Clinical Significance of Positive Immunoblotting but Negative Immunofluorescence for Antimitochondrial Antibodies in Patients with Liver Diseases Other than Primary Biliary Cirrhosis

The serum reaction to anti-2-oxo-acid dehydrogenase complex (2-OADC) enzymes, the antigens recognized by antimitochondrial antibodies (AMA), can be detected by immunoblotting in patients with liver diseases other than primary biliary cirrhosis (PBC), who are negative for AMA by conventional indirect immunofluorescence. Whether the presence of anti-2-OADC is related to PBC or represents preclinical PBC in such patients is obscure at present. We examined the immunoreactivity of AMA by immunofluorescense, immunoblotting, and enzyme inhibition assay in serum samples from 59 patients with liver diseases other than PBC and 71 healthy subjects. We also examined the clinical course of the patients in whom a positive result was obtained to elucidate whether such reaction was a "true" or "false" phenomenon. None of the 130 sera was positive for AMA by indirect immunofluorescence or for anti-pyruvate dehydrogenase complex (PDC) by enzyme inhibition assay. However, seven of 71 (10%) sera from healthy subjects contained weak IgG class antibody to PDC-E2 (four sera) or E2 subunit of branched-chain oxo-acid dehydrogenase complex (BCOADC-E2) (three sera). Of the 59 sera from patients with liver diseases other than PBC, four (7%) reacted against 2-OADC by immunoblotting. Of these, three sera were from patients with chronic hepatitis C virus (HCV) infection, and contained IgG class autoantibody to BCOADC-E2. The serum reactivity to BCOADC-E2 detected by immunoblotting in these three patients diminished after absorption with recombinant BCOADC-E2 fusion protein. During the 3-5 year follow-up period, AMA by immunofluorescence and anti-PDC activity by enzyme inhibition assay were always negative in these three patients. The other one serum was from patient with alcoholic cirrhosis, and contained IgM class autoantibody to E3 binding protein (E3-BP). This patient did not develop PBC during the following 2 years. Our results showed that anti-2-OADC antibodies could be detected in some patients with liver diseases other than PBC, and even in healthy individuals. The clinical significance of the presence of these serum reactions is obscure at this stage, but the production of anti-BCOADC-E2 may be linked to the presence of HCV in certain patients, Further prospective studies of larger population should clarify whether anti-2-OADC reaction can precede the clinical development of PBC.     

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…     

Frequency and Significance of Antibodies to Soluble Liver Antigen/Liver Pancreas in Variant Autoimmune Hepatitis

Background: Antibodies to soluble liver antigen/liver pancreas are highly specific markers of autoimmune hepatitis. Aims: Determine the frequency and clinical significance of these antibodies in the variant syndromes. Methods: Antibodies to soluble liver antigen/liver pancreas were determined in 28 patients with variant forms, including 10 with cryptogenic chronic hepatitis and 18 with cholestatic variants. One hundred and seventy-two patients with classical autoimmune hepatitis were similarly tested. Results: Seven of the 28 patients with variant forms had the antibodies, and this frequency was not statistically different than that in classical disease (25 vs. 12%, p = 0.08). Antibodies were most common in patients with cryptogenic chronic hepatitis (40%). Seropositive patients were indistinguishable from seronegative patients with variant forms, and they responded as well to corticosteroid therapy as patients with autoimmune hepatitis. Relapse after corticosteroid withdrawal invariably occurred in the seropositive patients whether with variant or classical disease, and HLA DR3 was more common in the seropositive patients with variant forms than in normal subjects (60 vs. 15%, p = 0.03). Conclusions: Antibodies to soluble liver antigen/liver pancreas occur commonly in the variant forms of autoimmune hepatitis and identify patients that closely resemble classical disease. Seropositivity is associated with relapse after corticosteroid withdrawal and HLA DR3. The antibodies may be surrogate markers of a genetic propensity to relapse that is independent of clinical phenotype.     

Levels of BAFF in Serum in Primary Biliary Cirrhosis and Autoimmune Diabetes

Levels of the B-cell activating cytokine BAFF are increased in serum in various autoimmune disease, and particularly Sjögren's syndrome in which there is evident B-lymphocyte proliferation. Studies in two autoimmune disease in which B-cell proliferation is less evident, primary biliary cirrhosis (PBC), and adult-onset Type 1 diabetes, showed serum levels of BAFF to be mostly in the normal range. A single raised level among eight sera tested in one patient studied with autoimmune hepatitis (AH) coincided with a relapse of the disease. Increased levels of BAFF in human sera, indexing a potent antigenic drive on B-cell production and survival in some autoimmune diseases, may mark only particular stages in the evolution of such diseases.     

Liver schistosomiasis and primary biliary cirrhosis

Summary The case of a patient suffering from liver schistosomiasis diagnosed by liver biopsies is presented. There were also lesions in the liver suggestive of non-suppurative destructive cholangitis and the serum-immunological studies strengthened the diagnosis of primary biliary cirrhosis. The eo-existence of these two affections and their possible interrelationship are discussed. The authors further discussed some clinical, biochemical, immunological and pathological aspects of these two conditions and evoked the eventuality of an auto-immune type of hepatic affection triggered by the presence of the schistosome eggs.The authors are grateful to Dr. A. Cruchaud for the immunological studies, to Mrs. M. Loup and Miss C. L. Seignemartin for secretarial services, and Mr. E. Denkinger for the photographs.     

The Hepatitic/Cholestatic "Overlap" Syndrome: An Italian Experience

Background: Patients with hepatitic and cholestatic autoimmune liver disease ("overlap syndrome") represent a diagnostic and therapeutic challenge. Aim: To evaluate the prevalence of the "hepatitic/cholestatic overlap" in a large series of consecutive patients with cholestatic autoimmune liver disease. Methods: We re-evaluated the diagnosis of 235 patients with autoimmune liver disease, including 70 with type 1 autoimmune hepatitis (AIH), 142 with primary biliary cirrhosis (PBC), and 23 with primary sclerosing cholangitis (PSC), using the revised International Autoimmune Hepatitis Group (IAIHG) scoring system. Anti-mitochondrial, anti-nuclear, anti-smooth muscle, anti-liver kidney microsomal type 1, anti-liver cytosol type 1, perinuclear anti-neutrophil nuclear and anti-soluble liver antigen antibodies were evaluated in each patient. Results: Ten patients (3 with a previous diagnosis of PBC and 7 of PSC) scored as "probable" or "definite" AIH. These patients did not have a specific autoantibody profile. Conclusions: Among patients with PBC, the occurrence of a PBC/AIH overlapping syndrome is rare (2.1%), whereas among patients with PSC an overlap between PSC and AIH is frequent (30.4%). Whether patients with the hepatitic/cholestatic overlap syndrome would benefit from a combination therapy with immunosuppression and ursodeoxycholic acid remains to be established.     

Microchimerism in Human Health and Disease

During pregnancy some cells traffic between the fetus and the mother. Recent investigative work indicates a low level of fetal cells commonly persists in the maternal circulation for years, or even indefinitely, after pregnancy has been completed. The term microchimerism refers to one individual harboring DNA or cells at a low level that derive from another individual. Chronic graft-versus-host disease (cGvHD) shares similarities with some autoimmune diseases and is an iatrogenic form of chimerism, occurring as a complication of hematopoietic stem cell transplantation. The HLA genes of the donor and the host are known to be of central importance to the development of cGvHD. When also considered in light of the female predilection to autoimmunity, these series of observations led to the hypothesis that microchimerism and HLA genes of host and non-host cells are involved in some autoimmune diseases. The hypothesis can also apply to men, children, and women who have not been pregnant because there are other sources of microchimerism. Persistent microchimerism can follow a blood transfusion, or can occur from transfer between twins in utereo. Additionally, maternal cells have recently been found to persist in her immune competent progeny. A number of studies have investigated a potential role of microchimerism in human diseases including systemic sclerosis (SSc), primary biliary cirrhosis (PBC), Sjögren's syndrome, polymorphic eruption of pregnancy, myositis, and thyroid disease. While some studies lend support to the concept that microchimerism is involved in the pathogenesis of selected autoimmune diseases, studies also indicate microchimerism is not uncommon in other human conditions and in healthy individuals.     

Patients with Insulin-dependent Diabetes but not those with Non-insulin-dependent Diabetes have Anti-sulfatide Antibodies as Determined with a New ELISA Assay

Background: In sera from newly diagnosed insulin-dependent diabetes mellitus patients (IDDM type 1) autoantibodies occur against different antigen determinants often shared with neural tissues. The role of these autoantibodies in the disease process is not yet clarified but they can be used as a diagnostic tool in the detection of IDDM patients. Methods: We have analysed the occurrence of sulfatide autoantibodies in serum from patients with type 1 diabetes (n = 20), individuals with pre-type 1 diabetes (n = 6), patients with type 2 diabetes (n = 32) and controls (n = 43). The method used for the determination of the autoantibodies was a newly developed microtitre-ELISA assay utilizing a complex of sulfatide-albumin as the ligand. Results: The new assay procedure for serum sulfatide autoantibodies showed good reproducibility. The total (day-to-day) imprecision based on analyses of three different serum samples with positive titres varied between 11 and 14% during an assay period of 6 months. None of the controls (0/43) had positive titres of sulfatide antibodies. Of the patients with type 1 diabetes, 85% displayed positive titres of anti-sulfatide antibodies while none of the type 2 patients did so. All individuals with pre-type 1 diabetes had positive titres of sulfatide antibodies. Conclusions: We conclude that sulfatide autoantibodies in serum can be reproducibly assayed by the newly developed microtitre-ELISA procedure. Elevated titres of sulfatide autoantibodies are a constant finding in newly diagnosed type 1 patients.     

几种临床疾病的血小板相关抗体检测结果分析

目的 :探讨血小板相关抗体在血小板减少症中的临床应用。方法 :选择健康者、血小板低于正常的体检者、血小板减少性紫癜(ITP)、再障 (AA)、肝脏疾患、系统性红斑狼疮 (SLE)患者等采用ELISA法定量测定血小板抗体PAIgG、PAIgM、PAIgA ,并分析ITP患者治疗前后血小板相关抗体的变化。结果 :血小板低于正常的体检者血小板抗体的阳性率 14 .9% ,ITP、AA、SLE等患者血小板相关抗体阳性率与正常组比较有显著差异 ,分别为 78.9%、3 1.2 %、62 .5 % ,肝病者血小板相关抗体与正常组无差异。ITP患者治疗前后三种血小板抗体均有非常显著性变化 (P <0 .0 1)。结论 :血小板相关抗体测定在自身免疫性疾病引起的血小板减少的诊疗中有重要意义     

The Prevalence of Anti-acetylcholinesterase Antibodies in Autoimmune Disease

A robust and precise enzyme linked immunosorbent assay (ELISA) with proven sensitivity and specificity has been employed to detect human antibodies (allogenic/autogenic) to human acetylcholinesterase (AChE). The sensitivity of the method has been established using mouse monoclonal antibodies (0.8?ng/ml) and uniquely, human sera positive for anti-Yt^a allogenic antibodies, to one phenotypic form (most common) of human AChE. The latter was also used as the positive human control to ensure functionality of the assay. The ELISA method was used to establish a normal distribution curve for absorbance values employing sera from healthy blood donors Subsequently, the ELISA was employed to investigate the prevalence of anti-AChE antibodies in patients with confirmed autoimmune disease and patients with non-autoimmune thyroid disease (diseased control). The results indicate that there is not a high prevalence of anti-AChE antibodies in patients with confirmed autoimmune disease. The lack of anti-AChE autoantibodies in patients' with clinically apparent Graves' ophthalmopathy, mitigates against there being a causal role of such antibodies in Graves' associated eye disease.     

Diagnostic Value of Anti-Nuclear Antibodies: Results From Korean University-Affiliated Hospitals

BackgroundUnnecessary and inappropriate laboratory testing accounts for a significant portion of waste in health care utilization. The aim of this study was to examine the diagnostic value of the anti-nuclear antibody (ANA) test by examining the rate of ANA associated rheumatic disease (AARD) diagnosis among ANA tested and ANA positive subjects and positive predictive value (PPV) of ANA test leading to AARD diagnosis in different ANA titers and different subsets of patients in 5 hospitals affiliated with a university.MethodsWe retrospectively extracted data from all subjects who were tested for ANA from year 2010 to 2019. Those who were first evaluated at or referred to rheumatology were further evaluated with extraction of data including ANA titer and ultimate diagnosis. PPVs for ANA test were evaluated after stratification according to clinically relevant key parameters, such as patient age (younger < 65 years vs. older), sex, and requesting department.ResultsFrom 2010 to 2019, A total of 94,153 patients were tested for ANA, of which 13,600 (14.4% of the total) were positive. AARD was diagnosed in only 0.69% among all ANA tested patients and 4.74% among ANA positive patients. The AARD diagnosis rate of ANA positive patients varied widely from 0.1% to 8.7% by requesting department. Using cutoff values above 1:320 yielded PPVs of 15.6 and 7.8% for all AARs and systemic lupus erythematosus. The PPV was significantly higher in young age (< 65 years) and in women, and when it was requested from internal medicine vs other departments.ConclusionAARD was diagnosed in less than 1% of all ANA tested patients in university-affiliated hospitals. This result shows that careful consideration before ordering the screening ANA is needed to improve the utility of the test for providers and patients and to reduce health costs spurred by unnecessary testing and its consequences.     

Association of Single Nucleotide Polymorphisms of the Interleukin-10 Promoter Gene and Susceptibility to Primary Biliary Cirrhosis: Immunogenetic Differences in Italian and Japanese Patients

Several lines of data suggest that genetic factors play an important role in the onset and/or progression of primary biliary cirrhosis (PBC). Since PBC is an autoimmune disease, it is reasoned to assume that genes encoding cytokines may confer susceptibility to disease. Amongst these factors, interleukin-10 (IL-10) has received significant attention. The promoter region of IL-10 gene has three single nucleotide polymorphisms (SNPs) at positions &#109 1082, &#109 819 and &#109 592. To elucidate the association of the three SNPs of IL-10 promoter region with susceptibility of PBC in two different genetic populations, 159 unrelated patients with PBC (94 Italian and 65 Japanese) and 143 local controls (72 Italian and 71 Japanese) were enrolled. SNPs were determined using allele-specific PCR/RFLP. In Italian PBC patients, the frequency of homozygosity for G/G at position &#109 1082 was significantly higher than that of local controls (p <0.041, OR=2.44, 95% C.I.; 1.02-5.86). The frequencies of haplotype GCC in PBC patients, possibly linked to higher IL-10 production, were also significant higher than local controls (p <0.033). However, in Japanese population, there were no significant differences in the three SNPs and haplotypes between PBC patients and controls. Excessive production of IL-10 may play an important role in some populations in modulating the onset of PBC. Further, immunogenetic studies of PBC should take into account ethnic and geographic variations; this makes such studies in heterogeneous population, like the USA, more difficult.     

Relationship between Tetrahydrobiopterin and Portal Hypertension in Patients with Chronic Liver Disease

Tetrahydrobiopterin (BH4) is an essential cofactor in NO synthesis by endothelial nitric oxide synthase (eNOS) enzymes. It has been previously suggested that reduced intrahepatic BH4 results in a decrease in intrahepatic NO and contributes to increased hepatic vascular resistance and portal pressure in animal models of cirrhosis. The main aim of the present study was to evaluate the relationship between BH4 and portal hypertension (PHT). One hundred ninety-three consecutive patients with chronic liver disease were included in the study. Liver biopsy, measurement of BH4 and hepatic venous pressure gradient (HVPG) were performed. Hepatic fibrosis was classified using the Laennec fibrosis scoring system. BH4 levels were determined in homogenized liver tissues of patients using a high performance liquid chromatography (HPLC) system. Statistical analysis was performed to evaluate the relationship between BH4 and HVPG, grade of hepatic fibrosis, clinical stage of cirrhosis, Child-Pugh class. A positive relationship between HVPG and hepatic fibrosis grade, clinical stage of cirrhosis and Child-Pugh class was observed. However, the BH4 level showed no significant correlation with HVPG or clinical features of cirrhosis. BH4 concentration in liver tissue has little relation to the severity of portal hypertension in patients with chronic liver disease.

Graphical Abstract

相似文献   

Usefulness of Non-invasive Markers for Predicting Significant Fibrosis in Patients with Chronic Liver Disease

The purpose of this prospective study was to verify and compare the strengths of various blood markers and fibrosis models in predicting significant liver fibrosis. One hundred fifty-eight patients with chronic liver disease who underwent liver biopsy were enrolled. The mean age was 41 yr and male patients accounted for 70.2%. The common causes of liver disease were hepatitis B (67.7%) and C (16.5%) and fatty liver (9.5%). Stages of liver fibrosis (F0-4) were assessed according to the Batts and Ludwig scoring system. Significant fibrosis was defined as ≥F2. Sixteen blood markers were measured along with liver biopsy, and estimates of hepatic fibrosis were calculated using various predictive models. Predictive accuracy was evaluated with a receiver-operating characteristics (ROC) curve. Liver biopsy revealed significant fibrosis in 106 cases (67.1%). On multivariate analysis, α2-macroglobulin, hyaluronic acid, and haptoglobin were found to be independently related to significant hepatic fibrosis. A new predictive model was constructed based on these variables, and its area under the ROC curve was 0.91 (95% confidence interval, 0.85-0.96). In conclusion, α2-macroglobulin, hyaluronic acid, and haptoglobin levels are independent predictors for significant hepatic fibrosis in chronic liver disease.     

原发性胆汁性肝硬化患者抗线粒体抗体亚型检测的临床意义

为探讨原发性胆汁性肝硬化(PBC)患者血清抗线粒体抗体(AMA)亚型抗M2、M4、M9抗体的临床意义,采用间接免疫荧光法检测96例PBC患者血清AMA水平,采用酶免疫斑点法检测AMA-M2、M4、M9抗体水平,应用全自动生化分析仪检测ALT等生化指标。结果表明96例PBC患者中,荧光法AMA阳性率为84.4%,抗M2抗体为81.3%;斑点法抗M2抗体阳性率为72.9%,抗M4抗体为44.8%,抗M9抗体为18.8%,抗M2和M4抗体同时阳性阳性率为43.8%,抗M2和M9抗体同时阳性阳性率为16.7%,抗M2、M4和M9抗体同时阳性阳性率为13.5%,抗M4抗体阳性的PBC患者ALT、AST和IgM水平明显高于抗M4抗体阴性(P<0.05),抗M9抗体阳性的PBC患者ALT、AST和IgG水平明显低于抗M9抗体阴性(P<0.05)。AMA-M2抗体的检测对PBC患者有诊断意义,抗M4抗体和抗M9抗体的检测对于PBC患者的病情判断有意义。     

Changing Trends in Liver Cirrhosis Etiology and Severity in Korea: the Increasing Impact of Alcohol

BackgroundChronic hepatitis B is the most common cause of liver cirrhosis in South Korea. However, alcoholic liver disease has shown an increasing trend. Although the clinical implications surrounding liver cirrhosis have been changing over the years, few studies have recently examined cirrhosis epidemiology. Therefore, we aimed to investigate changes in liver cirrhosis etiology and severity in Korea.MethodsWe retrospectively reviewed 16,888 records of cirrhotic patients from six tertiary hospitals in Korea from 2008 to 2017. Continuous and non-continuous variables were processed via linear and Poisson regression, expressed as beta (B) coefficients and as exponentiated values of coefficients (Exp[B]), respectively.ResultsChronic hepatitis B showed a decreasing trend (Exp[B] = 0.975, P < 0.001), whereas alcohol showed an increasing trend (Exp[B] = 1.013, P = 0.003), occupying the most common etiology in 2017. The Child-Turcotte-Pugh (CTP) score and decompensated liver cirrhosis prevalence did not change over the 10-year period. The incidence of variceal bleeding, severe ascites, hepatic encephalopathy, and spontaneous bacterial peritonitis significantly decreased from 12.3% to 7.7%, 7.8% to 4.1%, 1.0% to 0.5%, and 1.9% to 1.1%, respectively (P < 0.05 for all). In the subgroup analysis, the chronic hepatitis B group showed improving CTP scores (B = −0.025, P < 0.001) and decreasing decompensated liver cirrhosis rates (Exp[B] = 0.977, P = 0.016), whereas the alcohol group demonstrated increasing CTP class C (Exp[B] = 1.031, P = 0.005) and model for end-stage liver disease scores (B = 0.081, P = 0.005) over 10 years.ConclusionThe chronic hepatitis B group exhibited improved results, whereas the alcohol group still presented poor liver functions and outcomes. Future national policies and systematic approaches addressing the incidence, prevention, and treatment of alcoholic liver cirrhosis are indispensable.     

慢性丙型肝炎病毒感染者血清自身抗体检测的临床研究

目的 观察丙型肝炎病毒感染者血清中自身抗体的检测结果,探讨自身免疫在丙型肝炎病毒感染中的意义.方法 采用回顾性分析方法,对226例慢性丙型肝炎病毒感染者和137例慢性乙型肝炎患者血清进行ANA、anti-AMA、anti-Ro-52等自身抗体的检测,并探讨HCV-RNA含量、生化指标、年龄及性别、干扰素治疗后应答等与自身抗体变化的关系.结果 丙型肝炎感染者226例中有78例出现自身抗体阳性,检出率为34.5%（78/226）;明显高于慢性乙肝组的7.3%（10/137）（χ^2=34.396,P＜0.05）,其中ANA阳性69例,占30.5%.抗核抗体以低滴度为主.另外还检测到anti-AMA、anti-Ro-52等自身抗体;150例丙肝病毒复制指标HCV-RNA（＋）的患者,自身抗体检出率为40.7%,76例丙肝病毒复制指标HCV-RNA（-）的患者,自身抗体检出率为22.4%,两者总检出率差别有统计学意义（χ^2=7.473,P＜0.05）;自身抗体阴性、阳性者之间ALT、AST、TBIL数值分别为（65.1±24.4）U/L、（47.4±22.7）U/L、（17.2±8.2）μmol/L和（132.2±49.3）U/L、（100.7±35.2）U/L、（35.5±14.7）μmol/L,差异有统计学意义（t值分别为16.012,14.843,和9.000,均P＜0.05）;丙型肝炎自身抗体的检出率与年龄有关,而与性别无关;丙型肝炎患者经干扰素抗病毒治疗后,虽然自身抗体阳性组干扰素应答率73.9%（17/23）高于自身抗体阴性组的54.2%（26/48）,但差异无统计学意义（χ2=0.975,P＞0.05）.结论 丙型肝炎病毒感染可诱发机体自身免疫反应,产生多种自身抗体,尤其HCV-RNA阳性患者更为突出,自身抗体检出率与年龄明显有关,检测自身抗体对丙型肝炎的诊断治疗具有指导意义.