流行性出血热特异性循环免疫复合物的动态观察和分析

应用双抗体夹心ＥＬＩＳＡ检测了８８例流行性出血热患者２２８份血清中特异性循环免疫复合物，并与胶固素－ＥＬＩＳＡ检测循环免疫复合物的结果进行了比较。结果表明，胶固素－ＥＬＩＳＡ检出循环免疫复合物１８３份（８０．２６％），抗原特异性ＥＬＩＳＡ检出ＩｇＧ型循环免疫复合物１８８份（８２．４５％），ＩｇＭ循环免疫复合物２０５份（８９．９１％），ＩｇＡ型循环免疫复合物１４７份（６４．４７％）。在各类特异性循环免疫复合物中，ＩｇＭ型在各病期检出率最高，ＩｇＧ型次之，ＩｇＡ型最低。在发病初期，各类特异性循环免疫复合物检出率均较高，极期达高峰，随病情缓解而渐降低。对特异性循环免疫复合物检出率与流行性出血热患者病情程度相关性的观察表明，轻型患者检出率较中，重型患者为低，而中、重型间检出率差别不明显。此外，轻型患者特异性循环免疫复合物动态变化明显，极期达高峰后很快下降，而中、重型患者检出率下降趋势较缓慢。上述结果直接证实了流行性出血热循环免疫复合物的形成是特异性病毒抗原刺激机体免疫应答的结果，并构成了流行性出血热免疫发病机制中的重要环节。     

用ELISA检测流行性出血热特异性免疫复合物的初步研究

为检测流行性出血热特异性循环免疫复合物,建立了抗原特异性双抗体夹心ELISA技术。以兔抗EHF IgG包被反应板,使待检标本内循环免疫复合物通过暴露的EHF病毒抗原位点而被结合到固相上,再以辣根过氧化物酶标记羊抗人IgG进行示踪,从而检出EHF’特异性循环免疫复合物。在228份EHF患者血清中,用胶因素ELISA检出非特异性循环免疫复合物183份,抗原特异性ELISA检出特异性循环免疫复合物188份。本方法特异性强,重复性及稳定性较好,可应用于临床检测。     

豚鼠C1q在检测人CIC中的应用

本文首次报道了以优球蛋白沉淀法提取的豚鼠C_1q替代人C_1q,作固相酶联免疫吸附试验检测人循环免疫复合物的结果。用豚鼠C_1q和一定条件下的不同浓度的AHG,作固相酶联免疫试验,制订了检测人循环免疫复合物的标准曲线。通过对24例正常人、11例SLE病人、28例乙型肝炎病人和21例流行性出血热病人的检测,发现正常人与病人的循环免疫复合物水平间,经统计学处理有显著性差异。检出的阳性率分别是4.2％,90.9％,100％和47.6％。该方法简便,灵敏度高,成本低,且不受混淆IgG的影响。     

流行性出血热病人特异性循环免疫复合物的检测

应用ＥＬＩＳＡ（双抗体夹心法）检测了３３例４～７病日流行性出血热（ＥＨＦ）病人血清中特异性循环免疫复合物（ＣＩＣ）水平；并比较了联合抗过敏加输新鲜血浆疗法治疗前后的血清特异性ＣＩＣ消长情况。结果表明，ＥＨＦ病人可检出ＥＨＦ病毒抗原的特异性ＩｇＥ型和ＩｇＧ型ＣＩＣ。联合抗过敏加输新鲜血浆治疗后的病人血清中特异性ＩｇＥ型和ＩｇＧ型ＣＩＣ均明显降低。     

EHFV微量细胞病变中和试验方法的建立及其在中和抗体检测中的应用

本文报道了一种新的流行性出血热病毒中和抗体检测方法即微量细胞病变中和试验。该法利用流行性出血热病毒在Vero细胞上繁殖时可引起细胞病变的特性，将32－100CCID50病毒液与等量待检出血清混合，37℃中和1．5小时后种入微量培养板孔中的细胞悬液中，培养6－8天后即可判定结果，用该法检测流行性出血热病人血清和灭活疫苗免疫机体后和中和抗体，获得了与空斑减少中和试验法基本一致或者更加敏感的结果。该法简单，特异、准确、敏感、快速、它的推广应用对加我国流行必出血热的血清分型和流行病学调查及疫苗制备毒株的筛选等基础研究工作均有较大的意义。     

流行性出血热患者外周血中性粒细胞内免疫复合物的研究

本文首次报导了采用双标记免疫荧光技术,从流行性出血热(EHF)患者外周血中性粒细胞(NPL)内检出免疫复合物(IC)。在所检测的18例患者外周血涂片中,均见到NPL内有IC存在,IC呈点状或片状分布于细胞浆中。计数IC~+-NPL(以EHFV-Ag~+-NPL和人IgG~+-NPL表示),发现其检出率因病期而异,在发热期检出率可分别达79-93％和40-70％,随病程进展而下降,至少尿期和恢复期,IC~+-NPL已极少或完全消失。这表明,NPL参与吞噬和清除循环免疫复合物(CIC)的活动,其释出的活性物质和裂解产物又可能造成其后EHF病变过程。     

联合抗过敏加输新鲜血浆治疗流行性出血热后期病人537例疗效观察

流行性出血热免疫学发病机理是Ⅰ型和Ⅲ型变态反应联合作用。Ⅰ型变态反应参与ＥＨＦ早期发病过程，随病程进展特异性抗体水平上升，与相应抗原结合形成免疫复合物（ＩＣ）演变成以Ⅲ型变态反应为主的致病机制。ＩＣ沉积于肾脏等组织引起免疫损伤，导致病损加重，病情恶化。在此基础上，应用联合抗过敏加输新鲜血浆疗法治疗５３７例ＥＨＦ后期病人，获显著疗效，使病死率由原综合液体疗法的８．８２％下降到０．５５％。     

联合抗过敏加输新鲜血浆治疗流行性出血热后期病人537例疗…

流行性出血热免疫学发病机理是Ⅰ型和Ⅲ型变态反应联合作用。Ⅰ型变态反应参与ＥＨＦ早期发病过程，随病程进展特异性抗体水平上升，与相应抗原结合形成免疫复合物（ＩＣ）演变成以Ⅲ型变态反应为主的致病机制。ＩＣ沉积于肾脏等组织引起免疫损伤，导致病损加重，病情恶化。在此基础上，应用联合抗过敏加输新鲜血浆疗法治疗５３７例ＥＨＦ后期病人，获显著疗效，使病死率由原综合液体疗法的８．８２％下降到０．５５％。     

流行性出血热患者血中SOD、LPO与红细胞免疫粘附功能的变化

流行性出血热患者血中ＳＯＤ、ＬＰＯ与红细胞免疫粘附功能的变化古宏标，刘燕，何浩明广东药学院生物技术研究室广州５１０２２４近年来在对流行性出血热（ＥＨＦ）发病机理的研究中发现，患者血中早期即出现特异性免疫复合物，并沉积于红细胞、血小板、肾小球和肾小管基...     

国内医讯

流行性出血热病毒的直接损伤作用同济医科大学协和医院传染病学专家罗端德教授等进行的流行性出血热发病机理研究结果证实,流行性出血热病毒对人体细胞有直接损伤作用,早期抗病毒治疗可降低病死率。流行性出血热的发病机理是人体免疫损害作用,但部分患者应用免疫抑制治疗后,早期仍出现肾脏和血管损伤,使病死率增加。罗端德等采用体外培养人体骨髓细胞作为感染细胞,分别应用扫描电镜、透射电镜、免疫电镜和膜脂流动性测定等进行检测。结果发现,骨髓细胞感染流行性出血热病毒后,细胞形     

流行性出血热IgM抗体检测(抗体捕捉ELISA法)

本文报道应用IgM-抗体捕捉酶联免疫吸附法(IgM antibody-capture ELISA,或称反向间接酶联免疫吸附法,Reverse indirect ELISA)检测流行性出血热(Epidemic hemorrhagic fever,EHF)患者血清特异性IgM抗体的结果。该方法能特异、敏感地测出EHF患者发病第2天的血清IgM抗体,阳性率为87.5(7/8),第4病日阳性率达95％(57/60),而病程第7天的38例患者血清全部阳性。15例非EHF发热病人及10例正常人血清无1例阳性,并经2-MB试验与加热试验证明了其特异性。另外,研究表明,对于类风湿因子(RF)干扰所致假阳性反应,本实验所采用的阴性抗原平行对照设计能成功地予以校正而排除。     

Characterization of circulating immune complexes in heart disease

Circulating immune complexes (CIC) were characterized for the content in IgG, IgA, IgM, C3 and C4 in patients with heart disease. The levels of IgG, IgM and C4 in PEG-precipitates of patients' sera were significantly higher than those found in controls, and the precipitation profile was similar to that of rheumatoid arthritis patients. Differences were observed in the composition of CIC: IgM was highest in association with myocarditis, and C3 predominated in cases of valvular disease. Complex-bound C4 was significantly higher in patients with myocardial infarction which developed pericarditis either early or late in the evolution of the disorder. Antimyocardial antibodies could be detected in sera and in corresponding PEG-precipitates. The bulk of the data suggests that CIC might play a pathogenetic role in various heart diseases.     

Significance of circulating immune complexes in pulmonary tuberculosis

In the present study we have tried to demonstrate circulating immune complexes (CIC) in sera from patients with pulmonary tuberculosis (TB) by three techniques; latex agglutination; 3.5% PEG precipitation and determination of optical density at 280 nm and RIA of CIC using bovine spermatozoa. About 40 normal control sera and 100 TB patients sera were investigated for the presence of CIC. Seventeen per cent cases of pulmonary TB were positive by latex agglutination while none of the control was positive. Levels of CIC as detected by PEG precipitation and RIA were significantly elevated in patients as compared to normal controls. While IgG, IgA and IgM were elevated in the CIC of patients, IgM immunoglobulins were detected only in patients and not in controls. Detection of CIC may at times be useful in diagnosis, prognosis and therapeutic monitoring of disease processes, but it is the characterization of immune complexes (IC) and identification of the specific components of these complexes which holds the greatest potential for better understanding of disease mechanisms. CIC were precipitated using 3.5% PEG from sera of patients suffering from TB. The specific anti-TB antibody component of complex was determined using S. aureus protein A as a solid phase, Anti-BCG antibody and 125I-labelled TB antigen. The specific TB antigen component of the IC was dissociated thermally from TB antibody and assayed by a radioimmunoassay technique developed in our laboratory. Patients were classified into two groups. Those those sputum was positive for Mycobacterium tuberculosis by smear and/or culture and those whose sputum was negative. The TB antigen concentrations of CIC was higher 19.1 +/- 2.3 ng/ml (mean +s.e.) in sputum positive cases, and 9.9 +/- 1.9 ng/ml in sputum negative cases as compared to 2.2 +/- 0.3 ng/ml in controls. Patient groups were significantly different from controls as well as from each other (P less than 0.001). Anti-TB antibody ratios were 11.7 +/- 1.48, 5.1 +/- 1.5 and 0.6 +/- 0.1 in sputum positive, sputum negative and controls. The significance of differences between the groups was P less than 0.001. The effect of treatment administered over a period of 12 weeks or more was evaluated. It was observed that in patients with persistent demonstration of M. tuberculosis in the sputum, the TB antigen and TB antibody levels of CIC were consistently high. In patients who responded to anti-tubercular drugs the TB antigen levels decreased progressively while TB antibody levels remained high.(ABSTRACT TRUNCATED AT 400 WORDS)     

Diagnosis of Colorado tick fever virus infection by enzyme immunoassays for immunoglobulin M and G antibodies.

An immunoglobulin M (IgM) capture enzyme immunoassay technique was adapted for the detection of antibody to Colorado tick fever virus in sera from 84 individuals for whom diagnosis had been confirmed by virus isolation or neutralization test. Titers were compared with those for IgG and neutralizing antibodies in these Colorado tick fever cases. IgM antibody titers were higher than neutralizing antibody titers, but neither appeared until 1 to 2 weeks after the onset of illness. Neutralizing antibodies were detected earlier than IgM antibodies, and both were detected with greater frequency than IgG antibodies. Late-convalescent-phase sera contained both neutralizing and IgG antibodies, but IgM was all but undetectable by 2 months after onset. Although the neutralization test may remain the serological test of choice, the enzyme immunoassay for IgM antibody offers a simple and more rapid method of serodiagnosis; the enzyme immunoassay is, however, less sensitive than the neutralization test. Furthermore, because there was a sharp decline in IgM antibody after 45 days, the presence of IgM antibody in a single serum sample provides a basis for the presumptive serodiagnosis of recent Colorado tick fever virus infection.     

Detection and partial characterization of immune complexes in patients with rheumatoid arthritis plus Sjogren's syndrome and with Sjogren's syndrome alone.

In order to characterize the immune complexes detected in patients with Sjogren''s syndrome (SS) and with rheumatoid arthritis (RA), the sera of 19 patients with SS alone and 11 with SS plus RA were examined. Elevated quantities of circulating immune complexes (CIC) were detected in 67% by the C1q-binding assay (C1q-BA), 73% by the C1q-solid phase (C1q-SP) assay, 43% by the monoclonal rheumatoid factor solid phase assay (mRF-SP) and 33% by the mRF-inhibition assay (MRF-Inh). Elevated concentrations of IgM RF were detected in 83% and of IgG RF in 73% of the sera by radioimmunoassay. Strong correlations existed between RF of the IgM and IgG classes and both the C1q-BA and the C1q-SP. Three lines of evidence indicated that RF were important components of the immune complexes detected by these radioimmunoassays. These results indicated that in those patients with RA plus SS, as well as those with SS alone, both IgM and IgG RF made substantial contributions to immune complexes detected both by C1q-BA and C1q-SP.     

用免疫转印技术检测流行性出血热病人特异性抗体的探讨

采用免疫转印技术检测了147份临床诊断为EHF病人血清中的特异性IgG抗体,结果133份血清可检出针对分子量在68和66kd左右两种病毒多肽的抗体,检出率为90.4％,略高于免疫荧光试验(88.4％),两种抗特异性多肽抗体的几何平均滴度分别为荧光法的4倍和2.1倍。本法具有特异性强、敏感性高和重复性较好等特点,并能检出针对不同抗原亚基成份的抗体,可作为EHF特异性诊断的方法之一。     

Autoantibodies and circulating immune complexes in sera from patients with hepatitis B virus-related chronic liver disease.

Sera from 56 patients with hepatitis B virus-related chronic liver disease (CLD) and 30 normal individuals as controls were examined by enzyme immunoassay (EIA) for the presence of autoantibodies directed against actin, tubulin, myosin, double-stranded (ds) DNA, polymerized human albumin, thyroglobulin, and trinitrophenyl coupled to bovine serum albumin (TNP-BSA). Patients with CLD had consistently elevated levels of IgG, IgA, and IgM antibodies directed against all the panel antigens. The percentage of patients with autoantibodies of the IgA class was particularly high: respectively, 88 and 78% of the patients had strikingly high levels of anti-actin and anti-TNP-BSA IgA autoantibodies. High amounts of IgA and IgG antibodies to polymerized albumin as well as IgG to thyroglobulin were also detected. Circulating immune complexes (CIC) were isolated from patients' sera and their autoantibody activities were tested on the same antigen panel. The autoantibodies thus detected were of the same class and possessed the same activities, although at higher values than those present in the homologous sera. These results indicate that, regardless of their origin, autoantibodies are present in high amounts in the sera of these patients. Moreover, autoantibodies participating in the formation of CIC might play a pathological role.     

Performance of immunoglobulin G (IgG) and IgM enzyme-linked immunosorbent assays using a West Nile virus recombinant antigen (preM/E) for detection of West Nile virus- and other flavivirus-specific antibodies

Focus Technologies developed an indirect immunoglobulin G (IgG) enzyme-linked immunosorbent assay (ELISA) and a mu-capture IgM ELISA for the detection of West Nile virus (WNV)-specific antibodies based on a WNV preM/E protein recombinant antigen. Normal and disease state serum panels were used to assess the performance characteristics of the two WNV ELISA kits. Totals of 807 and 1,423 sera were used to assess the IgG ELISA and IgM ELISA kits, respectively. The Focus Technologies IgG ELISA had a sensitivity of 97.6% and a specificity of 92.1% (excluding non-WNV flavivirus sera). The comparative method for WNV IgG may lack sensitivity in detecting IgG in early WNV infection, so the specificity of the Focus IgG ELISA may be higher than 92.1%. When sera from patients either infected with or vaccinated against other flaviviruses were tested on the WNV IgG assay, 35% of the sera reacted as positive for WNV IgG. Yellow fever and Japanese encephalitis vaccinees were less reactive in the IgG ELISA than St. Louis and dengue fever patients. The Focus Technologies IgM ELISA had a sensitivity and a specificity of 99.3% (excluding the non-WNV flavivirus sera). The overall cross-reactivity for the IgM ELISA to flavivirus sera was 12%, with 31% of St. Louis encephalitis patients found to be WNV IgM positive and no yellow fever vaccinees found to be WNV IgM positive. In a selected population of 706 sera, 15 false-positive WNV IgM sera were identified. The use of a background subtraction method for the IgM ELISA eliminated all 15 false-positive results, giving a specificity of 100% for the Focus IgM ELISA.