弓形虫抗体免疫印迹试剂盒的研制

目的 研制敏感、特异的检测弓形虫IgM和IgG抗体免疫印迹试剂盒。 方法 收集人工感染RH株弓形虫速殖子昆明系小鼠的腹腔液，提取弓形虫胞质蛋白，采用十二烷基磺酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）分离弓形虫可溶性抗原，并经电泳转印至硝酸纤维膜，以无毒灵敏的四甲基联苯胺（TMB）为底物分别检测30份弓形虫IgM和28份IgG阳性血清，40份健康人血清。通过比较抗原制备方法和使用剂量、封闭剂、洗涤和稀释剂、工作浓度、作用时间以及反应带出现率，选择最佳实验条件，以敏感性、特异性、Youden指数以及稳定性作为试剂盒评价标准。 结果 用免疫印迹试剂盒检测30份弓形虫IgM和28份IgG阳性血清，敏感性分别为90.0%（27/30）和85.7%（24/28），40份健康对照者血清的弓形虫IgM和IgG抗体均为阴性，特异性均为100%，Youden指数分别为0.9和0.86。试剂盒于4℃ 保存约6个月的检测结果一致。 结论 该免疫印迹试剂盒敏感性和特异性均较高，且操作简便、快速。     

弓形虫培养上清体外对CD4+CD25+T细胞的影响

目的探讨体外培养条件下弓形虫培养上清对CD4+CD25+T细胞的影响。方法用弓形虫培养上清和自小鼠脾脏分离的CD4+CD25+T细胞共同孵育，Annexin-v染色法检测CD4+CD25+T细胞的凋亡情况，淋巴细胞增殖实验检测CD4+CD25+T细胞对CD4+CD25-T细胞的抑制功能。结果  用弓形虫培养上清孵育10 h的小鼠脾脏CD4+CD25+T细胞有(36. 90±0.36)%出现凋亡，和RPMI-1640孵育组相比，凋亡率上升了（13. 60士2.15）%。与RPMI-1640孵育组相比，用弓形虫培养上清处理5、10 h的小鼠脾脏CD4+CD2 5+T细胞对CD4+CD25 -T细胞的抑制功能下降(P<0.01)。结论  弓形虫培养上清中可能存在某些成分能够诱导CD4+CD25+T细胞出现凋亡，并使CD4+CD25+T缅胞对CD4+CD25-T细胞的抑制功能下降。     

Immunological response and protection of mice immunized with plasmid encoding Toxoplasma gondii glycolytic enzyme malate dehydrogenase

Toxoplasma gondii Malate dehydrogenase (TgMDH) plays an important role as part of the energy production cycle. In this investigation, immunological changes and protection efficiency of this protein delivered as a DNA vaccine have been evaluated. Mice were intramuscularly immunized with pTgMDH, followed by challenge with virulent T. gondii RH strain, 2 weeks after the booster immunization. Compared to the control groups, the results showed that pTgMDH has stimulated specific humoral response as demonstrated by significant high titers of total IgG and subclasses IgG₁ and IgG_2a, beside IgA and IgM, but not IgE. Analysis of cytokine profiles revealed significant increases of IFN‐γ, IL‐4 and IL‐17, while no significant changes were detected in TGF‐β1. In cell‐mediated response, both T lymphocytes subpopulations CD4⁺ and CD8⁺ were positively recruited as significant percentages were recorded in response to immunization with TgMDH. Significant long survival rate, 17 days, has been observed in the TgMDH vaccinated group, in contrast with control groups which died within 8–9 days after challenge. These results demonstrated that TgMDH could induce significant immunological responses leading to a considerable level of protection against acute toxoplasmosis infection.     

A serological Survey of Brucella spp., Salmonella spp., Toxoplasma gondii and Trichinella spp. in Iberian Fattening Pigs Reared in Free‐Range Systems

Zoonotic agents such as Brucella spp., Salmonella spp., Toxoplasma gondii and Trichinella spp., all considered high‐risk zoonotic pathogens by the European Food Safety Agency (EFSA), may cause no symptoms of infection in free‐range pigs yet still have a significant public health impact. A serological survey was therefore performed to determine the history of occurrence of these pathogens in such pigs in southern Spain. A total of 709 serum samples were collected at abattoir from pigs from 79 farms and analysed for specific antibodies against the above pathogens using commercially available ELISA kits. Encysted Trichinella spp. larvae were also sought following the artificial digestion method of diaphragm pillar muscle. The results showed Salmonella spp. to be widely distributed among the sampled herds [73.42%, 95% confidence interval (CI₉₅) 65.6–81.78] and Toxoplasma gondii to be present in over half (58.23%, CI₉₅ 47.33–69.07). The seroprevalence of Brucella spp. was very low (3.8%, CI₉₅ 0.18–7.42), and antibodies against Trichinella spp. were not detected. No encysted Trichinella spp. larvae were microscopically detected.     

艾滋病患者并发弓形体病误诊分析

目的为提高艾滋病合并弓形虫感染的认识和弓形虫形态学识别能力,避免误诊。方法文献复习艾滋病合并弓形虫感染患者的临床特征,弓形体病与黑热病的鉴别要点,以病原体图像比较利杜体、弓形虫形态特点。结果病例诊断为艾滋病合并弓形体病。结论误诊的主要原因是将骨髓中弓形虫速殖子误判为利杜体。     

弓形虫慢性感染小鼠的系统性细胞免疫应答规律

弓形虫感染可激发宿主产生Th1型免疫反应,因此研究弓形虫疫苗尤其需要关注系统性T细胞免疫应答与保护力的关系。本研究被弓形虫Pru株包囊口服感染BALB／c小鼠,成功建立了弓形虫慢性感染小鼠模型。慢性感染小鼠的脾细胞用弓形虫可溶性抗原刺激后可以产生特异性增殖。进一步分析发现弓形虫特异性CD3＋T、CD4＋T和CD8＋T细胞都可以产生IFN-γ。本研究建立的弓形虫慢性感染小鼠模型可以产生较强的系统性T细胞免疫应答,为弓形虫疫苗研制的效果评价提供重要参考。     

对宫内感染弓形虫出生子代远期发育研究

目的探讨先天性弓形虫感染对儿童学习和记忆的远期影响,为先天性弓形虫病防治和康复提供指导。方法选建保健册孕妇分娩、现读中学的儿童作为研究对象。应用酶联免疫吸附法(ELISA)检测孕妇外周血弓形虫IgM、CAg。对弓形虫IgM、CAg阳性孕妇,检查新生儿脐血弓形虫IgM、CAg,根据婴儿出生时脐血弓形虫检测结果分感染阳性组(感染组孕妇分娩儿IgM或CAg阳性)、感染阴性组(感染组孕妇分娩儿IgM和CAg阴性)和非感染组(非感染组孕妇分娩儿);选择愿意合作感染阳性组60例,分治疗和不治疗各30例,比较干预效果。采用简明精神状况调查量表(MMSE)、汉诺塔测试(TOH-1—TOH-d—TOH-s—TOH-2)和威斯康星卡片分类测试(WCST-1—WCST-d—WCST-s—WCST-2)进行神经心理学检查。结果在MMSE测试中,阳性未治组在回忆能力、计算力以及总分测试中得分显著低于非感染组(P<0.05)。学习前,在TOH和WCST测试中,阳性未治组与非感染组比较、阳性未治组与治疗组比较差异均有统计学意义(P均<0.05)。学习后,阳性治疗组、阳性未治组TOH得分与学习前差异无统计学意义(P>0.05),阳性未治组W...     

白细胞介素家族在弓形虫感染中的作用

白细胞介素（IL）在清除宿主机体细胞内寄生原虫弓形虫的过程中发挥着重要作用，如破坏染虫细胞，构成免疫网络、调节免疫平衡等。现对其中重要的几类IL进行综述，旨在为弓形虫感染的治疗提供科学依据。     

泰安市七种疾病患者弓形虫感染调查

目的：了解泰安地区特殊病种人群的弓形虫感染情况，探讨其之间的相互关节。方法：对本地区住院的7种疾病261例患者采用酶联免疫吸附试验进行弓形虫特异性IgG、IgM抗体和CAg的检测。结果：总阳性率为28.74%，其中三者的新时期阳性率分别为19.54%、6.51%和2.68%，三项均阳性的3例，阳性率为1.15%。7种疾病中肺癌的阳性率为40.00%，葡萄膜炎、白血病、精神病、心肌炎及心肌病、脑膜炎和肝炎的阳性率依次为33.33%、30.00%、29.41%、30.00%、13.33%和7.50%。结论：弓形虫感染在本地区特殊病种人群中较为普遍，其中以肺癌、葡萄膜炎、白血病、精神病为最常见，与患者的性别、年龄、等因素无显著关系，但与职业及猫、狗等动物的接触有关。