Immunoglobulin- and Hepatitis B Surface Antigen-Specific Circulating Immune Complexes in Chronic Hepatitis B Virus Infection

For assessing the role of circulating immune complexes (CIC) in chronic hepatitis B virus (HBV) infection, CICs containing IgM, IgG, and HBsAg were determined by C1q and conglutinin (K) assays in 216 patients with chronic HBV infection and 54 healthy controls. The concentration of each type of CIC in patients is higher than in controls (P= 0.0001). CIC is a common feature of chronic HBV infection with 95.8% of cases having at least one abnormal test result. At least one type of HBsAg-CIC is positive in 54.2% of patients. HBsAg-CIC positivity is associated with HBeAg positivity (P= 0.0001), higher aminotransferase levels (P< 0.002), and younger age (P= 0.001). IgG-CIC or IgM-HBsAg-CIC correlates with higher aminotransferase activity (P= 0.001). In conclusion, HBsAg-CIC correlates with HBV replication. IgG-CIC and/or IgM-HBsAg-CIC correlate with disease activity. Immune-mediated injury may play a role in the pathogenesis of chronic HBV infection.     

Tissue Localization of Australia Antigen Immune Complexes in Acute and Chronic Hepatitis and Liver Cirrhosis

In a significant percentage of examined cases of fulminant hepatitis, subacute hepatitis, chronic aggressive hepatitis, liver cirrhosis and chronic persistent hepatitis, Australia (hepatitis-associated) antigen (Au HAA) was identified in the liver and in extrahepatic locations. The several immunofluorescent patterns of Au HAA localization in hepatocytes strongly suggested various stages of Au HAA accumulation and release. Deposits of a mixture of immunoglobulins G and M and occasionally β1C-globulin were found in the cytoplasm of Au HAA containing hepatocytes, on their plasma membranes, on or in the nuclei, in the cytoplasm of Kupffer cells and, rarely, in the sinusoids. The accompanying tissue changes were hepatocellular degeneration and necrosis. These intra- and extracellular complexes of Au HAA and immunoglobulins displayed strong affinity for guinea pig complement in the immunohistochemical complement fixation reaction. When tested by immunodiffusion in agar, IgG dissociated from these complexes by potassium thiocyanate (KSCN) treatment showed anti-Au HAA specificity. In fulminant hepatitis neither Au HAA nor immunoglobulins and complement were found in the liver. In chronic aggressive hepatitis and subacute hepatitis the amount of the Au HAA immune complexes identified in the liver was approximately inversely proportional to the extent and severity of the parenchymal lesions. In liver cirrhosis and chronic persistent hepatitis there was a positive correlation between the amount of the Au HAA immune complexes found in the liver and the degree of hepatocellular damage. The deposits of Au HAA, identified in extrahepatic locations including germinal centers of lymph nodes and spleen, kidney glomeruli and blood vessel walls, were as a rule accompanied by deposits of IgG, IgM, β1C-globulin and fibrin. All these deposits showed strong affinity for guinea pig complement in the immunohistochemical reaction of complement fixation. Germinal center activation, chronic membraneous glomerulonephritis, panarteritis and simple arteriolar hyalinosis were found at sites of localization of these deposits.     

Increased IgM Class Circulating Immune Complexes in Acute Hepatitis A Virus Infection

For assessing the role of circulating immune complexes (CIC) in acute hepatitis A, IgM- and IgG-specific CIC were determined, by C1q and conglutinin (K) assays, in 205 patients with acute hepatitis A and 60 healthy controls. The concentration of each type of CIC in patients was higher than healthy controls (P= 0.0001). CIC was a common feature of acute hepatitis A with 95.6% of cases having at least one abnormal test result. The prevalence of abnormal IgM class CIC was significantly higher than IgG class CIC. There were significantly inverse correlations between levels of IgM class CIC and interval between onset of symptoms and patient presentation. The prevalence of abnormal IgM CIC was higher in patients with higher alanine aminotransferase (P= 0.001) and patients with jaundice (P= 0.0002). In conclusion, IgM class CIC is the predominant CIC in acute hepatitis A and correlated with disease activity. CIC may play a role in the pathogenesis of acute hepatitis A.     

Circulating Immune Complexes in Systemic Scleroderma

Circulating immune complexes were detected by the immunoelectrophoretic method in 18 of 29 (62 per cent) of patients with systemic scleroderma. The presence of immune complexes did not correlate with that of antinuclear antibodies to dsDNA, DNP, RNP, and Sm. The mean levels of immunoglobulins G, A, and M as well as of C3 were significantly higher in patients with systemic scleroderma than in blood donors.     

Circulating Immune Complexes in Hashimoto's Thyroiditis

Circulating immune complexes (IC) were determined in sera from 41 patients with Hashimoto's thyroiditis by a polyclonal rheumatoid factor (pRF) assay based on the inhibition of the agglutination of IgG-coated latex particles. Elevated levels of IC were found in 63% (26/41) of the sera. There was a significant correlation (Rho = 0.91, P < 0.001) between results obtained before and after treatment of sera with dithiothreitol (DTT). By precipitation with 2.5% polyethylene glycol (PEG) before pRF inhibition assay, the activity of IC was found in only 7% (3/41) of the sera. Size chromatography studies of the sera showed the inhibitory activity predominantly in the intermediary region. When found in the IgM-region the activity was not reduced by DTT. By use of a polyethylene glycol complement consumption test (PEG-CC) the occurrence of IC was 10% (4/41). It was not possible to find any correlation between the detectable IC and the presence of microsomal, thyroglobulin, or thyroid-stimulating antibodies. Based on our studies the sizes of IC seemed to be heterogeneously distributed and the majority were not precipitated by PEG (2.5%, final concentration). The antibodies involved in the formation of complexes seemed to be of IgG or IgA classes. HLA-D typing of the patients showed a non-significant association between HLA-Dw5 and low levels of IC while the presence of HLA-Dw4 was significantly associated with a high level of IC (P < 0.05).     

Circulating Immune Complexes in Systemic Scleroderma

Circulating immune complexes were detected by the immunoelectrophoretic method in 18 of 29 (62 per cent) of patients with systemic scleroderma. The presence of immune complexes did not correlate with that of antinuclear antibodies to dsDNA, DNP, RNP, and Sm. The mean levels of immunoglobulins G, A, and M as well as of C3 were significantly higher in patients with systemic scleroderma than in blood donors.     

A Survey for Circulating Immune Complexes in Patients with Acute Myocardial Infarction

A new assay for the detection of circulating C1q-binding immune complexes (IC) is described. The assay makes use of solid-phase C1q and iodinated soluble protein A, extracted from the cell wall of Staphylococcus aureus. In a model system the assay could detect heat-aggregated IgG down to a concentration of about 50 ng/ml. This method and three other assays, previously described, were used to survey the appearance of IC activity in sera from hospitalized patients with acute myocardial infarction. Depending on the assay system used, from 56% to 66% of the patients investigated were found to develop circulating IC. The earliest appearance of circulating IC was noted 5 days after infarction. The highest incidence of positive reactions and the strongest reactions occurred 2 to 3 weeks after hospitalization; thereafter the IC positiveness tapered off, and all patients were negative 6 weeks after infarction.     

确证TRFIA检测乙肝病毒表面抗体结果的研究

建立简易时间分辨免疫荧光法(TRFIA)检测乙型肝炎病毒表面抗体(抗-HBs)的中和试验方法,验证TRFIA检测结果.收集乙型肝炎病毒表面抗原(HBsAg)异常值血清,确定TRFIA中和抑制试验中和比为0.25-1.26ng/mL∶1mIU/mL,对抗-HBs的定量结果进行确认.结果显示,使用TRFIA定量检测的结果与...     

IgE-Containing Circulating Immune Complexes in Churg-Strauss Vasculitis

In five patients with vasculitis, hypereosinophilia, and elevated serum IgE levels a diagnosis of Churg-Strauss syndrome was established. To identify a possible role of IgE in pathogenic mechanisms leading to the vasculitis, we performed a sequential precipitation of the patients' sera with different concentrations of polyethylene glycol (PEG) 6000. Using a radio immunosorbent test, we tested the precipitates obtained for IgE. Considerable amounts of IgE were traced in the serum precipitates of all patients, especially after the second precipitation step (4.0% PEG). In contrast, no IgE-containing precipitates were detectable in sera from patients with different allergic diseases and high IgE serum levels. Together with an increase in C3d serum levels and the failure to demonstrate C1q-binding material in patients' sera, these data suggest the involvement of IgE-containing immune complexes in the pathogenesis of Churg-Strauss vasculitis, activating the complement via the alternate pathway.     

A Valuable Antigen Detection Method for Diagnosis of Acute Hepatitis E

Hepatitis E virus (HEV) is a serious public health problem. The commonly used tests that are specific for current HEV infection diagnosis include the detection of anti-HEV IgM and HEV RNA. Here, we report an improved enzyme-linked immunosorbent assay (ELISA) method for HEV antigen detection with a linear range equivalent to 6.3 × 10³ to 9.2 × 10⁵ RNA copies per ml. The monoclonal antibody (MAb) 12F12, a high-ability MAb that binds HEV virus, was selected as the capture antibody from a panel of 95 MAbs. The positive period of HEV antigenemia in infected monkeys using this test was, on average, 3 weeks longer than previously reported and covered the majority of the acute phase. The positive detection rates of IgM, RNA, and new antigen from the first serum samples collected from 16 confirmed acute hepatitis E patients were 81% (13/16), 81% (13/16), and 100% (16/16), respectively. In three patients, the initial serum specimens that tested negative for IgM, despite the presence of symptoms of acute hepatitis and elevated alanine aminotransferase (ALT) levels, were positive for HEV antigen and HEV RNA. In contrast, the serum samples of the three RNA-negative patients were antigen positive (and IgM positive), possibly due to the degradation of HEV nucleic acids. Our results suggest that this new antigen detection method has acceptable concordance with RNA detection and could serve as an important tool for diagnosing acute hepatitis E.     

乙型肝炎病毒表面抗原定量检测及临床意义分析

乙型肝炎病毒表面抗原（HBsAg）定量检测是临床治疗乙型肝炎的一个非常重要的指标,它不仅能区分乙型肝炎病程的各种分期,而且对药物治疗效果及预后有良好的预测作用.本文从乙型肝炎病毒表面抗原的结构、定量检测的方法学以及在临床中的应用等方面做一综述分析.     

乙型肝炎患者免疫功能的检测及其临床意义

研究乙型肝炎患者外周血T淋巴细胞亚群、NK细胞和血清免疫球蛋白的变化及临床意义.采用流式细胞仪检测150例乙肝患者和30名健康者(对照组)外周血T淋巴细胞亚群(CD3 CD4 、CD3 CD8 )、NK细胞,免疫散射法检测血清免疫球蛋白(IgG、IgM、IgA)变化.结果表明各临床类型乙肝患者NK细胞降低,与对照组比较有显著性差异(P<0.01);慢性乙型肝炎组、慢性重型乙型肝炎组、肝硬化组外周血CD3 CD4 、CD3 CD8 T细胞均下降,其中慢性重型乙型肝炎组外周血CD3 CD8 与对照组比较有显著性差异(P<0.01),急性乙型肝炎组外周血T细胞亚群变化不明显(P>0.05);各临床类型乙型肝炎患者血清免疫球蛋白IgG、IgA随着病情的进展逐渐升高,与对照组比较有显著性差异(P<0.01).因此认为慢性乙肝患者存在细胞免疫和体液免疫功能紊乱,免疫功能的检测对乙肝的诊断、治疗及预后的判断有着一定临床意义.     

慢性乙型肝炎患者e抗原消失的临床特点

目的 探讨慢性乙型肝炎患者使用核苷类似物抗病毒治疗e抗原消失后HBV DNA、转氨酶等指标的变化特点。方法 选取2014年12月~2017年6月四川省中西医结合医院门诊109例慢性乙型肝炎患者,在服用核苷类似物抗病毒的治疗过程中出现e抗原消失,检测并分析治疗前后e抗原抗体系统的变化与HBV DNA关系、ALT异常率、血清HBV DNA水平与HBeAg相关性分析。结果 HBV DNA转小二阳时及转后半年、1年与治疗前比较,差异有统计学意义（P＜0.05）;转小二阳后半年与刚转小二阳时比较,差异有统计学意义（P＜0.05）;转小二阳后1年与刚转小二阳时比较,差异无统计学意义（P＞0.05）。治疗前和治疗后的ALT异常率比较,差异有统计学意义（P＜0.05）,转小二阳时以及转后的半年、1年ALT异常率相比,差异均无统计学意义（P＞0.05）。血清HBV DNA水平与HBeAg呈正相关关系（r=0.879,P＜0.001）。结论 大三阳患者在治疗过程中,发生e抗原的消失的状态是不稳定的,仍然可能会出现病毒DNA的复制,转氨酶的异常。     

慢性乙肝患者血清前Pre-S1抗原检测及其与肝功能关系的研究

目的 检测慢性乙型肝炎病毒患者前S1(Pre-S1)抗原,探讨其与肝功能的关系.方法 收集270例慢性乙型肝炎病毒患者血清,采用ELISA方法检测Pre-S1抗原;采用日立7170全自动生化分析仪检测丙氨酸氨基转移酶(ALT)和天冬氨酸氨基转移酶(AST).结果 270例慢性乙型肝炎病毒患者中,Pre-S1抗原阳性中ALT异常检出率为91.1%,AST异常检出率为93.0%,Pre-S1抗原中阴性中ALT异常检出率为28.3%,AST异常检出率为35.4%,两组结果有显著性差异(P<0.05).结论慢性乙型肝炎患者中,Pre-S1抗原可作为判断乙型肝炎病毒(HBV)感染,复制及对肝细胞损伤的另一个指标之一.     

Immune Mechanisms and the Action of Interferon in Chronic Hepatitis B Virus Infection

Cytotoxic T lymphocytes can eliminate cells infected with hepatitis B virus. A defect of T lymphocyte-mediated cytolysis seems to exist in chronic hepatitis B infection. T cell-mediated lysis is dependent on HLA antigens of the infected host and this may explain the increased or decreased frequency of particular HLA antigens in chronic carriers of hepatitis B virus. This virus may decrease the concentration of HLA antigens expressed on liver cells. Interferon increases the HLA antigen expression on T lymphocytes and hepatocytes, thereby enhancing T lymphocyte-mediated lysis of infected liver cells and elimination of the hepatitis B virus.     

Antigen Display,T-Cell Activation,and Immune Evasion during Acute and Chronic Ehrlichiosis

How spatial and temporal changes in major histocompatibility complex/peptide antigen presentation to CD4 T cells regulate CD4 T-cell responses during intracellular bacterial infections is relatively unexplored. We have shown that immunization with an ehrlichial outer membrane protein, OMP-19, protects mice against fatal ehrlichial challenge infection, and we identified a CD4 T-cell epitope (IA^b/OMP-19_107-122) that elicited CD4 T cells following either immunization or infection. Here, we have used an IA^b/OMP-19_107-122-specific T-cell line to monitor antigen display ex vivo during acute and chronic infection with Ehrlichia muris, a bacterium that establishes persistent infection in C57BL/6 mice. The display of IA^b/OMP-19_107-122 by host antigen-presenting cells was detected by measuring intracellular gamma interferon (IFN-γ) production by the T-cell line. After intravenous infection, antigen presentation was detected in the spleen, peritoneal exudate cells, and lymph nodes, although the kinetics of antigen display differed among the tissues. Antigen presentation and bacterial colonization were closely linked in each anatomical location, and there was a direct relationship between antigen display and CD4 T-cell effector function. Spleen and lymph node dendritic cells (DCs) were efficient presenters of IA^b/OMP-19_107-122, demonstrating that DCs play an important role in ehrlichial infection and immunity. Chronic infection and antigen presentation occurred within the peritoneal cavity, even in the presence of highly activated CD4 T cells. These data indicated that the ehrlichiae maintain chronic infection not by inhibiting antigen presentation or T-cell activation but, in part, by avoiding signals mediated by activated T cells.Major histocompatibility complex class II (MHC-II)-restricted CD4 T cells are well known to contribute to immunity during many intracellular bacterial infections. Less is known, however, regarding when and where during infection MHC-II/peptide antigens are available to generate and maintain protective T-cell immunity. Some intracellular bacteria can modulate antigen presentation to CD4 T cells (7, 30, 49), although many intracellular bacteria elicit both acute and chronic CD4 T-cell responses, even in the absence of sterilizing immunity (25, 29, 33, 48, 50). How CD4 T-cell responses are initiated and maintained during both acute and chronic infection is, therefore, an ongoing question that is relevant to a number of bacterial, viral, and parasitic infections. Knowledge of T-cell antigen presentation will aid in understanding how, during acute infections, the quantity and quality of the antigens presented serve to regulate T-cell differentiation and memory development. During chronic infections, persistent antigen display likely regulates effector T-cell populations that are essential for the maintenance of immunity. Several studies have addressed where and when T-cell MHC/peptide ligands are presented during viral and bacterial infections. For example, during Listeria monocytogenes infection, early antigen presentation was sufficient to drive the full program of CD8 T-cell expansion and contraction (8). Antigen presented by MHC-I and MHC-II proteins has been shown to persist following influenza and vesicular stomatitis virus infections (15, 44), and it likely is responsible for the maintenance of effector T-cell responses during chronic infections (52).Ehrlichiosis is an emerging infectious disease of both humans and animals and is caused by obligate intracellular rickettsiae of the genus Ehrlichia. Our studies have focused on Ehrlichia muris, which first was identified in Eothenomys kageus mice in Japan and in Hemaphysalis flava ticks (47). Although the bacterium was discovered in mice, E. muris is closely related to ehrlichiae that cause human and animal diseases, in particular, Ehrlichia chaffeensis, the etiologic agent of human monocytotropic ehrlichiosis (32). Unlike E. chaffeensis, E. muris causes a low-level persistent infection in mice (31, 47). Moreover, E. muris infection generates protective immunity against another closely related but highly virulent ehrlichia, first identified in Ixodes ovatus ticks in Japan, known as ehrlichia from Ixodes ovatus (IOE) (37). It is not certain why E. muris, but not E. chaffeensis or low-dose IOE, can generate protective immunity against high-dose IOE infection. However, we have demonstrated that immunity to IOE can be achieved in the absence of classical CD4 T cell-mediated helper functions but not in the absence of B cells (4), suggesting that B cells and/or antibodies play a critical role in protective immunity. Nevertheless, CD4 T-cell responses are generated during infections by E. muris and other ehrlichiae and likely contribute to immunity, especially during chronic infection. For example, we have shown previously that CD4 T cells contribute to immunoglobulin class switching during E. muris infection, even though some class switching does occur in the absence of these cells (4).In the present study, the display of MHC/peptide antigen complexes in various tissues and anatomical locations has been monitored during acute and chronic E. muris infection. Our findings reveal that MHC-II/peptide antigen presentation and associated T-cell responses are regulated temporally and spatially during acute infection. Although dendritic cells (DCs) usually are considered the primary antigen-presenting cells (APCs) involved in antigen presentation to naive T cells (13, 28), the ehrlichiae are largely monocytotropic, and it was not known whether or when DCs harbor ehrlichiae and/or present MHC/peptide antigens to T cells. We show that CD11c-positive DCs harbor viable ehrlichiae and present specific antigen during the early phase of infection. Although chronic antigen presentation occurs in the peritoneal cavity and likely is responsible for chronic CD4 T-cell activation, the ehrlichiae nonetheless are able to avoid elimination while residing in macrophages. These findings have important implications for our understanding of how the ehrlichiae and related pathogens evade the cellular immune response during chronic infection.     

Levels of Schistosoma mansoni Circulating Antigen in Chronic Hepatitis C Patients with Different Stages of Liver Fibrosis

The goal of this study was to determine the levels of S. mansoni antigen in different liver fibrosis stages with chronic hepatitis C (CHC) Egyptian patients. A total of 174 CHC patients showing HCV-NS4 antigen and HCV- RNA in their sera were included. S. mansoni antigen was detected in serum using Western blot and ELISA. The levels of interferon-γ (IFN- γ) were determined using ELISA. The 50 kDa S. mansoni antigen discriminated patients infected with S. mansoni from healthy individuals with 0.93 area under curve (AUC), 92% sensitivity, and 97% specificity. The level of S. mansoni antigen (μg/ml) was significantly (P < 0.0001) increased with the progression of liver fibrosis stages (26.9 ± 17.5 in F1, 42.1 ± 25.2 in F2, 49.8 ± 30.3 in F3 and 62.2 ± 26.3 μg/mL in F4 liver cirrhosis), 26.9 ± 17.59 in significant fibrosis (F2–F4); 51.2 ± 27.9 in advanced fibrosis (F3–F4). A significant correlation (r = 0.506; P < 0.0001) was shown between the levels of the S. mansoni antigen and the HCV-NS4 antigen. In conclusion, the presence of S. mansoni antigen in different liver fibrosis stages of CHC patients confirming that concomitant schistosome infection aggravates liver disease.     

The Detection of Circulating Immune Complexes and IgG and IgM Rheumatoid Factors in Normal Human Pregnancy*

ABSTRACT: The development of immune complexes (IC) and rheumatoid factors (RF) during normal, uncomplicated pregnancy is a controversial issue. Discrepancies in previous reports are due most likely to the different methods used to detect both IC and RF. Using four sensitive radioimmunoassays for immune complexes employing both Clq and mRF and employing sensitive and specific radioimmunoassays for the detection of IgM-RF and IgG-RF, we examined the sera of 35 normal subjects in their third trimester of pregnancy. Immune complex concentrations as measured by four assays were not increased during gestation. However, both IgM-RF and IgG-RF were significantly elevated even though the concentrations of immunoglobulins M and G were virtually identical to the controls. These observations provide further insight into the immunological changes associated with pregnancy.     

Highly Sensitive Detection of Hepatitis B Virus Surface Antigen by Use of a Semiautomated Immune Complex Transfer Chemiluminescence Enzyme Immunoassay

The performance of hepatitis B surface antigen (HBsAg) screening assays is continuously improved to reduce the risk of transfusion-associated hepatitis B. In this study, a semiautomated immune complex transfer chemiluminescence enzyme immunoassay (ICT-CLEIA) for the detection of HBsAg, which is as sensitive as hepatitis B virus (HBV) DNA PCR, was developed; the ICT-CLEIA assay performance was compared with the performance of the Architect HBsAg QT assay and HBV DNA PCR. The specificities in the initial assay and after retesting were 99.50% (1,988/1,998 samples) and 99.95% (1,997/1,998 samples), respectively. The analytical detection limit was determined to be 0.2 mIU/ml using the 2nd International WHO HBsAg standard, and the cutoff value (0.5 mIU/ml) of the ICT-CLEIA assay was 8.0 standard deviations (SD) above the mean of the HBsAg-negative specimens. The ICT-CLEIA assay could detect HBsAg even in the presence of anti-HBs antibodies and demonstrated a 23.6-day-shorter window period using commercially available HBsAg seroconversion panels than the Architect HBsAg QT assay. Furthermore, the monitoring of the viral kinetics by the ICT-CLEIA assay and the HBV DNA PCR produced very similarly shaped curves during both the HBsAg seroconversion and reverse seroconversion periods. Therefore, the ICT-CLEIA assay may be useful not only for an earlier detection of HBV reactivation but also for the monitoring of hepatitis B patients.