羊脑多头蚴抗原的等电聚集电泳分析

采用等电聚焦电泳（ＩＥＦ）技术,对自然感染绵羊的脑多头蚴的头节可溶性怕、囊壁可溶抗原和囊液抗原进行了分析。ＩＥＦ电泳后,分别用考马斯蓝Ｒ－２５０法染蛋白质,ＰＡＳ法染多、醛酸α－萘酯－坚固蓝法染酯酶曲同工酶、Ｎｉｌｅ‘ｓ蓝法洒脂。蛋白质染一头节可溶性抗原显带４２条,等电点４．８９－８．７２;囊壁可溶性抗原显带５０条,ｐｌ４．８０－８．６８,囊液抗原显带３９第,ｐＩ４．８０－９．８８。要色后头节可溶     

猪囊尾蚴特异性单克隆抗体的制备及其识别抗原成分的鉴定

通过SDS-PAGE方法对猪囊尾蚴囊液抗原成分进行了分离鉴定,并分别分离纯化了其中的16 000和10 000特异性蛋白质成分;将16 000和10 000纯化蛋白质成分分别做成佛氏佐剂苗,分组免疫BALB/c小鼠,超免后,无菌取脾脏制备脾细胞,分别与NS0骨髓瘤细胞进行融合,经阳性筛选和特异性鉴定获得2株分泌猪囊尾蚴特异性单抗的杂交瘤细胞,命名为TSCF-1611H12B8和TSCF-1012G5B5。免疫学鉴定结果表明,单抗TSCF-1611H12B8和TSCF-1012G5B5与猪细颈囊尾蚴、旋毛虫、住肉孢子虫和蛔虫抗原间不存在交叉反应;单抗TSCF-1611H12B8识别囊液中16 000和10 000蛋白质抗原条带,而单抗TSCF-1012G5B5识别囊液中的10 000蛋白质条带。     

应用猪囊虫匀浆抗原间接血凝试验检测猪囊虫病抗体的研究

从囊虫匀浆浸提液中分离出一种抗原性与特异循环抗原相同的成分,以这种抗原成分通过间接血凝试验检测病、健猪的血清抗体,结果病猪抗体阳性率为93.2％(96/103),健猪阴性(?)合率为100％。     

绵羊肺腺瘤病毒NM株囊膜蛋白的二级结构及其B细胞抗原表位预测

利用生物信息学预测绵羊肺腺瘤病毒(OPAV)NM株囊膜蛋白优势抗原表位,为寻找用于绵羊肺腺瘤病诊断和预防的候选抗原表位提供参考。参照OPAV-NM株env基因序列(登录号:JQ837489),应用Gamier-Robson方法、SOPMA方法和网络服务器PSIPRED、Predictprotein预测OPAV-NM株囊膜蛋白的二级结构。用Kyte-Doolittle方法预测蛋白质的疏水区和亲水区,用Emini方法预测蛋白质表面可能性,以Jamson-Wolf方法预测蛋白的抗原指数,利用Bcepred、IEDB、ABCpred网络服务器预测囊膜蛋白的抗原性,然后综合评价囊膜蛋白的B细胞抗原表位。结果表明,囊膜蛋白(ENV)二级结构丰富,α螺旋和β折叠所占比例相对较多,也具有多处转角及无规则卷曲区域,提示OPAV-NM囊膜蛋白具有较多的优势抗原表位区段。这些优势抗原表位分别为第52～76,101～110,118～165,182～196,201～215,281～305,309～332,335～356,461～476,530～574位氨基酸,其中第461～476位和第530～574位的部分氨基酸序列位于胞外区,且第530～574区段位于en-OPAV和ex-OPAV的囊膜氨基酸序列的差异区,该段氨基酸序列对绵羊肺腺瘤病毒疫苗和诊断方法的研究有重要意义。     

旋毛虫病免疫诊断抗原的筛选研究

本文报道了旋毛虫肌幼虫匀浆抗原(HO抗原),排泄-分泌物抗原(ES抗原),可溶性杆细胞颗粒相关抗原(S_3抗原),葡聚糖凝胶G200柱层第一峰(FP)和第二峰(SP)抗原,亲和层析提纯抗原(PAW)的制备及其用于ELISA和Dot-ELISA诊断旋毛虫病的价值。证实亲和层析堤纯抗原的特异性高、敏感性强,可用于旋毛虫病的免疫诊断和流行病学调查。     

猪囊虫病提纯抗原炭凝诊断法

为了提高抗原的敏感性和特异性,曾采用硫酸铵盐析法、葡聚糖凝胶过滤法、电泳分离法和液氮冷冻法提纯囊液抗原,但在炭凝诊断上均未取得满意效果。而减压分镏法(真空分镏法)提纯的囊液抗原,其炭凝反应的敏感性、特异性都有明显提高。     

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。     

猪囊尾蚴抗原成分及其诊断价值分析

猪囊尾蚴病是一种重要的人畜共患寄生虫病。囊尾蚴病的快速诊断是囊尾蚴病治疗和预防的前提,免疫学诊断方法是近年来的囊尾蚴病临床诊断中的重要辅助手段,高特异性诊断抗原的筛选为近年来的研究热点。囊尾蚴抗原组成复杂,包括囊液抗原、头节抗原、囊壁抗原、循环抗原和排泄分泌抗原等。其中的囊液抗原特异性较好,可用于免疫学诊断,而排泄分泌抗原诊断囊尾蚴的价值尚待进一步研究。     

不同宿主源棘球蚴包囊液抗原的SDS—PAGE分析

本文利用ＳＤＳ－ＰＡＧＥ对不同宿主源棘球蚴囊液抗原的多肽组成进行了分析和比较,旨在为免疫诊断抗原的分离、鉴定和纯化奠定基础,为细粒棘球绦虫种内变异和株的鉴定提供参考指标。结果表明,在还原条件下,绵羊棘球蚴囊液抗原共有多肽带１９条,其中６６和５９ＫＤ多肽带为主带,４０、３４．５、３３、２４．５和１４ＫＤ多肽带次之。牛源囊液抗原共有多肽带１２条,６６和５９ＫＤ多肽带也为主带,２４．５ＫＤ多肽带次之。人源囊液抗原共有多肽带１３条,６６、４０、２０．５和１４ＫＤ多肽带为主带,５９ＫＤ多肽带近于缺乏,分子量在５９和２４ＫＤ之间的带明显偏少,而７１、６９、６８和２０．５ＫＤ多肽带为自身特有。３种不同宿主源棘球蚴囊液抗原其多肽组成均很复杂,羊、牛源囊液抗原的ＳＤＳ－ＰＡＧＥ图谱较相似,而人源与此差异明显。初步认为绵羊和牛源囊液抗原在免疫诊断中具有可替代性。     

兔豆状囊尾蚴囊液蛋白的纯化与抗原性鉴定

为了找出抗原性强、免疫原性好的兔豆状囊尾蚴抗原蛋白,本研究对兔豆状囊尾蚴的头节、体节和囊膜等结构性抗原与囊液中代谢性抗原进行了比较,发现囊液代谢性抗原的免疫效果优于结构性抗原。通过SDS-PAGE和微孔超滤分离法对囊液蛋白进行纯化,再用纯化的蛋白免疫小鼠,制备高免血清,并对免疫鼠的免疫器官进行病理学和免疫组织化学检查。结果证明用从囊液中纯化的63 000蛋白免疫效果最好。用纯化的63 000蛋白免疫小鼠可获得高效价抗血清,小鼠免疫器官活化,巨噬细胞、滤泡树突细胞、淋巴细胞和浆细胞等各种免疫细胞增多,用免疫组织化学染色,免疫器官中的CD4、CD8T细胞和CD20B细胞呈强阳性反应。总之,从囊液中分离纯化的63 000蛋白抗原性强、免疫原性,可用于对豆状囊尾蚴病的免疫学诊断。     

Comparative immunoelectrophoretic analysis of Echinococcus granulosus, Taenia hydatigena and Taenia pisiformis cyst fluid antigens by hyperimmune rabbit sera.

Cyst fluid antigens of Echinococcus granulosus, Taenia hydatigena and T pisiformis were examined by electrophoresis using homologous and heterologous hyperimmune rabbit sera to these antigens. While arc 5 forming antibodies were identified in sera from rabbits immunised with E granulosus and T hydatigena cyst fluids, antibodies responsible for forming precipitating antigen B band were detected in rabbit antisera to E granulosus, T hydatigena and T pisiformis antigens. T hydatigena cyst fluid appears to contain antigen similar to E granulosus antigen 5 and probably antigen B while T pisiformis cyst fluid has mainly an antigen close to hydatid antigen B.     

斑点金标渗滤法检测猪囊虫病

以亲和层析原理为基础,用猪囊虫纯化抗原作为检测用抗原,胶体金直接标记抗原,建立了斑点金标渗滤法诊断猪囊虫病的方法。其中囊虫抗原与胶体金溶液结合的最佳pH值为7.5,最佳浓度为52 mg/mL,检测时血清的最佳稀释度为1∶10。建立的斑点金标渗滤法检测猪囊虫病血清均为阳性,正常猪及其他病猪血清均显示为阴性,整个检测时间只需要5 min左右。试验结果显示,该方法操作简便、快速、特异性好,适用于猪囊虫病的临床诊断和流行病学调查。     

Protection against chicken leucocytozoonosis provided by immunization with spleen homogenate infected with Leucocytozoon caulleryi

Protection against chicken leucocytozoonosis was assessed in chickens immunized with spleen homogenates from chickens that had received sporozoites of Leucocytozoon caulleryi 7 or 13 days previously. Chickens immunized with the homogenate were challenged with sporozoites of L. caulleryi and observed for changes in clinical signs, parasitemia, serum-soluble antigen, and antibody responses. In chickens immunized with either the 7-day or 13-day homogenate, clinical signs and parasitemia were moderate, mild or absent. This was the case both after immunization with the homogenate and after sporozoite challenge. Immunization with spleen homogenates demonstrated protection against chicken leucocytozoonosis.     

Detection of rabbit haemorrhagic disease virus antigen in tissues by immunohistochemistry

Formalin fixed liver, spleen, kidney, heart, lung, duodenum and appendix tissues from nine rabbits, experimentally infected with rabbit haemorrhagic disease virus (RHDV), were investigated for evidence of RHDV antigen by the direct avidin-biotin peroxidase complex immunohistochemical method. In all the rabbits examined, RHDV antigen was detected in degenerative and necrotic hepatocytes of the liver tissues. The area involved coincided with histopathological lesions on serial liver sections. The RHDV antigen was expressed in the cytoplasm of the hepatocytes, suggesting that RHDV replicated in these cells. RHDV antigen was also detected in the spleen. The results of immunohistochemistry were supported by the demonstration of RHDV protein by Western blot analysis and of RHDV particles by protein A-gold immunoelectron microscopy in the liver homogenate from all the rabbits that were examined.     

异源性抗原抗猪囊尾蚴感染的研究

本文报道了泡状带绦虫活化六钩蚴的超声裂解抗原可以诱导猪体产生抗猪带绦虫攻击感染的交叉保护作用。猪囊尾蚴匀浆抗原也使猪体产生了较强的抗猪带绦虫攻击感染的保护作用。泡状带绦虫六钩蚴超裂抗原免疫组与猪囊尾蚴匀浆抗原免疫组的保护情况是相似的,这表明异源免疫也可使猪体产生较好的抗猪囊尾蚴感染的免疫。由于制备异源性抗原的泡状带绦虫能够从狗的体内获得,因此在体外培养猪带绦虫未获成功之前可以解决从人体获取猪带绦虫的困难。     

Quantitative estimation of amino acids in cysticercus of taenia hydatigena

Cyst membranes of Cysticercus tenuicollis, the larval stage of Taenia hydatigena, contained 14 amino acids, namely lysine, histidine, arginne, aspartic acid, threonine, serine, glutamic acid, proline, glycine, alanine, isoleucine and leucine. Methionine was present in traces. In comparison, the cyst fluid was deficient in histidine, arginne, threonine, methionine and valine and the scolices did not have threonine, serine, glutamic acid and methionine. The membrane contained large quantities of amino acids in comparison to the other components of the cyst.     

布氏姜片虫排泄分泌抗原的提取及其免疫化学特性研究

利用体外无菌培养技术，制备了布氏姜片虫的排泄分泌抗原(ES)。经梯度PAGE和SDS-PAGE分别分离出7条蛋白质区带和9条多肽区带，其中包括1条糖蛋白和1条糖脂蛋白，说明布氏姜片虫ES含复杂的蛋白组分。免疫电泳、免疫扩散即迹试验表明，布氏姜片虫ES含有较多与虫体抗原相同的抗原成分，也含有少数与虫卯抗原相同的抗原成分。     

Fasciola hepatica: studies on vaccination of rats and mice with a surface antigen prepared from fluke homogenate by means of a monoclonal antibody

T1 tegumental antigen was isolated from a homogenate of eight- to 10-week-old Fasciola hepatica using a T1-specific monoclonal antibody bound to sepharose in an antibody-affinity column. Rats and mice were vaccinated with T1 antigen in Freund's complete adjuvant, and control groups received equivalent amounts of non-T1 antigen (eluted from the antibody-affinity column) or ovalbumin. On completion of the immunisation programme, serum samples were collected for ELISA and IFA testing. The animals were challenged by oral infection with F hepatica metacercariae or, for several vaccinated rats, by intraperitoneal transplantation of live adult flukes. At autopsy, worm-burden and liver damage was assessed for each animal and the condition of transplanted flukes was examined. Comparison of test and control groups of animals showed that neither T1 nor non-T1 antigens provided significant protection against challenge, although specific antibody responses against the appropriate sensitising antigen were engendered. Flukes transplanted to the peritoneal cavity of immunised rats survived without damage, although they became encased in hollow fibrous capsules of host origin. The results lend support to the pre-existing concept that glycocalyx turnover by discharge of T1 secretory bodies at the apical surface of migrating flukes provides an efficient means of protection for the parasite against host immunity.     

Delayed-type hypersensitivity test for assessing tick-immune status of cattle in Zambia

Delayed-type hypersensitivity skin reactions were used to assess the tick resistance status of Tonga calves in Zambia. The antigen used in the tests was a homogenate of unfed nymphal Rhipicephalus appendiculatus which had been shown to give protective immunity in guinea pigs to adult female R appendiculatus. There was a significant negative correlation between the intensity of the reactions and the total number of ticks (Amblyomma variegatum, R appendiculatus, Hyalomma truncatum, Boophilus decoloratus and Rhipicephalus species) on the animals.