片形吸虫分泌排泄抗原和虫体抗原的分析

用牛源肝片吸虫（南京）、牛源大片吸虫（广西）和羊源肝片吸虫（实验感染）制备可溶性虫体抗原（BA）和和分泌排泄抗原（ES），以SDS－PAGE电泳分析比较，结果显示，3株虫体可溶性虫体抗原至少有20条以上的主要蛋白条带，分子量集中于10－90KD之间，它们之间无显著差异，而3株分泌排泄抗原蛋白成分较简单，明显可分的条带不超过5条，分子量集中于10－30KD之间，且均拥有26－28KD的蛋白成分，提示该蛋白应为片形吸虫ES抗原的主要免疫成分。     

小肠结肠耶氏菌O:9血清型与布氏杆菌M_5株共同抗原的免疫印迹分析

应用SDS-聚丙烯酰胺凝胶电泳和免疫印迹技术,对经超声波打碎的小肠结肠耶氏菌O:9血清型和布氏杆菌M5株全菌体蛋白成分进行了分子量测定及抗原分析。结果表明,小肠结肠耶氏菌O:9血清型Y15株与布氏杆菌M5株存在一条发生交叉反应的蛋白质共同抗原带,其分子量为11400。Y15株与M5株有多个分子量相同的条带,但不发生交叉反应。Y15株与M5株皆有多个各自特异的条带。     

泰氏锥虫抗原制备及实验室诊断研究

应用体外培养的泰氏锥虫制备可溶性抗原Ⅰ、抗原Ⅱ和代谢抗原.经测定,其蛋白含量每毫升分别为6.5mg、7.4mg和7.1mg.薄层等电聚焦电泳测定结果表明,抗原Ⅰ出现22条区带,抗原Ⅱ21条区带,代谢抗原28条区带,对照的伊氏锥虫琼脂免疫扩散抗原14条区带.经分析,泰氏锥虫抗原和伊氏锥虫抗原有4条区带在同一迁移率上.琼脂免疫扩散反应中,3种泰氏锥虫抗原均与相应免疫兔血清发生沉淀反应,抗原Ⅰ出现1条致密沉淀线,抗原Ⅱ和代谢抗原出现2～3条沉淀线,抗原效价为1:4～16.免疫电泳显示了类似的结果,抗原Ⅰ与免疫兔血清出现1条弧形沉淀线,抗原Ⅱ和代谢抗原与免疫兔血清出现了3条弧形沉淀线.间接血凝试验结果表明,泰氏锥虫自然感染牛血清效价为1:20～40,免疫兔血清为1:1280～5120.所制泰氏锥虫抗原对伊氏锥虫和媾疫锥虫血清也能很好地发生交叉反应,3份伊氏锥虫病马血清和3份伊氏锥虫人工感染兔血清血凝效价分别为1:10～40和1:8～1024;5份媾疫马血清有4份血凝效价为1:20～320.4份环形泰勒虫病牛血清均为阴性.     

胰吸虫与肝片形吸虫成虫抗原的酶联免疫印迹分析

用SDS-PAGE和酶联免疫印迹试验(ELIB)分析胰吸虫成虫(EP）及其与肝片形吸虫成虫(Fh)交叉／共同抗原的分子量和免疫活性。SDS-PAGE结果显示，胰吸虫和肝片形吸虫成虫抗原的多肽均达30余条，EP抗原分子量在123kd以下，主带4条；Fb抗原分子量在83kd以下，主带4条；两者具相同分子量的多肽6条。ELIB结果显示，抗原与同源抗血清呈现颜色较深、数量较多的区带，而与异源抗血清则呈现数量不等的交叉／共同反应区带，表明两虫之间存在交叉／共同抗原性多肽。     

肌旋毛虫功能性抗原实验研究

本实验应用生化萃取技术和体外人工培养技术,制备了肌旋毛虫匀浆(HO)抗原和排泄—分泌物(ES)抗原,并对两种不同来源抗原的部分生化特性和免疫学特性进行了比较。HO抗原通过SephadexG—200又分为第一峰(FP)和第二峰(SP)。在5～20％的聚丙烯酰胺梯度凝胶电泳和薄层聚丙烯酰胺凝胶等电聚焦电泳上,ES和FP抗原出现许多条电泳区带。ES、FP和HO抗原对小鼠的保护力(肌幼虫减少率)分别为78％,76％和46％。     

旋毛虫感染小鼠对p46 000重组抗原的抗体应答

分别以旋毛虫肌幼虫ES抗原和p46000重组蛋白作为抗原，对小鼠人工感染旋毛虫后的抗体应答进行了ELISA检测。结果表明，以肌幼虫200条／只经口感染小鼠后，肌幼虫ES抗原在感染后9d可检出抗体，并于感染后35～42d达到最高水平；应用重组抗原检测时，感染后10d可检出抗体，抗体水平略低于用ES抗原，但是其消长规律基本一致．而且与阴性血清相比差异明显；抗体在117d后仍维持于较高水平。     

伊氏锥虫和布氏锥虫动基体DNA酶切电泳比较

限制性内切酶ＭｂｏＩ，ＤｄｅＩ，Ｈｉｎｆｉ和ＴａｑＩ对布氏锥虫ＫＤＮＡ进行酶切后，电泳中均显示出多条ＤＮＡ区带，其总Ｋｂ数约等于２０ｋｂ，而各限制酶对伊氏锥虫ＫＮＤＡ消化后均显示出１至２条区带，总和约１ｋｂ明显区别于布氏锥虫。     

多头绦虫抗原特性的研究──多头绦虫节片抗原及原头节ES抗原的SDS-PAGE分析

为了探明多头绦虫成虫节片抗原及多头蚴原头节排泄分泌（ES）抗原的多肽组分，以供今后在免疫诊断与预防脑多头蚴病的应用，本试验应用SDS-PAGE首次分析了多头绦虫成虫节片抗原及多头蚴原头节ES抗原的多肽组分。应用12．5％凝胶的连续系统，垂直板型电泳，考马斯亮兰R－250染色的结果表明，成虫节片抗原共有16条多肽带，其分子量范围29～154kD，其中主带4条，分别为73、88、128、134kD，原头节ES抗原共6条多肽带，分子量范围33～132kD，其中主带2条，分别为33和108kD。本试验初步阐明多头绦虫成虫节片抗原和原头节ES抗原的多肽组分，为进一步研究多头绦虫抗原的特性奠定了基础。     

旋毛虫ES抗原特异性成分的分离和鉴定

在旋毛虫病的免疫学诊断中,目前常用的抗原是虫体粗制抗原(CWE)、排泄—分泌抗原(ES)及其纯化物。研究表明,CWE易出现假阳性反应,其诊断准确率不高;而ES易出现假阳性反应较少,特异性较强,因此愈来愈受到国内外学者的重视。但在ES中究竟存在几种特异性蛋白成分?它们的分子     

布氏姜片虫对哺乳动物的易感性

作者研究了布氏姜片虫对各种哺乳动物的易感性。小白鼠、大鼠、猴和狗均有抵抗力,几内亚猪仅有部分的易感性。然而,发现青年兔(6～8周龄)对布氏姜片虫有易感性,可作为理想的实验动物。     

Proteins and antigens of Pasteurella multocida serotype 1 from fowl cholera.

The protein profiles of Pasteurella multocida serotype 1 isolates and the response of chickens to serotype 1 antigens were investigated using SDS-PAGE. Patterns obtained with Coomassie blue staining of soluble protein extracts were similar. The major difference between isolates was the position of one of the major proteins in the 34-38 kDa region. When chickens were experimentally infected with a clinical isolate of P. multocida serotype 1 various proteins were recognised by immunoblotting, including one with a relative molecular weight of 34 kDa; however, no reactions were observed in the region where LPS is known to migrate. When these infection sera were used in an EIA with purified LPS obtained from Heddleston serotype 1 type strain (X-73) they reacted strongly. Serum used for serotyping isolates in the gel diffusion precipitin test recognised many antigens in common with sera from infected birds, but some antigens were specific to typing sera.     

用几种电泳方法分析牦牛肝片吸虫不同部位抗原

应用ＳＤＳ－ＰＡＧＥ、等电聚焦电泳（ＩＥＦ）及双向电泳三种电泳方法分析牦牛肝片吸虫成虫四种抗原（头抗原－ＨＡ、体抗原－ＢＡ、体表抗原ＳＡ、分泌排泄抗原－ＥＳ）。ＳＤＳ－ＰＡＧＥ结果表明，ＢＡ、ＨＡ、ＳＡ分子量在１２～１００ｋＤ之间，ＥＳ在１２．３～２６ｋＤ之间，蛋白质染色ＢＡ、ＨＡ、ＳＡ、ＥＳ分别显示２５、２４、１８、９条带；ＩＥＦ分析肝片吸虫分别得３４、３３、２８、２２条带，主要由酸性蛋白质组成，主要谱带在ｐＩ４．２０～６．５５内；双向电泳分析ＢＡ、ＥＳ多肽斑点为６９、３０个     

多头绦虫抗原特性的研究──应用IHA观察绵羊免疫及感染后的抗体消长规律

为了观察绵羊用多头蚴抗原免疫及感染后的抗体消长规律,为羊脑多头蚴病的免疫预防和免疫诊断提供依据,本试验应用多头蚴原头节可溶性抗原、囊壁可溶性抗原、囊液粗抗原致敏绵羊红细胞对绵羊免疫3次及虫卵攻击感染后的血清抗体进行间接血凝试验（IHA）检测。结果表明,原头节抗原免疫组、囊壁抗原免疫组、囊液抗原免疫组及原头节ES抗原免疫组在首次免疫后1周,抗体滴度迅速升高,第3次免疫后1周达到峰值,虫卵感染后开始下降,到感染后30周接近正常水平。多头蚴3种抗原对同种抗原免疫组血清检测敏感性、特异性优于其它抗原,原头节免疫组、囊壁免疫组、囊液免疫组抗体水平明显高于原头节ES抗原免疫组。     

肌旋毛虫免疫亲和层析抗原的制备及其特性

以Sepharose 4B为载体,兔抗旋毛虫多克隆抗体为配体,纯化了旋毛虫的S_3抗原,得到亲和层析抗原.本实验结果表明,存在于S_3抗原中的主要抗原成分,同样存在于亲和层析抗原中,亲和层析抗原的分子量范围在103000～40000 u之间,等电点范围在pH4.7～8.8之间.亲和层析抗原经转印后,在硝酸纤维素膜上至少有7条显色带,其中分子量为48000 u,50000u,58000u和87000u的4种蛋白的抗原性较强,而以分子量为48000u的蛋白抗原性最强.     

Trichinella antigens: a review

This paper presents a review of the Trichinella antigens within the context of species variation. As with other parasites, Trichinella antigens can be classified according to their localisation as surface, excretory/secretory (ES) and somatic components. Surface antigens are mainly constituents of the outer cuticle although secretions from inner parts of the body wall as well as from the oesophagus can temporarily accumulate in the surface. ES antigens come mainly from the excretory granules of the stichosome, whereas somatic constitutive antigens come from the internal parts of the worms. ES products are considered very important from an immunological point of view as they are easily targeted by the immune system, whereas parasite death is required for exposure of somatic products. Some of the antigenic components have been characterised chemically. Phosphorylcholine is an important hapten that modulates the immune responses in Trichinella infections. Glycoproteins are the major components of surface and excretory/secretory products. A 43-kDa glycoprotein has been regarded as a good candidate for diagnosis and vaccination purposes. Recently some glycans have received special attention either as relevant epitopes or as parasite evasion strategies.     

Vaccination of Lewis rats against Mycoplasma arthritidis-induced arthritis.

The nature of Mycoplasma arthritidis antigens responsible for eliciting protective immunity in rats was studied by inoculation of rats with mycoplasmal components that had been subjected to a variety of physical and chemical treatments. All inocula tested induced good protection against development of clinical illness, as assessed by changes in body weight and appearance of joint swelling and/or temporary hind limb paralysis. Although all preparations stimulated development in inoculated rats of high titer of antimycoplasmal antibodies measured by ELISA, the complement-fixation antibody response was poor and, in some cases, lacking altogether. This indicated that completion-fixation antibodies may not be involved in protecting rats against M arthritidis-induced illness. Protective antigens were stable to heat (100 C for 10 minutes), formalin, and denaturation by sodium dodecyl sulfate (SDS). Inoculation with membrane and soluble cytoplasmic fractions was protective, as was inoculation with 5 M arthritidis fractions separated according to molecular weight by SDS-polyacrylamide gel electrophoresis (SDS-PAGE). For this latter experiment, rat antisera obtained after vaccination, but prior to challenge exposure, were tested by immunoblot analysis against electrophoretically separated M arthritidis membrane proteins. Interestingly, all antisera from these rats recognized antigens migrating far outside the molecular weight range of the cell fractions with which rats were inoculated. This indicated either that the protective antigens may be composed of numerous antigenically related subunits that separated by SDS-PAGE into a variety of molecular weight ranges or that a few major antigens may exist in several forms or phases within a given population of M arthritidis.     

PAPS免疫微球快速诊断猪旋毛虫病的研究

应用新型的聚醛化聚苯乙烯（ＰＡＰＳ）载体微球与最佳的旋毛虫抗原共价交联制备成特异性强、敏感性高、重复性好的快速诊断试剂,应用于猪旋毛虫病的生前诊断。我们对旋毛虫各个发育期的虫体和分泌排泄抗原进行了分析研究,并以同一蛋白质浓度（０．６ｍｇ／ｍＬ）的成虫,新生幼虫、肌幼虫、成虫ＥＳ、肌幼虫ＥＳ、肌幼虫Ａ峰、肌幼虫Ｂ峰等七种抗原分别与ＰＡＰＳ载体微球共价交联制成各种快诊试剂,然后进行效果比较试验,其结果     

Proteolytic enzymes from Trichinella spiralis larvae.

Trichinella spiralis larvae infect their hosts by the penetration of small intestine enterocytes. The exact mechanism of penetration is unknown, but the presence of proteolytic enzymes is suspected. In this study, whole worm extracts and excretory-secretory (ES) components were obtained and their proteolytic enzymes examined. Enzymes from worm extracts were capable of hydrolysing azocoll, a general protease substrate in a wide range of pH (2-8), with maximal activity at pH 5. Trichinella spiralis larval enzymes were sensitive to metalloprotease and serine protease inhibitors. Three proteases were identified in worm extracts at molecular weight (MW) 48, 54 and 62 kDa by incorporating a gelatine substrate into a standard or a modified sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) set-up, in which we used low SDS concentration in gel and electrophoresis buffer (0.01%). Intact larvae incubated in a medium containing azocoll showed azocollytic activity. Subsequent analysis of ES products by modified SDS-PAGE in gels containing gelatine demonstrated the presence of three protease of apparent MW 33, 62 and 230 kDa.