Tetanus toxoid as an antigen for delayed cutaneous hypersensitivity

We studied the usefulness of tetanus toxoid (TT) as a skin test antigen in assessing cellular immune function. Hospitalized patients were skin tested with four antigens, and the response rates between these antigens were compared. Candida and mumps antigens resulted in significantly more positive responses than did TT or PPD. The response rate to the TT significantly declined in older patients, suggesting these persons may not be adequately immunized against tetanus. We found TT is not as useful as Candida or mumps antigens in the evaluation of cellular immune function in a hospitalized population but it may have some usefulness in the evaluation of energy. Negative skin test results may convert to positive after patients receive an immunizing dose of TT, as shown by our data. In addition, there were several patients with a positive response to TT and negative responses to the other common skin test antigens.     

马尔尼菲青霉菌素的制备和应用

目的探讨马尔尼菲青霉菌素的制备方法,并评价以马尔尼菲青霉菌素皮肤试验来诊断播散性马尔尼菲青霉菌病的价值。方法将新西兰大白兔随机分为对照组、马尔尼菲青霉菌组和白色念珠菌组,每组20只。马尔尼菲青霉菌组感染马尔尼菲青霉菌,白色念珠菌组感染白色念珠菌,对照组不作任何处理。按真菌抗原提取法制备马尔尼菲青霉菌素并进行部分改良,用Lowry法测定马尔尼菲青霉菌素中纯蛋白衍生物的含量;将马尔尼菲青霉菌素皮内注射于3组新西兰大白兔耳部内侧皮肤,并于注射后24、48、60h观察皮肤红肿反应。结果获得的马尔尼菲青霉菌素原液纯度高,特异性好;抗原效价测定结果显示马尔尼菲青霉菌素中纯蛋白衍生物含量为68mg·L-1,且具有良好的安全性。马尔尼菲青霉菌组和白色念珠菌组各时间点皮肤红肿反应平均直径比较,差别有统计学意义(P<0.001)。对照组未出现皮肤反应。注射后24h,马尔尼菲青霉菌组皮肤红肿反应全部为阳性,白色念珠菌组有2只为阳性。马尔尼菲青霉菌素皮肤试验方法对马尔尼菲青霉菌病进行早期诊断的敏感性为100.0%,特异性为90.0%,阳性预测值为90.9%,阴性预测值为100.0%。结论对真菌抗原提取法进行部分改良可获得具有良好安全性的马尔尼菲青霉菌素;马尔尼菲青霉菌素皮肤试验方法简便,结果稳定,可用于马尔尼菲青霉菌病的辅助诊断。     

单克隆抗体Mab03.2C1-C2靶抗原性质的初步研究

目的分析抗白念珠菌芽管胞壁外膜抗原单抗Mab03.2C1-C2靶抗原的性质,探讨该抗原用于实验室检测的可行性.方法制备单克隆抗体,通过SDS-PAGE及Western Blotting法分析该单抗所识别的抗原的分子量.体外诱导形成不同长度的芽管,分析其表面靶抗原的形成情况.不同的生化方法处理芽管,采用间接免疫荧光方法,对单抗靶抗原表位性质进行分析.取口腔念珠菌病患者的真菌涂片标本,用IIF方法观察体外诱导培养之芽管与体内白念芽管表面抗原分布异同.结果体外诱导30 min后白念珠菌芽管表面方可检测到靶抗原.体内白念菌丝与体外诱导菌丝表面靶抗原分布不同.靶抗原表位可能位于N-糖链上.结论Mab03.2C1-C2靶抗原为白念菌丝形成过程中生成的新成分,可能参与白念致病过程.靶抗原为糖蛋白,抗原性显著,对临床应用于系统性白念珠菌感染的检测有意义.     

白念珠菌免疫显性抗原的分离纯化及鉴定

目的：纯化和鉴定白念珠菌免疫显性抗原。方法：用产胶过滤性分离纯化白念珠菌免疫显性抗原，用酶联免疫法和免疫印迹法鉴定抗原性。结果：获得的两种白念珠菌免疫显性抗原达到电泳纯：p38蛋白和p28蛋白。经SDS－PAGE测定其分子量分别为38.1-kDa和28.1-kDa,Lowry法测定其含量分别为0.582mg/ml和1.345mg/ml。酶联免疫法和免疫印迹法显示p38和p28都能和小鼠抗白念珠菌血清反应。结论：初步证实p38和p28为白念珠菌免疫显性抗原，初步建立了分离纯化白念珠菌免疫显性抗原的基本方法。     

Humoral and cell-mediated immune response to crude antigens of Dermatophilus congolensis during experimental infection of rabbits.

Rabbits were infected with Dermatophilus congolensis and tested for humoral immune response by indirect haemagglutination and for cell-mediated immune response to crude antigens of D. congolensis. Lymphocyte transformation and macrophage migration inhibition assays were used as in vitro correlates of cell-mediated immune response while cutaneous delayed hypersensitivity was used in vivo. Endo-antigen and whole cell antigen were found to significantly induce cell-mediated immune response. In contrast, humoral responses were found to be more significantly induced by exo-antigen. A biphasic immune response was revealed by the lymphocyte transformation test.     

用PCR技术直接检测白色念珠菌DNA的实验研究

目的建立和评价直接用PCR从血标本中检测白色念珠菌核酸的方法。方法用红、白细胞裂解液、破壁酶和真菌DNA提取盒直接处理血标本,得到的微量靶DNA用白色念珠菌种特异性引物进行扩增。结果在白色念珠菌制备的人血标本中检测出靶DNA,其敏感性达10个孢子/ml以下,从处理标本到报告结果仅需6h。结论用PCR法可特异、敏感地从血标本中直接检测白色念珠菌核酸,有助于临床快速诊断深部白色念珠菌感染。     

Analysis of the Candida albicans - specific T-cell response and oropharyngeal Candida colonization in a cohort of HIV-1-infected patients

To investigate Candida epidemiology and immunologic correlates of protection in HIV-1 infected patients, we analyzed oral Candida colonization in correlation to the Candida-specific T-cell response measured by g-IFN ELISPOT using different Candida (C.) albicans strains. In 16/46 patients (13 asymptomatic, 3 with oral thrush), but in 0/28 controls, Candida (13 C. albicans, 1 C. lusitaniae, 1 C. krusei, 1 C. parapsilosis) was isolated. Candida specific T-cells were detected more frequently in controls (20/28) than in HIV-1+ subjects (16/46, p= 0.03). We observed a significant association of higher CD4 cell numbers with both detection of Candida specific T-cells and lack of oral Candida colonization, but there was no significant correlation of oral Candida colonization to the detection of Candida specific T-cells, viral load or antiretroviral therapy. Thus, local mucosal immunity seems to be more important in the pathogenesis of Candida colonization than circulating Candida specific T-cells. The pathogenic C. albicans strain K24122 was less frequently recognized by patients (6/46) than the laboratory adapted strain SC5314 (14/46, p= 0.03), whereas a similar recognition of both strains was observed in healthy controls. This indicates an impaired Candida-specific T-cell repertoire in HIV+ patients that could increase the risk of immune evasion by C. albicans.     

树突状细胞与白念珠菌感染的研究进展

白念珠菌是一种机会性致病菌,宿主对白念珠菌的识别和免疫应答是其免疫系统与白念珠菌相互作用的结果。树突状细胞在抗白念珠菌感染免疫中起重要作用,可以通过产生不同的细胞因子诱导CD4+T细胞向不同类型的Th细胞分化,从而产生不同的免疫应答,最终影响白念珠菌感染的结局。本文对白色念珠菌的易感性、免疫应答机制及树突状细胞与白念珠菌的联系进行了综述,为进一步研究白念珠菌等感染性疾病的发病机制和治疗提供了新思路。     

两种方法检测白色念珠菌对氟康唑的体外药敏结果比较及耐药原因分析

目的 比较纸片扩散(K-B)法和微量肉汤稀释(MIC)法检测18株临床分离白色念珠菌和4株标准白色念珠菌对氟康唑体外药敏试验的检测结果,并分析白色念珠菌对氟康唑的耐药原因。方法 根据K-B法和MIC法检测白色念珠菌对氟康唑体外药敏试验,比较两种方法结果的一致性;通过甲基四氮盐(XTT)减低法测量白色念珠菌生物膜的形成及其与氟康唑耐药的关系。结果 K-B法和MIC法检测白色念珠菌对氟康唑的体外药敏实验结果,经比较二者无明显差异,对氟康唑耐药的白色念珠菌其OD值远远高于对氟康唑敏感的白色念珠菌的OD值。结论 K-B法和MIC法对于检测白色念珠菌对氟康唑的体外药敏试验结果具有较高的一致性;白色念珠菌对氟康唑的耐药与其形成生物膜有重要相关性。     

小鼠念珠菌感染模型和遗传因素研究

白色念珠菌是能感染人类皮肤和粘膜的真菌.当宿主的免疫系统受到抑制时,可导致念珠菌病.建立动物实验模型对研究不同类型的人类念珠菌病非常必要,如系统性和皮肤粘膜的念珠菌感染模型.目前,最为常见的是系统性念珠菌动物模型.而能准确反映人类皮肤粘膜念珠菌感染的动物模型仍未稳定建立.值得庆幸的是,特异性基因剔除小鼠(Specific-gene knock out mice)的出现为研究念珠菌与宿主的免疫应答提供了更为直接和准确的模型.     

恒河猴Mamu-A*01基因与SIV/SHIV感染相关的研究进展

SIV/SHIV感染的恒河猴是研究艾滋病及艾滋病药物筛选、疫苗评价较理想的动物模型.MHC在细胞免疫中起着关键作用,研究表明,MHC-I类分子的多态性与SIV/SHIV感染者的疾病进展有着明显的关联作用,Mamu-A*01是恒河猴中的一种MHC-I类分子,它可以呈递特定的病毒蛋白片段到细胞的表面,从而激发CTL反应.国外发现Mamu-A*01阳性的猴艾滋病恒河猴会出现疾病进展缓慢,存活时间长等特征.本文就恒河猴Mamu-A*01基因与SIV/SHIV感染相关的研究进展做一综述,以期进一步加深对MHC在疫苗研究中的作用的了解,并促进更行之有效地对HIV/AIDS疫苗进行评价.     

前列腺素E_1对家免细胞免疫反应的影响

<正> 已经证实,前列腺素E_1(PGE_1)对免疫反应具有调节作用,尤其在体液免疫方面已有若干报道,但对细胞免疫反应的影响如何,研究较少。为此,我们在过去有关的两项实验基础上,重点观察了体内注射PGE_1对家兔细胞免疫反应的影响,实验过程中还对家兔淋巴细胞转化试验(简称淋转)方法作了一些改进,一并介绍如下。     

念珠菌变应性鼻炎的sIgE检测

目的：对念珠菌变应性鼻炎患者进行血清sIgE检测。方法：用RAST ELA法(Rabioallergosorbent test)以及CAP System对60例念珠菌皮试阳性的变应性鼻炎患者血清中sIgE作了检测。结果：RAST ELA法和CAP System sIgE阳性率分别为56．7％和61．7％,二者高度相关。结论：念珠菌皮试阳性患者中约40％并非真正的念珠菌过敏者。     

口服伊曲康唑在人角质层抗念珠菌活性的研究

目的通过测定伊曲康唑(口服给药)在人角质层的抗念珠菌活性,探索一种与体内环境相似的药敏试验新方法。方法将21例健康志愿者用数字表法随机分为2组,分别口服伊曲康唑200 mg,1次/d(Qd)×7 d和200 mg,2次/d(B id)×7 d。于服药前、服药后1、4、7、10、14、21、28、35 d制备皮肤角质层剥离条,将白念珠菌、光滑念珠菌、近平滑念珠菌和热带念珠菌孢子接种其上,培养48 h,过碘酸-雪夫(PAS)染色,采用角质层真菌计量生物学分析法,并用计算机辅助图像分析测定真菌覆盖的面积。结果2种剂量的伊曲康唑在皮肤角质层对4种念珠菌均有明显的抗真菌活性。2种剂量的伊曲康唑对白念珠菌、近平滑念珠菌和热带念珠菌的抗真菌效果的差异无统计学意义(P>0.05);而对于光滑念珠菌,伊曲康唑200 mg B id×7 d在人角质层的抗真菌活性明显优于200 mg Qd×7 d(P<0.01)。结论角质层真菌计量生物学分析法是介于体外试验与体内疗效之间的一种新方法的有效尝试。     

泛力克油剂抗白色念珠菌的体内外研究

为探讨泛力克油剂对白色念珠菌的作用，在体外将泛力克油剂用琼脂板孔中滴加法对白色念珠菌行抑菌试验，在体内以泛力克油剂局部外用治疗念珠性阴道炎２８例。结果表明：泛力克油剂无论在体内或体外都是一种高效的抗真菌药，其抗真菌效果明显优于达克宁霜、克霉唑软膏和无极膏（Ｐ＜００１）。     

耳念珠菌不同条件下生长特性及体内毒性

目的 探讨耳念珠菌在不同条件下的生长特性、代谢特点及体内毒性特征。方法 以白色念珠菌为参照,测定并比较耳念珠菌与白色念珠菌在常规培养条件、不同pH值和不同盐含量下的生长情况;利用微量生化反应管测定两种念珠菌糖醇发酵情况;以四周龄裸鼠为实验动物,用随机数字表法分为3组,耳念珠菌感染组、白色念珠菌感染组和生理盐水对照组,5只/组。耳念珠菌感染组和白色念珠菌感染组取0.3 mL已调至5×109 cfu/mL菌液,对裸鼠进行灌胃处理,建立胃肠道感染模型,对照组用等量生理盐水灌胃,取肝脏、肾脏、肠、粪便和血液等标本,通过真菌培养和组织病理学变化分析耳念珠菌的体内分布和毒性特征。结果 耳念珠菌的对数生长期约为8~24 h,约在24 h后进入生长稳定期,最适生长的pH值范围为5~7,生长模式与白色念珠菌相同;耳念珠菌在浓度为5%和10%的NaCl培养基中生长情况优于白色念珠菌,能发酵葡萄糖、蔗糖、海藻糖和山梨醇;从裸鼠的粪便、血液、肝脏和肾脏组织中分离出耳念珠菌,肝脏载菌量最多为5.7 log10 cfu/g,其可造成肝组织和肠组织病理学改变,但病变程度比白色念珠菌弱。结论 耳念珠菌在弱酸或中性条件下最适宜生长,具有高耐盐性,在免疫抑制状态下感染机体后,存在突破肠屏障入血及随血流播散并导致组织病变的可能。     

鸡卵黄乙型肝炎特异性转移因子口服液的制备及其免疫活性鉴定

目的制备鸡卵黄乙肝病毒特异性转移因子（EYHBV-STF）口服液,并鉴定其活性,探索一种新型、有效治疗乙肝的免疫调
节剂。方法采用乙肝疫苗免疫母鸡,从卵黄中提取乙型肝炎病毒特异转移因子（HBV-STF）,进行理化特性等检测。采用白细
胞粘附抑制试验检测转移因子的免疫活性,将其制成口服液,经小鼠灌胃后,迟发型皮肤超敏反应试验鉴定其体内特异性细胞
免疫活性,免疫脏器指数鉴定其非特异性免疫活性。同时分别与经注射或口服途径的EYHBV-STF组、传统方法制备的猪脾乙
肝特异性转移因子组、猪脾非特异性转移因子组及未经免疫的鸡卵黄提取液组进行比较。结果EYHBV-STF口服液各项理化
特性及安全实验指标符合TF生物制品标准,体外实验能明显抑制正常小鼠白细胞黏附,体内实验能显著增强小鼠足垫肿胀度
反应,提示该制品可将供体的乙肝抗原特异性皮肤迟发型超敏反应转移给受体,并能够显著提高免疫脏器指数（P<0.01）,与注
射EYHBV-STF、传统方法制备的注射、口服PHBV-STF 对照组比较,各项实验指标结果相近（P>0.05）。结论口服
EYHBV-STF与注射途径同样具有依赖乙肝抗原的特异性细胞免疫活性,可明显增强正常小鼠特异性细胞免疫功能。
     

化疗药物对白念珠菌粘附及播散的影响

吕药等对播散性念珠菌病的影响。方法采用体外实验，观察几种抗癌药和抗生素对白念 菌粘附上皮细胞的影响；同时对经环磷酰胺处理的小鼠静脉接种白念 珠菌，观察抗癌药物对血管内皮细胞的作用及其与白念 珠菌播散的关系。     

Pregnancy estrogen drives the changes of T-lymphocyte subsets and cytokines and prolongs the survival of H-Y skin graft in murine model

Background Estrogen as well as CD4＋Foxp3＋ regulatory T cells were shown to have a protective role not only in maintaining maternal-fetal tolerance but also against autoimmune diseases. We aimed to investigate whether the pregnancy levels of estrogen are enough to induce transplant tolerance as to maintain fetal-maternal tolerance. Methods We established H-Y skin graft transplantation in C57BL/6 ovariectomized mice that reconstituted with estrogen. Subsequently, consecutive daily estrogen injection was administrated. Tregs and the cytokines in the peripheral blood were detected by flow cytometry and ELISA pre- and post-transplant. Results The results indicated that pregnancy levels of estrogen could promote Tregs in secondary lymphoid organs and peripheral blood （P 〈0.05） but not thymus （P 〉0.05）. The estrogen-treated recipients accepted H-Y skin grafts for more than 35 days （median survival time （MST）： （44.0_＋1.2） days） compared with estrogen-untreated mice （MST： （23.0_＋1.6） days） （P 〈0.05）. It was also observed that estrogen up-regulated the expression of Foxp3, but did not affect CD3＋CD8＋ effector T-cells in non-transplant mice. While in the presence of H-Y antigens, the expression of Foxp3 was more significant and CD3＋CD8＋ effector T cells were decreased significantly （P 〈0.05）. Meanwhile, the up-regulated IL-10 and IL-4, and down-regulated IFN-v could be observed （P 〈0.05）. Conclusions Pregnancy levels of estrogen may promote the conversion of peripheral Tregs in secondary lymphoid organs, but show no effect on the natural Tregs production, differentiation and maturity in central lymphoid organs. Furthermore, pregnancy levels of estrogen could significantly prolong the survivals of H-Y skin grafts by the expansion of TreQs, suppression of CD3＋CD8＋ effector T-cells and immune shift towards Th2 cvtokines.