Status of hepatitis B virus DNA in alcoholic liver disease: a study of a large urban population in the United States

Two reports have shown hepatitis B virus DNA in serum and liver tissue in alcoholic liver disease with negative serum HBsAg, suggesting a pathogenetic role for hepatitis B virus. We studied hepatitis B virus DNA in serum and liver from three groups of alcoholic patients; (Group 1) 50 patients without liver disease, (Group 2) 108 patients with alcoholic liver disease and (Group 3) five patients with alcoholic liver disease and hepatocellular carcinoma. Serum was tested for HBsAg, anti-hepatitis B core and anti-hepatitis B surface by radioimmunoassay and hepatitis B virus DNA by direct spot hybridization. Liver tissue from Groups 2 and 3 (113 patients) was examined by Southern blot analysis using 32P-labeled hepatitis B virus DNA clone from pBR322. Controls were 21 patients with chronic hepatitis B virus (14 patients with chronic active hepatitis, seven patients with cirrhosis and hepatocellular carcinoma). Serum and tissue were analyzed for hepatitis B virus DNA. Hepatitis B virus DNA was not detected in either serum or liver tissue in any of the 163 patients (Groups 1 to 3). In contrast, among the controls, hepatitis B virus DNA was present in the serum of 15 of the 21. Tissue DNA in those with chronic active hepatitis revealed 10/14 with free hepatitis B virus DNA, two with integrated sequences and two with no viral sequences. All seven patients with hepatocellular carcinoma had integrated viral DNA sequences in the tumor tissues. From these results, it appears that hepatitis B virus does not play a role in the pathogenesis of alcoholic liver disease.     

Liver disease associated with duck hepatitis B virus infection of domestic ducks.

The liver disease associated with duck hepatitis B viremia was investigated in naturally infected ducks from Chi-tung county in China and in both naturally and experimentally infected ducks from the United States. Liver and serum specimens of adult Chinese ducks were examined for duck hepatitis B virus (DHBV) DNA by dot and gel blot hybridization. DHBV was found in serum and (in episomal form only) in livers of 6 of 11 birds exhibiting various degrees of chronic hepatitis. In 1 bird with hepatocellular carcinoma, DHBV DNA was detected at the limit of assay sensitivity and in another not at all, contrasting with findings in humans and woodchucks. In work with California Pekin and Khaki Campbell ducks, known amounts of DHBV were injected into the egg 10 days before, or into ducklings 1 day after, hatching and the livers were examined 6 weeks later. The majority of the injected ducklings had viremia detectable by hybridization 1 or 2 weeks after injection. The presence but not the amount of viremia correlated with incidence and degree of hepatitis, determined under code. The most severe instances of hepatitis, all in Pekin ducks, resembled the hepatitis in adult Chinese ducks of Chi-tung county. Severe and moderate hepatitis were found only in indoor-caged injected animals with viremia and in some uninjected birds without viremia that had been kept in outdoor flocks. The latter hepatitis, as some hepatitis in adult Chinese ducks, may not be related to DHBV. Mild and insignificant hepatitis were also found in injected and noninjected ducklings, some of which had the vertically transmitted spontaneous viremia previously described. The good correlation of experimentally induced viremia with incidence and severity of hepatitis in the Pekin duckling provides a simple, rapid, and relatively inexpensive model to study the relation of lesions to hepatitis B family infection in nonprimates.     

拉米夫定联合泛昔洛韦抗鸭乙型肝炎病毒的实验研究

观察核苷酸类似物拉米夫定联合泛昔洛韦体内抗鸭乙型肝炎病毒（DHBV）的作用。方法 采用重庆麻鸭乙型肝炎动物模型,用拉米夫定联合泛昔洛韦口服治疗4周,停药观察1周,检测用药前后血清中的DHBV DNA、DHBsAg及血清转氨酶（ALT、AST）、肝组织HE染色病理。并以单用拉米夫定、泛昔洛韦、阿昔洛韦作对照。结果 拉米夫定联合泛昔洛韦用药后能使血清中DHBV DNA含量总体水平显著降低（P＜0．01）,停药1周后DHBV DNA较用药4周时DHBV DNA含量回升现象不明显。用药前后清血DHBsAg的吸光度值（490nm)的变化与DNA含量改变相似;此外,肝脏病理检查及治疗4周、停药1周后血清转氨酶检测未发现联合用药对鸭肝组织有明显的毒性损害。结论 拉米夫定联合泛昔洛韦连续用药4周在鸭体内有抗鸭乙型肝炎病毒的作用,且停药后DHBV DNA无明显“反跳”,二者用药有协同作用。     

IFN-γ increases efficiency of DNA vaccine in protecting ducks against infection

AIM: To detect the effects of DNA vaccines in combination with duck IFN-γ gene on the protection of ducks against duck hepatitis B virus (DHBV) infection.METHODS: DuIFN-γ cDNA was cloned and expressed in COS-7 cells, and the antiviral activity of DuIFN-γ was detected and neutralized by specific antibodies. Ducks were vaccinated with DHBpreS/S DNA alone or coimmunized with plasmid expressing DuIFN-γ. DuIFN-γmRNA in peripheral blood mononuclear cells (PBMCs) from immunized ducks was detected by semi-quantitative competitive RT-PCR. Anti-DHBpreS was titrated by enzyme-linked immunosorbent assay (ELISA). DHBV DNA in sera and liver was detected by Southern blot hybridization, after ducks were challenged with high doses of DHBV.RESULTS: DuIFN-γ expressed by COS-7 was able to protect duck fibroblasts against vesicular stomatitis virus (VSV) infection in a dose-dependent fashion, and antiDuIFN-γ antibodies neutralized the antiviral effects. DuIFN-γin the supernatant also inhibited the release of DHBV DNA from LMH-D2 cells. When ducks were co-immunized with DNA vaccine expressing DHBpreS/S and DuIFN-γ gene as an adjuvant, the level of DuIFN-γ mRNA in PBMCs was higher than that in ducks vaccinated with DHBpreS/S DNA alone. However, the titer of anti-DHBpreS elicited by DHBpreS/S DNA alone was higher than that co-immunized with DuIFN-γ gene and DHBpreS/S DNA. After being challenged with DHBV at high doses, the load of DHBV in sera dropped faster, and the amount of total DNA and cccDNA in the liver decreased more significantly in the group of ducks co-immunized with DuIFN-γ gene and DHBpreS/S DNA than in other groups.CONCLUSION: DHBV preS/S DNA vaccine can protect ducks against DHBV infection, DuIFN-γ gene as an immune adjuvant enhances its efficacy.     

IFN-γ increases efficiency of DNA vaccine in protecting ducks against infection

AIM: To detect the effects of DNA vaccines in combination with duck IFN-gamma gene on the protection of ducks against duck hepatitis B virus (DHBV) infection. METHODS: DuIFN-gamma cDNA was cloned and expressed in COS-7 cells, and the antiviral activity of DuIFN-gamma was detected and neutralized by specific antibodies. Ducks were vaccinated with DHBpreS/S DNA alone or co-immunized with plasmid expressing DuIFN-gamma. DuIFN-gamma mRNA in peripheral blood mononuclear cells (PBMCs) from immunized ducks was detected by semi-quantitative competitive RT-PCR. Anti-DHBpreS was titrated by enzyme-linked immunosorbent assay (ELISA). DHBV DNA in sera and liver was detected by Southern blot hybridization, after ducks were challenged with high doses of DHBV. RESULTS: DuIFN-gamma expressed by COS-7 was able to protect duck fibroblasts against vesicular stomatitis virus (VSV) infection in a dose-dependent fashion, and anti-DuIFN-gamma antibodies neutralized the antiviral effects. DuIFN-gamma in the supernatant also inhibited the release of DHBV DNA from LMH-D2 cells. When ducks were co-immunized with DNA vaccine expressing DHBpreS/S and DuIFN-gamma gene as an adjuvant, the level of DuIFN-gamma mRNA in PBMCs was higher than that in ducks vaccinated with DHBpreS/S DNA alone. However, the titer of anti-DHBpreS elicited by DHBpreS/S DNA alone was higher than that co-immunized with DuIFN-gamma gene and DHBpreS/S DNA. After being challenged with DHBV at high doses, the load of DHBV in sera dropped faster, and the amount of total DNA and cccDNA in the liver decreased more significantly in the group of ducks co-immunized with DuIFN-gamma gene and DHBpreS/S DNA than in other groups. CONCLUSION: DHBV preS/S DNA vaccine can protect ducks against DHBV infection, DuIFN-gamma gene as an immune adjuvant enhances its efficacy.     

The relationship between duck hepatitis B virus infection and duck liver diseases in Qitong, China

The relationship between duck hepatitis B virus (DHBV) infection and duck liver diseases was analysed by spot and gel blot hybridization in sera and liver tissues. One hundred and forty ducks were obtained from Qitong county in China. The DHBV-infected rate was 65.7% and the incidence of duck liver diseases was 70%. The state of DHBV DNA was free in the hepatocytes. The detection rate of DHBV DNA in liver was higher than in serum. The results showed that Qitong duck liver diseases are closely correlated to DHBV infection. The incidence of Qitong duck liver diseases and the DHBV infection rate were different in various species of ducks. Qitong ducks have several hepatic pathological changes as seen in human beings. Therefore, Qitong ducks are most appropriate as an experimental model for human HBV infection.     

Duck hepatitis B virus and liver diseases

The presence of duck hepatitis B virus in serum was studied in 61 ducks (24 from Chi-tung county, China, 20 from Changchun, China, and 17 from Chiba, Japan) with relation to liver disease. None of the 37 ducks from Chiba and Changchun was positive for duck hepatitis B virus as assayed by electron microscopy, endogenous deoxyribonucleic acid-polymerase activity, and hybridization with duck hepatitis B virus deoxyribonucleic acid. No liver disease was seen in these ducks. In contrast, viruslike particles were present in the serum of 12 of 24 (50%) ducks from Chi-tung, China. The presence of duck hepatitis B virus in serum was indicated by the hybridization spot test and deoxyribonucleic acid-polymerase activities. A variety of liver diseases including chronic hepatitis and cirrhosis were seen in the livers of a majority of the ducks from Chi-tung. One duck hepatitis B virus-positive duck had multicentric hepatocellular carcinoma with underlying cirrhosis. Comparison of serum duck hepatitis B virus markers and liver disease in the affected flock revealed a tendency for seronegative ducks to have advanced liver diseases. Duck hepatitis B virus infection may be used as an experimental model to test various hypotheses concerning the pathogenesis of hepatitis B virus-associated liver disease in humans.     

Enhanced duck hepatitis B virus gene expression following aflatoxin B1 exposure

Epidemiological studies have suggested synergistic interactions between chronic hepatitis B virus (HBV) infection and aflatoxin B1 (AFB1) exposure in the etiology of hepatocellular carcinoma (HCC), although the molecular mechanisms of their interactions are still not understood. The aim of this study was to use the Pekin duck model to investigate the impact of AFB1 exposure on duck hepatitis B virus (DHBV) replication during the early stages of virus-carcinogen interactions. Six-week-old chronic DHBV-carrier or uninfected ducks were exposed to AFB1 for 5 weeks or treated with dimethylsulfoxide (DMSO) as a control. Animals were observed for 6 to 13 weeks after AFB1 treatment to study the influence of AFB1 exposure on DHBV replication and liver pathologies. Histological analysis showed more marked changes in the livers of AFB1-treated ducks, and these were enhanced by DHBV infection. A significant increase in serum and liver DHBV DNA level was observed in AFB1-treated ducks as compared with DMSO-treated controls. In addition, viral RNAs, in particular the pregenomic RNA that is the template of viral replication, and intrahepatic DHBV DNA replicative intermediates, were significantly increased by AFB1 treatment. Moreover, an overexpression and accumulation of DHBV large envelope (L) protein was observed in the hepatocytes of AFB1-exposed animals. The in vitro study has further confirmed an increase in intracellular viral DNA and in virus release in AFB1-treated primary duck hepatocytes. Taken together, our results indicate that AFB1 exposure leads to an increase in virus gene expression associated with intrahepatic accumulation of DHBV L protein and enhanced liver pathology.     

Hepatocellular carcinoma in woodchuck hepatitis virus-infected woodchucks: presence of viral DNA in tumor tissue from chronic carriers and animals serologically recovered from acute infections

During long-term studies of the natural history of woodchuck hepatitis virus infection, five cases of histologically confirmed, primary hepatocellular carcinoma were observed in a total of 92 woodchucks which had recovered, by analysis of viral serologic markers (WHsAg-, anti-WHc+, anti-WHs+), from experimental acute woodchuck hepatitis virus infections 20 to 30 months prior to the detection of hepatocellular carcinoma. No hepatocellular carcinoma was observed in 167 uninfected controls at least 3 years of age and held in the same laboratory environment. Southern blot hybridization analysis of liver tissue taken from four of these recovered woodchucks revealed the presence of low levels (0.1 to 0.3 copies per cell) of integrated woodchuck hepatitis virus DNA in hepatocellular carcinoma (four of four animals) and nonneoplastic tissue (three of four animals). Similarly, hepatocellular carcinoma tissue obtained from two wild-caught, naturally infected and serologically recovered woodchucks also contained low levels of integrated woodchuck hepatitis virus DNA. Liver tissues from another 27 of these 92 recovered woodchucks (without hepatocellular carcinoma) were examined for woodchuck hepatitis virus nucleic acids 13 to 31 months following experimental woodchuck hepatitis virus infection. Nonreplicating woodchuck hepatitis virus DNA was present in the liver of eight (30%) and in the peripheral blood lymphocytes from eight (30%) of these 27 animals. These results were in marked contrast to the analysis of woodchuck hepatitis virus DNA in the liver tissue of chronic woodchuck hepatitis virus carriers (20 experimentally infected and nine naturally infected). In these animals, high levels of replicating woodchuck hepatitis virus DNA (up to 2,000 copies per cell) were observed in all hepatocellular carcinoma and nonneoplastic liver tissue. Integrated woodchuck hepatitis virus DNA was found in eight of 60 individual hepatocellular carcinomas detected in 29 chronic carriers, 15 to 40 months postinfection. Integrated woodchuck hepatitis virus DNA was present in the nonneoplastic tissue from four of these 29 chronic woodchuck hepatitis virus carriers.     

鸭乙型肝炎病毒体液免疫血清学指标的系统建立与应用

目的：建立简便,特异的鸭乙型肝炎病毒（DHBV）感染及免疫血清学检测系统。方法：制备、纯化检测所需同的抗DHVB前S区单克隆抗体腹水,DHBsAg和rDHBcAg , 对随机筛选的80份感染血清及实验感染1日龄重庆麻鸭系列血清分别进行DHBsAg 和抗－DHBc,抗－DHBs检测。结果：PCR方法检测出阳性66份,阴性14例,选用PCR阳性的66份标本,经HDBsAg ELISA检测出58份阳性,斑点杂交检出阳性份数为62份。与DHBVRCR相比较,灵敏度分别为87．9％和93．9％,而8只实验感染鸭仅2只在感染后第3周血清抗－DHBc阳性,抗体效价为1：10,至第4周一已为阴性,抗－DHBs则在所有标本均为阴性,结论：DHBV感染及免疫血清学酶联免疫方法的建立,为进一步研究DHBV感染后腹制规律及体风免疫应答状况奠定了基础。     

State of hepatitis B virus DNA in hepatocytes of patients with hepatitis B surface antigen-positive and -negative liver diseases.

Using the Southern blot technique and cloned hepatitis B virus (HBV) DNA as a probe, we studied the state of HBV DNA in the liver of 13 patients with hepatocellular carcinoma, 17 patients with chronic hepatitis, and 2 patients with acute hepatitis. The hybridization results were compared with the serological and immunohistological data. Integration of HBV DNA in cellular DNA of the liver from patients with hepatocellular carcinoma was demonstrated. In two patients from which tumorous and nontumorous liver tissue samples were available the integration patterns were different. In one patient with hepatitis B e antigen (HBeAg)-positive early hepatocellular carcinoma, free viral DNA was present in the liver. In some patients with HBeAg-negative chronic hepatitis, without tumor, integration of HBV DNA in cellular DNA was also demonstrated. This suggests that HBV is not the only factor involved in the development of a tumor. In patients with HBeAg-positive chronic hepatitis, free viral DNA was detected in the liver. In the two acute hepatitis patients analyzed, the restriction endonuclease patterns strongly suggested HBV DNA integration. Therefore, viral DNA integration seems to occur early in infection. Whatever the form of the disease, discrete bands were observed, suggesting the existence of limited and specific integration sites in host cellular DNA. The presence of integrated or free DNA sequences has implications for antiviral therapy. In addition, detection of HBV DNA in the liver is another sensitive viral marker that could be useful for diagnostic purposes.     

Studies on duck hepatitis B virus: Identification and epidemiological survey in Taiwan

The presence of duck hepatitis B virus (DHBV) in domestic ducks in Taiwan was confirmed by DNA polymerase assay, Southern blot analysis and electron microscopy. To investigate the epidemiology of this virus, a total of 1274 serum samples were collected from 30 duck farms from different areas of Taiwan and studied by spot hybridization and/or DNA polymerase assay. The positive rates varied among different strains of ducks: 16% in 243 Pekin ducks, 12% in 392 Chinese common domestic ducks, 4% in 196 Muscovy, 25% in 292 Taiwan Kaiyas and 13% in 151 mule ducks. The positive rate was much higher in the younger ducks; it was highest (30.7%) in ducklings under 1 month of age, followed by ducks aged 1–12 months (11.8%), and lowest in those ducks older than 1 year (7.7%). It was concluded that the prevalence of DHBV infection in domestic ducks in Taiwan is generally high, and that the infected ducks may serve as an animal model for human hepatitis B virus infection which is also prevalent in Taiwan.     

State of hepatitis B viral DNA in the liver of patients with hepatocellular carcinoma and chronic liver disease

In order to clarify the relationship between the integration of hepatitis B virus (HBV) DNA and human hepatocellular carcinoma (HCC), the states of HBV DNA in liver tissues were examined by the Southern blot hybridization procedure. Integration of HBV DNA was found in 12 of 25 HCC cases and 5 of 12 cirrhotic cases. Of these 17 cases, 11 were positive for serum hepatitis B virus surface antigen (HBsAg) and the remaining six were positive for more than one of the serum HBV-related antibodies. In all three cases of chronic hepatitis and 18 controls, integration of HBV DNA could not be detected. Free viral DNA was found in 12 of 15 cases with serum HBsAg. One patient without serum HBsAg also had free viral DNA in the liver. Integration of HBV DNA could be observed in HCC cases positive for serum HBsAg at the highest frequency. However, there was one HCC case from an HBsAg carrier in whom integration of HBV DNA might not have a causal effect on malignant transformation, because integrated HBV DNA could be detected only in a non-tumor region. Since integrated HBV DNA could not be detected in 13 of 25 HCC cases, other etiologic factors than the integration of HBV DNA must also be taken into consideration for HCC.     

Experimental duck hepatitis B virus infection: pathology and evolution of hepatic and extrahepatic infection

Seventy, 1-day-old ducklings inoculated intraperitoneally with duck hepatitis B virus and 30 controls have been studied over a 2-year period. Infection with duck hepatitis B virus occurred in all inoculated ducks, although this was not associated with clinical morbidity. Duck hepatitis B virus DNA was first detected in liver on Day 3, in pancreatic acinar cells on Day 4, serum on Day 6, splenic red and white pulp on Day 7 and in the renal glomurulus on Day 14, using a combination of dot, Southern blot and in situ hybridization techniques. Peak levels of circulating virus, as determined by DNA polymerase levels, occurred 1 to 4 weeks postinoculation. Mild degrees of portal inflammation were seen in sections of liver tissue in both infected and control ducks. However, moderately severe inflammatory changes were present in 8 of 22 infected birds compared with 0 of 18 controls (p less than 0.025). Appearance of this inflammatory infiltrate 6 weeks postinoculation coincided with a decrease in levels of duck hepatitis B virus DNA in hepatocytes and within the pancreatic acinar cells. At the same time, duck hepatitis B virus DNA became increasingly localized to the splenic germinal centers, and viral DNA was first detected in pancreatic islet cells. No histological changes accompanied the extra-hepatic tissue infection. The sequence and significance of duck hepatitis B virus infection in liver and extra-hepatic tissues is discussed in relation to the pathogenesis of hepatitis B virus infection in man.     

Liver orcein stain and viral DNA in duck hepatitis B virus infection in Chinese ducks and experimentally infected Japanese ducklings

Liver sections were stained with orcein, and duck hepatitis B virus was identified in sera and livers by the hybridization technique in 106 ducks (44 Chinese ducks, 15 Japanese ducks and 47 Japanese ducklings). Orcein-positive hepatocytes were found in 18 of 38 (47%) duck hepatitis B virus DNA seropositive ducks, and only in 3 of 68 (4%) seronegative ducks. The three ducks were all from a heavily infected flock in southern China. Serial analyses of viral DNA by Southern blot and spot hybridizations in experimentally infected Japanese ducklings revealed a dissociation or a time gap between the amount of viral DNA in serum and the emergence of orcein positive hepatocytes. Orcein-positive hepatocytes were generally associated with prolonged presence of viral infection for at least 4 to 6 months. These findings support the clinical hypothesis that the presence of orcein-positive hepatocytes indicates persistent rather than acute infection. Since orcein-positive hepatocytes have been seen in infection with hepatitis B, woodchuck hepatitis, ground squirrel and duck hepatitis B viruses, accumulation of orcein-positive material in liver cells may be one of the common properties these viruses share. This stain may be utilized for screening new hepatitis B virus-like viruses.     

乙型肝炎肝纤维化麻鸭球结膜微循环及血液流变学特点

目的:观察鸭乙型肝炎肝纤维化动物模型的球结膜微循环及血液流变学特点。方法:用鸭乙型肝炎病毒(DHBV)阳性血清反复攻击建立鸭乙型肝炎肝纤维化动物模型,并检测球结膜微循环、血液流变学指标,肝纤维化指标、肝脏病理组织学变化。结果:用DHBV反复攻击造模后,其病理组织学改变具有肝脏细胞炎症,肝纤维组织增生,血清肝纤维化指标升高(P<0.01),球结膜微循环明显障碍(P<0.01),血液流变学改变(P<0.01)。结论:乙型肝炎肝纤维化麻鸭具有明显的微循环障碍及血液流变学改变。     

Inflammation of the liver causes mutations in duck hepatitis B virus genome

To investigate whether hepatitis causes mutation in the viral genome, DNA sequences in the pre-core region of duck hepatitis B virus (DHBV) DNA were analyzed in both ducks with hepatitis and without hepatitis. Five DHBV carrier ducks were injected with DHBV particle proteins purified from duck serum with Freund’s complete adjuvant (FCA) intrahepatically from 14 day posthatch for 9 weeks (immunized group). Serum was drawn at the end of the 1st and 4th week after the 1st injection of DHBV particle protein and ducks were killed at the end of the 9th week to obtain the liver. Another five ducks without treatment were used as controls. All ducks of the immunized group showed moderate to severe hepatitis at the 9th week. All ducks in the immunized group showed one mutation except one duck that showed two mutations only at the 9th week. Mutations were observed in the 5th, 13th, 21st, 22nd, and 28th codon of the precore region. All of them were point mutation at the 3rd base in the triplets. The frequency of mutation was different in each duck from 20% to 60% but not 100%. There was no mutations in ducks in control group. These results suggest that hepatitis causes mutation in the pre-core lesion genome of duck hepatitis B virus.     

Protective and therapeutic effect of DNA-based immunization against hepadnavirus large envelope protein

BACKGROUND & AIMS: Studies in the murine model suggest that injection of DNA encoding hepatitis B virus structural proteins is promising for the induction of a specific immune response. We used the duck hepatitis B virus (DHBV) model to study the protective and therapeutic effects of naked DNA immunization against hepadnaviral large envelope protein. METHODS: A pCI-preS/S plasmid expressing the DHBV large protein was used for intramuscular immunization of ducks. The humoral response was tested by enzyme-linked immunosorbent assay, immunoblotting, neutralization, and in vivo protection tests. For DNA therapy, DHBV-carrier ducks received four injections of this plasmid. Viremia was monitored for 10 months; thereafter, liver biopsies were performed. RESULTS: Immunization with pCI-preS/S plasmid induced a specific, long-lasting, neutralizing, and highly protective anti-preS humoral response in uninfected animals. After pCI-preS/S treatment, a significant and sustained decrease in serum and liver DHBV DNA was observed for carrier ducks compared with the controls. CONCLUSIONS: DNA immunization against DHBV large protein results in a potent and protective anti-preS response in the duck model. The results of long-term follow-up of DNA-treated chronically infected ducks are promising and show the usefulness of this model for the study of genetic immunization in chronic hepatitis B therapy.     

Ampligen及其与Ganciclovir和Coumermycin Al联合应用对血清中鸭乙型肝炎病毒抑制效果

Ａｍｐｌｉｇｅｎ是一种免疫调节剂或干扰素诱导剂。单独和联合Ｇａｎｃｉｃｌｏｖｉｒ和／或ＣｏｕｍｅｒｍｙｃｉｎＡｌ治疗了鸭乙型肝炎病毒（ＤＨＢＶ）阳性鸭模型。结果表明,上述三种治疗方。案对鸭血清中ＤＨＢＶ均有抑制作用,但治疗结束ＤＨＢＶ回升到治疗前水平。任何一种方案对血清循环ＤＨＢＶ表面抗原（ＤＨＢｓＡｇ）均未产生明显抑制效果。本研究结果为人乙型肝炎治疗时合理使用抗病毒药物和免疫调节、诱导剂提供了重要参考。