旋毛虫肌幼虫排泄分泌抗原诊断早期旋毛虫病的研究

目的评价旋毛虫肌幼虫排泄分泌(excretory-secretory,ES)抗原诊断早期旋毛虫病的价值。方法应用旋毛虫肌幼虫ES抗原Western blot对旋毛虫感染后6~11d的小鼠血清及19d的早期旋毛虫病人血清进行检测,并与感染后35d的晚期旋毛虫病人和其他寄生虫病人血清的检测结果进行比较。结果 Western blot分析显示,肌幼虫ES蛋白中的2条蛋白带(41.5、55kDa)可被旋毛虫感染后7~11d的小鼠血清识别,6条蛋白带(41.5、44.1、45、55、61和65.2kDa)能被早期和晚期旋毛虫病人血清识别,阳性反应率均为100%(15/15);这6条蛋白带与裂头蚴病人和健康人血清无交叉反应,但与其他寄生虫病(血吸虫病、并殖吸虫病、华支睾吸虫病、棘球蚴病及囊尾蚴病)患者血清具有明显的交叉反应(19.12%~38.23%)。结论旋毛虫肌幼虫ES抗原中的41.5kDa~65.2kDa蛋白带可与旋毛虫感染早期血清反应,但与其他蠕虫病患者有明显的交叉反应。     

旋毛虫肌幼虫特异性抗原的鉴定

目的 寻找诊断旋毛虫病的特异性抗原。方法 应用SDS—PAGE和Western blot对旋毛虫肌幼虫可溶性抗原中的蛋白组分进行研究。结果 旋毛虫肌幼虫可溶性抗原经SDS—PAGE后显示29条蛋白带，分子量范围在112—12kDa之间，其中65、43、42、31、30、20、17、16kDa为主带。112、110、108、102、97、95、65、63、58、55、53、49、45、43、42kDa蛋白组分与并殖吸虫感染的大鼠及患者血清、华支睾吸虫病、血吸虫病及囊尾蚴病患者血清均具有明显的交叉反应带；24—20kDa蛋白组分只与旋毛虫感染的大鼠、小鼠及患者血清反应，而不与其它寄生虫感染的动物或患者血清、正常大鼠和小鼠及正常人血清发生交叉反应。结论旋毛虫肌幼虫可溶性抗原中24—20kDa蛋白组分为旋毛虫肌幼虫的特异性抗原，可用于旋毛虫病的免疫学诊断及血清流行病学调杏。     

旋毛虫肌幼虫排泄分泌物中特异性诊断抗原的研究

目的　寻找旋毛虫肌幼虫排泄分泌(ES)物中的特异性诊断抗原。　方法　应用SDSPAGE和Western印迹对旋毛虫肌幼虫体外培养18、30h后的ES抗原中的蛋白组分进行研究。　结果　旋毛虫肌幼虫培养18、30h后得到的ES抗原组分大致相同,两种ES抗原中主要蛋白带的分子量为112、110、108、97、53、49、45、42、35、23和16kDa。18hES抗原中的102、97、95和53kDa以及30hES抗原中的53、49、45和43kDa均与并殖吸虫病、华支睾吸虫病、日本血吸虫病及囊尾蚴病患者血清发生明显的交叉反应。ES抗原中的23kDa蛋白组分只与旋毛虫感染的大鼠、小鼠及患者血清反应,而不与上述其它寄生虫感染者、正常大鼠和小鼠及正常人血清发生交叉反应。　结论　旋毛虫肌幼虫ES抗原中的23kDa蛋白组分为旋毛虫肌幼虫的特异性抗原,可用于旋毛虫病的血清学诊断及血清流行病学调查。     

旋毛虫成囊前期幼虫抗原保护性免疫的研究

目的比较旋毛虫成囊前期幼虫虫体抗原、排泄分泌抗原和表面抗原对小鼠产生的免疫保护作用。方法分别用旋毛虫成囊前期幼虫虫体抗原、排泄分泌抗原、表面抗原免疫小鼠,同时设佐剂组和阴性对照组,间隔7d免疫1次,共3次。末次免疫后7d,每只小鼠用200条旋毛虫感染期幼虫经口进行攻击感染。感染后7d和30d分别检查各组小鼠肠道成虫数和肌幼虫数;用ELISA测血清中抗旋毛虫肌幼虫IgG抗体。结果虫体抗原、排泄分泌抗原、表面抗原和佐剂组的成虫减虫率分别为84．89％、89．73％、85．65％、2．57％;肌幼虫减虫率分别为71．71％、80．98％、73．66％、5．60％。排泄分泌抗原组、表面抗原组的成虫减虫率（P均〈0．05）及肌幼虫减虫率（P均〈0．01）均高于虫体抗原组。各免疫组小鼠血清IgG抗体滴度明显升高,虫体抗原组、排泄分泌抗原组和表面抗原组的几何平均倒数滴度分别为2798．89、3474．51、2984．83,分别为阴性对照组（459．32）的6．09、7．56、6．50倍。结论旋毛虫成囊前期幼虫虫体抗原、排泄分泌抗原和表面抗原均能诱导宿主产生较强的抗攻击感染保护力。成囊前期幼虫的排泄分泌抗原显示出更强的免疫原性。     

旋毛虫抗原的国际分类和种间的抗原变异

对旋毛虫抗原的分类 ,传统上是按照抗原的来源将其分为虫体抗原、表面抗原、排泄 -分泌 (ES)抗原及杆细胞颗粒相关抗原 ,但以前报道的多种旋毛虫抗原是相似的或相同的 ,在多数情况下是因分析技术的不同而导致了抗原分子量的细小差异 ,故一些学者曾试图根据旋毛虫抗原产生的免疫反应将其进行分组或分类 ,但需要制定统一的分类和命名标准〔1,2〕。鉴于此 ,国际旋毛虫病委员会 (InternationalCom missiononTrichinellosis ,ICT)于 1990年在荷兰的Bilthoven组织了旋毛虫抗原国际讨论会。由不同研究小组提供针对旋毛虫成虫、新生幼虫和肌幼虫…     

旋毛虫三种抗原作为免疫诊断抗原的比较研究

用旋毛虫成虫可溶性抗原,成虫排泄分泌抗原和肌肉幼虫抗原作为免疫诊断抗原,采用ＥＬＩＳＡ方法检测,观察各自在旋毛虫病免疫不诊断中的敏感性和特异性。结果表明;抗原的敏感性由强到弱依次是肌肉幼虫抗原,成虫排泄分泌抗原和成虫可溶性抗原;特异性由高到低分别是成虫排泄分泌抗原,肌肉幼虫抗原和成虫可溶性抗原。     

旋毛虫成囊前期幼虫排泄分泌抗原的研究进展

在旋毛虫感染过程中,成囊前期幼虫（PEL）是旋毛虫侵入宿主肌肉的侵入期。旋毛虫PEL比成囊期幼虫（EL）在宿主体内约早2周出现。成囊前期幼虫抗原（PELA）对旋毛虫病的早期免疫学诊断具有较高的敏感性。而旋毛虫排泄分泌抗原具双重免疫功能,即有良好的抗原性和免疫原性。因此,本文就旋毛虫成囊前期幼虫ES抗原研究进展做一综述。     

旋毛虫成虫三种抗原SDS-PAGE及Western-blot比较研究

目的研究大理猪源旋毛虫(Trichinella spiralis)成虫虫体可溶性抗原、表面抗原和排泄分泌抗原3种抗原的蛋白组分及其之间的差异,比较分析其免疫反应性。方法以十二烷基硫酸钠-聚丙烯酰胺凝胶电泳和蛋白印迹对成虫3种抗原进行蛋白组分及其免疫反应性分析。结果十二烷基硫酸钠-聚丙烯酰胺凝胶电泳结果:虫体可溶性抗原显示29条蛋白带,分子量范围在112kDa~10kDa之间,其中主带13条;表面抗原显示4条主要蛋白带,分子量分别为50、48、40、34kDa;排泄分泌抗原显示17条蛋白带,分子量范围在120~14kDa之间,其中主带6条,分子量分别为120、64、43、40、35、33kDa。蛋白印迹结果:虫体可溶性抗原出现13条反应条带,其中分子量为81、40、37、33、30kDa的条带着色明显;表面抗原出现4条反应条带,其分子量分别为50、48、40、34kDa;排泄分泌抗原显示7条反应带,其中43、40、27、18kDa着色明显。结论旋毛虫成虫虫体可溶性抗原与排泄分泌抗原较表面抗原蛋白组分复杂,3者主带部分属低分子量区;免疫反应性的强度表面抗原和排泄分泌抗原高于虫体可溶性抗原,说明前两种抗原是旋毛虫成虫刺激机体产生免疫应答的主要靶抗原。     

抗旋毛虫肌幼虫ES抗原单克隆抗体的制备与鉴定

目的制备分泌抗旋毛虫肌幼虫排泄分泌(ES)抗原单克隆抗体(McAb)杂交瘤细胞株,并对其及分泌的McAb进行鉴定。方法用旋毛虫肌幼虫ES抗原免疫BALB/c小鼠,取其脾细胞与SP2/0细胞融合,筛选分泌特异性、高滴度McAb的杂交瘤细胞株,制备腹水,进行纯化。ELISA间接法、夹心法和竞争法分别测定McAb滴度、相对亲和力和相加效应,琼脂双扩散试验鉴定免疫球蛋白类型,检测McAb与其他寄生虫抗原的交叉反应。电镜观察杂交瘤细胞超微结构,计数染色体数目及观察杂交瘤细胞分泌McAb的稳定性。结果筛选出2株(B2C12和E11F11)分泌抗旋毛虫肌幼虫ES抗原McAb杂交瘤细胞株。电镜显示,杂交瘤细胞具有肿瘤细胞和浆细胞的特征,胞浆内可见大量分泌颗粒;杂交瘤细胞染色体数目平均为98.6条,均能稳定分泌McAb(属IgM)。B2C12株培养上清液和腹水McAb滴度分别为1∶1280和1∶2.048×104,E11F11株为1∶640和1∶1.024×104,其中前者的相对亲和力较后者稍高。2株McAb识别ES抗原不同的抗原决定簇;且不与其他寄生虫抗原发生交叉反应。结论获得2株抗旋毛虫肌幼虫ES抗原杂交瘤细胞株,均能稳定分泌高滴度、特异性的IgM类McAb,并识别不同的抗原决定簇,为研制旋毛虫病诊断试剂盒和制备基因工程抗体奠定了基础。     

旋毛虫感染期幼虫抗原的特异性分析

旋毛虫病的免疫诊断假阳性率高，与其它寄生虫病存在交叉反应。国内外学者发现旋毛虫幼虫可溶性抗原与线虫、吸虫、绦虫以及原虫等寄生虫病患者血清发生交叉反应［１］。为了阐明交叉反应的机理，提高诊断的特异性和敏感性，本实验用ＳＤＳ－ＰＡＧＥ分析了旋毛虫感染期幼...     

Isolation of human scFv antibody fragments against ABO blood group antigens from a phage display library

BACKGROUND AND OBJECTIVES: Phage display has proven very useful for the isolation of antibodies against a number of antigens. We used this technology to isolate scFv antibody fragments against A and B red blood cell antigens. MATERIALS AND METHODS: Phages from a phage display library were selected using unmodified red blood cells as a source of antigen. Bound phages were absorbed onto cells lacking the antigen of interest and used to infect Escherichia coli cells. Phages were rescued and assayed for specificity by enzyme-linked immunosorbent assay (ELISA). RESULTS: After several rounds of panning and subtraction on red blood cells, one anti-A and one anti-B human scFv antibody fragments were isolated. CONCLUSION: Isolation of anti-A and anti-B scFv antibody fragments on whole cells is an alternative method of obtaining antibodies against native cell-surface antigens.     

Immunogenicity of cellular and acellular antigen preparations from a methylcholanthrene-induced mouse sarcoma

Summary Inbred mice were immunized with various cellular and subcellular preparations from a syngeneic, methylcholanthrene-induced sarcoma to induce transplantation immunity against a subsequent challenge of viable cells from the same tumor. Only with conventional immunization procedures using radiation-attenuated tumor cells or viable tumor cells in admixture with bacillus Calmette-Guérin was transplantation immunity obtained. Neither native soluble tumor extracts prepared using several extraction methods, nor cross-linked preparations of these extracts gave rise to transplantation immunity. With few exceptions, however, the immunizing preparations resulted in the production of tumor-specific humoral antibody. The manner in which tumor antigens are presented to the host as well as the experimental system (animal strain; tumor) used appear to play an important role in the development of transplantation immunity against tumors.

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Abbreviations BCG Bacillus Calmette-Guérin - ENM extranuclear membrane - FCA Freund's complete adjuvant - GA glutaraldehyde - PBS phosphate-buffered saline - NP 40 Nonidet P 40 detergent Supported by the Landesamt für Forschung, Ministerium für Wissenschaft und Forschung des Landes Nordrhein-Westfalen, Düsseldorf     

Biological characterization of human bone tumors

Summary A total of 45 cases of bone tumors and tumor-like lesions were studied in order to determine the expression of an HLA-DR antigen by the monoclonal antibody 910-D-7, and its possible correlation with histology, using the indirect immunoperoxidase method on frozen sections. The pattern of antigen expression was nearly constant for the individual cell types, though varying in intensity, and did not depend on the biological behavior of the respective lesions. No clear correlation could be established between antigen expression and cell maturation. Although the biological significance of antigen expression in these tumors is not yet understood, it is clear that here, too, the were presence of an HLA-DR antigen cannot be interpreted as a sign of malignant transformation.Supported by grant RO 648/1-1 of Deutsche ForschungsgemeinschaftFellow of Deutscher Akademischer Austauschdienst     

Predictors of loss of hepatitis B surface antigen in HIV-infected patients

AIM:To study factors associated with loss of hepatitis B surface antigen (HBsAg) in patients co-infected with human immunodeficiency virus (HIV) and hepatitis B virus (HBV).METHODS:We retrospectively reviewed the medical records of 5681 patients followed up at two New York City HIV clinics from January 1999 to May 2007.Clinical and laboratory parameters including baseline and follow-up HIV viral loads,CD4 cell counts,alanine transaminase levels,demographics,presence of hepatitis C infection,and treatment with highly active antiretroviral therapy dually active against both HIV and HBV infection,were analyzed to determine factors associated with loss of HBsAg.RESULTS:Three hundred and fifty five patients (355/5681,6.84%) were co-infected with HIV and HBV and were evaluated.Of these,226 patients with more than 12 mo follow-up were included in further analysis to determine factors associated with loss of HBsAg in the long-term follow-up.In the univariate analysis,baseline CD4 cell count was associated with loss of HBsAg (P=0.052).Cox regression analysis revealed that loss of HBsAg was associated with baseline CD4 cell count > 500 cells/mm3 (P=0.016,odds ratio:76.174,95% confidence interval:2.233-2598.481).CONCLUSION:Our study showed an interesting association of loss of HBsAg in HIV-HBV co-infected patients with higher CD4 cell count,suggesting that T-cell cytolytic activity against HBV may still be effective in clearing HBV infection.     

Identification of soluble and membrane antigenic markers of acquired toxoplasmosis by immunoblot

The overall performance of quantitative assays in the detection of anti‐Toxoplasma IgG is satisfactory, but discrepancies between assays are not uncommon especially when IgG concentrations are close to the limit of detection of the tests. The purpose of our study was to identify soluble and membrane antigens extracted from Toxoplasma gondii tachyzoites by immunoblot to select the most relevant antigenic bands to be used for qualitative serodiagnosis of acquired toxoplasmosis. We selected five relevant bands (98, 36, 33, 32 and 21 kDa) with soluble antigens and four relevant bands (42, 35, 32 and 30 kDa) with membrane antigens which gave high sensitivity and/or specificity in immunodiagnosis. The association on the same blot of at least three of the five relevant bands in the soluble antigen immunoblot showed the highest sensitivity/specificity (97·4%/99·0%, respectively). Our results indicate that immunoblot using soluble tachyzoite extract with simultaneous detection of at least three of the five bands (98, 36, 33, 32 and 21 kDa) represents a valuable test for serodiagnosis of acquired toxoplasmosis and should be further evaluated as a confirmatory test for sera which give discrepant results in quantitative assays.     

Immunophenotyping of acute myeloid leukaemia: Relevance of analysing different lineage-associated markers

Summary The immunophenotype of 135 previously untreated patients with FAB defined acute myeloid leukaemia (AML) was studied at diagnosis. The panel of reagents included monoclonal antibodies (MoAb) recognising myeloid-associated determinants (CD11, CD13, CD14, CD33 and others) as well as MoAb directed towards lymphoid antigens (CD7, CD10, CD19) and TdT. The results indicate that CD13 and/or CD33 are consistently expressed in AML and only rarely in ALL blasts (131/135 +ve cases, versus 4/130 in ALL). Lymphoid antigen expression was rarely detected when CD10 and CD19 were investigated in AML (0.9% and 2% +ve cases, respectively), whereas significant positivities were found for TdT and CD7 (20% and 10% respectively). Concerning FAB subtypes, two new MoAb (LAM3 and LAM7) proved very useful in the specific recognition of AML with monocytic features. The phenotype CD13+ and/or CD33+, CD9+, HLA-DR– was found to be almost exclusive for M3 AML. The response to induction chemotherapy was analysed in CD7+ and in TdT+ patients. In the latter group a statistically significant lower response rate was found with respect to TdT-ve-AML patients.     

弓形虫抗原组分提取及免疫原性观察

以不同剂量的弓形虫粗制与纯化抗原经皮下注入小白鼠，３５天后检查血清，在高剂量粗制抗原组显示有免疫学改变。以０．２ｍｌ弓形虫感染小鼠后新鲜腹水感染试验鼠，亦显示试组具保护力，可延长小鼠的存活时间。但总的看来，保护力不强，尤其是低剂量粗制抗原及纯化抗原均无保护力。     

汉族类风湿关节炎患者HLA—DR和—DQ基因分型研究

为探讨ＨＬＡ－ＤＲ和－ＤＱ等位基因与我国汉族类风湿关节炎（ＲＡ）的相关性，采用聚合酶链反应－限制性内切酶片段长度多态性分析（ＰＣＲ－ＲＦＬＰ）方法对汉族人群中３５例ＲＡ患者和１００名健康人的ＤＲ和ＤＱ位点进行ＤＮＡ定型分析。结果发现，ＤＲ４频率在正常人为２４．０％，在ＲＡ患者为５１．４％（Ｐ＜０．０１，ＲＲ＝３．３０）；ＤＲ４亚型位点分析发现２组均以ＤＲＢ１＊０４０５占多数，但ＤＲＢ１＊０４０５频率在ＲＡ组为３１．４％，高于正常人的１０．０％（Ｐ＜０．０１）。ＤＱ４频率在全部ＲＡ患者为３７．１％，在ＤＲ＋４ＲＡ患者为７２．２％，均高于全部正常人的１０．０％（Ｐ＜０．０１）和ＤＲ＋４正常人的４１．６％（Ｐ＝０．０３７６）；ＤＲ＋４－ＤＱ＋４ＲＡ患者的病情重于ＤＲ－４患者。结果提示，ＤＲ４、主要是ＤＲＢ１＊０４０５与ＲＡ相关，ＤＱ４可增加ＤＲ４对ＲＡ的易感性，ＤＲ＋４－ＤＱ＋４单倍型是ＲＡ病情严重程度的标志。     

The roles of latency-associated antigens in tuberculosis vaccines

Tuberculosis (TB) is a re-emerging disease and is caused by Mycobacterium tuberculosis (M. tuberculosis). TB is currently one of the leading causes of morbidity and mortality, worldwide. The only available vaccine against TB infection, Bacillus Calmette–Guérin (BCG), fails to adequately protect against reactivation of latent infections in adults. Furthermore, recently developed subunit vaccines, which are in various stages of clinical trials, are all prophylactic vaccines based on proteins expressed in replicating stage of M. tuberculosis and they are not preventive of reactivation of latent TB infection. Thus, an appropriate subunit post-exposure vaccine needs to be developed to control all forms of TB infection. To produce a multi-stage subunit vaccine, scientists should combine the early secreted M. tuberculosis antigens with latency antigens. For this purpose, some latency proteins are known which could be important antigens in the production of specific humoral and cellular immune responses in latent M. tuberculosis infected individuals. Several studies have evaluated the immunogenicity of these proteins in improving the TB vaccines. The present study is a comprehensive review of several studies on the role of the latency antigens in the development of TB vaccines. Overall, the studies indicate that the latency-associated antigens including the resuscitation-promoting factors, the Dormancy of survival regulon (DosR) proteins and the starvation stimulant proteins are potential candidates for the development of subunit vaccines against TB infection.     

蜱抗原免疫接种诱导宿主抵抗力的研究

本文选用中华硬蜱雌性成虫中肠抗原、卵抗原对家免进行人工免疫接种，在按常规方法免疫接种三次后，用中华硬蟀成虫进行感染（叮咬），分别观察中肠抗原接种组、卵抗原接种组、佐剂对照组和空白对照组中华硬蜱的吸血量、生殖情况。结果显示．中华硬蜱叮咬中肠抗原免疫接种兔后其吸血量、产卵量均较对照组显著降低，蜱的吸血量较佐剂和空白对照组分别下降48．56％和54．65％，产卵量下降52.8％和57.9％，而中华硬蜱叮咬卵抗原免疫接种组兔后其吸血、生殖能力较对照组无显著性差异（P＜0．05）。实验表明中华硬碑中肠抗原可有效诱导宿主产生特异性抵抗力，并对该现象进行了分析讨论。