ENHANCED ANTIBODY ACTIVITY IN SERUM DEPLETED OF ANTIGEN

Presence of antigen and antibodies in a sample may interfere with the antibody, as well as with antigen detection assays. In such a situation, avidity of the probing antigen or antibody plays the key role in the assay. In the present study, using monoclonal antibodies against a mycobacterial antigen, lipoarabinomannan, patient serum is depleted of mycobacterial antigen by capture immunoradiometric assay and this antigen-depleted serum is tested for anti-lipoarabinomannan antibodies by inhibition immunoradio-metric assay. It is observed that serum, after depletion of antigen, revealed enhanced antibody activity compared to initial levels. It, therefore, appears that the avidity of the probing monoclonal antibody may detach the antigen from the loosely attached complexes and renders the complexed antibody free, thus increasing the reactive antibody molecules in the serum.     

Preparation of Horseradish Peroxidase Hydrazide and Its Use in Immunoassay

Abstract

Preparation of horseradish peroxidase (HRP) hydrazide that is HRP linked to adipic acid dihydrazide (HRP-ADH) and its use in enzyme immunoassay (EIA) is described. In this new strategy, horseradish peroxidase was conjugated to adipic acid dihydrazide using a carbodiimide coupling method. The resulting HRP-ADH was then coupled to cortisol-21-hemisuccinate (Cortisol-21-HS) to prepare enzyme conjugate. The prepared cortisol-21-HS coupled ADH-HRP (Cortisol-21-HS-ADH-HRP) enzyme conjugate was used for the development of an enzyme linked immunosorbent assay (ELISA) for direct estimation of cortisol. To the cortisol antibody coated microtiter wells, standard or serum samples (50 µL), along with cortisol-21-HS-ADH-HRP enzyme conjugate (100 µL) were incubated for 1 h at 37°C. Bound enzyme activity was measured by using tetramethyl benzidine/hydrogen peroxide (TMB/H₂O₂) as substrate. The sensitivity of the assay was 0.05 µg/dL and the analytical recovery ranged from 92.9 to 101.7%.     

The inductive effect of interleukin-4 on IgG4 and IgE synthesis in human peripheral blood lymphocytes

Using murine monoclonal antibodies against human IgG subclasses, specific and sensitive ELISAs assay to quantify the four human IgG subclasses in cell culture supernatants were established. The effect of human recombinant interleukin-4 (IL-4) on the regulation of IgG subclasses by normal peripheral blood lymphocytes was investigated. In addition to the enhancement of IgE synthesis, IL-4 preferentially induced IgG4 synthesis in vitro, whereas IL-4 had no effect on IgG1, IgG2, and IgG3 synthesis. IL-4-induced IgG4 production was blocked in a dose-dependent manner by recombinant interferon-gamma and anti-human IL-4 monoclonal antibody. Collectively, this data indicates that IL-4 plays an important regulatory role in both IgG subclass and IgE synthesis.     

自身甲状腺球蛋白抗体阳性时甲状腺球蛋白测定方法的评估

应用单克隆固相放免法及双抗体法测定了自身免疫性甲状腺球蛋白抗体(TG-Ab)阳性病人血清中甲状腺球蛋白(TG)浓度。34例慢性淋巴细胞性甲状腺炎病人,TG-Ab阳性及TG-Ab阴性病人的TG值,分别为9.5±4.2ng/ml和10.9±3.7ng/ml(固相法);3.75±0.71ng/ml和4.25±1.1ng/ml(双抗体法)。50例Grave’s病人测定值分别为30.5±18.9ng/ml和41.5±20.5ng/ml(固相法);20.88±10.5ng/ml和27.5±12.5ng/ml(双抗体法)。38例甲状腺癌病人中,无转移灶者,两种方法测定值未见明显差异;8例有转移灶病人中,TG-Ab阳性者其TG值明显低于TG-Ab阴性者(p<0.01)。我们对10例慢性淋巴细胞性甲状腺炎TG-Ab阳性者进行了回收试验,固相法平均为81.6±14.0ng/ml,双抗体法平均为70.8±14.1ng/ml。我们认为,应用单克隆抗体固相法测定的TG值明显高于双抗体法,其回收率也高于双抗体法,但是在TG-Ab阳性病人血清中,TG测定值较TG-Ab阴性者为低,这可能由于在TG-Ab存在的情况下,体内TG代谢清除率增快而使TG浓度下降。因此,对TG-Ab阳性病人TG测定,仍有待于进一步探讨。     

肺癌患者血清NSEIRMA的临床意义

为研究NSE在肺癌诊断、疗效监测和预后观察的临应意义及应用价值,本文对129例肺癌病人和54例肺部良性疾病患者,应用免疫放射分析法(IRMA)检测NSE的血清含量。肺癌病人平均血清水平30.1±30.9ng/ml,良性病人平均13.4±6.2ng/ml(p<0,01),两组差异显著。肺癌病人平均阳性检出率55％,其中肺小细胞癌病人阳性率是87.5％(27/32),鳞癌和腺癌阳性率分别为47.5％和41.9％,良性病人假阳性率为35.2％。测定灵敏度为55％,特异性65.5％,准确率58.0％。肺癌病人治疗后的NSE平均水平从治疗前的62.6土50.9ng/ml降至19.4±17.6ng/ml,11例治疗有效病人血清水平均有不同程度下降,2例治疗无效者,血清NSE水平增高。提示NSE对肺癌诊断有一定的临床价值,尤其对肺小细胞癌的诊断、疗效鉴定及预报复发意义更大。     

抗戊型肝炎病毒IgG的检测

应用ＥＬＩＳＡ方法对１９９０年沈阳一起暴发流行的戊型肝炎病人及对照人群７９份血清进行了抗戊型肝炎病毒（ＨＥＶ）ＩｇＧ的检测。显性感染病人与非疫区正常对照人群抗－ＨＥＶＩｇＧ检出率分别为５５．３２％（２６／４７）和２０．００％（２／１０），两者存在显著性差异（ｘ２＝４．１２，Ｐ＜０．０５）。显性感染病人急性期，病后８个月血清抗－ＨＥＶＩｇＧ检出率分别为１００．００％（８／８）和谐４１．９４（１３／３１），前者明显高于后者（ｘ２＝６．８３，Ｐ＜０．０１）。８例病人急性期和病后４个月血清抗体平均滴度分别为０．６８４和０．２３４，抗体滴度下降２．９２３倍，存在显著性差异（Ｔ＝２．２３９，Ｐ＜０．０５）。显性感染病人和ＳＧＰＴ升高无症状人群急性期血清间抗－ＨＥＶＩｇＧ检出率、抗体平均摘度存在明显差异（Ｐ＜０．０５，Ｔ＞１．９６０，Ｐ＜０．０５）。     

IgG-KAPPA-PRODUCING PRIMARY PLASMACYTOMA OF THE THYROID GLAND WITH PREOPERATIVE SERUM M PROTEIN

A case of primary plasmacytoma of the thyroid gland which occurred in a 63-year-old woman is reported. Histologic and ultramicroscopic examination revealed that the excised thyroid tumor was plasmacytoma superimposed on lymphocytic thyroiditis. Immunohistological study showed that the tumor cells produced intracytoplasmic immunoglobulin (IgG-kappa). Electropho-retic and immunoelectrophoretic studies disclosed the presence of monoclonal immunoglobulin (IgG-kappa) in samples of the patient's serum which had been obtained preoperatively. After completion of irradiation therapy to the neck following tumor removal, the serum monoclonal immunoglobulin disappeared. The patient is currently alive and well without any evidence of the tumor three years after surgery.     

QUANTITATION OF HYALURONIC ACID IN SERUM WITH AUTOMATED MICROPARTICLE PHOTOMETRIC AGGLUTINATION ASSAY

ABSTRACT

A microparticle photometric agglutination assay to quantitate hyaluronic acid (HA) in serum has been developed. The principle of the method is that hyaluronic acid binding protein covalently sensitized on microparticle surface initiates the particles to agglutinate in the presence of HA. By measuring the time-course of transmittance changes at 800 nm, due to the particles’ agglutination, HA concentration is estimated as a function of the magnitude of agglutination in a fully automated immunochemistry analyzer. The analytical range for HA was found to be between 10 and 1200 ng/mL. The precision (CV) was between 3.0 and 8.4% in the intra-assay (n = 10), and 4.8 and 8.9% in the inter-assay (n = 3). The lower limit of detection was 10 ng/mL. The deviation of the linearity study indicated within 8% of expected values, and the analytical recovery was between 96.5 and 106.3%. The correlation coefficient against HA plate ELISA was 0.989 with a slope of 1.01 (n = 31).     

sIL—2R在肺癌患者围手术期血清中的表达

应用ＥＬＩＳＡ双抗体夹惦法对８例胸部良性病变和４４例肺癌患者围手术期血清ｓＩＬ－２Ｒ水平测定，结果显示：肺癌患者组术前血清ｓＩＬ－２Ｒ水平明显高于胸部良性病变组；手术切除组术后３０ｄ，ｓＩＬ－２Ｒ水平明显下降；胸探查组较术前升高；肺癌患者各组治疗前后其ｓＩＬ－２Ｒ水平均有显著差异。     

不同程度脑外伤后甲状腺激素水平的临床观察

按脑外伤后10小对GCS评分，随机选择轻，中、重度脑外伤病例各30例．分别测其入院时，第一周内各天及第14天早晨甲状腺激素水平，发现甲状腺激素水平与病情及其变化明显相关；1．T3下降最快，最明显，且病情越重下降越显著；2．T4持续下降低于正常值提示预后不良；3．rT2随病情加重而升高，但恢复较快；4．FT3，FT4变化基本与T3，T4相同，但不如其显著；5．TSH早期可升高，危重组低于正常值。     

以低效价抗-HBe血清制备乙型肝炎e系统酶免疫试剂盒

国内目前普遍采用酶免疫法(ELISA)检测乙型肝炎e系统,其所使用的试剂盒,由于受原材料的质量及来源所限(需琼脂糖双扩法ID≥1:16高效价抗-HBe血清,即高效价血源),往往不能保证供应,且反应模式有时欠佳,不易判定,结果不稳。因此,     

Detection by ELISA of immunoglobulin G subclass-specific antibody responses in rotavirus infections in children

IgG subclass-specific antibody responses to a human subgroup 2 rotavirus were studied in 26 children by ELISA by use of monoclonal antibodies specific to the four human IgG subclasses. One hundred twenty-nine serum samples were obtained before, during, and after an episode of rotavirus-induced diarrhoea in these patients. When these sera were investigated, an increase in IgG1 and IgG3 subclass-specific antibodies was detected in all 26 patients. IgG3 antibodies reached a peak concentration 1 week after rotavirus was detected in faecal samples and then progressively declined over the following months, whereas the peak concentration of IgG1 subclass antibodies was found 2 months later and seemed to persist thereafter. IgG2 rotavirus-specific subclass antibodies were never found and IgG4 subclass antibodies were detected only in sera from seven of the 26 patients.     

转移程度不同的小鼠肝癌细胞株对腹腔巨噬细胞免疫功能的影响

目的 研究转移程度不同的小鼠肝癌细胞株对腹腔巨噬细胞功能的影响.方法 在两株转移程度不同的小鼠腹水型肝癌模型中,取荷瘤小鼠腹腔巨噬细胞,检测其在干扰素γ(IFN-γ)和脂多糖(LPS)刺激下产生一氧化碳(NO)和肿瘤坏死因子α(TNF-α)的水平,并检测其活化后的杀伤能力.进一步采用夹心酶联免疫吸附测定(ELISA)方法,研究不同荷瘤小鼠腹水中IFN-γ与转化生长因子β1(TGF-β1)的水平,并用相应抗体封闭TGF-β1后,检测活化巨噬细胞产生NO能力及杀伤活性.结果 经IFN-γ和LPS活化后,荷瘤小鼠腹腔巨噬细胞分泌NO和TNF-α能力明显低于正常巨噬细胞,杀伤能力下降.高转移性肝癌小鼠巨噬细胞分泌NO水平和杀伤能力均低于低转移性小鼠,但其分泌TNF-α量较高.此外,荷瘤小鼠腹水含较高水平IFN-γ与TGF-β1,不同转移程度荷瘤小鼠IFN-γ水平接近,但高转移性肝癌小鼠腹水含更多TGF-β1,而且TGF-β1的封闭可导致与肿瘤细胞共培养的巨噬细胞分泌NO的能力部分恢复.结论 肿瘤细胞可以通过分泌TGF-β1等抑制性因子下调巨噬细胞的活性和免疫功能.肿瘤的转移程度可能与其分泌免疫抑制因子的能力相关.     

卵巢癌患者血清及腹水白细胞介素6测定的临床意义

选择未接受治疗的原发性卵巢上皮癌３３例，良性卵巢上皮种瘤１２例及正常妇女行输卵管吻合术者１０例作为研究对象，结果ＩＬ－６依赖性细胞株ＭＨ６０－ＢＳＦ２增殖反应ＭＴＴ法测定血清及腹水（或腹腔液）中ＩＬ－６水平。结果表明，卵巢患者血清及腹水中的ＩＬ－６水平明显高于良性卵巢上皮肿瘤及正常对照组，临床晚期（Ⅲ－Ⅳ）者ＩＬ－６水平明显高于临床早期（Ｉ－Ⅱ者），但良性组与正常对照组之间无差异，提示ＩＬ－６水平     

Enzyme-linked immunosorbent assay for IgG antibodies to Epstein-Barr virus-associated early antigens and viral capsid antigen

An enzyme-linked immunosorbent assay has been developed for the detection of IgG antibodies to Epstein-Barr virus-associated early antigens and late antigens including the viral capsid antigen. The antibody titers of human sera determined in this way correlate well with those by indirect immunofluorescence. ELISA was more sensitive than the IF method. The assays described may be used for rapid and sensitive diagnosis of EBV-related diseases. In addition, the ELISA will be useful for the determination of antibody titers to isolated EBV-associated antigens, e.g., purified components of the EA complex.     

老年Graves病患者血清心肌钙蛋白的动态观察

目的：探讨老年Ｇｒａｖｅｓ病患者血清中心肌钙蛋白Ｉ（ＣＴｎＩ）含量测定的临床价值。方法：用放射免疫分析法（ＲＩＡ）测定３７例老年及老年前期Ｇｒａｖｅｓ病患者及３０例健康老年人血清甲状腺素（Ｔ４）、三碘腺原氨酸（Ｔ３）、促甲状腺素（ＴＳＨ）、游离Ｔ３（ＦＴ３）、游离Ｔ４（ＦＴ４）及ＣＴｎＩ的含量。结果：观察组患者血清ＣＴｎＩ含量显著高于对照组；观察组年龄与ＣＴｎＩ含量之间无显著相关性。观察组患者缓解     

连续监测格林-巴利综合征患儿血清IgG、IgA和IgM含量的意义

大量实验证明,格林-巴利综合征(GBS)病人体内存在着针对某些微生物的特异性抗体和抗自身周围神经组织的抗体,且随病情的好转,抗体滴度逐步下降,