试剂、大提质粒pNL4-3︰VSV-G为4︰1时,转染效率最高,包装的假病毒滴度最高。     

人骨形态蛋白-2可调控系统在大鼠骨髓间充质干细胞的表达

目的构建携带四环素真核诱导表达系统(Tet-on)调控的人骨形态发生蛋白2(h BMP-2)慢病毒载体转染大鼠骨髓间充质干细胞(BMSCs),为骨缺损修复提供可控的成骨活性细胞。方法设计目的基因h BMP-2引物,将扩增纯化的目的基因h BMP-2定向克隆至携带Tet-on的慢病毒GV347载体上并测序鉴定产物。h BMP-2-GV347质粒、空载的GV347质粒分别与辅助质粒共感染293T细胞以收获慢病毒浓缩液。用h BMP-2-GV347慢病毒转染大鼠BMSCs,得到h BMP-2阳性表达细胞,探索转染最佳感染复数(MOI)。用CCK8法比较BMSCs转染前后的增殖活性;强力霉素(DOX)诱导打开Tet-on,real-time PCR、Western Blot和酶联免疫吸附试验(ELISA)检测转染后各组BMSCs中BMP-2蛋白及RNA的表达情况。结果 PCR后得到目的基因h BMP-2,定向克隆至携带Tet-on系统调控的GV347质粒上,经酶切后电泳、测序结果证实成功构建携带可调控系统的h BMP-2-GV347质粒。h BMP-2-GV347质粒与空载GV347质粒分别感染293T细胞后获得h BMP-2-GV347慢病毒浓缩液。在DOX诱导下,h BMP-2-GV347慢病毒载体在293T细胞内表达BMP-2蛋白。h BMP-2-GV347慢病毒载体转染最佳MOI值为9,且转染后的BMSC细胞增殖能力较普通BMSC强,并在DOX浓度为10μg/ml诱导下稳定表达BMP-2蛋白和RNA。结论本研究成功构建携带Tet-on系统调控h BMP-2慢病毒表达载体,转染BMSCs后在DOX调控下可持续高效表达BMP-2蛋白。     

丙型肝炎患者血清中针对高变区1-N端模拟7肽的抗体反应研究

目的检测丙型肝炎病毒(HCV)高变区1(HVRl)模拟7肽与HCV患者血清的反应特点。方法采用酶联免疫吸附试验。结果(1)7肽与450份HCV抗体阳性血清中243份(54％)反应阳性．91份正常血清中5份(5．5％)反应阳性，90份乙型肝炎患者血清中6份(6．7％)反应阳性。(2)7肽与113份长期血清进行反应，0～3月组48份HCV抗体阳性血清中35份(73％)反应阳性，4～6月组10份血清中4份(40％)反应阳性，6月以上组55血清中41份(75％)反应阳性。结论(1)我们设计的HCV高变区1模拟7肽与HCV患者血清反应具有特异性，且反应范围广。(2)此模拟7肽与HCV患者血清反应性随时间变化差异不明显。     

活化T细胞核因子-2可介导高迁移率族蛋白B1促进白细胞介素-2转录表达

目的 观察活化T细胞核因子-2(NFAT2)在高迁移率族蛋白B1(HMGB1)促进白细胞介素-2(IL-2)转录表达中的介导作用.方法 将构建好的HMGB1和NFAT2质粒单独或共同转染人胚胎肾细胞株293T,同时转染IL-2报告基因,并在此基础上应用小分子干扰RNA(siRNA)质粒对内源性及外源性NFAT2表达进行特异性抑制,观察对IL-2报告基因活性的影响.结果 在293T细胞中,内源性转染实验促使IL-2报告基因活性增加到17.81±1.49,但随着siRNA质粒转染剂量从0 μg/孔逐渐增加到4.8μg/孔,IL-2报告基因活性在内源性抑制实验中降低最小到1.42±0.17,在外源性抑制实验中活性最高为54.16±5.49,受抑制后降低最小到2.97±0.34.结论 NFAT2介导HMGB1促进IL-2报告基因转录表达.     

丙型肝炎患者血清中针对高变区1-N端模拟7肽的抗体反应研究

目的　检测丙型肝炎病毒 (HCV)高变区 1(HVR1)模拟 7肽与HCV患者血清的反应特点。方法　采用酶联免疫吸附试验。结果　 (1) 7肽与 4 5 0份HCV抗体阳性血清中 2 4 3份 (5 4 % )反应阳性 ,91份正常血清中 5份 (5 .5 % )反应阳性 ,90份乙型肝炎患者血清中 6份 (6 .7% )反应阳性。(2 ) 7肽与 113份长期血清进行反应 ,0～ 3月组 4 8份HCV抗体阳性血清中 35份 (73% )反应阳性 ,4～ 6月组 10份血清中 4份 (4 0 % )反应阳性 ,6月以上组 5 5血清中 4 1份 (75 % )反应阳性。结论　 (1)我们设计的HCV高变区 1模拟 7肽与HCV患者血清反应具有特异性 ,且反应范围广。 (2 )此模拟 7肽与HCV患者血清反应性随时间变化差异不明显。     

腺病毒介导人VEGF_(165)和ANG-1双基因转染大鼠BMSCs及对其增殖的影响

[目的]探讨携带人血管内皮细胞生长因子165(VEGF165)和血管生成素-1(ANG-1)双基因的腺病毒表达载体pAd-VIA转染大鼠骨髓基质干细胞(BMSCs)的体外表达及对BMSCs增殖的影响。[方法]将本室构建的编码人VEGF165和ANG-1双基因的腺病毒质粒pAd-VIA在QBI-293A细胞内进行包装和扩增,体外转染大鼠BMSCs,通过绿色荧光蛋白(GFP)表达、Western-blotting、酶联免疫吸附分析(ELISA)方法检测外源基因的表达,利用MTT法检测MOI(multiplicity of infection)=50、100、200、400 pfu/cell不同浓度腺病毒转染后对BMSCs增殖活性的影响。[结果]重组腺病毒质粒pAd-VIA在QBI-293A细胞内成功包装和扩增,体外转染BMSCs后,有大量的GFP表达,Western-blotting检测显示,转染组与VEGF165和ANG-1抗体结合,在45 KD和14.4 KD左右出现印迹条带;ELISA结果显示:转染组第1、2、3 d,上清中的VEGF165浓度和ANG-1浓度持续增加,未转染组均未检测到外源基因的表达,差异...     

构建携带hTERT基因的慢病毒重组载体及其包装和表达

目的构建含hTERT基因的慢病毒重组载体,进行病毒包装并检测hTERT在293T细胞中的表达。方法PCR扩增hTERT．插入慢病毒载体,并与包装质粒转染293T细胞,进行病毒包装,并测定其滴度。Western—Blot检测慢病毒液感染293T细胞后hTERT的表达。结果成功构建了含hTERT的慢病毒重组载体,滴度为2．79×10^7Tu／mL,Western-B10t检测显示慢病毒液感染的293T细胞中hTERT基因高表达。结论含hTERT的慢病毒重组载体成功构建并包装后能够顺利感染293T细胞．并阳性表达目的基因。     

Epidemiology and clinical significance of chronic hepatitis-related viruses infection in hemodialysis patients from Taiwan

BACKGROUND/AIMS: A novel DNA virus which was designated TT virus (TTV) in 1997 was considered a possible hepatitis-related virus, like hepatitis C (HCV), hepatitis B (HBV) and GB virus C/hepatitis G viruses (GBV-C/HGV). In the present study, the molecular epidemiology and clinical significance of TTV, GBV-C/HGV and HCV infection in hemodialysis patients from Taiwan are investigated. METHODS: Sera of 85 patients on maintenance hemodialysis were tested for alanine aminotransferase (ALT), hepatitis B surface antigen (HBsAg), second-generation HCV antibody (anti-HCV), anti-envelope protein 2 antibody (anti-E2) and RNA of GBV-C/HGV, HCV RNA and TTV DNA. Sera of patients with positive TTV DNA, GBV-C/HGV RNA or HCV RNA were tested for viruses 2 years later. RESULTS: Seven (8.2%) 29 (34.1%), 21 (24.7%), 12 (14.1%) and 9 (10.6%) hemodialysis patients were positive for HBsAg, Anti-HCV, HCV RNA, GBV-C/HGV RNA and anti-E2, respectively. TTV DNA was positive in 46 (54.1%) patients. Neither clinical nor virological factors were associated with TTV viremia. The ALT level was significantly elevated in HCV RNA-positive individuals than -negative ones (34.5 vs. 12.5%, p < 0.05). TTV DNA, GBV-C/HGV RNA and HCV RNA remained detectable in sera of 31 (86.1%), 6 (50%) and 21 (100%) patients collected 2 years after first diagnosis of viremia. CONCLUSION: Among Taiwanese hemodialysis patients, TTV infection is highly prevalent. No clinical or virological factor was observed to be significantly associated with TTV infection. The ALT abnormality was mainly attributable to HCV but not TTV infection in Taiwanese hemodialysis patients.     

Hepatitis G virus exposure in dialysis patients

Background Hepatitis G virus (HGV) is a blood-borne virus. The predominant route of its transmission is parenteral. The aim of this study was to assess the frequency of HGV exposure in haemodialysis (HD) and continuous ambulatory peritoneal dialysis (CAPD) patients in Iran. Methods This study was performed in a major dialysis centre in Tehran, Iran. The study cohort consisted of 77 patients on HD and 13 patients on CAPD. The presence of anti-HGV envelope protein E2 (anti-E2) in the blood serum, as determined by means of an ELISA assay, indicated HGV exposure. All patients were also screened for hepatitis B surface antigen (HBsAg), hepatitis B surface antibody (anti-HBs) and hepatitis C antibody (anti-HCV). In patients who tested positive for anti-E2, HGV RNA was detected by RT-PCR using primers derived from the NS5A region of the viral genome. Results In total, 3.89% of the HD patients and none of the CAPD patients tested positive for anti-E2. None of the patients tested positive for HGV RNA. The mean age of the anti-E2-positive patients was 53.3 ± 26.5 years, with 66.66% having previously received blood transfusion. The mean duration of dialysis of the anti-E2-positive patients was 68 ± 64 months. Co-infection with HCV or HBV was not observed in the anti-E2 positive patients. Conclusion The rate of exposure to HGV was low among the dialysis patients in our study. The appearance of anti-E2 was accompanied by clearance of serum HGV-RNA. No relationship was noted between HGV exposure and age, sex, history of blood transfusion, time on dialysis and HCV or HBV markers.     

HCV infection in hemodialyzed patients: incidence and correlation with dialytic age.

Hemodialyzed patients mean an high-risk population for hepatitis C infection. Our work was performed to evaluate the incidence of hepatitis C virus (HCV) infection in 51 hemodialyzed patients; the presence of anti-HCV antibodies was studied using the EIA and RIBA test of 1st and 2nd generation. 18 patients (35.29%) showed anti-HCV antibodies with the 1st test; 27 patients (52.94%) showed the presence of anti-HCV antibodies using 2nd generation test. The incidence of test positivity is not related to blood transfusions while it is strictly related with dialytic age. All HCV seropositive patients show antibodies against the c22-3 protein of "virus core". The presence in serum of anti-c22-3 antibodies means that in these patients, there is viral replication.     

TT virus infection in haemodialysis patients.

BACKGROUND: The recent discovery of a new parenterally transmitted DNA virus called TT virus (TTV) led us to investigate its prevalence in haemodialysis patients, a high-risk group for blood-borne infection, and to evaluate its role in liver disease. Moreover, we compared the TTV prevalence with the prevalence of other hepatitis virus coinfections. METHODS: Serum samples of 78 patients on maintenance haemodialysis were tested for TTV-DNA, hepatitis G virus (HGV)-RNA, anti-E2, anti-hepatitis C virus (HCV) and HCV-RNA. TTV-DNA was detected by semi-nested PCR using the primers from open reading frame 1 (ORF). HGV-RNA was detected by PCR using specific primers for the NS3 and the 5'-UTR genome regions while anti-E2 were checked by an enzyme immunological test. Anti-HCV was tested by the second generation Chiron RIBA HCV test system. HCV-RNA was evaluated by nested PCR with primers directed to the highly conserved 5' non-coding region of the HCV genome. RESULTS: TTV prevalence in our patients was 19% (15/78) while the prevalence of HCV and HGV infection proved to be 20 and 15.4%, respectively. Among TTV positive patients HGV co-infection was present in five cases (33%), HCV in six cases (39.9%), while HBV co-infection was not present in any of the patients. Only three patients proved positive for all three viruses. ALT levels were normal in most cases (13/15; 86%). In particular, patients with TTV infection alone showed normal ALT levels and HCV coinfection was found in the two patients with moderate ALT increases. CONCLUSIONS: TTV prevalence in haemodialysed patients is significant though the real clinical impact is still unclear. However, we must keep in mind that the epidemiological relevance of TTV infection is probably underestimated due to the impossibility in detecting the corresponding antibody.     

Epidemiology of hepatitis C in a population of hemodialysis patients.

A search for antibodies against hepatitis C virus (HCV) was performed in 185 patients on chronic hemodialysis by means of 1st and 2nd generation ELISA tests. Immunoblot assays were performed on positive sera. This study shows a 38% prevalence of HCV-positive patients in our dialysis population according to the 2nd generation ELISA test which shows a higher specificity and sensitivity when compared to the 1st generation one (38 vs. 20%). A correlation was found between the prevalence of HCV-positive patients and how long they had been on dialysis and how many blood transfusions they had received.     

Monoclonal antibody HCV-AbXTL68 in patients undergoing liver transplantation for HCV: results of a phase 2 randomized study.

A randomized, double-blind, dose-escalation study evaluated the safety and efficacy of hepatitis C virus (HCV)-Ab(XTL)68, a neutralizing, high-affinity, fully human, anti-E2 monoclonal antibody, in 24 HCV-positive patients undergoing liver transplantation. HCV-Ab(XTL)68 or placebo was administered at doses from 20-240 mg as 2-4 infusions during the first 24 hours after transplantation, followed by daily infusions for 6 days, weekly infusions for 3 weeks, and either 2 or 4 weekly infusions for 8 weeks. Serum concentrations of total anti-E2 obtained during daily infusions of 120-240 mg HCV-Ab(XTL)68 were 50-200 microg/mL above concentrations in the placebo group. Median serum concentration of HCV RNA dropped below baseline in all groups immediately after transplantation. On day 2, median change from baseline in HCV RNA was -1.8 and -2.4 log in the 120-mg and 240-mg groups, respectively, compared with -1.5 log with placebo. The difference was lost after day 7 when the dosing frequency was reduced. The coincidence of increases in anti-E2 with decreases in HCV RNA concentration indicate that the dose-related changes in HCV RNA concentration were a result of HCV-Ab(XTL)68 administration in the 120- and 240-mg groups. The overall incidence of nonfatal serious adverse events was higher with placebo (60%) vs. all active treatments combined (42%). In conclusion, HCV-Ab(XTL)68 may decrease serum concentrations of HCV RNA in patients after liver transplantation. Studies evaluating more frequent daily dosing at doses >120 mg are necessary to investigate sustained viral suppression in this population.     

维持性血液透析患者感染乙型和丙型肝炎的分析

目的为了评价血液透析（血透）患者乙型和丙型肝炎（HBV、HCV）感染状态及对临床情况和肝功能的影响。方法对62例血透患者应用ELISA法和RT-PCR法检测抗-HCV和HCVRNA，采用斑点杂交法和固相放免法检测HBV标志，并检测肝功能和血浆蛋白电泳。结果62例患者中，抗-HCVIgM阳性27例（43．6％），抗-HCVIgG阳性29例（46．8％），HCVRNA阳性34例（54．8％），三项任一项阳性37例（59．7％），5例（8．1％）HBsAg阳性，其中HBeAg和HBVDNA阳性3例。结论向透患者中HCV感染严重，临床情况及预后差，检测血浆蛋白和电泳较肝功能酶学能更好地作为肝炎诊断和反映病情的指标。     

High prevalence of hepatitis G virus (HGV) infection in renal transplantation

Introduction: The newly discovered (1995) hepatitis G virus (HGV) is an RNA virus from the Flaviviridae family with 85% genomic homology to GB virus C (GBV-C). We studied the prevalence of HGV infection among a cohort of 398 renal transplant recipients (PT), all of whom had previously received blood transfusions, been grafted between August 1984 and December 1991, and been treated by cyclosporin A (CsA) as the main immunosuppressant. Subjects and methods. According to hepatitis C virus (HCV) antibody status, and after exclusion of 28 HBs antigen-positive recipients, this cohort had previously been divided into an HCV +ve subgroup (106 RTR; 62M vs 44F; 29 French vs 77 non-French) and an HCV -ve subgroup (264 RTR; 181M vs 83F, 196 French vs 68 non-French). We randomly selected 27 RTR in the HCV+/HBV- subgroup (14M vs 13F, 10 French vs 17 Italians) and 27 RTR in the HCV-/HBV- subgroup (19M vs 8F, 18 French vs 9 Italians) for HGV screening. The detection of HGV RNA sequences in serum (viraemia) was done by double nested RT-PCR using specific primers chosen in the 5' non-coding genomic region. The serum detection of specific antibodies (anti-E2) was done by ELISA test. All sera (at time of liver biopsy or at last follow-up) were tested in duplicate. Results: The prevalence of HGV viraemia was 26% (14/54) in the whole group and in both HCV +ve and -ve subgroups (7/27). The prevalence of HGV infection (viraemia+ and/or antiE2 antibodies+) was 44% (24/54) in the whole group and in both HC +ve and -ve subgroups (12/27). In addition, the prevalence was similar in males vs females and in French vs foreigners recipients (mostly Italians). In the HCV +ve subgroup, the seven HGV viraemia-positive patients who previously had liver biopsies disclosed chronic active hepatitis in four (mean Knodell score 5.75) and normal livers in three, with only one case of elevated ALT (CAH 5). In the HCV- subgroup, non of the seven HGV+ viraemic patients had elevated ALT and liver biopsy was not performed. Conclusion: HGV infection prevalence is high (44%) in RTR, but clearly independent of HCV status and/or the geographical origin of the recipients. This data indicates a different epidemiology as compared to our HCVs previous experience.