慢性乙肝患者抗病毒治疗前后特异性T细胞免疫观察

目的 了解抗病毒治疗前后慢性乙型肝炎患者特异性T淋巴细胞对HBV抗原蛋白免疫应答的变化及其特征.方法 收集17例慢性乙型肝炎患者抗病毒治疗前及治疗后1个月、3个月的外周血单个核细胞,以HBV特异性抗原蛋白HBsAg、HBcAg和HBeAg为刺激物,酶联免疫斑点法检测其分泌IFN-γ产生斑点的情况.同时对血清HBV DNA和HBsAg、HBeAg等病毒学指标及谷丙转氨酶(ALT)等生化学指标进行榆测并分析其相关性.结果 治疗前,所有患者ALT、总胆红素(TBiL)均高于正常上限,17例患者HBV DNA均大于104拷贝/ml;治疗1个月后,ALT复常率为35.3%,9例患者HBV DNA降为检测下限以下;治疗3个月后,ALT复常率为58.8%,有11例患者HBV DNA降为检测下限以下.抗病毒治疗前、治疗1个月、治疗3个月患者针对HBV特异性蛋白总的T细胞反应阳性率分别为64.7%、76.5%和82.4%,其差别无统计学意义.不论治疗前后,患者对HBeAg的特异性T细胞反应频率和平均反应强度最高;治疗后,对3种蛋白的特异性T细胞反应频率和平均反应强度各有不同程度的增加,其中以对HBcAg蛋白的平均反应强度的增强最明显,治疗前和治疗3个月,治疗1个月和治疗3个月之间的差别都有统计学意义.患者对HBcAg蛋白的特异性T细胞反应平均反应强度与病毒载量有明娃负相关,与血清ALT无明显相关性.结论 本研究结果提示抗病毒治疗后,患者对HBV的特异性T细胞免疫应答有所增强,这种改变可能与HBV DNA的下降有关,检测HBV特异性T细胞反应对丁解患者的免疫状态有重要的意义.

Abstract：

Objective To explore the responses of antigen-specific T cells stimulated by hepatitis B virus(HBV)-specific proteins in chronic hepatitis B patients accepting antiviral therapy. Methods Seventeen patients with chronic hepatitis B (CHB) accepting antiviral therapy were included in this study. The peripheral blood monocular cell ( PBMC) were separated from the whole blood collected at the three different time of before and one and three months after accepting antiviral therapy. ELISPOT assay was used to detect the frequency and strength of secreting IFN-γ cells of PBMC stimulated by HBsAg, HBcAg and HBeAg. HBV virus loading, HBsAg, HBeAg, ALT and AST in serum were detected at the same time. Results After three months therapy, ALT, TBiL were improved in all patients, and HBV DNA level were dropped and undetectable in 11 cases. The rates of T cell response in patients to HBV specific proteins were 64. 7% , 76. 5% and 82. 4% at the time of before and one and three months after accepting antiviral therapy, respectively. The frequency of responses of antigen-specific T cells stimulated by HBcAg was higher than that stimulated by HBsAg or HBeAg, and the frequency was enhanced after antiviral therapy. The average response magnitude was expressed as spot forming cells (SFC) per million input cells. SFC of T cell responses to HBcAg was also higher than to HBsAg or HBeAg. There was no significant difference in SFC of T cell responses to HBsAg or HBeAg at the time of before and after antiviral therapy, but there were significant difference in SFC of T cell responses to HBcAg at the time of before and after antiviral therapy. SFC of T cell responses to HBcAg was negatively associated with HBV DNA, and no associated with level of ALT in serum. Conclusion The responses of antigen-specific T cells were improved in CHB patients accepting antiviral therapy which associated with the decrease of HBV DNA. It suggested to investigate HBV specific T cell responses was important.     

幽门螺杆菌黏附素A有效T和B联合抗原表位噬菌体展示和抗原性测定

Objective To analyze and determine the efficient T- and B-combined (T/B) antigenic epitopes in Helicobacter pylori adhesin A. Methods Recombinant HpaA (rHpaA) was expressed for immunizing rabbit to generate antiserum. T- and B-cell epitopes in HpaA molecule were predicted by using bioinformatic technique. The segments to encode T/B combined epitope peptides were amplified by PCR and the phage display systems of T/B combined epitopes were subsequently constructed. PEG/NaCl precipitation method was applied to extract the recombinant phage PⅢ (rPⅢ) that displayed T/B combined epitopes. By using either commercial IgG against whole-cell of Helicobacter pylori or rHpaA antiserum as the primary antibody, the T/B combined epitopes displayed in rP Ⅲ s were screened and identified by Western blot and ELISA. MTT was applied to determine the proliferation of rHpaA-immunized mouse splenocytes after stimulation of the different recombinant rPⅢ proteins. Results In the HpaA molecule there were five T/B combined epitopes: HpaA10, HpaA37, HpaA79, HpaA116 and HpaA143. All the T/B combined epitopes were successfully displayed on the surface of PⅢ protein of phage M13. The results of Western blot, ELISA and MTT confirmed that HpaA116 was the predominant antigenic epitope, both HpaA37 and HpaA79 were the efficient antigenic epitopes. However, HpaA10 and HpaA143 were identified as ineffective antigenic epitopes. Conclusion The phage display systems of T/B combined epitope peptides of H. pylori adhesin A have been successfully generated in this study. HpaA37 and HpaA79, especially HpaA116 are the efficient T/B combined antigenic epitopes in HpaA of H. pylori.     

幽门螺杆菌黏附素A有效T和B联合抗原表位噬菌体展示和抗原性测定

Objective To analyze and determine the efficient T- and B-combined (T/B) antigenic epitopes in Helicobacter pylori adhesin A. Methods Recombinant HpaA (rHpaA) was expressed for immunizing rabbit to generate antiserum. T- and B-cell epitopes in HpaA molecule were predicted by using bioinformatic technique. The segments to encode T/B combined epitope peptides were amplified by PCR and the phage display systems of T/B combined epitopes were subsequently constructed. PEG/NaCl precipitation method was applied to extract the recombinant phage PⅢ (rPⅢ) that displayed T/B combined epitopes. By using either commercial IgG against whole-cell of Helicobacter pylori or rHpaA antiserum as the primary antibody, the T/B combined epitopes displayed in rP Ⅲ s were screened and identified by Western blot and ELISA. MTT was applied to determine the proliferation of rHpaA-immunized mouse splenocytes after stimulation of the different recombinant rPⅢ proteins. Results In the HpaA molecule there were five T/B combined epitopes: HpaA10, HpaA37, HpaA79, HpaA116 and HpaA143. All the T/B combined epitopes were successfully displayed on the surface of PⅢ protein of phage M13. The results of Western blot, ELISA and MTT confirmed that HpaA116 was the predominant antigenic epitope, both HpaA37 and HpaA79 were the efficient antigenic epitopes. However, HpaA10 and HpaA143 were identified as ineffective antigenic epitopes. Conclusion The phage display systems of T/B combined epitope peptides of H. pylori adhesin A have been successfully generated in this study. HpaA37 and HpaA79, especially HpaA116 are the efficient T/B combined antigenic epitopes in HpaA of H. pylori.     

Immune responses to six synthetic peptides of capsid protein with sera from HIV-1 infected individuals

Many B cell epitopes within p24 of human immunodeficiency virus type 1 （HIV-1） were identified, while most of them were determined by using murine monoclonal antibodies reacting with overlapping peptides of p24. Therefore these epitopes may not represent the actual epitopes recognized by the HIV-1 infected individuals. In the present study, immune responses of 67 HIV-1 positive sera from Yunnan Province, China to five peptides on p24 of HIV-1 and one of HIV-2 were analyzed. All of 67 sera did not recognize peptide GA-12 on HIV-1 and peptide AG-23 on HIV-2, which indicated that GA-12 was not human B cell epitope and AG-23 did not cross-react with HIV-1 positive serum. Except 13 sera （19.4%）, all remaining sera did not recognize peptides NI-15, DR-16, DC-22 and PS-18, which indicated that these four peptides represented B cell linear epitopes of HIV-1 p24 in some HIV-1 infected individuals but not the immuno-dominant epitopes in most individuals.     

Study on Antibodies to Liver Antigens in Chinese Patients with Different Liver Diseases

Autoimmune liver diseases include autoimmune hepatitis(AIH), primary biliary cirrhosis (PBC) and primary scle rosing cholangitis (PSC). The examination of autoimmuneantibodies is important to diagnose these kinds of diseases.But up to the present, due to the limited knowledge of au toantigens and immature method of examination, especiallyof some autoantibodies which are not only present in au toimmune diseases but found in 10% 15% of the patientswith viral hepatitis or other immune…     

肺炎支原体P1黏附因子主要抗原表位的表达及其临床应用研究

目的 探索肺炎支原体(Mycoplasma pneumoniae,Mp)P1蛋白作为抗原成分在临床血清抗体检测中的应用.方法 通过生物信息学分析P1基因主要抗原决定簇,选择其分值较高的部位进行原核表达、纯化,并进行Western blot鉴定,用纯化后的蛋白免疫小鼠,评价其免疫原性,同时用其作为抗原,采用ELISA方法检测肺炎支原体患儿血清抗体并与全菌体抗原进行比较.结果 成功构建了pET-28C-P1-534原核表达载体,经表达、纯化后获得了相对分子质量为26×103的融合蛋白;Western blot检测其能与Mp阳性血清发生特异性反应;利用纯化的重组蛋白免疫小鼠,能诱导小鼠产生特异性免疫应答,抗体滴度可达 1∶2000;用此蛋白作为抗原成分检测患儿血清,其敏感性和特异性分别为85.00%及97.67%.结论 重组表达的P1-534蛋白具有良好的免疫原性,作为新的抗原成分其敏感性和特异性优于全菌体抗原,为肺炎支原体抗原成分及其蛋白保护作用的研究奠定了基础.

Abstract：

Objective To study the application of P1 adhesin protein epitopes in diagnosis of Mycoplasma pneumoniae(Mp) infected patient. Methods The major epitope(P1-534) of P1 adhesin protein were predicted by ProPred and ANTIGENIC according to its primary structure. The high value fragment was cloned into a constructed recombinant vector. The gene was induced to express fusion protein in E. coli host strain BL21(DE3) and the fusion protein was identified by Western blot. BALB/c mice were immunized with purified protein to test its immunogenicity. Then the purified protein was used as antigen to test the serum of Mp infected patient by ELISA, and compared with the Mp whole cell antigen. Results The P1-534 protein was successfully expressed and purified. ELISA data showed that P1-534 protein could elicit high levels of IgG in immunized mice, the sensitivity and specificity of P1-534 were determined to be 85.00% and 97.67%, while the Mp whole cell antigen were 72.50% and 74.42%. Conclusion The results conformed that the recombinant epitope has certain immunogenicity,and its sensitivity and specificity are better than Mp whole cell antigen. P1-534 protein can be used as an antigen for immunodiagnosis of Mp infection.     

慢性乙肝患者血清趋化因子RANTES水平与肝功能生化指标等的相关性研究

Objective To investigate the level of the serum chemokine RANTES and its correlation with serum biochemical indices of liver function test, HBeAg and HBV DNA load in patients with chronic hepatitis B.Methods 144 patients with chronic hepatitis B (observed group) and 18 normal cases (control group) were enrolled in this study. The serum level of chemokine RANTES was detected with an ABC-ELISA assay. Statistical analysis was performed on the software of SPSS13.0. Results The serum chemokine RANTES level in the observed group (3930.12 ng/ml±2856.96) ng/ml was significantly higher than that in the control group (329.46 ng/ml±152.23) ng/ml. The results from the observed group indicated the positive correlation of serum RANTES level with indices of liver function test, including ALT (r=0. 197, P=0.018), AST(r=0.239, P=0.004) and TBil (r=0.316, P=0.001), but did not with PTA (r=-0.078, P=0.357). Neither difference of serum chemokine RANTES level between HBeAg-positive group and HBeAg-negative group nor that between high HBV DNA load group (≥105 copies/ml) and low HBV DNA load group (< 105 copies/ml) were statistically significant (P=0.407 and 0.185, respectively). Conclusions Serum chemokine RANTES level in patients with chronic hepatitis B elevates significantly and is not affected by HBeAg or HBV DNA load. Its positive correlation with indices of liver function test indicates that RANTES might play an important rule in the pathogenesis of chronic hepatitis B.     

慢性乙肝患者血清趋化因子RANTES水平与肝功能生化指标等的相关性研究

Objective To investigate the level of the serum chemokine RANTES and its correlation with serum biochemical indices of liver function test, HBeAg and HBV DNA load in patients with chronic hepatitis B.Methods 144 patients with chronic hepatitis B (observed group) and 18 normal cases (control group) were enrolled in this study. The serum level of chemokine RANTES was detected with an ABC-ELISA assay. Statistical analysis was performed on the software of SPSS13.0. Results The serum chemokine RANTES level in the observed group (3930.12 ng/ml±2856.96) ng/ml was significantly higher than that in the control group (329.46 ng/ml±152.23) ng/ml. The results from the observed group indicated the positive correlation of serum RANTES level with indices of liver function test, including ALT (r=0. 197, P=0.018), AST(r=0.239, P=0.004) and TBil (r=0.316, P=0.001), but did not with PTA (r=-0.078, P=0.357). Neither difference of serum chemokine RANTES level between HBeAg-positive group and HBeAg-negative group nor that between high HBV DNA load group (≥105 copies/ml) and low HBV DNA load group (< 105 copies/ml) were statistically significant (P=0.407 and 0.185, respectively). Conclusions Serum chemokine RANTES level in patients with chronic hepatitis B elevates significantly and is not affected by HBeAg or HBV DNA load. Its positive correlation with indices of liver function test indicates that RANTES might play an important rule in the pathogenesis of chronic hepatitis B.     

慢性乙肝患者血清趋化因子RANTES水平与肝功能生化指标等的相关性研究

Objective To investigate the level of the serum chemokine RANTES and its correlation with serum biochemical indices of liver function test, HBeAg and HBV DNA load in patients with chronic hepatitis B.Methods 144 patients with chronic hepatitis B (observed group) and 18 normal cases (control group) were enrolled in this study. The serum level of chemokine RANTES was detected with an ABC-ELISA assay. Statistical analysis was performed on the software of SPSS13.0. Results The serum chemokine RANTES level in the observed group (3930.12 ng/ml±2856.96) ng/ml was significantly higher than that in the control group (329.46 ng/ml±152.23) ng/ml. The results from the observed group indicated the positive correlation of serum RANTES level with indices of liver function test, including ALT (r=0. 197, P=0.018), AST(r=0.239, P=0.004) and TBil (r=0.316, P=0.001), but did not with PTA (r=-0.078, P=0.357). Neither difference of serum chemokine RANTES level between HBeAg-positive group and HBeAg-negative group nor that between high HBV DNA load group (≥105 copies/ml) and low HBV DNA load group (< 105 copies/ml) were statistically significant (P=0.407 and 0.185, respectively). Conclusions Serum chemokine RANTES level in patients with chronic hepatitis B elevates significantly and is not affected by HBeAg or HBV DNA load. Its positive correlation with indices of liver function test indicates that RANTES might play an important rule in the pathogenesis of chronic hepatitis B.     

慢性乙肝患者血清趋化因子RANTES水平与肝功能生化指标等的相关性研究

Objective To investigate the level of the serum chemokine RANTES and its correlation with serum biochemical indices of liver function test, HBeAg and HBV DNA load in patients with chronic hepatitis B.Methods 144 patients with chronic hepatitis B (observed group) and 18 normal cases (control group) were enrolled in this study. The serum level of chemokine RANTES was detected with an ABC-ELISA assay. Statistical analysis was performed on the software of SPSS13.0. Results The serum chemokine RANTES level in the observed group (3930.12 ng/ml±2856.96) ng/ml was significantly higher than that in the control group (329.46 ng/ml±152.23) ng/ml. The results from the observed group indicated the positive correlation of serum RANTES level with indices of liver function test, including ALT (r=0. 197, P=0.018), AST(r=0.239, P=0.004) and TBil (r=0.316, P=0.001), but did not with PTA (r=-0.078, P=0.357). Neither difference of serum chemokine RANTES level between HBeAg-positive group and HBeAg-negative group nor that between high HBV DNA load group (≥105 copies/ml) and low HBV DNA load group (< 105 copies/ml) were statistically significant (P=0.407 and 0.185, respectively). Conclusions Serum chemokine RANTES level in patients with chronic hepatitis B elevates significantly and is not affected by HBeAg or HBV DNA load. Its positive correlation with indices of liver function test indicates that RANTES might play an important rule in the pathogenesis of chronic hepatitis B.     

Increase of chemokine interferon-inducible protein-10 (IP-10) in the serum of patients with autoimmune liver diseases and increase of its mRNA expression in hepatocytes

To clarify the role of IP-10 in autoimmune liver diseases, we studied the serum levels of IP-10 in 14 patients with autoimmune hepatitis (AIH), 23 patients with primary biliary cirrhosis (PBC), and 65 patients with chronic viral hepatitis (20 type B and 45 type C). The hepatic expression of IP-10 mRNA and the correlation between the serum levels of IP-10 and clinical parameters were also evaluated. In addition to 20 healthy controls, 16 rheumatoid arthritis (RA) patients were included as an extrahepatic inflammatory disease. The serum level of IP-10 was significantly (P < 0.02) higher in patients with AIH, PBC, and chronic hepatitis B and C than in healthy controls, and it was significantly correlated (P < 0.05) with the serum levels of aspartate aminotransferase and alanine aminotransferase in patients with AIH, PBC, and chronic hepatitis B and C. The serum level of IP-10 was not elevated in RA patients. After successful treatment of AIH and chronic hepatitis C, the serum level of IP-10 decreased to the same level as in healthy volunteers. As we previously showed in cases with chronic hepatitis B or C, in situ hybridization in both AIH and PBC cases demonstrated the expression of IP-10 mRNA in hepatocytes around focal or lobular necrosis surrounded by infiltrating mononuclear cells, whereas IP-10 mRNA was not expressed in areas around the damaged bile ducts in PBC cases. The present results suggest that IP-10 is specifically produced by hepatocytes in inflammatory areas irrespective of the aetiology of hepatitis, and that IP-10 may help to recruit T cells to the hepatic lesions in autoimmune liver diseases as well as in chronic viral hepatitis.     

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.     

Epitopes of calreticulin recognised by IgA autoantibodies from patients with hepatic and coeliac disease

Calreticulin (CRT) was identified as a frequent target of serum autoantibodies (Ab) in various diseases, but anti-CRT Ab of IgA isotype were described only in coeliac (CLD) and some hepatic diseases. Employing ELISA with recombinant CRT we found significantly higher (P<0.001) levels of IgA anti-CRT Ab in sera of patients with primary biliary cirrhosis (PBC) (77.6+/-8.9 AU/mean+/-SE), autoimmune hepatitis (AIH) (105.1+/-9.2 AU) and alcoholic liver cirrhosis (ALC) (193.5+/-21.0 AU) relative to healthy controls (38.6+/-2.0 AU). The levels of IgG anti-CRT Ab in sera of patients with PBC (59.5+/-3.4 AU), AIH (89.7+/-7.9 AU) and ALC (86.4+/-6.2 AU) were also significantly increased (P<0.001) when compared with controls (38.5+/-2.1 AU). Pepscan technique with decapeptides of CRT (each overlapping by eight amino acids) revealed antigenic epitopes of this molecule recognised by IgA Ab of almost all tested patients-KGKNVLINKD and QVKSGTIFDNFL. We also identified disease specific antigenic epitopes on CRT molecule, predominantly recognised by IgA Ab of patients suffering from a particular disease: GGYVKLFPNS and YVKLFPNSLD in AIH (83%, 92% of patients), GLQTSQDARF and EQRLKEEEED in CLD (both 75%) and ASKPEDWDER in ALC (67%). Identification of disease specific CRT epitopes contributes to clarification of autoreactivity against this molecule.     

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。     

Detection of anti-nuclear antibodies in liver diseases by indirect immunofluorescence method and enzyme-linked immunosorbent assay]

Detection of anti-nuclear antibodies (ANA) is essential for diagnosing autoimmune diseases including autoimmune liver diseases. An indirect immunofluorescence (IIF) method with a cell line (HEp-2) derived from human laryngeal carcinoma has been used as a standard substrate. Recently, an enzyme-linked immunosorbent assay (ELISA) using multiple solid-phase antigens has been developed. We assayed sera from 272 cases of chronic liver diseases, 91 cases of healthy subjects and studied clinical significance of ANA. The sensitivity of IIF method in detection of ANA (fluorescence-ANA: FANA) and that of ELISA (ELISA-ANA: EANA) were 19.2% and 17.3% in chronic hepatitis B (CH-B), 16.7% and 17.3% in chronic hepatitis C (CH-C), 84.2% and 50.9% in primary biliary cirrhosis (PBC), 100% and 85.7% in autoimmune hepatitis (AIH) and 15.4% and 18.7% in healthy subjects. The sensitivity of EANA was considerably lower than that of FANA in PBC and AIH, but the sensitivity was the same in CH-C, CH-B, and healthy subjects. Because the solid-phase target antigens do not include nuclear antigen components recognized only by patients with PBC or AIH, ELISA can not detect all the species of ANA. This accounts for the low sensitivity of EANA in PBC and AIH. In conclusion, the current EANA is useful for screening of ANA, but FANA should be performed when PBC or AIH is suspected.     

IgA Anti-b2GPI Antibodies in Patients with Autoimmune Liver Diseases

INTRODUCTION: Recently, we reported a high prevalence of immunoglobulin G and/or immunoglobulin M anticardiolipin antibodies (aCL) in patients with autoimmune liver diseases, namely, autoimmune hepatitis (AIH), primary biliary cirrhosis (PBC), and primary sclerosing cholangitis (PSC), which were independent of the respective isotypes of antibodies against beta2-glycoprotein I (anti-b2GPI). Immunoglobulin A (IgA) aCL and IgA anti-b2GPI are the least studied of the three specific isotypes either in antiphospholipid syndrome (APS) or in other conditions. METHODS: Therefore, we investigated the prevalence and clinical significance of IgA anti-b2GPI and IgA aCL by enzyme-linked immunosorbent assays in another set of Caucasian patients with autoimmune liver diseases (59 AIH, 96 PBC, and 37 PSC). The disease controls group consisted of 50 hepatitis C virus (HCV) patients, 50 hepatitis B virus (HBV), 30 alcoholic liver disease (ALD), 30 non-alcoholic steatohepatitis (NASH), and 110 healthy controls. RESULTS AND DISCUSSION: IgA anti-b2GPI prevalence was higher in AIH (50.8%) compared to PBC (p = 0.005), PSC (p = 0.008), NASH (p = 0.004), ALD (p = 0.01), and HCV (p = 0.002). The titers were also significantly higher in AIH compared to any other group of the study. IgA aCL prevalence was higher in AIH (33.9%) compared to PBC (p = 0.005), PSC (p = 0.014), NASH (p = 0.001), ALD (p = 0.004), and HCV (p < 0.001). IgA anti-b2GPI or IgA aCL were not associated with APS features in patients with liver autoimmunity. Of note, IgA anti-b2GPI and IgA aCL were associated with clinical and biochemical markers of disease severity in AIH and PBC. We demonstrated a high prevalence and high titers of IgA anti-b2GPI in patients with AIH compared to any other liver disease of the study. CONCLUSION: IgA anti-b2GPI and IgA aCL were associated with the severity and biochemical activity of AIH and PBC, but long-term prospective studies are needed to address whether this new finding is of clinical importance in AIH and PBC patients.     

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.     

cDNA microarray analysis of autoimmune hepatitis, primary biliary cirrhosis and consecutive disease manifestation

While autoimmune hepatitis (AIH) and primary biliary cirrhosis (PBC) usually have distinct clinical manifestations, some patients present with features of both conditions. Using cDNA microarrays, we analyzed and compared gene expression profiles in 8 patients with AIH, 9 with PBC, 8 with chronic hepatitis C (CHC), 8 with non-alcoholic steatohepatitis (NASH) and 9 with normal livers. We subsequently applied this method to a tissue sample from a 61-year-old woman with overlapping features of both AIH and PBC. A 61-year-old woman was admitted to our hospital for evaluation of elevated serum alkaline phosphatase. A liver biopsy showed accumulation of mononuclear cells around the bile duct cells, a feature characteristic of chronic non-suppurative destructive cholangitis (CNSDC). Three years later, her serum alanine aminotransferase (ALT) level had increased, and a liver biopsy demonstrated evidence of a severe form of hepatitis. A cDNA microarray analysis of both biopsies identified the molecular events associated with her altered histology. The expression profile of this patient, which was originally different from that of the other PBC patients, changed to an AIH pattern. Our results suggest that this patient has characteristics of both AIH and PBC.     

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   

Autoimmune hepatitis associated with bile duct injury resembling chronic non-suppurative destructive cholangitis

Autoimmune hepatitis (AIH) and primary biliary cirrhosis (PBC) are representative autoimmune liver diseases in which hepatocytes and intrahepatic bile ducts, respectively, are selectively damaged by autoimmune mechanisms. Bile duct injury and loss is characteristic of PBC and chronic non-suppurative destructive cholangitis (CNSDC), in particular, is a histological hallmark of PBC. In this report, we present an unusual case of AIH accompanied by CNSDC-like bile duct injury in a 46-year-old woman. The patient's serum aminotransferase level was abnormally high. The serum levels of alkaline phosphatase, gamma-GTP and IgG were also elevated, but the IgM level was within normal limits. The titer of antismooth muscle antibody (SMA) was 1:80, while antinuclear autoantibody (ANA) and the M2 fraction of antimitochondrial antibody (AMA) were both negative. Liver biopsy disclosed CNSDC-like bile duct injuries and severe interface hepatitis and lobular hepatitis with perivenular zonal necrosis were observed. The aggregate score of the International Autoimmune Hepatitis Group corresponded to the category of probable AIH. Immunohistochemically, histocompatibility leukocyte antigen-DR, which is aberrantly expressed in the damaged bile ducts of PBC, was not found in the injured bile ducts of this case. Laboratory data were much improved by treatment with prednisone, but ursodeoxycholic acid was not effective. Although the possibility of an overlapping syndrome of AIH- and AMA-negative PBC could not be excluded, this case was diagnosed as AIH with CNSDC-like bile duct lesions.