HBV特异性T细胞治疗慢性乙型肝炎的研究

为寻求一种治疗慢性乙型肝炎的新方法,应用ＨＢＶ特异性Ｔ细胞输注治疗慢性乙型肝炎９例,结果显示,在疗程结束后,患者的ＨＢｓＡｇ,ＨＢｅＡｇＨＢＶＤＮＡ的含量均有所下降,其中ＨＢｅＡｇ下降较明显,Ｐ〈０．０１,２例ＨＢｃＡｇ阳性患者,ＨＢｃＡｇ阴转,ＣＤ＾＋３,ＣＤ＾＋４,ＣＤ＾＋４／ＣＤ＾＋８ＮＫ活性上升,ＣＤ＾＋８,ｓＩＬ－２Ｒ较治疗前后所回复,表明特异性Ｔ细胞治疗慢性乙型肝炎有一定疗效,远期疗效     

Phenotyping of Intrahepatic and Peripheral Blood Lymphocytes in Patients with Chronic Hepatitis C

The host immune responses have been suggested toplay a role in liver injury occurring in patients withchronic hepatitis C. In order to explore therelationship between the relative proportions ofintrahepatic and peripheral blood lymphocytes (IHL, PBL),the levels of viremia, and the histological hepatitisactivity score, three-color fluorescence-activatedcytometric analysis was performed for 36 patients with chronic hepatitis C and six control subjectswithout chronic hepatitis. The liver biopsy wasperformed before any antiviral therapy. Each liverspecimen was divided into two parts: one forhistological examination and one for immunological analysis.Tricolor CD45 was used to improvelymphogating. Fluorescein isothiocyanate-or phycoerythrin-conjugated monoclonal antibodies withspecificity for CD3, CD4, CD8, and CD20 (lymphocyte subpopulations),for CD69 (activated lymphocytes), and for CD16/56(natural killer cells) were used. The livers of patientswith chronic hepatitis C contained a greater proportion of CD4⁺ lymphocytes that exhibitedmarked expression of CD69 than in control subjects (20.7± 7.3% vs 10.2 ± 4.6%, P = 0.027).Moreover, in patients with chronic hepatitis C, theproportion of CD4⁺ IHL correlated with the histological hepatitisactivity evaluated by the Knodell score (r = 0.48, P =0.004). No correlation was found between the percentageof CD4⁺ IHL and the level of viremia ortransaminase activities. Our findings clearly indicate thata cellular immune response does take place inHCV-infected livers and could thus contribute to theoutcome of hepatitis C virus infection.     

Hepatitis B Virus Genomes of Chronic Hepatitis Patients Do Not Contain Specific Mutations Related to Acute Exacerbation

To determine the specific viral variants associated with acute exacerbation of chronic hepatitis from hepatitis B virus (HBV) infection, we analyzed the complete nucleotide sequences of the HBV genome in serial serum samples from two chronic active hepatitis patients who seroconverted from HBeAg to anti-HBe. HBV DNA was amplified by polymerase chain reaction (PCR) and sequenced. A 1896 precore stop codon mutant (G to A at nt 1896) coexisting with the wild sequence was found in both patients prior to seroconversion from HBeAg to anti-HBe. Core promoter mutations at nucleotide positions 1762 (A to T) and 1764 (G to A) were found in both patients throughout the observation period. Mutations were observed in the HBV genome of the two patients at different time points, and there was no correlation between the mutations and liver disease or DNA polymerase levels. The nucleotide divergence rate and the composition of quasispecies in the HBV sequence at the time of acute exacerbation were almost the same as were found at other time points. These results suggest that acute exacerbation does not appear to be caused by a characteristic HBV species. The multiple factors that cause generalized HBV replication activation may contribute to acute exacerbation.     

Changes of Intrahepatic Localization of Hepatitis B Core Antigen in HBeAg Positive Patients with Chronic Active Hepatitis B Treated with Interferon

The intrahepatic distribution of hepatitis B core antigen (HBcAg) was studied in 14 HBeAg-positive patients with chronic active hepatitis B who were treated with interferon. Patients received 5 to 6 million units daily of human lymphoblastoid interferon or human diploid fibroblast interferon for 28 days. Intrahepatic HBcAg was detected by the indirect immunoperoxidase technique. Before interferon therapy, the intrahepatic HBcAg was detected almost equally in both the nucleus and cytoplasm. The expression of nuclear HBcAg decreased significantly (p less than 0.001) after interferon therapy, irrespective of the outcome for DNA polymerase and/or HBeAg/anti-HBe status. The expression of cytoplasmic HBcAg decreased significantly (p less than 0.05) only in the patients who lost DNA polymerase after treatment, whereas it increased in patients with DNA polymerase and/or HBeAg in the serum. These findings suggested that a shift of intrahepatic HBcAg from the nucleus to the cytoplasm occurred in HBeAg-positive patients with chronic active hepatitis B receiving interferon therapy.     

慢性乙型肝炎拉米夫定治疗后的HBeAg早期血清转换

目的:分析拉米夫定与中成药五灵丸治疗慢性乙型肝炎的临床疗效。方法:对25例成人慢性乙型肝炎患者进行为期1年的拉米夫定的开放性临床治疗,头3个月加用五灵丸。治疗期间定期进行血清ALT、HBeAg、抗-HBe及HBV DNA检测,停药后随访半年。结果:治疗期间共有12例患者(48％)发生HBeAg转阴,并出现抗-HBe;在HBeAg血清转换后的2～4w,8例患者抗-HBe随后消失;在治疗的头3个月随着HBV DNA水平下降及消失(<420copies/ml),所有患者ALT水平逐渐降至正常。结论:治疗头3个月是应用拉米夫定治疗患者HBeAg发生血清转移的关键。这种早期转换的可能解释之一是慢性乙肝患者存在对拉米夫定特别敏感的亚型或种群。     

Kidney Transplantation in Patients with Chronic Hepatitis B Virus Infection

The impact of hepatitis B virus (HBV) infection on the long-term outcome of kidney transplant patients is controversial. A total of 34 chronic hepatitis B surface antigen (HBsAg) carriers among 143 renal allograft recipients were identified in this study (mean follow-up period: 5.6 ± 3.3 years; range: 1–13 years). During the follow-up, one HBsAg-positive recipient with preexisting cirrhosis died of liver failure, and seven (21%) others developed serious HBV-related complications (four fulminant hepatitis, two hepatocellular carcinoma, one cirrhosis), and four died. Although HBsAg-positive recipients had a higher rate of liver-related complications and deaths than HBsAg-negative recipients did, there were no significant differences in the long-term graft and patient survival between the two groups. The survival rates, liver-related complications, and deaths in HBsAg-positive allograft recipients and 28 HBsAg-positive uremic patients under dialysis were similar. In conclusion, HBV infection is not a contraindication to kidney transplantation. However, pretransplant candidates should be warned of potentially serious liver-related complications.     

Quasispecies and Pre-Existing Drug-Resistant Mutations of Hepatitis B Virus in Patients with Chronic Hepatitis B

Background/Aims

To investigate pre-existing hepatitis B virus (HBV) quasispecies and the genotypic evolution of several variants.

Methods

From six patients with lamivudine (LAM) failure, serum samples at pretreatment, 6 months of LAM therapy, and virologic breakthrough were obtained. One hundred clones with HBV inserts in each patient were sequenced at each time point. Pretreatment serum samples were also analyzed from six patients who achieved good responses to LAM therapy.

Results

Among the six patients with LAM failure, the analysis of 100 clones from patient 1 revealed the substitutions L180M in 1% of clones and V173L in 2% of clones. Patient 2 had substitutions of L80V, W153Q, and L180M. In patient 3, mutations conferring resistance to adefovir at V84I (5%), I169L (1%), and N236H (7%) and entecavir at S202G (2%) were detected. Patient 4 had mutations at T128N (1%), I169L (1%), V173L (2%), A181V (1%), and Q215H (1%). In patient 5, M204V/I was detected in 1% and 2% of clones, respectively. L80I and V173L were also identified in patient 6. In the six patients who responded to LAM, the degree of overall quasispecies was less than those with LAM failure.

Conclusions

Various HBV quasispecies associated with drug resistance existed before treatment, and the quasispecies dynamically changed through LAM therapy.     

Hepatitis B e Antigen Seroconversion: A Critical Event in Chronic Hepatitis B Virus Infection

Background  

Replication of hepatitis B virus (HBV) is the primary driver of disease progression and clinical outcomes in patients with chronic hepatitis B (CHB), but other factors, such as hepatitis B e antigen (HBeAg) status, also influence disease course. The importance of HBeAg seroconversion is underscored by current CHB treatment guidelines that recommend limiting the duration of antiviral therapy in HBeAg-positive patients who achieve seroconversion.     

Intrahepatic Expression of C-C Motif ligand 5 in Patients with Chronic Hepatitis B

Background: C-C motif ligand 5 (CCL5) is reported to play a key role in acute and chronic liver diseases. However, the association between CCL5 and chronic hepatitis B (CHB) remains to be explored. We aimed to investigate the CCL5 expression in the liver tissues of CHB patients and compared the CCL5 expression among CHB patients with different stages of liver inflammation and fibrosis. Methods: Liver tissue specimens from 51 CHB patients who underwent liver biopsy and twelve healthy liver donors were included in the present study. CCL5 expression in the liver tissues was analyzed using immunohistochemistry. The hepatic inflammation grades and fibrotic stages of CHB patients were assessed by the Scheuer classification system. Results: Livers of CHB patients exhibited significantly accumulated CCL5+ cells when compared to those of healthy controls (42.80 ± 4.37 vs. 7.25 ± 0.99/HPF, P < .001). CHB patients with higher hepatic inflammation grades had more CCL5+ cells in their livers than those with lower grades (P < .05). However, the numbers of CCL5+ cells were not correlated with the fibrotic stages in CHB patients (r = .073, P = .61). The number of CCL5+ cells in the liver tissues of CHB patients was positively correlated with alanine transaminase levels (r = .278, P = .041) and aspartate aminotransferase levels (r = .328, P = .009). Conclusions: CHB patients have a significant accumulation of CCL5+ cells in the liver, and CCL5 may play a pathological role in hepatic inflammation of CHB.     

Sterilized Hepatitis B (Surface) Antigen for Production of Specific Antisera

The combined treatment with beta-propiolactone and ultraviolet rays described for the sterilization of HBsAg-positive sera does not interfere with the purification procedures for the production of HBs antigen. The immunogenicity of the HBs antigen is retained.     

急慢性丙型肝炎患者进行乙型肝炎病毒标志物检测的临床意义

采用ＥＩＡ、ＰＣＲ技术对急、慢性丙肝患者血清标本进行乙型肝炎病毒标志物、丙型肝炎不同功能区抗体和ＨＢＶＤＮＡ、ＨＣＶＲＮＡ检测。结果：排除甲、戊型肝炎的急、慢性丙肝的乙肝感染率的７４．６７％和５９．０９％均以ＨＢｃＡｂ和ＨＢｓＡｂ为主,其他出现,有乙肝三内肝ＡＬＴ值较高。ＨＢＶＤＮＡ检出率较低,且与ＨＣＶＲＮＡ极少同时检出;急、慢性丙肝ＨＣＶＲＮＡ检出率有乙肝旮明显偏低。有乙肝感染时急性丙肝中Ｅ     

乙、丙型肝炎病毒在慢性肝炎、肝硬化患者胃黏膜表达的临床研究

目的:探讨乙、丙型肝炎病毒(HBV、HCV)的泛嗜性.方法:选择慢性乙、丙型肝炎(慢肝组)28例、肝炎肝硬化(肝硬化组)44例,共72例作为研究对象.受检者常规胃镜检查,取胃窦幽门周围3cm以内活体组织两块,除普通病理检查外,分别做乙型肝炎病毒表面抗原(HBsAg)、乙型肝炎病毒核心抗原(HBcAg)、丙型肝炎病毒抗原(HCVAg)免疫组化法检测.结果:慢肝组有不同程度的胃黏膜慢性炎症者达92.9%(26/28)、肝硬化组达95.5%(42/44),排除年龄影响因素外,慢肝组以单纯慢性炎症为多,而肝硬化组以伴萎缩和肠化者为多.慢肝组与肝硬化组患者分别有53.6%(15/28)、81.8%(36/44)胃黏膜HBVAg阳性,其中HBsAg、HBcAg双阳性31例.在51例患者胃黏膜HCVAg检测中有33例(占64.7%)阳性表达、66.7%(22/33)与HBcAg同时表达.肝硬化组HBVAg及HBsAg、HBcAg双阳性者均高于慢肝组(P值均＜0.05).结论:HBV、HCV在慢性及肝硬化患者胃黏膜表达明显,应重视其在胃黏膜病变发病中的作用,并加强防护措施.     

慢性肝病患者肝组织中庚型肝炎病毒抗原的定位研究

应用抗HGV NS5单克隆抗体(McAb),以辣根过氧化物酶与抗辣根过氧化物酶法(PAP)检测了166例慢性肝病患者(CLD)肝组织中庚型肝炎病毒抗原(HGV-Ag)。结果显示,慢性非甲～戊型肝炎、慢性丙型肝炎、慢性乙型肝炎、肝硬变和肝细胞癌患者阳性检出率分别为28.57％、22.45％、14.28％、18.52％和15.38％。HGV-Ag阳性肝细胞多呈散在、局灶和弥漫性三种形式分布,HGV-Ag染色阳性颗粒可见于细胞浆中,HGV-Ag NS5的检出率与HBV、HCV感染有关。提示HGV NS5抗原在慢性肝病患者肝细胞中表达,且HGV NS5的表达与血清HGV RNA水平关系较为密切,HGV感染可能在慢性肝病发生中起一定病原学作用。     

Hepatitis B Virus Vaccine for Patients with Hepatitis C Virus Infection

Summary Hepatitis C is a disease with varying rates of progression. The role of hepatitis B virus (HBV) as a cofactor in the development of hepatitis C virus (HCV)-related cirrhosis and hepatocellular carcinoma (HCC) has been suggested and the use of HBV vaccine in all HCV-infected patients has been advocated. This review presents the implications of HBV and HCV coinfection and addresses the issues of HBV vaccine immunogenicity and safety in patients with chronic HCV infection. Received: December, 1999 · Revision accepted: July 1, 2000     

A Pilot Study of Long-Term Weekly Interferon-β Administration for Chronic Hepatitis B

Interferon-β was given weekly for 24 wk, at a dose of 3 million units, intravenously, to 10 patients with chronic hepatitis B who were serologically positive for HBsAg and HBeAg. Their condition was followed for 6 months after the end of therapy. Both serum hepatitis B virus-associated DNA-polymerase activity and alanine aminotransferase level became significantly lower during therapy and during the 6 months after the end of therapy than at the beginning of therapy. In five of 10 patients, the seroconversion from HBeAg positive to anti-HBe positive had occurred by 6 months after the end of therapy, and in four of these five patients, serum alanine aminotransferase level became normal. Weekly interferon-β administration over 6 months seems effective in inducing seroconversion and in normalizing serum alanine aminotransferase level.