Detection of TT virus in lymph node biopsies of B-cell lymphoma and Hodgkin's disease,and its association with EBV infection

Human TT virus (TTV) recently isolated from the serum of a patient with post-transfusion hepatitis does seem to have only hepatopathic effect. The virus can also infect the serum, peripheral blood mononuclear cells (PBMC) and bone marrow cells (BMC ). Additional evidence has indicated that TTV is also present in the serum of people with hematopoietic malignancies. A significant increase in the incidence of lymphoma has recently been observed worldwide. We have investigated the presence of TTV DNA in lymph node biopsies of Italian patients affected with the most common lymphoma types in Western Countries: follicular lymphoma (FL), diffuse large B-cell lymphoma (DLBCL) and nodular sclerosis Hodgkin's disease (NS-HD). The possible role of a co-infection with Epstein-Barr virus (EBV) has also been investigated. DNA was extracted from 73 paraffin-embedded and 38 snap-frozen tissue specimens. From these, only 67 samples (29 paraffin-embedded and 38 snap-frozen tissues) from a total of 56 patients, were suitable for PCR analysis. TTV and EBV were detected by PCR using primers from two different conserved region in TTV and EBV genomes respectively. TTV DNA was detected in 30.0-50.0% of FL, 30.8% of DLBCL and 30.0-50.0% of NS-HD cases, depending on the primers used. All cases of non-specific reactive lymphoid hyperplasia (RLH), used as a putative control, were negative. The two major TTV genotypes circulating in Italy (G1 and G2) were detected in the analysed lymphoid neoplasms. EBV DNA was detected in 40.0% of FL, in 72.7%of DLBCL, in 80.0% of SN-HD and in 40.0% of RLH cases. EBV co-infection was found in 90% of TTV positive cases. The in situ hybridization assay was performed in TTV positive frozen samples. The significant prevalence of TTV DNA in lymphocytes circulating in the lymph nodes of both B-cell lymphomas and HD reported herewith suggests an implication of TTV infection in the development of these lymphoproliferative disorders.     

新型病毒TTV传播规律及相关性分析

目的：探讨输血后新型病毒（TTV）传播规律及相关的研究，为预防和阻断TTV的传播提供依据。方法：采用套式PCR检测孕产妇静脉血、脐血和婴儿足跟血以及乳汁的TTV DNA，并对其阳性妇女所生的婴儿随访1年，以进行相关性调查分析。结果：102例孕产妇静脉血TTV DNA阳性检出率为53．9％（55／102）；106例脐血TTV DNA阳性检出率为17．0％（18106）；52例婴儿（均为TTV DNA阳性妇女所生）1周内足跟血阳性检出率为15．4％（8／52）；55例TTV DNA阳性妇女的乳汁，有40例TTV DNA阳性检出率为72．7％（40／55）；对55例TTV DNA阳性妇女所生的57例婴儿，出生后随访1年，除2例外有55名婴儿呈阳性结果。结论：TTV母婴传播率很高，可达96．5，且宫内感染率达15．4％。故认为应尽早制订相应的预防和阻断措施，有效控制宫内感染和监测阳性妇女乳汁中TTV DNA的含量，以降低婴儿感染率。     

The response of hepatitis C virus and TT virus to high dose and long duration interferon-alpha therapy in na?ve chronic hepatitis C patients.

To investigate responses of hepatitis C virus (HCV) and TT virus (TTV) to high dose and long duration interferon-alpha (IFN-alpha) therapy (540 million units in 36 weeks) and factors associated with the viral clearance, sera of 165 Taiwanese na?ve chronic hepatitis C patients were tested for alanine aminotransferase, HCV RNA levels, HCV genotypes and TTV DNA. With 41.8% of TTV DNA prevalence, TTV viremia was significantly associated with history of blood transfusion (P<0.01). After IFN therapy, HCV complete response was achieved in 60 (36.4%) patients and significantly associated with lower pretreatment levels of HCV RNA (P<0.01) and HCV genotype non-1b (P<0.05). Fifty-three patients with concurrent TTV infection were evaluated for TTV response. TTV sustained clearance was achieved in 24 (48%) patients and significantly associated with loss of TTV DNA at the end point of treatment. In conclusion, concurrent TTV infection is highly prevalent, related to blood transfusion and independent of HCV infection. After high dose and long duration IFN-alpha therapy, HCV and TTV clearance are achieved among more than one-third and around one-half patients. HCV RNA levels and HCV genotypes are predictors for HCV response and no clinical factors are observed to be associated with TTV clearance.     

病毒性肝炎TTV DNA阳性者的临床分型和肝组织学观察

目的：了解输血传播病毒（TTV）感染的临床特点和肝组织形态学特征,探讨TTV与肝损伤的关联性,方法：对93例血清TTV DNA阳性病毒性肝炎患者的临床资料及其中10例单一TTV笔DNA阳性慢性非甲－瘐型肝炎（HNA－G）的肝组织光镜和电镜检查结果进行分析,结果：各型病毒性肝炎中的均有一定比例的TTVDNA阳性,以慢性肝炎,慢性重型肝炎,肝硬化多见,可呈T TV＋HAV,TTV＋HBV,TTV＋HEV,TTV＋HAV,TTV＋HBV＋HCV,TTV＋HBV＋HEV,TTV＋HBV＋HGV重叠感染,与HBV重叠感染可有慢性重型肝炎,肝硬化之临床类型,半日一TTV DNA阳性慢性肝炎患者存在肝细胞变性,坏死和肝组织淋巴细胞浸润,纤维明显增生,结论：TTV DNA阳性慢性肝炎患者在肝细胞变性,坏死和肝组织淋巴细胞浸润,纤维明显增生,结论：TTV在肝脏慢性损伤中可能起一定作用,可引起慢性肝炎并与肝硬化和慢性重型肝炎的发生相关联。     

肝炎患者血清TT病毒核酸检测及分析

目的：证实在不明原因的肝炎患者血清中一种新型病毒（ＴＴＶ）在存在,分析其感染特征。方法：用地高辛素标记基因探针,斑点杂交法检测未知和已知病原的肝炎患者７５例,正常对照１７例。结果：９２例血清ＴＴＶ ＤＮＡ总检出率为２３．９１％（２２/９２）。其中非甲～非戊型肝炎病人组检出率为３５％（７/２０）,甲～戊肝组２１．８１％（２/５）,肝硬化中１５．７９％（３/１９）。结论：ＴＴ病毒可能是导致病毒性肝炎的一种     

不同人群输血传播病毒感染的检测和传播途径分析

目的 了解深圳地区不同人群TTV感染状况,探讨TTV传播途径。方法建立套式聚合酶链反应方法．对临床病毒性肝炎病人、血液透析病人、静脉吸毒者、性病患者和献血者人群进行TTV DNA的PCR检测,并对部分阳性株进行序列测定。结果非甲-戊型肝炎病人、血透患者和反复输血病人TFVDNA阳性率分别为41.7％、40.5％和35．7％．明显高于甲-丙型肝炎病人（17．7％,P〈0．01）;静脉吸毒者中TTV DNA阳性率高达39．3％,明显高于献血者人群（20．4％。P〈0．05）：ALT异常献血员中TTV DNA检出率为明显高于无偿献血员人群（P〈0.05）;性病患者生殖道分泌物中TTV DNA检测出率为15．8％。2株TTV分离株与TTV标准株（AB008394）同源性在92．5％以上。结论TTV感染与肝炎有关。可能是非甲-度型肝炎的病原之一;TTV除了经血源途径传播外。性传播可能成为重要传播途径之一。     

温州市吸毒人群输血传播病毒感染的检测分析

目的 :了解我市静脉吸毒者输血传播病毒 (TTV)感染情况。方法 :采集静脉吸毒者血液标本 ,分离血清 ,用半巢式聚合酶链式反应 (PCR)检测TTV。结果 :共检测 30 6例 ,检出TTV阳性 2 3例 ,阳性率为 7 5 2 %。吸毒时间≤ 1a者TTV阳性率较低 ,为 4 31% ,而 >1a者阳性率较高 ,为 14 4 3% ,两者比较差异有显著性 (χ2 =9 77,P <0 0 1)。不同性别和不同血型者的TTV感染率比较差异无显著性 (P >0 0 5 )。结论 :温州地区静脉吸毒者是TTV感染的高危人群 ,应采取控制措施预防和减少TTV在吸毒人群中的传播。     

107例静注海洛因成瘾者TT病毒感染状况

目的：调查贵州地区静注海洛因成瘾者TT病毒感染状况。方法：以公布的TTV第1读码区序列设计两对寡核苷酸引物，用套式聚合酶链反应（nPCR)检测107例静注海洛因成瘾者血清中TTVDNA，以62列正常人为对照。同时用酶联免疫法测丙肝病毒抗体（抗HCV），逆转录套式聚合酶链反应（RT－nPCR)测庚肝病毒核糖核酸（HGVRNA）。结果：正常人TTVDNA阳性率6．45％，海洛因成瘾者25．23％。两组相比，差异有显著意义（P＜0．005）。成瘾者TTVDNA阳性率在男、女组间无差异，与注毒时间长短无相关（P＞0．05）。27例TTVDNA阳性成瘾者中，11例为单纯TTV感染，16 重叠HCV或HGV感染，其中TTV、HCV、HGV三重感染8例，TTV重叠HCV5例，重叠GCV3例。结论：贵州地区静注海洛因成瘾者中有TTV感染存在，感染率高低与性别、注毒时间长短无关。TTV可以单独感染，但与HCV、HGV的重叠感染率较高。     

输血传播病毒在不同人群中的感染状况及致病性分析

目的探讨TTV在不同人群中的感染状况及其致病性。方法对454例人员,采用微板核酸杂交ELISA法检测TTV;血清酶测定ALT、AST和TBILI。结果454例标本中TTV阳性检出率为5.95%(27/454)。其中在肝病组检出率为3.92%(6/153),分别为肝硬化7.69%(1/13),肝癌0%(0/16),重症慢性乙肝0%(0/22),急性黄疸型乙肝11.11%(1/9),未分型急性黄疸型肝炎0%(0/2),乙肝病毒携带者4.40%(4/91)。在非肝病组检出率为6.89%(21/301),分别为新生儿高胆红素血症0%(0/25),恶性肿瘤9.76%(4/41),献血员11.0%(11/100),体检人员4.44%(6/135)。献血员和体检人员与各种疾病患者的感染率无明显差异(P>0.05);TTV感染者与未感染者的肝功能状况无明显差异(P>0.05);在肝炎患者中,感染TTV者和未感染TTV者的肝功能状况无明显差异(P>0.05)。结论在一般人群和献血员中TTV感染呈“无症状”的“携带状态”,在多种疾病患者中TTV感染无明显加重病情的作用。     

贵州省首例急性输血传播病毒性肝炎报告

目的 检测贵州省是否存在输血传播病毒（ＴＴＶ）感染。方法 对１５０人次急慢性肝炎、肝硬化病人的血清用酶联免疫试验进行抗－ＴＴＶ检测。结果 发现１例抗－ＴＴＶ阳性。为确认结果的可靠性,对该患者不同时间采取的两份血清复检,其结果相同,确诊为急性ＴＴＶ性肝炎。经抗病毒药物等治疗,病人痊愈出院。结论 贵州省存在ＴＴＶ感染。     

新疆静脉注射毒品者输血传播病毒的检测及部分核苷酸序列分析

目的 :调查新疆维吾尔自治区 (新疆 )静脉注射毒品者中输血传播病毒 (TTV)的感染状况及基因型别。方法 :采用巢式PCR技术检测 10 2例汉族、维吾尔族、回族静脉注射毒品者血清中TTVDNA ,以 38例正常体检者为对照 ,比较静脉注射毒品者中TTVDNA阳性率是否与共用注射器有关。将一例维吾尔族TTV阳性扩增产物插入pGEM -TEasy载体 ,测序并序列分析。结果 :静脉注射毒品者TTVDNA阳性率为 32 35 % ,显著高于正常体检者 (5 2 6 % ) ,P <0 0 5。静脉注射毒品者中汉族、维吾尔族、回族TTVDNA阳性率分别为 32 6 1% (15 4 6 )、35 14 % (13 37) %、2 6 32 % (5 19) ,各民族之间差异无显著性 (P >0 0 5 )。静脉注射毒品者TTVDNA阳性率与共用注射器有关 (P <0 0 5 )。一例维吾尔族TTVDNA部分核苷酸测序 ,与日本株 (AB0 0 8394 )比较同源性为 98% ,属于G1a型。结论 :静脉注射毒品者是TTV感染的高危人群。一例维吾尔族TTV核酸序列与日本株 (AB0 0 8394 )相比有高度同源性 ,属于同一基因型别     

地高辛标记探针原位杂交检测肝外组织中经血传播病毒DNA

目的 探讨血清单一经血传播病毒（ＴＴＶ）阳性患者肝外组织中ＴＴＶＤＮＡ的表达。方法 采用地高辛标记ＴＴＶＤＮＡ探针原位杂交技术检测免疫组织化学检测肝组织中丙肝病毒ＮＳ３、庚肝病毒ＮＳ５等为阴性的５例尸检肝外组织标本。结果 ３例（３／３）胰腺组织、２例（２／２）睾丸组织、１例（１／１）卵巢组织、２例（２／３）心肌组织、３例（３／５）肾组织检出阳性信号。杂交阳性信号主要为胞核型，仅在部分胰腺组织中出现     

输血传播病毒感染状况调查

目的　调查河南省不同人群输血传播病毒(TTV)的感染状况。方法　采用酶联免疫法 (ELISA)和套式聚合酶链反应(PCR)法对 321例正常人群、正常献血员、急性肝炎、慢性肝炎和肝癌患者血清进行了抗 -TTVIgG和TTVDNA检测。结果　以上人群TTV感染率分别为 5. 0%、0、11. 25%、17. 24%、26. 42%、34. 18%;混合感染中TTV合并HBV、HCV感染率最高,分别为 32. 20%和 30. 51%。结论　河南省不同人群均有TTV的感染;TTV与各型肝炎均有混合感染;ELISA法检测抗-TTVIgG与PCR法检测TTVDNA结果有较好的一致性。     

对南京地区各型肝炎患者TTV感染情况的调查

目的 了解南京地区肝炎患者TTV的感染情况。方法 用巢式聚合酶链反应对390例各型肝炎患者进行TTVDNA检测。结果 TTV在154例非甲～庚型肝炎患者和236例甲～庚型肝炎患者中的阳性率分别为11.69％(18/154)和12.71％(30/236);48例TTV阳性者中9例有输注血制品史。结论 TTV感染可能与部分非甲～庚型肝炎有关,TTV感染可能存在输血外的其他传播途径。     

N-acetylphenylisoserinates of Lactarius sesquiterpenoid alcohols--cytotoxic, antiviral, antiproliferative and immunotropic activities in vitro

In the present study a new group of derivatives of both paclitaxel and Lactarius sesquiterpenes, the N-acetylphenylisoserinates of Lactarius sesquiterpenoid alcohols, was tested for antiviral, cytotoxic, antiproliferative and immunotropic activities in vitro. Among the 13 compounds tested two, isolactarorufin 8-[ N-acetyl-(2' R,3' S)-3'-phenylisoserinate] and 3-O-ethylfurandiol 8-[ N-acetyl-(2' R,3' S)-3'-phenylisoserinate] inhibited Herpes simplex virus type 1 (HSV-1) replication at low cytotoxic concentrations. Selectivity indices were: > 12.2 and > 106.4, respectively. Both compounds decreased also the number of cell divisions. Mitotic indices of the cells submitted to these compounds were: 96/2000 and 48/1000, respectively, in comparison with control (169/2000). It seems that the antimitotic action of the compounds may be associated with their antiviral activity. Moreover, isolactarorufin 8-[ N-acetyl-(2' R,3' S)-3'-phenylisoserinate] inhibited PHA-induced T lymphocyte proliferation.     

Synthesis and antiviral activity of various 3'-azido, 3'-amino, 2',3'-unsaturated, and 2',3'-dideoxy analogues of pyrimidine deoxyribonucleosides against retroviruses

Various 3'-azido, 3'-amino, 2',3'-unsaturated, 2',3'-dideoxy, and 5-substituted analogues of pyrimidine deoxyribonucleosides have been prepared and tested against Moloney-murine leukemia virus (M-MULV), a mammalian T-lymphotropic retrovirus in vitro. Among these compounds, the 3'-azido analogues of thymidine, 2'-deoxy-5-bromouridine, and 2'-deoxy-5-iodouridine, the 2',3'-unsaturated analogue of thymidine and and 2'-deoxycytidine, and 2',3'-dideoxycytidine were found to be most active, with ED50 values of 0.02, 1.5, 3.0, 2.5, 3.7, and 4.0 microM, respectively. These active compounds were nontoxic to the host SC-1 cells up to 100 microM concentration. The 3'-azido analogues of thymidine and 2'-deoxy-5-bromouridine were also tested in vitro against HTLV-III/LAV/AAV ("AIDS" virus) and found to be significantly active, with ED50 values of 0.23 and 2.3 microM, respectively. The structure-activity relationships are discussed.     

IL-6、IL-12、IL-33在丙型肝炎患者中的表达及其临床意义

目的探讨丙型肝炎患者血清中IL-6、IL-12及IL-33含量的变化及其临床意义。方法ELISA法检测30例丙型肝炎患者、30例丙型肝炎病毒携带者及30例健康对照者血清中IL-6、IL-12及IL-33的表达水平。结果相比丙型肝炎病毒携带者及健康对照者,丙型肝炎患者血清中IL-6、IL-12及IL-33的表达水平显著升高（P〈0．01）,且丙型肝炎患者血清中IL-6、IL-12及IL-33的含量显著高于丙型肝炎病毒携带者（P〈0．01）;经治疗后丙型肝炎患者血清中IL-6、IL-12及IL-33的含量显著降低（P〈0．01）。结论IL-6、IL-12及IL-33介导的免疫应答参与了丙型肝炎的发病,在丙型肝炎病毒感染的致病机制中有-定的临床价值。     

Synthesis and antiviral evaluation of some beta-L-2', 3'-dideoxy-5-chloropyrimidine nucleosides and pronucleotides

The synthesis and in vitro anti human immunodeficiency virus (HIV) and anti-hepatitis B virus (HBV) activities of some unnatural beta-L-nucleoside enantiomers related to the anti-HIV compound 2', 3'-dideoxy-3'-fluoro-5-chlorouridine (beta-D-3'Fdd5ClU) are reported. In contrast to beta-D-3'Fdd5ClU, beta-L-3'Fdd5ClU and the other L-congeners were devoid of significant anti-HIV effects, but beta-L-2',3'-dideoxy-5-chlorocytidine (beta-L-dd5ClC) and beta-L-2', 3'-dideoxy-3'-fluoro-cytidine (beta-L-3'FddC) showed a distinct anti-HBV activity. Three mononucleoside phosphotriester derivatives with S-pivaloyl-2-thioethyl (t-BuSATE) groups as biolabile phosphate protective groups were also synthesized. The bis(t-BuSATE) derivative of beta-D-3'Fdd5ClU retained anti-HIV activity in thymidine kinase deficient (TK(-)) CEM cells.     

Inhibition of hepatitis B virus production by modified 2',3'-dideoxy-thymidine and 2',3'-dideoxy-5-methylcytidine derivatives. In vitro and in vivo studies.

The effect of analogues of both 2',3'-dideoxy-3'-fluorothymidine (FddThd) [2',3'-dideoxy-3'-fluorouridine (FddUrd), 2',3'-dideoxy-3'-fluoro-5-chlorouridine (FddClUrd), 2',3'-dideoxy-3'- fluoro-5-bromouridine (FddBrUrd) and 2',3'-dideoxy-3'-fluoro-5-bromovinyluridine (FddBVUrd)] and 2',3'-dideoxy-3'-fluorocytidine (FddCyt) [2',3'-dideoxy-3'-fluoro-5-fluorocytidine (FddFCyt), 2',3'-dideoxy-3'-fluoro-5-chlorocytidine (FddClCyt), 2',3'-dideoxy-3'-fluoro-5-methylcytidine (FddMeCyt), 2',3'-dideoxy-3'-fluoro-5-ethylcytidine (FddEtCyt), 2',3'-dideoxy-3'-chloro-5-methylcytidine (ClddMeCyt), 2',3'-dideoxy-3'-amino-5-methylcytidine (AmddMeCyt), 2',3'-dideoxy-3'-azido-5- methylcytidine (AzddMeCyt) and arabinosyl-5-methylcytosine (AraMeCyt)] were tested for their potential antiviral activity in vitro using the human hepatoblastoma cell line, Hep G2 2.2.15, which was transfected with a vector containing hepatitis B virus (HBV). It was found that FddThd, FddMeCyt, FddEtCyt, ClddMeCyt, AmddMeCyt and AraMeCyt display cytostatic activity at concentrations (CD50 values) between 0.54 (FddMeCyt) and 3.93 microM (FddEtCyt), while FddUrd, FddClUrd, FddBrUrd, FddBVUrd, FddCyt, FddFCyt, FddClCyt and AzddMeCyt do not affect cell growth at concentrations of up to 25 microM. Among the thymidine analogues tested, FddThd is the most effective antiviral agent: at a concentration of 0.03 microM a more than 90% reduction of HBV DNA synthesis was measured. On the other hand, the antiviral indexes displayed by FddClUrd, FddBrUrd and FddBVUrd are higher than tht of FddThd; FddUrd was completely inactive. The most powerful antiviral agents in the group of cytidine analogues tested in vitro were FddMeCyt (more than 90% reduction of HBVDNA synthesis at 0.10 microM) and ClddMeCyt (0.10 microM); FddClCyt, FddEtCyt, AmddMeCyt and AraMeCyt were of intermediate activity. None of the negligible antiviral activity was determined for FddUrd, FddCyt, FddFCyt and AzddMeCyt. FddThd and FddMeCyt displayed in vivo an antiviral effect in the duck/duck HBV (DHBV) animal system. Administration of 10 or 20 mg/kg (total daily dose) of FddThd and 5 or 10 mg/kg of FddMeCyt (i.m. daily) to ducks infected with DHBV for 12 days blocked virus production. Termination of treatment with FddThd of infected animals led to reappearance of the virus in the serum though at lower levels. The in vitro and the in vivo data suggest that FddThd and FddMeCyt might be promising antiviral agents for the treatment of infection caused by HBV in humans.