Anti-Saccharomyces cerevisiae antibodies (ASCA) and autoimmune liver diseases

Antibodies to the baker's yeast Saccharomyces cerevisiae (ASCA), recently proposed as a serological marker of Crohn's disease, have also been detected in other autoimmune disorders. The aim of this study was to determine prevalence and clinical significance of ASCA in autoimmune liver disease. The presence of IgG and IgA ASCA was evaluated using a commercially available immunoassay in 215 patients with autoimmune liver disease (primary biliary cirrhosis, PBC, 123 cases; autoimmune hepatitis, AIH, 67 cases; primary sclerosing cholangitis, PSC, 25 cases), 48 with inflammatory bowel disease and 19 healthy blood donors. Anti neutrophil cytoplasmic antibodies with the perinuclear pattern (p-ANCA) were assessed by indirect immunofluorescence in PSC patients. The main clinical and biochemical parameters between ASCA-positive and negative patients were analysed and compared. ASCA are predominant in Crohn's disease (70%); among liver patients, PSC and AMA-negative PBC show the highest ASCA prevalence (53% and 44%). In PBC ASCA correlate with higher levels of circulating IgA (P < 0.05). In PSC the detection of either ASCA or p-ANCA is neither associated with any clinical or biochemical feature, nor with an underlying inflammatory bowel disease. ASCA can not be considered an additional serological marker of autoimmune liver disease, but the possibility of detecting such a reactivity in autoimmune liver disorders should be considered; their correlation with elevated IgA in PBC suggests that ASCA may be an indirect sign of enhanced mucosal immunity; in PSC patients neither ASCA nor p-ANCA predict the occurrence of a concomitant inflammatory bowel disease.     

自身免疫肝病患者抗中性粒细胞胞浆抗体的检测及临床意义

探讨抗中性粒细胞胞浆抗体(anti-neutrophil cytoplastic antibodies,ANCA)对自身免疫肝病的临床意义。应用间接免疫荧光法和斑点法检测149例自身免疫性肝病患者[自身免疫性肝炎(autoimmune hepatitis,AIH)患者57例,原发性胆汁性肝硬化(primary biliary cirrhosis,PBC)患者42例,不明原因肝损患者50例]的ANCA和抗可提取性核抗原抗体(extract-able nuclear antigen,ENA),进而用ELISA法分析60例ANCA阳性的自身免疫肝病患者的ANCA抗原谱。以200例健康献血员为正常对照。结果ANCA在AIH、PBC、不明原因肝损中的阳性率分别为81%、40%、30%,其中非典型性pANCA的阳性率依次为70%、40%、28%。AIH组与PBC组及AIH组与不明原因肝损组间非典型性pANCA的阳性率有显著性差异(P<0.01),但PBC组与不明原因肝损组间差异不显著(P>0.05)。各疾病组ANCA抗原谱如下:AIH组中,乳铁蛋白3%阳性,MPO 11%阳性,组织蛋白酶G和BPI的阳性率分别为11%、17%;PBC组中,弹性蛋白酶和组织蛋白酶G阳性率均为5%,BPI的阳性率为16%;不明原因肝损组中,BPI阳性率为17%。大多数自身免疫性肝病患者非典型性pANCA为阳性(28%~70%),且伴有特征性自身抗体。注重非典型性pANCA的检测对明确诊断自身免疫肝病及其分类有很大的帮助。     

Decreased serum DNase1-activity in patients with autoimmune liver diseases

Deoxyribonuclease1 (DNase1) is involved in chromatin degradation of apoptotic cells. Its deficiency results in accumulation of self-DNA, which in turn may induce inflammation and autoimmunity. We assessed for the first time serum DNase1-activity in a large consecutive cohort of treatment-naïve patients with autoimmune liver diseases (ALD). DNase1-activity was determined by single radial enzyme-diffusion (SRED) at diagnosis of 224 patients with autoimmune hepatitis (AIH), 249 with primary biliary cirrhosis (PBC) and 36 with primary sclerosing cholangitis (PSC). Sera from 146 patients with chronic hepatitis B or C, 140 with nonalcoholic fatty liver disease/nonalcoholic steatohepatitis (NAFLD/NASH) and 114 healthy individuals served as disease and healthy controls. Available serum samples during remission from 50 AIH and 39 PBC patients were also investigated by paired analyzes. DNase1-activity was significantly lower in AIH, PBC and PSC compared to viral hepatitis (p?p?p?=?0.03), NAFLD/NASH (p?p?p?p?1400?mg/dl; p?p?p?p?=?0.008). In PSC, DNase1-activity was inversely associated with alkaline phosphatase (ALP) (p?p?p?相似文献   

In situ mass spectrometry of autoimmune liver diseases

Primary biliary cirrhosis (PBC), primary sclerosing cholangitis (PSC) and autoimmune hepatitis (AIH) are the major forms of autoimmune liver diseases each characterized by the destruction of a specific liver cell type and the presence of differing auto-antibodies. We took a proteomic approach utilizing in situ matrix-assisted laser desorption/ionization mass spectrometry (MALDI MS) to obtain profiles directly from liver samples of patients with PBC, PSC, AIH and controls. The ability to precisely localize the region for acquisition of MALDI MS allowed us to obtain profiles from bile ducts, inflammatory infiltrates and hepatocytes from each biopsy sample. Analysis tools developed to identify peaks and compare peaks across diseases and cell types were used to develop models to classify the samples. Using an initial set of testing samples from PBC patients and controls, we identified unique peaks present in bile ducts, inflammatory infiltrates and hepatocytes that could classify samples in a validation cohort with 88-91% accuracy. Interestingly, profiles of PSC and AIH did not differ significantly from PBC. Identification of proteins in these peaks may represent novel autoantigens or effector molecules. These findings illustrate the potential of a proteomic approach to autoimmune diseases with in situ MALDI MS.     

实验性变态反应性脑脊髓炎大鼠单核细胞趋化蛋白-1的表达与临床评分的相关性

目的：探讨实验性变态反应性脑脊髓炎（EAE）大鼠脑和脊髓中单核细胞趋化蛋白-1（MCP-1）的表达及其与临床评分的关系。方法：将Wistar大鼠分为正常组、佐剂（CFA）组和EAE组,取脑和脊髓制成石蜡切片,进行HE染色和MCP-1mRNA的原位杂交,并与各项临床指标比较。结果：EAE组的体重减轻、MCP-1 mRNA表达的阳性细胞百分数与正常大鼠、佐剂组相比显著性增加。EAE组大鼠MCP-1 mRNA的表达呈动态性变化,其MCP-1 mRNA先于临床症状高表达,并随临床评分的升高而升高,与临床评分呈正相关。结论：MCP-1是参与EAE发病的重要的炎性介质。     

Role of liver biopsy in autoimmune liver disease

This article will review histological aspects of three chronic liver diseases – autoimmune hepatitis (AIH), primary biliary cirrhosis (PBC) and primary sclerosing cholangitis (PSC) – in which autoimmune mechanisms are thought to be involved. The changing role of liver biopsy in the diagnosis and management of patients with autoimmune liver disease will also be discussed. In the case of autoimmune hepatitis, histological assessments remain important in establishing a diagnosis, identifying prognostic features and monitoring therapeutic responses. By contrast, for many patients with PBC and PSC a diagnosis can now be made on the basis of biochemical, serological and/or radiological findings alone and histological confirmation may not be required. Liver biopsy can still be used to assess disease severity in such cases and remains important in establishing a diagnosis in patients with atypical features (e.g. AMA-negative PBC or the small-duct variant of PSC). Liver biopsy is also increasingly used in the assessment of patients suspected to have “overlap syndromes” involving AIH and PBC or PSC.     

Intrahepatic expression of the co-stimulatory molecules programmed death-1, and its ligands in autoimmune liver disease

Liver-infiltrating T cells play an essential role in the immunopathogenesis of autoimmune liver disease. Programmed death-1 (PD-1) and its ligands, B7-H1/PD-L1 and B7-DC/PD-L2, are new CD28-B7 family members that are involved in the regulation of immune responses. The ligation of PD-1 inhibits T-cell receptor-mediated T cell proliferation and cytokine production, and PD-1-deficient mice develop various organ-specific autoimmune diseases. To investigate the expressions of PD-1 and its ligands in autoimmune liver disease, in particular autoimmune hepatitis (AIH) and primary biliary cirrhosis (PBC), immunohistochemical analysis was performed. Liver biopsy specimens obtained from 17 patients with AIH and PBC were studied. PD-1 was expressed on more than half of the liver-infiltrating T cells within the portal tract. Some of the intrahepatic T cells expressed B7-H1 in patients with AIH and PBC. B7-H1 and B7-DC were mainly expressed on some Kupffer cells (KC) and liver sinusoidal endothelial cells (LSEC) within the sinusoids and their expression was upregulated in autoimmune liver disease. These results suggest that the interaction of PD-1 on T cells with increased expression of B7-H1 and B7-DC on KC and LSEC might be involved in the downregulation of autoreactive lymphocytes and result in the regulation of pathogenesis in autoimmune liver disease.     

Immunoglobulin classes in biopsies of autoimmune liver disease

The immunological isotypes of plasma cell infiltrates in a series of consecutive liver biopsies from patients with chronic active hepatitis (CAH) and primary biliary cirrhosis (PBC) were determined immunohistochemically. The plasma cell infiltrate was more pronounced in PBC than in CAH. IgA and IgG isotypes predominated in CAH, and IgM and IgG in PBC. The expected predominance of kappa light chains was observed in every biopsy in PBC. However, in 8/14 CAH biopsies the plasma cells were predominantly lambda isotype. Lambda predominance was significantly associated with the presence of serum autoantibodies. These findings would suggest that different mechanisms operate in the pathogenesis of these two autoimmune liver diseases.     

肝病患者自身抗体特征性的研究

目的：观察抗肝抗原自身抗体在我国不同类型肝病患者中的存在状况；探讨自身免疫性肝脏疾病的自身抗体特征。方法：间接免疫荧光法初筛1412例肝功能异常血清，从中选择28例：①自身免疫性肝病组42例：初步诊断为AIH18例、PBC21例、PSC3例。②HAV组23例；③DBV组70例；④HCV组33例；⑤非甲—非戊型肝炎组60例。结合Western blot、酶免疫条带技术等分别检测ANA、AMA、SMA、LKM—1、LC—1、SLA/LP和AMA-M2亚型、dsDNA及ENA类多种抗体。结果：1412例中诊断AIH、PBC和PSC者分别为送检标本的12．7‰，14．9‰和2．1‰。28例血清中2例LKM—1阳性和2例SLA／LP阳性；按AIH的分型标准，自身免疫性肝病组属于I型AIH者14例(78％)，Ⅱ型2例(11％)，Ⅲ型2例(11％)；AIH患者ANA抗体未见特定的荧光类型。PBC患者AMA和M2全部阳性；其ANA以核膜型为主(7／14)。NonA—E组4例AMA和M2阳性，3例SMA高滴度阳性，4例出现SS-A、SS-B或dsDNA等抗体。结论：三型自身免疫性肝炎在中国都存在，肝抗原自身抗体和ANA及AMA的分型检测有助于自身免疫性肝病的诊断与治疗；少数非甲—非戊型肝炎应考虑自身免疫性肝病诊断。     

Expression of the interferon-inducible chemokine IP-10 (CXCL10), a chemokine with proposed anti-neoplastic functions, in Hodgkin lymphoma and nasopharyngeal carcinoma

Hodgkin lymphoma (HL) and nasopharyngeal carcinoma (NPC) are characterized by their association with Epstein-Barr virus (EBV) and an abundant infiltrate of reactive lymphoid cells. The presence of this lymphoid stroma may influence the effect of anti-viral immunotherapy. The interferon-inducible chemokine IP-10 has anti-neoplastic effects in several model systems mediated by T-cells expressing the CXCR3 chemokine receptor. Using in situ hybridization, it is shown that IP-10 is expressed in neoplastic cells of HL and correlates both with the mixed cellularity histotype and with EBV infection. IP-10 expression was also detected in tumour cells of most NPCs as well as in EBV-negative squamous cell carcinomas of the tongue. Thus, in carcinomas, IP-10 expression showed no correlation with EBV infection. Numerous CXCR3-positive lymphocytes were detected in the lymphoid stroma of HL and NPC, raising the possibility of a Th1-predominant immune response in these cases. In view of the proposed anti-neoplastic functions of IP-10 and CXCR3-positive lymphocytes, these findings are unexpected and raise the possibility that endogenous IP-10 expression in the context of human tumours may not exert the anti-tumour effects ascribed to it by in vitro experiments.     

不同肝病患者抗肝抗原自身抗体的研究

目的：观察我国不同类型肝病患者中几种抗肝抗原自身抗体的存在状况；探讨自身免疫性肝脏疾病的自身抗体特征。方法：由1412例标本中选择230例肝功能异常患者分为5组：①自身免疫性肝病组42例：自身免疫性肝炎（AIH）18例、原发性胆汁性肝硬化（PBC）21例、原发性硬化性胆管炎（PSC）3例。②HAV组23例；③HBV组70例；④HCV组35例；⑤非甲-戊型肝炎组60例。用间接免疫荧光、Western blot、酶免疫条带技术等分别检测抗核抗体（ANA）、抗线粒体抗体（AMA）、平滑肌抗体（SMA）、肝肾微粒抗体I型（LKM-1）、肝细胞胞溶质抗原I型（LC-1）、可溶性肝抗原（SLA）/肝胰抗原（LP）和AMA-M2亚型，以及SS-A、SS-B、dsDNA等多种抗体。结果：1412例中诊断AIH、PBC和PSC者分别为送检标本的1.27%，1.49%和0.21%。230例血清中2例LKM-1阳性和2例SLA/LP阳性，分别见于AIH和HCV感染者。PBC患者AMA和M2全部阳性；其ANA以核膜型为主（7/14）；AIH患者ANA抗体未见特定的荧光类型，而抗-Actin仅见于AIH者。非甲-戊组4例AMA和M2阳性，3例SMA高滴度阳性，4例出现SS-A、SS-B或dsDNA等抗体。结论：肝抗原抗体和ANA及AMA分型的检测有助于自身免疫性肝端正和重叠多种免疫性肝病的诊断；非甲-戊型肝炎诊断时应考虑自身免疫性疾病。     

Reduction in serum levels of antimitochondrial (M2) antibodies following immunoglobulin therapy in severe combined immunodeficient (SCID) mice reconstituted with lymphocytes from patients with primary biliary cirrhosis (PBC)

The effect of gammaglobulin treatment on autoantibody production was investigated in SCID mice reconstituted with human peripheral blood mononuclear cells (PBMC) obtained from patients with PBC. All reconstituted mice displayed the presence of human antimitochondrial antibodies (αM2Ab) of both IgG and IgM types before treatment with human immunoglobulin. Two weeks after i.p. injection of 20 ×10⁶ PBMC into SCID mice, i.p. treatment with various preparations of human immunoglobulin was initiated. In control animals treated with saline, serum levels of human αM2Ab of the IgG type increased with time, peaking around 4 weeks after reconstitution. In contrast, human IgG autoantibodies rapidly decreased in all animals treated with human IgG. Treatment with a human IgM preparation had no effect on serum levels of αM2Ab of the IgG type. The results may suggest that the pronounced reduction of specific IgG autoantibodies was due to an increased catabolism of human IgG, including the autoantibodies, in the gammaglobulin-treated mice. Although the production of human αM2Ab in reconstituted mice could be easily shown, PBC-specific liver lesions or bile duct destruction were not observed, irrespective of treatment protocol.     

中国人自身免疫性肝病相关性 CTLA-4基因多态性研究

目的　探讨细胞毒性 T细胞相关抗原 - 4 (cytotoxic T lymphocyte- associated antigen- 4 ,CTL A- 4 )基因启动子 - 318和第 1外显子区第 4 9位基因多态性与中国人自身免疫性肝炎 (autoimmunehepatitis,AIH)、原发性胆汁性肝硬化 (primary biliary cirrhosis,PBC)发病的相关性。方法　应用限制性片段长度多态性方法分析 6 2例 AIH和 77例 PBC患者外周血单核细胞基因组 DNA CTL A- 4启动子 -318T/ C、第 1外显子区第 4 9位基因 A/ G多态性 ,并与 16 0名正常对照比较。结果　 AIH组 CTL A- 4启动子 - 318位 T/ C基因型分布与对照组比较差异无显著性 ,但 C等位基因频率明显高于正常对照组 (P=0 .0 2 ,OR=2 .4 3)。 PBC患者 CTL A- 4第 1外显子区第 4 9等位基因分布与正常对照组比较差异非常显著(P=0 .0 0 6 ) ,PBC患者 G等位基因频率明显高于正常组 (P=0 .0 0 4 6 ,OR=1.8)。联合分析 CTL A- 4启动子与第 1外显子的基因多态性分布 ,虽然 AIH组和 PBC组 GG- CC型携带率均比正常人高 (AIH组 :32 .3% ,PBC组 :37.7% ,对照组 :2 2 .5 % ) ,但是统计学分析结果均显示两组患者与正常人差异无显著性。结论　 CTL A- 4启动子 - 318和第 1外显子区第 4 9位基因多态性可能与中国人 AIH、PBC易感性相关。     

Immunoglobulin-containing cells in liver biopsies of patients with chronic hepatitis and primary biliary cirrhosis

The numbers of IgA, IgM and IgG-containing cells were studied by means of an indirect immunoperoxidase technique and morphometry in liver biopsies of patients with primary biliary cirrhosis and chronic hepatitis, in whom serum immunoglobulin concentrations were also determined. In patients with primary biliary cirrhosis the absolute and relative number of IgM-containing cells in the liver was significantly higher, whereas the absolute and relative number of IgG-containing cells in the liver was significantly lower compared to patients with chronic hepatitis. IgM-containing cells in liver biopsies of patients with primary biliary cirrhosis correlated strongly with their serum IgM levels. It is concluded that determination of the pattern of immunoglobulin containing cells in liver biopsies may help in the differentiation of primary biliary cirrhosis from chronic hepatitis and that local production of IgM in the liver may contribute significantly to the high serum IgM levels in patients with primary biliary cirrhosis.     

原发性胆汁性肝硬化患者外周血单个核细胞中Tim-3 mRNA表达增高及其意义

为研究Tim-3是否参与了原发性胆汁性肝硬化(PBC)的发病机制,我们采用实时荧光定量逆转录-聚合酶链反应(RT-PCR)技术检测了38例PBC患者及30例健康者外周血单个核细胞(PBMC)中Tim-3mRNA的相对表达量,并分析Tim-3mRNA表达与PBC患者Mayo危险评分和碱性磷酸酶(ALP)之间的关系。结果表明,PBC患者外周血PBMC中Tim-3mRNA表达较健康对照组明显增高(P<0.01),且与Mayo危险评分呈正相关(r2=0.31,P<0.01),与血清ALP水平呈负相关关系(r2=0.37,P<0.01)。本研究表明Tim-3可能参与了PBC的发病机制,同时还是PBC的潜在标志物。     

Autoimmune hepatitis in primary Sjogren's syndrome: pathological study of the livers and labial salivary glands in 17 patients with primary Sjogren's syndrome

Although primary Sjogren's syndrome (pSS) is an autoimmune exocrinopathy, the involvement of liver has been reported. Because no study focusing on autoimmune hepatitis (AIH) in pSS has been published, the purpose of the present study was to perform a clinical and histological examination of the liver, focusing on AIH, in 17 pSS patients. The patients had liver enzyme abnormalities without hepatitis virus infection. In all cases, biopsied livers were examined, and in 10 cases biopsied labial salivary glands were also examined histologically. Based on the authors' diagnostic criteria for AIH in pSS, the liver diseases consisted of AIH (eight cases, 47%), primary biliary cirrhosis (PBC; six cases, 35%), non-specified chronic hepatitis (two cases, 12%) and acute hepatitis (one case, 6%). Lymphoplasmacytic infiltrate, with predominancy of CD3(+) T cells, was noted in both the liver and salivary glands in the patients with AIH. The patients with AIH with severe interface hepatitis had a good response to immunosuppressive therapy. The comparison of liver histology between the PBC with pSS group and the PBC without pSS group showed that the incidence of lymphoid non-suppurative cholangitis was higher in PBC with pSS. In conclusion, the present study offers new information on the relatively common occurrence, diagnostic criteria and treatment effects of AIH in pSS.     

Peribiliary capillary plexus around interlobular bile ducts in various chronic liver diseases: An immunohistochemical and morphometric study

Peribiliary capillary plexus (PCP) is a network of capillaries which arise from the hepatic artery surrounding the intrahepatic bile ducts. We immunohistochemically investigated the density of PCP around interlobular bile ducts in various chronic liver diseases including primary biliary cirrhosis (PBC) (n = 47), autoimmune hepatitis (AIH) (n = 12), chronic hepatitis B (n = 16), chronic hepatitis C (n = 19), liver cirrhosis due to hepatitis B virus (n = 13), liver cirrhosis due to hepatitis C virus (n = 20), alcoholic hepatitis (n = 20), alcoholic liver cirrhosis (n = 17), using human liver biopsies fixed in formalin and embedded in paraffin wax. PCP was immunohistochemically detected by an endothelial marker, the CD34 antigen. The number of PCP per duct was 1.21 +/- 0.18 in normal livers. Compared with normal liver, vasopenia was observed in PBC and AIH, the number in which was 0.93 +/- 0.34 (P < 0.0001) and 0.82 +/- 0.38 (P < 0.005) per duct, respectively. In contrast, increased number of PCP was observed in liver cirrhosis due to hepatitis B or C virus, alcoholic hepatitis, and alcoholic liver cirrhosis, the number in which were 1.59 +/- 0.37 (P < 0.005), 1.55 +/- 0.52 (P < 0. 02), 1.38 +/- 0.23 (P < 0.02) and 1.61 +/- 0.33 (P < 0.002) per duct, respectively. These data suggest that PCP may be destroyed in autoimmune liver diseases, including PBC and AIH, but PCP may proliferate in other inflammatory and alcoholic liver diseases.     

Changes of inflammatory cytokines/chemokines during ravidasvir plus ritonavir-boosted danoprevir and ribavirin therapy for patients with genotype 1b hepatitis C infection

This study investigated the safety and efficacy of ravidasvir (RDV) plus ritonavir-boosted danoprevir (DNVr) and ribavirin (RBV) regimens for treatment-naïve non-cirrhotic patients with hepatitis C virus (HCV) genotype 1b in mainland China. We also gained insight into HCV-host interactions during anti-HCV treatment. 16 patients with HCV and 10 healthy people enrolled the study. Three of 16 patients received 12-weeks' placebo treatment first and served as the placebo controls. All (n = 16) patients received 12-weeks' RDV plus DNVr and RBV treatment. The adverse effects (AEs), viral loads, alanine transaminase, and aspartate aminotransferase were recorded during study. We also performed multianalyte profiling of 48 cytokines/chemokines in 16 patients with HCV and 10 normal controls. Seventy-five percent patients treated with RDV plus DNVr and RBV experienced AEs. No death, treatment-related serious AEs or AEs leading to discontinuation were reported. The serum HCV-RNA levels remained extremely high in 3 placebo controls after treated with placebo. After RDV plus DNVr and RBV treatment, all patients achieved sustained virologic response (SVR) at posttreatment week 12, but 1 patient experienced viral relapse at SVR 24. The cytokine/chemokine expression pattern was markedly altered in patients with HCV as compared with healthy controls. The interferon-inducible protein-10 (IP-10) decreased after anti-HCV treatment, and dramatically increased in one patient with viral relapse. The regimen of RDV and DNVr plus RBV represents a highly safe and effective treatment option for HCV patients in mainland China. The IP-10 has the potential to be an indicator of innate immune viral recognition.