猪囊尾蚴病免疫和诊断抗原的分子生物学研究进展

猪囊尾蚴病是一种危害严重的人兽共患寄生虫病。猪囊尾蚴特异性和保护性抗原的研究是猪带绦虫病、猪囊尾蚴病免疫和诊断的基础。天然抗原来源有限,限制了其应用,而重组抗原的应用可解决质量控制和抗原来源的问题。该文就近年来猪囊尾蚴病免疫和诊断抗原的分子生物学研究进展进行了综述。     

猪囊尾蚴跨膜蛋白T24基因的克隆及序列分析

目的 寻找猪囊尾蚴病新的免疫学候选诊断分子.方法 设计合成引物,用PCR法从猪囊尾蚴cDNA文库中扩增出猪囊尾蚴跨膜蛋白T24基因编码序列,将其与线性克隆载体PMD-18T连接,分别进行酶切和PCR鉴定及DNA测序证实.结果 PCR法扩增出一条长度约678 bp的特异性片段,将重组质粒PMD-18T-T24作BamHⅠ和XhoⅠ双酶切,均能得到一个大小与PCR扩增产物一致的插入片段,对插入片段的测序结果表明,T24具有一个长度为678 bp的完整开放阅读框,编码225个氨基酸,理论分子量为23.91 kDa,与GenBank收录的猪囊尾蚴T24基因(编号为AY211879)具有高度的同源性(99.85%).结论 猪囊尾蚴跨膜蛋白T24编码基因克隆成功,为进一步的表达、鉴定及免疫诊断研究奠定了基础.     

毕赤酵母重组猪囊尾蚴抗原cC1的特征研究

目的 研究毕赤酵母重组猪囊尾蚴抗原cC1蛋白的特征变化。方法 对表达产物行SDS—PAGE,FPLC离子交换层析和分子筛，等电聚焦，脱磷酸反应，N端氨基酸测序，小鼠免疫接种。结果 毕赤酵母重组猪囊尾蚴抗原cC1蛋白的分子量较原核的大，易于纯化，等电点较理论值低，重组cC1已被磷酸化，N端携带了来自载体的4个氨基酸，免疫原性较原核的明显增强。结论 毕赤酵母重组猪囊尾蚴抗原cC1蛋白的特征较原核的有了明显变化，尤其是蛋白易于纯化及免疫原性的增强，表明毕赤酵母表达系统非常适合重组亚单位疫苗的生产制备。     

猪囊尾蚴病重组DNA疫苗pcDNA 3.1-TSO45W在小鼠体内诱导的体液免疫效应

目的观察基于六钩蚴期的猪囊尾蚴病基因疫苗TSO45W诱导的小鼠体液免疫应答。方法碱裂解法大量制备重组质粒pcDNA3.1-TSO45W及对照质粒pcDNA3.1。将45只雌性昆明小鼠(4~6周)随机分为3组,每组15只。A组(生理盐水组):每只小鼠肌肉注射生理盐水100μl;B组(空质粒pcDNA3.1对照组):每只小鼠肌肉注射空质粒100μg;C组(重组质粒pcDNA3.1-TSO45W组):每只小鼠肌肉注射重组质粒100μg。分别于免疫后2、4、6、8、10周以ELISA方法检测小鼠血清免疫球蛋白IgG,IgM及IgA含量。结果 pcDNA3.1-TSO45-4B免疫组小鼠血清IgG、IgM、IgA均于第4周开始升高,分别为(17.36±1.85)μg/ml、(9.25±1.78)μg/ml和(9.41±0.88)μg/ml,并分别于第8、第6、第8周达到最高,含量分别为(24.26±2.52)μg/ml、(10.69±0.52)μg/ml和(11.22±1.23)μg/ml,与空质粒对照组(B组)和生理盐水对照组(A组)比较差异均有统计学意义(P<0.05)。结论猪囊尾蚴保护性抗原TSO45WDNA疫苗能在小鼠体内诱导体液免疫效应。     

Childhood neurocysticerosis in South India

A total of 21 cases of childhood neurocysticerosis seen over five years (1985–89) at JIPMER hospital Pondicherry, are reported. Nine of these patients were males and twelve females. Their age ranged between 5 to 15 years. The presenting features were convulsive seizures (14), features of raised intracranial pressure (6) and meningoencephalitis syndrome (1). Diagnosis of neurocysticercosis was based on positive CSF serological tests (11), CT morphology (11), brain biopsy (1) and autopsy (1). Praziquantel therapy was given in 4 cases, 3 of them showed remarkable improvement in neurological status and one died of acute reaction.     

Cysticercosis as a major risk factor for epilepsy in Burundi,east Africa

PURPOSE: Human cysticercosis is a direct consequence of infection by Taenia solium larvae (Cysticercus cellulosae). Results of studies on the impact of neurocysticercosis on epilepsy in Africa are inconsistent. The objective was to evaluate the role of cysticercosis in epilepsy in Burundi. METHODS: A prevalent matched case-control design was used in the Kiremba area, Burundi, between March and April 2001. One case with epilepsy was matched to two control subjects, according to their age. Cases were subjects who had shown at least two unprovoked epileptic seizures within a 24-h time range and who lived in the Kiremba area. The control subjects also lived in Kiremba and had neither neurologic illness nor kinship with the people with epilepsy. Seropositivity for cysticercosis was the exposure variable. Three hundred twenty-four prevalent cases, with onset of epilepsy between 1950 and 2000, and 648 age-matched controls were included. RESULTS: This study found a link between cysticercosis infestation and the occurrence of epilepsy (odds ratio, 3.8; 95% confidence interval, 2.5-5.1). CONCLUSIONS: The study highlighted the importance of cysticercosis in the area of Kiremba, as 31.5% of the control subjects screened positive for this parasite. The attributable risk for cysticercosis was 50% (95% confidence interval, 42-57) in this population.     

Epilepsy and neurocysticercosis in Atahualpa: a door-to-door survey in rural coastal Ecuador

PURPOSE: To determine the prevalence of epilepsy and the role of neurocysticercosis in the occurrence of epilepsy in Atahualpa. METHODS: We used a door-to-door survey to detect subjects with epileptic seizures, to collect a blood sample for determination of anticysticercal antibodies, and to evaluate social characteristics of the population, including household pig ownership. Neurologists examined suspected cases and a sample of negative individuals. Then patients with epilepsy, as well as age- and sex-matched controls, underwent a head computed tomography (CT) and a scalp EEG. RESULTS: The questionnaire was answered by 2,415 of 2,548 residents of Atahualpa, and cysticercosis serology was performed in 1,687 consenting individuals. Cysticercosis seroprevalence was 145 (8.6%) per 1,686). Neurologic examination confirmed 24 patients with epilepsy (crude prevalence, 9.9 per 1,000 population, and 10.8 per 1,000 when adjusted to the United States population). After adjustment by age, sex, and pig raising, positive serology was strongly associated with epilepsy (odds ratio (OR), 4.16; 95% confidence interval (CI), 1.6-11.2). CT findings compatible with neurocysticercosis were found in five patients with epilepsy and also were more frequent than in controls, although this did not reach statistical significance (five of 19 vs. one of 19; p = 0.125, McNemar's test). Besides these five cases, three other patients with epilepsy had positive serology (one with a normal CT and two who did not have a CT). CONCLUSIONS: Neurocysticercosis is associated with one-third of cases of epilepsy in Atahualpa and may be a major contributory factor for the excess fraction of epilepsy seen in this population.     

用SDS—PAGE、Western blot和ELISA方法检测猪囊虫囊液中有效抗原的研究

用SDS-PAGE、Western blot和ELISA方法检测猪囊虫囊液中的有效抗原,结果猪囊虫囊液中有8条抗原区带被猪囊虫病患者血清中抗体识别,分子量分别为255、185、105、69、64、60、48、和41kDa。其中第至3第7条免疫活性较强,含量较高,为有希望用于研制猪囊虫病疫苗的抗原分子。     

囊虫病患者血清sICAM—1的检测

用ＥＬＩＳＡ法检测了２０例未经治疗的囊虫病患者血清中可溶性细胞间粘附分子１（ｓＩＣＡＭ１）的水平，以１５例健康人作对照。结果显示，患者血清中ｓＩＣＡＭ１含量为（２８２８０±６１８４）Ｕ／ｍｌ，极显著低于正常对照组（Ｐ＜００１）。提示未经治疗的囊虫病患者免疫功能低下