携带泛素-HBcAg融合基因的慢病毒表达载体构建及其体外诱导小鼠髓源性树突状细胞成熟

目的构建携带泛素-HBcAg融合基因的慢病毒表达载体,包装成重组慢病毒并观察其体外诱导小鼠髓源性树突状细胞（DC）成熟。方法 PCR扩增Ub-HBcAg融合基因,插入到慢病毒骨架质粒pWPXLd中,构建重组质粒pW-Ub-HBcAg。将构建的重组慢病毒质粒pW-Ub-HBcAg和包装质粒psPAX2、包膜质粒PMD2.G用脂质体共同转染293T细胞,获得携带Ub-HBcAg基因的重组慢病毒LV-Ub-HBcAg,并检测其在293T细胞中的表达。体外分离培养小鼠髓源性DC,加入重组慢病毒,流式细胞仪测定DC表面分子表达,ELISA测定DC培养上清中IL-12分泌水平。结果强制泛素化HBcAg融合基因的慢病毒表达载体经测序证实目的基因序列及插入方向均正确,W estern b lot能检测到目的蛋白在293T细胞中的表达。LV-Ub-HBcAg能上调DC表面分子的表达（CD86、CD80、MHC-Ⅱ类分子）,并且能促进DC分泌IL-12（139.2±10.75）pg/mL,明显高于LV-HBcAg组分泌的量（P〈0.01）。结论成功构建了携带强制泛素化HBcAg融合基因的慢病毒,转染293T细胞后能够稳定表达目的基因,并且能诱导DC分化、成熟,上调表面共刺激分子的表达,促进IL-12因子的分泌。     

负载GST抗原的树突状细胞疫苗联合CpG ODN抗日本血吸虫感染的保护性研究

目的研究负载GST抗原的树突状细胞(DC)疫苗联合非甲基化胞嘧啶鸟嘌呤二核苷酸寡脱氧核苷酸(CpGODN)免疫小鼠抗日本血吸虫感染的作用。方法将GST抗原纯化后负载树突状细胞株DC2.4,免疫荧光染色法检测GST的负载情况,并进行动物保护性实验。35只C57BL/6小鼠随机均分7组(每组5只),分别作免疫注射:A组为未处理的DC,B组为牛血清白蛋白(BSA)处理的DC,C组为GST负载的DC,D组为GST+CpGODN共刺激的DC,E组为CpGODN刺激的DC,F组为GST蛋白,G组为空白对照组。A～E各组DC经0.25%胰蛋白酶消化后用PBS调整密度至107/ml,每鼠皮下注射0.1ml,每次间隔2周,共免疫3次。F组首次每鼠免疫50μgGST蛋白加福氏完全佐剂,第2、3次分别免疫50μg、10μgGST蛋白加福氏不完全佐剂,均为皮下注射。各组于末次免疫后10d收集血清,ELISA方法分析血清中的特异性抗体。各组小鼠于末次免疫后2周每鼠经腹部感染尾蚴30±1条。6周后剖杀小鼠,计算减虫率。结果 DC经GST负载后可在荧光显微镜下观察到抗GST的特异荧光,表明抗原已被DC摄取。各组小鼠免疫后,F组抗体水平最高,为2.1270±0.4115,另外C组(0.5552±0.0789)和D组(0.7150±0.0523)的抗体水平均高于G组(0.2358±0.0889),差异有统计学意义(P0.05)。免疫后攻击感染,D组小鼠的减虫率最高为53.3%,其次为F组(24.0%)和C组(21.3%),但D组与两组比较差异无统计学意义(P0.05)。结论 CpGODN联合GST抗原负载的树突状细胞疫苗具有一定的抗日本血吸虫感染作用。     

带有HA标签的HBc蛋白真核表达质粒的构建及其在HEK293细胞中的表达

目的构建带有流感病毒血凝素9钛表位标签（HA标签）的乙型肝炎病毒（HBV）核心蛋白（HBc）的表达质粒,为进一步研究HBe蛋白基因的功能奠定实验基础。方法设计并合成扩增HBc蛋白全长的PCR引物,以野生型的HBV质粒pDolTHBV-1为模板,PCR扩增全长的HBc蛋白,大小为563bp;PCR产物经过Eco砒和XholI酶切后,与线性化的pcDNA3／HA载体连接后转化大肠杆菌,扩增、纯化获得所需质粒,以琼脂糖凝胶电泳、酶切鉴定以及基因测序鉴定其分子质量和插入片段的序列;构建的质粒转染HEK293细胞48h后,RIPA裂解细胞,Western blot检测HBc蛋白表达。结果纯化质粒的相对分子质量为5．9kb,酶切鉴定结果符合目的条带（563bp）大小,插入的寡核苷酸序列与野生型HBV基因序列完全相符,表达的融合蛋白大小为28kDa。结论成功构建了带有HA标签的HBc蛋白的真核表达质粒pcDNA3／HA—HBc蛋白,为进一步研究HBe蛋白的功能奠定了良好的实验基础。     

CpG-ODN对慢性乙型肝炎患者外周血树突状细胞表型和功能的影响

研究含非甲基化CpG基序的寡核苷酸(CpG-ODN)对慢性乙型肝炎患者(CHB)外周血树突状细胞(den- dritic cell,DC)表型和功能的影响。以CpG-ODN刺激CHB和健康者外周血单核细胞衍生的DC成熟,结果显示,与 non-ODN和PBS组比,CpG-ODN能明显提高CHB患者DC的HLA-DR、CD86表达,使其IL-12分泌增加,刺激T 细胞增殖的能力亦显著增强(P<0．05),但不能明显提高CD1a表达。因此,CpG-ODN与TNF-α一样能促进CHB 患者外周血DC成熟进瓶增强其抗原提呈功能;CHB患者的細胞因子环境可能是DC功能沉默的重要原因。     

树突状细胞在蠕虫感染免疫应答中的作用

树突状细胞是体内功能最强的专职抗原呈递细胞。寄生性蠕虫感染不但能促进树突状细胞分化成熟,诱导机体产生Th2型免疫应答,还能抑制树突状细胞正常功能,诱导免疫逃避。本文对树突状细胞在蠕虫感染免疫中的作用进行综述。     

人DC-SIGN真核表达载体的构建及在K-562细胞中的表达

目的构建含人DC-SIGN基因的真核表达载体,探讨DC-SIGN在K-562细胞中的表达,为研究丙型肝炎病毒(HCV)与DC-SIGN的相互作用奠定基础.方法分离人外周血单个核细胞,体外诱导分化为树突状细胞(DC);提取细胞总RNA并反转录为cDNA,设计上下游引物,利用PCR技术扩增DC-SIGN片段,连接入克隆载体pGEM-T easy;应用双酶切回收基因片段,定向克隆入真核表达载体pCDNA3.1;通过脂质体介导的基因转染技术将pCDNA3.1-DC-SIGN和空载体转入K-562细胞,应用G-418筛选稳定表达DC-SIGN的K-562细胞,以DC-SIGN单抗通过免疫荧光法检测K-562细胞的表达产物.结果 PBMC体外成功刺激分化为DC,含DC-SIGN基因的克隆载体pGEM-DC-SIGN经酶切、PCR及测序鉴定分析正确,pCDNA3.1-DC-SIGN经酶切、PCR鉴定分析正确,DC-SIGN可在K-562细胞表面稳定表达.结论 DC-SIGN可在K-562细胞中大量表达,为进一步研究DC-SIGN在HCV感染中的作用奠定了基础.     

恶性疟原虫ABRA基因真核表达重组质粒的构建及序列分析

目的 构建恶性疟原虫海南（FCC1／HN）株ABRA基因真核表达重组质粒pcDNA3-ABRA;测定ABRA基因序列,并了解FCC1／HN株与其它分离株ABRA序列的差异。方法 根据ABRA基因已知序列设计合成一对引物,用PCR技术从FCC1／HN株基因组的DNA中扩增ABRA基因,将ABRA基因定向克隆入真核表达载体pcDNA3,转化大肠杆菌DH5α感受态细胞;用切,PCR扩增鉴定筛选到的重 质粒阳性克隆。以正确的重组质粒为模板,用双脱氧链末端终止法测定ABRA基因序列,应用软件辅助分析ABRA序列及进行同生比较。结果 PCR扩增得到特异的FCC1／HN株ABRA基因;经双酶切及PCR鉴定表明获得正确的pcDNA3-ABRA重组质粒。测序表明,FCC1／HN株ABRA基因大小为2220bp,编码739个氨基酸。恶性疟原虫FCC1／HN株与Camp,3D7,FC27株ABRA基因编码氨基酸序列主要在C－端存在少量不同;FCR3株与以上4株ABRA基因编码氨基酸序列相比,只有少量的氨基酸替换。结论 从恶性疟原虫FCC1／HN株基因组DNA中获取ABRA基因,成功构建真核表达重组质粒pcDNA3-ABRA,并测定了FCC1／HN株ABRA基因的序列;FCC1／HN株与其它分离株ABRA基因有很高的同源性。     

含CpG基序寡核苷酸对慢性乙型肝炎患者外周血树突状细胞的免疫激活研究

目的 研究含非甲基化CpG基序的寡核苷酸（CpG-ODN）对慢性乙型肝炎患者（CHB）外周血树突状细胞（DC）表型和功能的影响。方法以CD14磁性分选微珠分离CHB患者外周血高纯度单核细胞；以重组人粒细胞巨噬细胞集落刺激因子（hGM—CSF）、重组人白细胞介素-4（hIL-4）诱导扩增DC；以CpG-ODN刺激DC成熟，并与肿瘤坏死因子（TNF）-n比较，评价其对DC表达表面分子人类白细胞组织相容性抗原（HLA）-DR、CD86、CD1a，分泌IL-12p70以及刺激同种T细胞增殖能力的影响。结果与non—ODN和磷酸盐缓冲液（PBS）组比较，CpG-ODN能明显提高CHB患者外周血DC表面分子HLA—DR、CD86的表达，使IL-12分泌增加，刺激同种异体T细胞增殖的能力亦显著增强（P=0．017和0．023），但不能明显提高CD1a的表达；CpG-ODN的上述刺激作用接近或略逊于hTNF-α，但差异无统计学意义（P〉0．05）。结论CpG-ODN与hTNF-α一样能够促进CHB患者外周血DC成熟进而增强其抗原提呈功能。     

CpG-ODN联合HBsAg对慢性乙型肝炎患者外周血树突状细胞表型和功能的影响

目的　研究含非甲基化CpG基序的免疫刺激寡核苷酸 (CpG- ODN)与重组HBsAg对慢性乙型肝炎患者 (CHB)外周血树突状细胞 (dendriticcell ,DC)表型和功能的影响。方法　以重组人GM CSF、IL 4自CHB患者和健康者外周血单个核细胞诱导扩增DC ;以CpG ODN和HBsAg单独或联合刺激DC ,并与TNF α比较 ,评价其对DC表达表面分子HLA DR、CD86、CD1a ,分泌IL 12p70以及刺激同种T细胞增殖能力的影响 ;同时检测血浆TGF- β、IFN- γ含量。 结果　与PBS组比 ,CpG- ODN单用或联合HBsAg均能明显提高CHB患者DC表面分子HLA DR的表达 ,使IL- 12分泌增加 ,刺激同种T细胞增殖的能力亦增强 ,CpG ODN联合HBsAg尚能明显提高CD1a的表达 ;CpG- ODN的上述刺激作用类似于TNF α ;CHB患者血浆TGF- β、IFN -γ含量明显高于正常对照。 结论　CpG -ODN与TNF α一样能够促进CHB患者外周血DC分化和成熟 ;CpG- ODN与HBsAg联合刺激能协同增强DC的特异性抗原递呈作用 ;CHB患者的细胞因子环境可能是DC功能沉默的重要原因。     

树突状细胞和调节性T细胞在自身抗原所致小鼠免疫耐受模型中的作用

目的:探讨树突状细胞(DC)及CD4 CD25 调节性T细胞在胰岛素自身抗原sc所诱导的小鼠胰岛素依赖性糖尿病(IDDM)的免疫耐受中的重要作用．方法:低剂量链脲佐菌素(STZ)(40 mg／kg)ip 连续5次在Balb／c小鼠体内建立IDDM模型,胰岛素(100 μg)与不完全弗氏佐剂(IFA,1:1)混合液sc 1次／wk,连续4 wk．模型建立后每周测定血糖,5 wk时处死动物,取胰腺进行病理组织学检查．分离骨髓DC前体及脾脏T淋巴细胞并进行体外培养．采用流式细胞术测定DC表型和CD4 CD25 调节性T细胞,以同种淋巴细胞刺激实验检测DC刺激淋巴细胞增殖功能．结果:胰岛素sc 4 wk后可明显降低小鼠的血糖,与模型对照组有极显著差异(13．79± 2．71 mmol/L vs 20．98±1．43 mmol／L,P<0．05), 胰岛内炎症细胞浸润减少,组织结构完整． IDDM模型建立后,小鼠骨髓来源树突状细胞CD11c表达为26．4％,DC分化异常,而正常小鼠CD11c表达为47．5％;混合淋巴细胞反应中DC刺激能力减弱,刺激指数分别为1．47± 0．01和1．32±0．01(刺激细胞和反应细胞比例分别为1:10和1:20),与正常小鼠相比,差别具有极显著性意义(P值均小于0．01)．脾脏 CD4 CD25 调节性T细胞减少到1．43％,而正常小鼠为5．09％．与此相反,胰岛素自身抗原连续应用后,不仅使血糖得到控制,表达 CD11c的树突状细胞数量增加,CD86和MHC- Ⅱ表面分子表达降低到26．6％和28．8％,刺激淋巴细胞反应的能力弱于正常DC,但强于模型小鼠的DC,刺激指数分别为2．30±0．06(1: 10)和2．17±0．02(1:20),CD4 CD25 调节性T 细胞数量上升到7．15％．结论:胰岛素sc可预防STZ所致小鼠IDDM的发生,自身抗原可以通过改善功能异常的树突状细胞．诱导CD4 CD25 调节性T细胞分化在模型小鼠体内建立免疫耐受．     

辅助性T细胞表位HBV表面抗原融合基因真核表达质粒的构建

为提高HBVDNA疫苗的免疫效率，将一个通用型辅助性T细胞表位基因引入HBV表面抗原基因的5‘末端，构建成真核表达质粒。PCR方法合成PADRE-HBsAg目的基因，产物与pMDl8T载体连接，经Hind Ⅲ，EcoR Ⅰ双酶切，再克隆入真核表达载体pcDNA3．1( )。酶切及测序鉴定，该真核表达载体构建成功，为进一步观察其特异性细胞和体液免疫反应奠定了基础。     

DC-SIGN在慢性乙型肝炎病毒感染时树突状细胞成熟活化中的作用

目的研究慢性HBV感染时,DC-SIGN在树突状细胞（DC）成熟和活化中介导的作用。方法将α-甘露糖苷酶抑制剂-基夫碱作用于HepG2.2.15细胞,收获上清中的高甘露糖型HBV颗粒,并于第5 d加入到外周血单核细胞衍生的DC培养基中,培养至第7 d,采用流式细胞仪检测DC表面CDla、CD83、CD80、CD86、HLA-DR分子的表达,MTT法检测DC刺激淋巴细胞增殖的能力,ELISA法检测DC分泌IL-12的水平。结果高甘露糖型HBV组,与天然的HBV组相比,DC表面CDla、CD83、CD80、CD86、HLA-DR分子的表达增加,分泌IL-12的水平升高,刺激同种异体淋巴细胞增殖的能力亦明显增强,且上述效应均可被DC-SIGN特异性抗体所阻断。结论 DC-SIGN识别高甘露糖型HBV后可以促进DC的成熟和活化,天然的HBV可能利用α-甘露糖苷酶参与的去甘露聚糖修饰来逃避DC-SIGN的识别,从而诱导DC功能的缺陷。     

HBV X基因-HCV C基因cDNA融合基因真核表达载体的构建及其表达

目的：构建HBV X-HCV C融合基因真核表达载体，并获得稳定表达该基因的HepG2细胞株。方法：双酶切质粒pXT1-X，得到完整的HBV X基因片段后，将其插入到质粒PBK-CMV和PBK-HCVC的相应酶切位点，得到重组质粒PBK-X和PBK-X-C；再将质粒RBK-CMV、PBK-X、PBK-HCV C和PBK-X-C分别导入肝癌细胞株HepG2中，G418筛选，RT-PCR、蛋白印迹鉴定HBV X和HCV C蛋白表达。结果：质粒PBK-CMV、PBK-X、PBK-HCV C和PBK-X-C在HepG2细胞中有稳定表达。结论：成功构建HBV X-HCVC融合基因真核表达载体，并获得稳定表达该基因的HepG2细胞株。     

Safety and immunogenicity of hepatitis B surface antigen‐pulsed dendritic cells in patients with chronic hepatitis B

Summary. The immune modulator capacity of antigen‐pulsed dendritic cells (DC) has been documented in patients with cancers and in animal models of chronic viral infections. Cancer antigen‐pulsed DC are now used for treating patients with cancer. But viral antigen‐pulsed DC are not used in chronic viral‐infected patients because safety of antigen‐pulsed DC has not been evaluated in these patients. DC were isolated from human peripheral blood mononuclear cells by culturing with human‐grade granulocyte‐macrophage colony stimulating factor and interleukin‐4. Human blood DC were cultured with hepatitis B surface antigen (HBsAg) for 8 h to prepare HBsAg‐pulsed DC. After immunogenicity assessment of HBsAg‐pulsed DC in vitro, five million HBsAg‐pulsed DC were administered intradermally to five patients with chronic hepatitis B (CHB) 1–3 times. HBsAg‐pulsed DC were immunogenic in nature because they produced significantly higher levels of interleukin‐12 and interferon‐γ compared to unpulsed DC (P < 0.05). Also, HBsAg‐pulsed DC induced proliferation of HBsAg‐specific T lymphocytes in vitro. CHB patients injected with HBsAg‐pulsed DC did not exhibit generalized inflammation, exacerbation of liver damage, abnormal kidney function, or features of autoimmunity. Administration of HBsAg‐pulsed DC induced anti‐HBs in two patients and HBsAg‐specific cellular immunity in 1 patient. This is the first study about preparation of antigen‐pulsed DC using human consumable materials for treating patients with CHB. Because HBsAg‐pulsed DC were safe for all patients with CHB and had immune modulation capacity in some patients, phase I and phase II clinical trials with antigen‐pulsed DC in CHB and other chronic infections are warranted.     

HBsAg-negative hepatitis B virus infections in hepatitis C virus-associated hepatocellular carcinoma

This study was conducted to evaluate reports that hepatitis B virus (HBV) DNA sequences can be found in the serum and/or tumour tissue from some hepatocellular carcinoma (HCC) patients who have no detectable hepatitis B surface antigen (HBsAg) in their sera. Such HBV infections would be highly atypical, because prospective studies have shown a clear succession of specific serologic markers during and after most HBV infections. As most HBsAg-negative HCC patients in Japan have hepatitis C virus (HCV) infections, the present study was conducted to determine whether some of these patients actually have unrecognized HBV infections. Thirty newly diagnosed HCC patients from Kurume, Japan, with antibody to the hepatitis C virus (anti-HCV) were studied. None of the 30 had HBsAg detectable in their serum. Of 22 for whom test results for antibodies to the hepatitis B core antigen (anti-HBc) and antibodies to HBsAg (anti-HBs) were available, 14 (64%) had anti-HBc and anti-HBs, four (18%) had anti-HBc alone, and four (18%) had no HBV markers. Nested polymerase chain reaction was used to detect the HBV surface (S), core (C), polymerase (P) and core promoter gene sequences in the HCC tissues and in the adjacent nontumorous liver tissues. HBV DNA was detected in HCC and/or adjacent nontumorous liver in 22 of 30 (73%) patients [detected in both HCC and nontumorous liver in 19/30 patients (63%)]. Among the 22 patients with detectable HBV DNA, more than one HBV gene was detected in 10 (46%). Among the four patients whose sera were negative for all HBV markers, three had HBV DNA in either HCC and nontumorous liver (two cases) or only in the nontumorous liver (one case); HBV DNA could not be detected in tissues from the fourth patient. In 18 of 21 (86%) patients with detectable HBV core promoter sequences, mutations at both nucleotides 1762 (A-GT) and 1764 (G-A) in the core promoter region were found. No deletions were detected in the core promoter gene region of the type reported to be associated with some cases of HBsAg-negative HBV infection. Thus, HBV DNA was detectable in 22 (73%) HBsAg-negative, anti-HCV-positive HCCs, including three (10%) who were also negative for anti-HBc and anti-HBs. HBV mutations at both nucleotides 1762 (A-GT) and 1764 (G-A) in the core promoter region were found in the majority of cases, mutations that have previously been reported in HBV that is integrated in HCC DNA. In serologic surveys to determine etiologic associations of HCC, patients such as those in this study would have been incorrectly designated as having 'HCV-associated HCC,' whereas the data in this study suggest that HBV could have played a role in the development of their HCCs.     

Monocyte derived dendritic cells retain their functional capacity in patients following infection with hepatitis C virus

Summary. Studies assessing the function of monocyte derived dendritic cells (MD‐DC) in individuals with hepatitis C virus (HCV) infection have shown conflicting results. Impaired MD‐DC function in chronic HCV infection would have important implications both for understanding the pathogenesis of HCV infection and in the use of autologous MD‐DC in vaccination strategies. We determined the allostimulatory capacity of MD‐DC in the same patient before and after HCV infection. Next, the phenotype, cytokine production and allostimulatory function of immature and mature MD‐DC in individuals with persistent HCV infection were compared directly with MD‐DC from healthy individuals. Finally, we assessed the ability of MD‐DC to prime autologous naïve peptide specific CD8+ T cells using HLA‐A2 class‐I tetramers. DCs retained the same allostimulatory capacity before and following the establishment of persistent HCV infection. The surface phenotype and the amount of interleukin (IL)‐10 and IL‐12(p70) produced during DC maturation did not differ between HCV‐infected individuals and healthy controls. Mature DCs from HCV‐infected individuals performed comparably in an allogeneic MLR compared with healthy individuals. Mature MD‐DC from HCV‐infected individuals stimulated the expansion of peptide specific naïve CD8+ T cells. MD‐DC from HCV‐infected and healthy individuals are phenotypically indistinguishable and perform comparably in functional assays.     

联合检测HBV前S1抗原和核心抗原的临床意义

探讨联合检测血清HBV前s1抗原（preSl）和核心抗原（HBcAg）（均为HBV核酸相关抗原，nucleic acids related antigen，HBV NRAg）的意义及临床价值。方法：采用ELISA法对393份HBsAg、HBV DNA双阳性的血清和612份HBsAg阴性血清进行HBV NRAg检测，所有标本均采用多区段巢式PCR确认阳性、阴性，采用荧光定量PCR法进行HBV DNA定量分析。结果：393份HBsAg、HBV DNA双阳性血清中，HBV NRAg阳性为382份，其阳性率为97．2％；612份HBsAg阴性的血清标本中，609份确认为HBV DNA阴性，其中检出2份HBV NRAg阳性，607份为阴性，其HBV NRAg的阴性率为99．7％（607／609），另3份HBsAg阴性血清HBV DNA阳性者，其HBV NRAg均为阳性。结论：联合检测preSl和HBcAg的HBV NRAg可作为临床HBV感染的筛选及判断HBV复制的有意义的补充项目。     

A class C CpG toll-like receptor 9 agonist successfully induces robust interferon-alpha production by plasmacytoid dendritic cells from patients chronically infected with hepatitis C

Summary.  Immunomodulators that induce local endogenous interferon-alpha (IFN-α) production by plasmacytoid dendritic cells (pDCs) may offer new strategies for the treatment of patients chronically infected with the hepatitis C virus (HCV). However, such an approach may be compromised if reports are true that IFN-α production by pDCs from patients with chronic HCV (cHCV) is profoundly impaired. To address the question of pDC dysfunction in cHCV more definitively, in the present study a panel of four prototypic synthetic agonists of toll-like receptor 7 (TLR7) or TLR9 were administered in vitro to pDCs purified from cHCV patients and from normal uninfected donors and their responses compared in terms of not only IFN-α production but also the global expression of other cytokines and phenotypic maturation. Plasmacytoid DCs from uninfected donors produced substantial levels of IFN-α in response to three of the four agonists and yet only one TLR9 agonist, a class C CpG oligodeoxynucleotide (ODN), induced robust IFN-α production by pDCs from cHCV patients. Proinflammatory cytokine production and phenotypic maturation in response to all four agonists was equivalent in infected and uninfected pDCs. These data point to a profound but selective defect in IFN-α production by pDCs from cHCV donors. Nonetheless, a class C CpG ODN successfully induced robust IFN-α production, suggesting that this class of TLR9 agonist may have utility as a future immunotherapeutic for the treatment of chronic HCV infection.     

Significance of hepatitis B virus surface antigen, hepatitis C virus expression in hepatocellular carcinoma and pericarcinomatous tissues

AIM： To investigate the correlation between hepatitis B virus surface antigen （HBsAg）, hepatitis C virus （HCV） expression in hepatocellular carcinoma （HCC）, the HAI score of the noncancerous region of the liver and the serum Alpha fetoprotein （AFP） level.
METHODS： The patterns of HBsAg and HCV in 100 cases of HCC and their surrounding liver tissues were studied on paraffin-embedded sections with immunohistochemistry, the histological status was determined by one pathologist and one surgeon simultaneously using the hepatitis activity index （HAIl score, and AFP was detected by radioimmunity. The study included 100 consecutive patients who underwent curative resection for HCC. Based on HBsAg and HCV expression, the patients were classified into 4 groups： patients positive for HBsAg （HBsAg group）, patients positive for HCV （HCV group）, patients negative for both HCV and HBsAg （NBNC group） and patients positive for both HBsAg and HCV （BC group）.
RESULTS： The BC group had significantly higher HAI scores than the other three groups. （BC 〉 HCV 〉 HBsAg 〉 NBNC）. HBV and HCV virus infection was positively correlated with HAI （rs = 0.39, P = 0.00011. The positive rate of AFP （85.7%） and the value of AFP （541.2 ng/mL） in the group with HBV and HCV co-infection were the highest among the four groups. The positive rate （53.3%） of AFP and the value of AFP （ 53.3 ng/mL） in the group with none-infection of HBV and HCV were the lowest. HBV and HCV virus infection was positively correlated with AFP（rs = 0.38, P = 0.0001）.
CONCLUSION： The AFP increase in patients with liver cancer was positively correlated with the infection of HBV and HCV. The-serum AFP elevation by the infection of HBV and HCV is one of mechanisms which lead to hepatocarcinogenesis, and the antivirus intervening treatment of hepatitis is significant for the prognosis of liver cancer. From our Spearman＇s rank correlation analysis, we can conclude that the severity of virally induced