A Series of Serological Tests for the Detection of Antigens and Specific Antibodies in Deep-Seated Candidosis: Experimental Aspects/Eine Gruppe von Antigen- und Antikörpertests zur Serodiagnose von tieflokalisierten Candida-Mykosen: Experimentelle Aspekte

Summary: We describe a series of six serological tests for the diagnosis of deep-seated candidosis. The array comprises two commercial tests (antigen test, Ramco Inc., and antibody test, Roche), as well as four enzyme immunoassays which have been developed in this laboratory: an antigen test for detection of Candida-proteinase, the corresponding assays for monitoring of anti-proteinase antibodies, and two assays for monitoring of IgG and IgM against heterogenous metabolic antigens of C. albicans. The highly sensitive and specific proteinase antigen-test tolerates samples with high concentration of serum proteins. Proteinase antigen was detected in 10 out of 11 normal mice after intravenous infection with C. albicans blastospores. The proteinase antigen peaked between the second and fourth day after infection. A rise in corresponding antibodies was observed in all animals. No proteinase antigen was detected in sera of healthy human individuals; anti-proteinase antibody titers in these sera amounted up to 1:8000. In related ELISAs, using metabolic fungal antigens, titer values of specific IgG and IgM amounted to 5120 and 1280, respectively. The six tests were carried out in an comparative study under diagnostic conditions, the results of which are the subject of a forthcoming communication. Zusammenfassung: Ein Satz von sechs serologischen Tests für die Diagnostik der tiefen Candida-Mykosen wird vorgestellt. Die Gruppe schließt zwei kommerziell vertriebene Testbestecke ein (Latex-Agglutinationstest zum Antigennachweis, Ramco Inc., und Hämagglutinationstest zum Antikörpernachweis, Roche). Vier weitere Enzymimmuntests wurden von uns entwickelt: Ein Antigentest zum Nachweis von sekretorischer Candida-Protease, ein entsprechender Test zum Nachweis von Antikörpem gegen Candida-Protease, und zwei Assays zum Nachweis von IgG-bzw. IgM-Antikörpem gegen heterogene metabolische Antigene von C. albicans. Der empfindliche spezifische Protease-Antigentest toleriert hohe Konzentrationen unspezifischer Serumproteine und kann deshalb auf Serumproben in geringer Verdünnung (z. B. 1:20) angewandt werden. Protease-Antigen war in 200 fach verdünnten Seren von 10 aus 11 intravenös infizierten NWNI-Mäusen nachweisbar. Die höchste Antigen-Konzentration trat zwischen dem 2. und 4. Tag nach Infektion auf; die Serum-Halbwertszeit von gereinigter Protease in der Maus betrug etwa 60 nun. Ein Anstieg korrespondierender Antikörper war in alien infizierten Tieren zu beobachten. Auch im Serum gesunder Probanden waren Antiprotease-Antikörper bis zu einem Titer von 1:8000 nachweisbar; der Protease-Antigentest fiel hingegen immer negativ aus. Die Titer von Antikörpern gegen metabolische Candida-Antigene erreichten in derselben Gruppe von Seren Werte von 1:5120 bzw. 1:1280. Die sechs Tests wurden unter diagnostischen Bedingungen verglichen; Ergebnisse dieser Studie sind Gegenstand einer weiteren Mitteilung.     

肾移植受者血清巨细胞病毒IgE和IgA抗体检测

采用间接ＥＬＩＳＡ检测２３名肾移植受者血清巨细胞病毒（ＣＭＶ）抗体，共检出１８名（７８％）活动性ＣＭＶ感染，其中１０名（４４％）为原发性感染。结果证实ＣＭＶ－ＩｇＥ和－ＩｇＡ具有较好的血清学诊断价值，优于ＣＭＶ－ＩｇＭ。     

应用单克隆抗体检测肺癌病人外周血中T细胞亚群

本文应用单克隆抗体检测33例原发性肺癌病人治疗前后外周血中T淋巴细胞亚群变化。结果提示:肺癌病人外周血中T细胞亚群明显有别于正常人,主要表现为T_H细胞降低和T_S细胞升高以及T_H/T_S比值明显倒置。短期观察结果表明,肺癌病人外周血中T_S细胞持续升高和T_H/T_S比值持续降低者,预后不良。临床检测T细胞亚群变化及它们的动态变化可作为判断宿主抗肿瘤免疫功能状态、病情发展和预后的敏感指标。     

间接血凝法检测抗Sm抗体和抗RNP抗体

应用间接血凝法测定抗Ｓｍ抗体和抗ＲＮＰ抗体。对不同疾病的检测结果表明，建立的该种方法与免疫电泳法结果无显著性差异（Ｐ＞０．０５）。对ＳＬＥ病４５例检测抗Ｓｍ抗体阳性率为１００％，抗ＲＮＰ抗体阳性率为９１％，抗Ｓｍ抗体可作为ＳＬＥ的特异性标志抗体，抗ＲＮＰ抗体在ＲＡ、ＰＳＳ、ＤＭ、ＭＣＴＤ病人中检出，其中ＭＣＴＤ病２３例，抗ＲＮＰ抗体阳性率高达９６％。且凝集效价高，可作为此病诊断的重要依据。间接血凝法具有操作简便、快速、敏感、特异性高、便于推广等特点，可代替免疫电泳法。     

噬菌体抗体库固相筛选条件的初步研究

目的:探讨噬菌体抗体库的固相筛选条件,为筛选方案的设计提供实验依据。方法:利用多种针对HEVNE2蛋白的特异性噬菌体人源抗体和非特异性噬菌体人源抗体,对噬菌体抗体与抗原的结合时间、抗原包被的浓度、洗涤强度和洗脱方式等多种筛选的条件进行初步探索。结果:阳性噬菌体抗体与抗原反应1min,就可较好结合,洗涤次数为20～30次、洗涤液的pH为5时,筛选得到的阳性率最高。包被抗原的浓度对筛选的阳性率没有明显影响,用10mg/L抗原竞争洗脱60min,可得到较高的阳性率。结论:噬菌体抗体库的筛选是一个非常复杂的过程,其中的各个条件之间有着密切的联系,应该根据具体情况进行调整。     

三种淫羊藿的总黄酮分极提取物对小鼠免疫功能的影响

观察了巫山淫羊藿、柔毛淫羊藿和粗毛淫羊获的总黄酮正丁醇提取物（代号Ａ１、Ａ２、Ａ３）和巫山淫羊藿总黄酮乙酸乙酯提取物（Ｂ１）对正常小鼠免疫功能的影响。结果；Ａ１可以明显促进脾脏抗体形成细胞的功能（Ｐ＜０．０５），Ａ２、Ａ３虽也有一定的促进作用，但无统计学意义；４种提取物在体外实验中均没有协同ＣｏｎＡ诱导牌细胞增殖作用，只有提取物浓度在１００μｇ／ｍｌ时，有一定抑制作用。４种提取物对２４小时培养脾细胞存活率无明显影响。     

A novel lamina lucida component of epithelial and endothelial basement membranes detected by LH39 monoclonal antibody.

The murine monoclonal antibody, LH39 was characterized in this study and appeared to bind to a novel basement membrane epitope. This antigen was expressed in the epithelial basement membrane of human tissue derived from all three germ cell layers and in basement membranes surrounding small blood vessels within the stroma of all organs examined. LH39 antigen could be first detected in fetal skin at the dermo-epidermal junction at 7 weeks estimated gestational age but was not present in the dermal vasculature until 16 weeks. When tested against tissue from a range of lower mammalian species, LH39 antigen appeared to be primate-specific. The epithelial basement membrane zone in organotypical cultures, where there is de novo synthesis of basement membrane components, contained abundant LH39 antigen in contrast to other basement membrane components, type IV collagen, laminin, and type VII collagen. Ultrastructural localization of LH39 epitope, using immunogold electron microscopy on unfixed freshly frozen tissue, was to the lamina lucida. No cross-reactivity could be detected between LH39 and laminin, fibronectin, and collagens I, III, IV, and V using the ELISA assay. In vitro studies with a range of proteolytic enzymes suggested that the antigen was non-collagenous in nature. LH39 precipitated a polypeptide with a molecular weight of 185 kD from extracts of metabolically labelled cultured keratinocytes, and polypeptides of 185 and 200 kD from the culture medium. The tissue distribution of LH39 antigen suggested that it may be an epitope within anchoring filaments. Potential applications of this antibody include the study of benign and malignant human vascular disorders, diseases and tumours associated with angiogenesis, epithelial neoplasms, and conditions of tissue regeneration and repair, such as wound healing.     

学报创刊15周年暨增刊通报

学报创刊１５周年暨增刊通报正值《中山医科大学学报》创刊１５周年之际，本刊编辑部于１９９５年６月１７日召开了“学报创刊１５周年总结会”。本刊编辑部主任、副主编关淡庄同志作了总结报告。首先感谢了学报的各位前辈为学报的创刊和发展所立下的汗马功劳，再次感谢了...     

南京口岸1987—1993年艾滋病监测报告

本文报道了南京卫生检疫局1987～1993连续7年对南京口岸重点人群进行艾滋病监测的情况．7年来南京局共监测各类标本43085份，在外籍留学生中检出HIV感染者2例，另外还检出4份进口人血丙种球旦白HIV抗体阳性．该局对检出的2侧HIV感染者及4份阳性进口人血丙种球旦白进行了游行病学调查和处理．并对艾滋病监测管理的有关问题进行了探讨．     

Immunological blockade of endogenous thyrotropin-releasing hormone impairs recovery from hyperglycemia in mice

Administration of antiserum to thyrotropin-releasing hormone (TRH) into the lateral cerebral ventricle of mice significantly attenuated recovery from hyperglycemia induced by treatment with 2-deoxyglucose but had no effect on the plasma glucose of saline-treated mice. TRH, injected centrally together with the anti-TRH antibody, reversed the effect of the antiserum and blocked the development of hyperglycemia. These findings suggest that activation of TRH neurons in the central nervous system may be a physiological event influencing recovery from hyperglycemia.