IDENTIFICATION OF IMMUNE-COMPLEX DEPOSITION AND T-LYMPHOCYTE IN FULMINANT B-VIRAL HEPATITIS WITH IMMUNOHISTOCHEMICAL TECHNIQUES

Cryostat sections of the liver representing fulminant B-viral hepatitis were investigated with antisera against human immunoglobulin G(IgG), immunoglobulin M(IgM), C3, C4, C1q, hepatitis B surface antigen (HBsAg), and T-lymphocytes using the immunofluorescence and immunoperoxldase techniques. The liver showed positive staining for IgG, IgM, C3, C4, C1q, and HBsAg in the viable and necrotic hepatocytes and Kupffer cells in a granular fashion. Furthermore, the cell membrane of lymphocytes present in the liver were positively stained with anti T-lymphocyte sera. The numbers of T-lymphocytes recognized were predominant both in portal tracts and within hepatic lobules over those of non-T-lymphocytes. It suggests that perhaps some of the end results of fulminant hepatitis inflammatory reactions may be mediated in part by T-lymphocytes.     

FULMINANT TYPE A VIRAL HEPATITIS IN A CHIMPANZEE

A case of a chimpanzee with fulminant hepatitis caused by spontaneous hepatitis A virus (HAV) infection was reported. The liver at autopsy revealed massive liver cell necrosis with mononuclear and polymorphonuclear cell infiltration. Aggregation of HAV-like particles (22–25 nm in diameter) were found within the vesicles of hepatocytes under the electron microscope. Immunofluorescent examination of the liver showed positive staining for HAV antigen, C₁q, C3, C4, immunoglobulin M (IgM), and immunoglobulin G (IgG) in the hepatocytes and/or Kupffer cells in a granular fashion. The anti-HAV antibody (IgM type) and circulating immune complexes were detected in the postmortem serum. The present study suggests the possibility that the deposition of immune complexes of HAV and anti-HAV antibody in the liver cell plays an important role in the pathogenesis of massive liver cell necrosis in fulminant type A viral hepatitis.     

Immunoglobulin and complement complexes in blood following infection with human immunodeficiency virus type 1.

Freely soluble and complexed plasma immunoglobulin A (IgA), IgG, IgM, C1q, C3, and factor B in 36 human immunodeficiency virus type I (HIV-1)-seronegative controls, 69 asymptomatic HIV+ subjects, and 117 individuals with symptomatic HIV-associated disease were characterized. Levels of free and complexed IgG and IgA, and to a lesser extent free C1q and complexed IgM, increased with HIV-1 infection. In stark contrast, both HIV+ groups showed three- to sixfold declines in complexed C3, C1q, and factor B levels. The asymptomatic HIV+ population showed declines in levels of C3-bound IgA, IgG2, and IgG4 complexes. The asymptomatic group showed reductions in C3-complexed IgM, IgA, IgG2, and IgG4 levels. HIV infection is associated with complement-deficient immune complexes.     

Immunoglobulin isotypes of anti-HBc and anti-HBe and hepatitis B virus (HBV) DNA elimination in acute hepatitis B

Antibodies to hepatitis B core antigen (anti-HBc) and e antigen (anti-HBe) were assayed in 46 sera from ten patients with acute hepatitis B utilizing immunoglobulin class- and subclass-specific enzyme immunoassays (EIAs). The sera were sampled 1 to 512 days after onset of hepatic symptoms. Four patients cleared HBsAg rapidly, within 24 days, and six patients cleared HBsAg slowly, within 27-74 days after the onset of symptoms. In three of the patients with rapid clearance of HBsAg, hepatitis B virus (HBV) DNA was not detected in sera tested during the first week after onset. The fourth patient was not tested until 12 days after onset and was then found to be negative for HBV DNA. In four of the patients with slow clearance of HBsAg, HBV DNA was present during the first week of illness. In the other two patients, HBV DNA was not detected in the first serum, 11 and 17 days after the onset of illness. Anti-HBc IgM and IgA1 were detected in all patients, with maximum titers shortly after onset. Anti-HBc IgG1 was present in all sera tested. Anti-HBc IgG2 was not detected in any of the sera. Anti-HBc IgG3 and IgG4 were detected in all patient sera, with IgG3 paralleling IgG1, and IgG4 mainly in sera long after onset. Anti-HBe IgG1, IgG3, and IgG4 were detected in three, two, and two patients, respectively. Anti-HBe IgG2, IgM, IgA1, or IgA2 was not found in any patient. The time required for maximum titer of anti-HBc IgG1 was shorter in the patients with rapid clearance of HBsAg.(ABSTRACT TRUNCATED AT 250 WORDS)     

Resolution of HBV infection occurs sooner than recovery of renal disease in adult serum HBsAg‐negative HBV–associated glomerulonephritis

Most cases of hepatitis B virus–associated glomerulonephritis (HBV‐GN) occur in children and present with serum HBsAg positivity. Few studies have investigated adult patients with HBV‐GN who are serum HBsAg‐negative. This study aimed to determine the clinical and pathological features of adult patients with HBV‐GN who are serum hepatitis B surface antigen (HBsAg)‐negative. Clinical, pathologic, and laboratory findings were collected and analyzed in a cohort of 27 adult patients with HBV‐GN who were serum HBsAg‐negative upon diagnosis. The study population included mostly men of middle age (40‐59 years). Clinically, patients presented with nephrotic syndrome. Serum immunoglobulin G levels were low, whereas serum immunoglobulin M, immunoglobulin A, complement C3 (C3), and complement C4 (C4) levels as well as liver and renal function tests were normal in most or all patients. Among the 27 patients, 21 tested positive for HBV antibodies. Membranous nephropathy was the dominant pathological form on kidney biopsy. In addition, only a few patients showed a “full house” staining pattern and renal immune deposit of complement C1q (C1q). Serum HBsAg‐negative HBV‐GN may represent a late stage of HBV infection. We recommend routine testing for HBV markers on renal biopsy in regions where HBV is prevalent, even when tests for serum HBV markers are negative.     

Spectrum of hepatitis E virus infection in India

A solid phase enzyme linked immunosorbent assay (ELISA) that detects IgM and IgG to hepatitis E virus (HEV) was used to study seroepidemiology in 40 healthy subjects and 227 consecutive patients with liver diseases in an endemic area. Fifty-two of the liver diseases patients (22.9 percent) had acute hepatitis E. In contrast, none of the 40 healthy subjects were positive for IgM anti-HEV, validating the ELISA assay. Twenty-three of 25 (92%) patients with epidemic non-A, non-B hepatitis were confirmed as having acute hepatitis E. Only 1 of the 10 patients with sporadic, fulminant hepatic failuire of non-A, non-B, non-C etiology was positive for IgM anti-HEV. Five (31.2%) of the 16 patients with acute hepatitis in HBsAg carriers were positive for IgM anti-HEV. One patient with acute hepatitis B wascoinfected with acute hepatitis E. Acute hepatitis was a disease of the adult population, with peak attack rates in the second and third decades of life. This disease was seen in only 4 (16%) of the 25 patients with acute viral hepatitis occurring below 14 years of age. Cholestasis was predominant in 25% of patients, enzyme elevation was monophasic, and all patients had clinical and biochemical recovery from the disease. The data suggest that the majority of patients with acute sporadic non-A, non-B, non-C hepatitis in India have hepatitis E. However, fulminant hepatic failure to sporadic nature is rarely from hepatitis E. © 1994 Wiley-Liss, Inc.     

1995至2000年北京地区散发性急性肝炎患者肝炎病毒抗体的检测与分析

目的 了解北京市地区散发性肝炎患者甲型、乙型、丙型和戊型肝炎病毒感染型别分布及重叠感染。方法 用EIA法检测1995斫2月至2000年12月北京地区散发性急性肝炎患者抗－HAVIgM、HBsAg／抗－HBcIgM、抗－HCVIgM/IgG和抗－HEVIgM/IgG。结果 214例散发性急性肝炎患者血清,抗甲、乙、内和戊型肝炎病毒IgM总阳性数155例,戊型肝炎76例。有9名患者检出2种肝炎病毒抗原或抗体阳性,其中在3名肝硬化患者和2名静脉吸毒者同时检测到HBV和HCV抗原或抗体,1例HBsAg阳性者检测到抗－HAVIgM,3例－HCVIgG阳性者中分别检测到2例抗－HBVIgM和1例抗－HEVIgM。肝炎病毒重叠感染的9名患者年龄在31岁至49岁之间。结论 北京地区散发性急性肝炎78％是由消化道传染的甲戊型肝炎病毒引起,戊型肝炎在四种肝炎中位居首位,其次为甲型肝炎、乙型肝炎和丙型肝炎。肝炎病毒重叠感染多见乙、丙型肝炎病毒合并感染或慢性乙、丙型肝炎患者合并甲型或戊型肝炎病毒感染。在我国对散发性病毒性肝炎的预防应引起高度重视。     

Immunoglobulin and hepatitis B surface antigen-specific circulating immune complexes in chronic hepatitis with hepatitis delta virus infection

IgM, IgG, and HBsAg containing circulating immune complexes (CIC) were determined, by conglutinin (K) and C1q assays, for assessing the role of CIC in hepatitis delta virus (HDV) infection in 54 HBsAg-negative controls and 85 HBsAg-positive patients with chronic hepatitis. The prevalence of HDV markers (HDV antigen and anti-HD) was 24.70% (21/85). CIC were a common feature of HDV infection with 95.24% of patients having at least one abnormal test resutlt. The prevalence of elevated IgM-K, IgG-K, IgM-C1q, and IgG-C1q CIC were 85.71, 85.71, 57.14, and 85.71%, respectively. The prevalence of IgM class CIC were statistically higher in patients with HDV infection than in those without (P = .001 for the K assay and P = .023 for the C1q assay). There was no difference in the prevalence of IgG class CIC. Patients with HDV infection also have significantly higher median levels of IgM K-CIC (P = .002), IgG K-CIC (P = .049), and IgG C1q-CIC (P = .008). In patients with HDV infection, there was positive correlation between IgM C1q-CIC and transminase levels (r = .519, P = .016 for AST; r = .500, P = .021 for ALT). There was no difference in the prevalence of HBsAg containing CIC between patients with HDV infection (76.19%) and those without (74.60%). In conclusion, IgM class CIC are the major CIC and correlate with disease activity in HDV infection. CIC may play a role in the pathogenesis of HDV infection.     

血清sP-selectin、sICAM-1和免疫球蛋白检测对乙肝肝硬化预后评估的意义

目的：旨在探讨血清sP-selectin、sICAM-1和免疫球蛋白检测对乙肝肝硬化预后评估的意义。方法：选取池州市人民医院于2014年1月~2016年12月收治的162例乙肝肝硬化患者作为研究对象,分别测定患者血清可溶性P-选择素(sP-selectin)、可溶性细胞间黏附分子-1(sICAM-1)、免疫球蛋白和相关血清指标。比较不同肝硬化患者和肝癌患者血清指标差异性,Pearson分析血清可溶性P-选择素(sP-selectin)、可溶性细胞间黏附分子-1(sICAM-1)、免疫球蛋白和相关血清指标相关性,logistic多因素回归分析肝硬化转肝癌危险因素,受试者工作特征（ROC）曲线评价OR>1参数对肝硬化转肝癌预测价值。结果：原发性肝癌患者sP-selectin、sICAM-1、IgM、IgG、IgA显著高于代偿期肝硬化和失代偿期肝硬化患者（P<0.05）。失代偿期肝硬化患者sP-selectin、sICAM-1、IgM、IgG、IgA显著高于代偿期肝硬化患者（P<0.05）。不同Child-Pugh分级肝硬化患者中,C级患者sP-selectin、sICAM-1、IgM、IgG、IgA显著高于B级和A级患者（P<0.05）。B级患者sP-selectin、sICAM-1、IgM、IgG、IgA显著高于A级患者（P<0.05）。多发癌灶患者和单发癌灶原发性肝癌患者sP-selectin、sICAM-1和免疫球蛋白表达差异无统计学意义（P>0.05）。Ⅱ期原发性肝癌患者sP-selectin、sICAM-1和免疫球蛋白显著高于Ⅰ期患者（P<0.05）。肝硬化患者sP-selectin、sICAM-1、免疫球蛋白之间均互为正相关性（P<0.05）。以肝硬化是否转原发性肝癌为因变量进行多因素Logistic回归分析,结果显示,ALT、AST、sP-selectin、sICAM-1、IgM、IgG、IgA均是肝硬化转原发性肝癌的危险因素。ROC曲线分析可得,sICAM-1和sP-selectin诊断肝硬化转肝癌敏感性和特异性均高于其他因素。结论：血清sP-selectin、sICAM-1对乙肝肝硬化预后评估具有较高临床诊断精度,对肝硬化转原发性肝癌的诊断特异度、敏感度高于单纯免疫球蛋白检测,故其联合检测诊断肝硬化预后值得临床推广。     

Chronic liver disease: the detection and characterization of circulating immune complexes.

Circulating immune complexes containing IgG, IgM and hepatitis B surface antigen (HBsAg) in sera from groups of patients with various liver diseases were detected by both the C1q and conglutinin solid phase assays. Elevated levels of antigen non-specific immune complexes were observed in sera from all groups and complexes containing IgG were present to a greater extent than were IgM-containing complexes. Higher levels of complexes were generally obtained using the conglutinin assay than the C1q assay and the two assays were shown to preferentially bind complexes of different size ranges and antigen-antibody ratios. Only sera from HBsAg-positive patients had complexes containing HBsAg, and although serum HBsAg titres and levels of HBsAg-containing complexes were correlated, the correlation coefficient was low. The mean levels of immune complexes and the frequency of positive sera varied between different disease categories, but there was little correlation between levels of the three types of complexes detected by the two tests. Assay of immune complexes in sequential serum samples from an individual patient revealed considerable variation in the levels of the three complex types, demonstrating that the measurement of complexes in single serum samples is of limited value in assessing the potential significance of circulating immune complexes in hepatitis B.     

Prevalence of hepatitis A, B, C and E virus markers among patients with elevated levels of Alanine aminotransferase and Aspartate aminotransferase in Phnom Penh (Cambodia) and Nha Trang (Central Vietnam)

In order to describe the respective part of viral hepatitis in liver diseases observed in Cambodia and Vietnam, ninety consecutive patients with Aspartate aminotransferase (AST) and Alanine aminotransferase (ALT) > or = 100 Ul/l were tested for hepatitis A, B, C and E markers in Phnom Penh and Nha Trang. The markers were IgM antibodies to hepatitis A virus (anti-HAV IgM), hepatitis B surface antigen (HBsAg), antibodies to hepatitis C virus (anti-HCVAb) and IgG antibodies to hepatitis E virus (anti-HEV IgG). Recruited patients were predominantly adults and male (sex ratio 76%). Among these patients, 81% were tested positive to at least one marker in Nha Trang and 79% in Phnom Penh. In Nha Trang, HBsAg was more frequent (73%) than anti-HCV Ab (9%) while in Phnom Penh both markers were closely similar (HBsAg: 41%, anti-HCV Ab: 39%). In both population samples, HBsAg was more prevalent among young people whereas anti-HCV Ab were only detected in adults. No case of acute HAV infection was diagnosed in Nha Trang while anti-HAV IgM were detected in 20% of Cambodian patients. Anti-HEV IgG were infrequent (2% in Nha Trang, 5.5% in Phnom Penh). Only one case was notified, a male Vietnamese patient probably suffering from acute hepatitis E. More studies would be useful to improve the control measures against viral hepatitis in the public health programs.     

Anti-HBc IgM bei akuter und chronischer Hepatitis-B-Virus-Infektion

Summary Hepatitis B core antigen (HBcAg) synthesized in E. coli was used for determination of immunoglobulin M class-specific antibodies against HBcAg. It was found that 98% of cases with acute hepatitis B surface antigen (HBsAg) positive hepatitis type B were anti-HBc immunoglobulin M (IgM) positive. Atypical hepatitis B was detected in 33% of anti-HBc-positive HBsAg-negative cases with acute hepatitis. Anti-HBc IgM was positive for 6 months in acute resolving hepatitis type B, whereas cases resulting in chronic hepatitis B remained anti-HBc IgM-positive for up to 900 days. Chronic HBsAg carriers with severe liver disease had anti-HBc IgM more often than individuals with minor liver damage; 83% of HBsAg-positive liver cirrhoses, 63% of chronic aggressive hepatitis, 50% of HBsAg-positive liver carcinoma, but only 17% of chronic persistent hepatitis or 7% of healthy blood donors were anti-HBc IgM-positive. Determination of anti-HBc IgM is useful in detecting atypical hepatitis B virus infections without HBsAg in serum and, with some restrictions, in discriminating acute and chronic hepatitis type B.

     

Studies on HBsAg binding with polymerised human serum albumin by ELISA

A simple and sensitive ELISA was developed to characterize the interaction between polymerised human serum albumin (pHSA) and HBsAg, using pHSA-coated polyvinylmicrotitre plates as solid phase and anti-HBs-coupled HRPO as the conjugate. The interaction was found to be specific and dependent on the size of albumin polymer. pHSA-binding activity (pHSA-BA) was studied in both HBsAg-negative and HBsAg-positive sera from various liver diseases including acute viral hepatitis, fulminant hepatitis, cirrhosis of liver, chronic active hepatitis, and healthy HBsAg carriers. pHSA-BA was detected only in HBsAg-positive sera. Analysis of HBsAg-positive sera indicated pHSA-BA in high proportions of patients sera as compared to sera from healthy HBsAg carriers. pHSA-BA was detected both in the presence and absence of HBe markers, though the mean BA was relatively high in presence of HBeAg. The effect of human serum immunoglobulins (IgG, IgA, and IgM) on the BA was investigated and a correlation between pHSA-BA and HBsAg-IgM complex positivity in sera was established. Finally, the probable role of human serum IgM in facilitating the binding process was discussed.     

乙型肝炎患者免疫球蛋白检测的作用分析

目的 探讨乙型肝炎患者免疫球蛋白检测的临床意义.方法 128例乙肝患者分为急性乙肝组、慢性乙肝组和慢性重型乙肝组,另随机选取同期我院体检健康者40名作为对照组.4组受试者均晨起取空腹静脉血,测定血清免疫球蛋白（IgG、IgA、IgM）.结果乙肝患者IgG、IgM、IgA水平明显高于对照组,差异有统计学意义（P＜0.05）.结论 免疫球蛋白的增多提示患者肝脏细胞的受损加重,在一定程度上也可辅助判定患者肝脏损伤的程度和病情的进展阶段.     

散发性戊型肝炎病毒感染的诊断

用基因工程重组的戊型肝炎病毒基因结构区第二码框架和第二读码框架具有免疫表位的嵌合抗原，建立了间接酶联免疫法，检测散发性急性肝炎病人血清中抗－ＨＥＶＩｇＧ和ＩｇＭ抗体。在４６例急性肝炎病人中出抗－ＨＥＶＩｇＧ抗体阳性７例，阳性率为１５．２２％，７例ＩｇＧ抗体阳性中，有５例ＩｇＭ抗体也阳性，占７１．４％。     

Radioimmunoassay detection of igm antibodies to the hbv-associated delta (δ) antigen: Clinical significance in δ infection

A sensitive radioimmunoassay was developed for specific detection of IgM antibodies to the hepatitis B virus-associated δ antigen. The test is based on the selective absorption of IgM by anti–IgM fixed on a solid phase. Transient primary IgM anti4 responses with no conversion to a secondary IgG response were observed in acute self–limited δ infection. IgM anti-δ was invariably found in hepatitis B surface antigen (HBsAg) carriers with active δ infection and liver disease, while it was absent in HBsAg-positive or negative individuals with anti-δ of IgG class but without liver damage or intrahepatic δ antigen. IgM anti-δ appears useful in defining the epidemiology of acute δ infection and in the serological diagnosis of active δ disease from nonpathogenic or past δ infection.     

The IgM antibody responses to the core antigen of hepatitis B virus

Little is known about the immunoglobulin class of antibodies to HBcAg. In the present study sera containing anti-HBc were fractionated by sucrose density-gradient centrifugation, and all serum fractions were tested against HBcAg by immunoelectro-osmophoresis. In addition selected fractions were examined by complement fixation test, immune adherence hemagglutination and immune electron microscopy. Anti-HBc activity in IgG serum fractions was demonstrated by all four techniques used, but HBcAg-specific IgM was detected only by immunoelectro-osmophoresis and by immune electron microscopy. In acute hepatitis B, HBcAg-specific IgM was detected for up to eight weeks after the onset of jaundice. It was also found transiently in two patients who developed chronic hepatitis B without an icteric episode and in one out of thirteen patients with HBsAg-positive chronic liver disease, but in none of eight healthy HBsAg carriers. The results suggested that HBc Agspecific IgM is formed transiently in response to primary HBV infection but is generally undetectable in established HBsAg carriers.     

IgM antibody to a hepatitis C virus core peptide (CP14) for monitoring activity of liver disease in patients with acute or chronic hepatitis C

Antibodies to the hepatitis C virus (HCV) core of various immunoglobulin classes were determined by enzyme immunoassays with three synthetic peptides, CP14 (amino acids 5-40 of the core protein), CP10 (5-23), and CP9 (39-74). In 135 patients with chronic type C liver disease, anti-CPU, anti-CP10, and anti-CP9 of IgG class were detected in 99%, 94%, 82%, respectively; those of IgM class in 86%, 69%, and 39%; and those of IgA class in 56%, 40%, and 4%. Thus anti-CP14 was more prevalent than anti-CPlO or anti-CP9 in every immunoglobulin class. The prevalence of IgM anti-CP14 was much higher (P < 0.001) in patients (116/135 or 86%) than in asymptomatic carriers of HCV (13/39 or 33%). In seven patients with acute hepatitis C, IgM anti-CP14 continued to decrease in two in whom hepatitis resolved, but increased in five in whom hepatitis once resolved and then exacerbated. IgM anti-CP14 was followed in 30 patients with chronic hepatitis C during 24 weeks while they received recombinant interferon α-2a. IgM anti-CP14 decreased remarkably within 8 weeks in all of them. Thereafter, it continued to decrease in nine patients who responded to interferon and lost HCV RNA from circulation, but started to increase in five non-responders who continued to have high titers of HCV RNA. In the remaining 16 patients in whom HCV RNA decreased once and then increased, IgM anti-CP14 continued to decrease till 20 weeks and then increased. These results indicate that IgM anti-CP14 reflects the activity of liver disease, and is useful in following the outcome of patients with acute hepatitis C and in monitoring the response to interferon in patients with chronic hepatitis C. © 1994 Wiley-Liss, Inc.     

New assays for quantitative determination of viral markers in management of chronic hepatitis B virus infection.

We performed a quantitative study of serum hepatitis B virus (HBV) markers, including new parameters such as pre-S1 antigen (Ag), pre-S2 Ag, and anti-HBx, in 88 chronic hepatitis B surface antigen (HBsAg) carriers. New IMx assays for HBsAg and immunoglobulin M (IgM) anti-HBc detection were also used. The population studied was composed of 65 chronic hepatitis cases (40 positive for hepatitis B antigen [HBeAg] and 25 positive for anti-HBe) and 23 anti-HBe-positive, asymptomatic HBsAg carriers. Serum HBsAg levels detected by IMx were higher in HBeAg-positive than in anti-HBe-positive HBsAg carriers (all patient subgroups included) and correlated with the serum HBV DNA level (P = 0.0001). Both pre-S1 and pre-S2 Ags were detected by enzyme immunoassays in almost all HBsAg carriers. Both pre-S1 and pre-S2 Ag titers correlated positively with the serum HBsAg concentration (P = 0.0001), but only the pre-S1 Ag titer correlated with the level of serum HBV DNA (P = 0.02). The detection of low levels of IgM anti-hepatitis B core (anti-HBc) antibodies by IMx was associated with the presence of liver disease (P = 0.05) but not with the level of viral replication. The prevalence of anti-HBx antibodies detected by the enzyme immunoassay was slightly, although not significantly, higher in patients with high levels of HBV DNA (greater than 100 pg/ml) than in patients without detectable HBV DNA (P = 0.16). In anti-HBe-positive chronic HBsAg carriers, the quantitative detection of serum HBV DNA, pre-S Ag titers, and IgM anti HBc allowed us to predict which patients suffered from chronic liver disease and/or supported viral replication (P < 0.05). In a follow-up study of eight patients undergoing antiviral therapy, the clearance of both pre-S1 Ag and HBV DNA was associated with a subsequent clearance of HBV. Therefore, the quantitative determination of HBV DNA, pre-S Ags, IgM anti-HBc may prove useful for the decision to use and the monitoring of antiviral therapy, especially in anti-HBe-positive HBsAg carriers.