被忽视的青少年和成人百日咳

百日咳长期以来被认为只发生于儿童。近30年来国外对百日咳再现开展的流行病学调查发现,与疫苗前时代不同,青少年和成人已成为百日咳的主要发病人群。目前,国内百日咳报告病例以婴儿为主,这与临床对青少年和成人百日咳认识不足有关。疫苗时代多数青少年和成人百日咳多表现为轻症,常未就诊;即使就诊,因缺乏典型症状和实验室检查等原因常被漏诊、误诊。因此,临床应高度关注青少年和成人百日咳问题,积极开展广泛深入的研究,认清百日咳表现的多样性,对可疑患者及时进行实验室检测以实现早诊断。只有加强青少年和成人百日咳病例管理,才能有效预防婴儿百日咳重症和死亡的发生。     

双抗夹心ELISA法在无细胞百日咳疫苗生产质控中的应用

实验中对无细胞百日咳疫苗的脱毒工序优化后,采用双抗夹心ELISA法来检测百日咳有效组分含量,同时采用效价试验方法来验证结果。ELISA法定量测定有效成分的结果和效价试验结果均证明达到《中国药典》三部2005版的要求。由于双抗夹心ELISA法特异性强,灵敏度高,适用于无细胞百日咳疫苗生产各个环节的质量控制。     

深圳市南山区健康人群百日咳抗体水平及疫苗免疫成功率监测

目的了解深圳市南山区健康人群百日咳免疫现状及疫苗免疫成功率。方法健康人群抗体水平监测按照<1岁、1岁~、2岁~、3岁~、4岁~、5岁~、10岁~、15岁~、≥20岁共9个年龄组,每个年龄组随机抽取50~60人共508人进行观察,采用ELISA检测其百日咳抗体水平,免疫成功率监测选定监测对象为50人,同一对象于基础免疫前和完成基础免疫后1个月进行抗体水平检测。结果健康人群百日咳抗体阳性率为44.3%,抗体几何平均浓度(GMC)为21.69 U/m L。免疫成功率的免疫前抗体检测结果为:阴性94.0%、临界4.0%、阳性2.0%,抗体GMC为3.39 U/m L;免疫后抗体检测结果:阴性38.0%、临界16.0%、阳性46.0%,抗体GMC为20.75 U/m L。结论深圳市南山区健康人群百日咳抗体阳性率和免疫成功率均不高,提示要提高百日咳疫苗的免疫成功率和接种率,必要时进行加强免疫。     

腺苷酸环化酶毒素在无细胞百日咳疫苗中的免疫保护效果评价

目的 利用小鼠百日咳感染模型评估在无细胞百日咳疫苗中加入腺苷酸环化酶毒素的C端结构域(RTX751),能否提升无细胞百日咳疫苗的免疫保护效果.方法 ①用腺苷酸环化酶毒素的C端结构域与百日咳毒素(pertussis toxin,PT)、丝状血凝素(filamentous hemagglutinin,FHA)、百日咳黏附素...     

腺病毒载体在疫苗研究中的应用

以病毒为载体的活疫苗为疾病预防和治疗研究提供了新手段。目前用于疫苗研究的病毒载体主要包括痘苗病毒载体、腺病毒载体、腺相关病毒载体、单纯疱疹病毒载体及逆转录病毒载体等。其中,重组腺病毒载体因其基因组大小适中,易于基因重组操作,繁殖滴度高,易于大量制备和保存,宿主范围广,转导效率高,安全性好,能刺激机体产生强烈的体液和细胞免疫反应等特点,而被广泛应用于重要感染性疾病及恶性肿瘤的疫苗研究。腺病毒载体在人免疫缺陷病毒(HIV)疫苗研究和临床试验中的成败更是备受关注。然而,与其他载体疫苗一样,机体对载体的免疫反应仍是阻碍腺病毒载体疫苗在临床中广泛应用的主要问题。那么,腺病毒载体解决这类问题的优势何在?我们简要综述腺病毒载体的特点及其在疫苗研究中的应用和存在的问题,为进一步优化和利用腺病毒载体在疫苗方面的研究提供参考。     

尿流动力学在儿童及青少年遗尿中的应用价值

目的:探讨和分析尿流动力学检查在儿童及青少年遗尿症中的临床价值.方法:对我院诊治的61例遗尿症儿童及青少年进行尿流动力学检查,根据检查结果,并结合其临床症状、体征及其它辅助检查,选择适当的个体化治疗方案.结果:其中有42例尿流动力学检查结果显示异常,占总例数的68.9％; 19例尿流动力学检查结果显示无异常发现,占总例数的31.1％.结论:膀胱敏感性增加、膀胱容量相对减小、膀胱逼尿肌无抑制性不稳定收缩、膀胱顺应性和尿道压减低、出现残余尿量是遗尿症儿童及青少年尿流动力学检查结果出现异常的常见原因,行尿动力学检查在一定程度上能为遗尿症儿童及青少年在诊疗方案的选择上提供临床指导和参考.     

国产层析填料在百日咳毒素和丝状血凝素纯化中的筛选及应用

目的 筛选适合的国产层析填料以满足对组分百日咳疫苗中百日咳毒素(pertussis toxin, PT)与丝状血凝素(filamentous hemagglutinin, FHA)的分离纯化。方法 筛选对PT与FHA结合能力较好且分离度高的国产层析填料,优化纯化工艺,通过3批纯化试验比较国产填料与进口填料的抗原回收率、目的抗原纯度以及对目的抗原的动态结合载量。结果 通过纯化试验筛选到国产填料SP resin-1与MMC resin-1并优化了纯化工艺。3批纯化试验显示,SP resin-1纯化百日咳抗原工艺稳定,FHA的纯度和回收率与Capto SP ImpRes差异无统计学意义(P>0.05);且与Capto SP ImpRes相比,SP resin-1对抗原的动态结合载量更高。3批PT精纯试验显示,国产填料MMC resin-1精纯PT的工艺稳定,PT的纯度和回收率均与Capto MMC差异无统计学意义(P>0.05),并且MMC resin-1对PT的载量高于Capto MMC。结论 试验筛选出纯化PT与FHA的国产填料SP resin-1与MMC resin-1,纯...     

核酸疫苗的特点、组成及在动物免疫中的应用

核酸疫苗是一种新型的基因工程疫苗,它将含有编码某种抗原蛋白基因序列的重组质粒作为疫苗直接导入机体细胞内,通过宿主细胞的转录系统合成抗原蛋白,诱导机体产生体液免疫和细胞免疫．从而使被接种机体获得相应的免疫保护而达到防病治病的目的。就核酸疫苗的特点、组成、免疫方式、目前存在的主要问题及在动物免疫中的应用情况进行综述。     

初免-加强免疫策略在疫苗研究中的应用

大多数传统疫苗的免疫保护机理主要是诱导能维持较长时间的体液免疫应答,而大量研究显示机体在对抗胞内感染微生物及一些病毒的感染时,细胞免疫应答是至关重要的。鉴于初始-加强(Prim e-Boost)免疫策略在诱导细胞免疫应答方面具备的优势,其显示了很好的应用前景,本文综述了Prim e-Boost免疫策略在疫苗研究中的应用并对其机制进行了初步的探讨。     

mRNA疫苗在疾病预防与治疗中的研究进展与展望

基于信使RNA(messenger RNA, mRNA)的核酸疫苗是近年来兴起的一种mRNA技术。mRNA疫苗比传统疫苗有许多优点,能够实现快速、经济、高效的生产。单个mRNA疫苗可以编码多种抗原,增强对特定病原体的免疫反应,提高疾病的治疗效率,以单一配方针对多种病原微生物或疾病。mRNA疫苗相关技术在新型冠状病毒肺炎疫情防控中被视作一种革命性的疫苗技术,以创纪录的速度完成研发并成功应用。由于mRNA自身稳定性差,新型递送系统的开发与应用至关重要。随着mRNA相关药理学的深入研究,mRNA疫苗的临床应用进入了一个崭新的阶段。近年来。mRNA疫苗在传染性疾病预防、肿瘤治疗等方面获得充分发展并取得了一定的研究成果,对其进行概述并进行一定程度的展望。     

Increase in intradermal vascular permeability caused by pertussis toxin from Bordetella pertussis

Rabbits that were injected intradermally with pertussis toxin (PT), produced from Bordetella pertussis, showed slight edema and erythema at the injection sites, but not hemorrhage nor necrosis. The edema lesions were stained blue by the intravenous injection of Pontamine Sky Blue 6B dye, suggesting that PT caused increased vascular permeability, similarly to the permeability factor (PF) of cholera toxin. The reaction of the PF of PT could be determined by measuring the diameter of the blue area. The diameter of the blue area bore a good linear relationship to the logarithm of the dose of PT. The activity of the PF was neutralized by anti-PT rabbit serum. Detoxification of PT with formalin did not increase the vascular permeability, but reverted pertussis toxoid showed a PF reaction in proportion to the reverted leukocytosis-promoting and histamine-sensitizing activities of PT. The supernate of a Bordetella pertussis culture also induced a PF reaction and the reaction could be made clear by heating the supernate at 56 C for 30 min, but the supernate of Bordetella bronchiseptica did not induce the reaction at all.     

Essential role of the consensus nucleotide-binding site of PtlH in secretion of pertussis toxin from Bordetella pertussis.

PtlH is a member of a specialized set of transport proteins that is essential for secretion of pertussis toxin (PT) from Bordetella pertussis. Previously, PtlH was shown to contain a consensus nucleotide-binding motif. Here, we demonstrate that introduction of plasmids containing mutant forms of ptlH, altered in the putative nucleotide-binding region, into a wild-type strain of B. pertussis resulted in inhibition of PT secretion. Thus, this region of PtlH appears to be essential for protein function. Moreover, the observed dominant negative phenotype suggests that PtlH either functions as a multimer or interacts with another component necessary for secretion of PT.     

Release of Outer Membrane Vesicles from Bordetella pertussis

The aim of the study reported here was to investigate the production of Bordetella pertussis outer membrane vesicles (OMVs). Numerous vesicles released from cells grown in Stainer-Scholte liquid medium were observed. The formation of similar vesicle-like structures could also be artificially induced by sonication of concentrated bacterial suspensions. Immunoblot analysis showed that OMVs contain adenylate cyclase-hemolysin (AC-Hly), among other polypeptides, as well as the lipopolysaccharide (LPS). Experiments carried out employing purified AC-Hly and OMVs isolated from B. pertussis AC-Hly⁻ showed that AC-Hly is an integral component of the vesicles. OMVs reported here contain several protective immunogens and might be considered a possible basic material for the development of acellular pertussis vaccines. Received: 22 October 1998 / Accepted: 17 December 1998     

Detection and subcellular localization of three Ptl proteins involved in the secretion of pertussis toxin from Bordetella pertussis.

The ptl locus of Bordetella pertussis contains eight open reading frames which are predicted to encode proteins (PtlA to PtlH) that are essential for secretion of pertussis toxin from the bacterium and which are members of a family of transport proteins found in other types of bacteria. We have detected PtlE, PtlF, and PtlG in immunoblots of extracts of B. pertussis by using antibodies raised to fusion proteins consisting of maltose-binding protein and the individual Ptl proteins. These proteins have apparent molecular weights similar to those predicted by DNA sequence analysis. Cell fractionation studies indicated that all three Ptl proteins are associated with the membranes of B. pertussis, suggesting that the Ptl proteins form a gate or channel which facilitates transport of pertussis toxin. Cell extracts of other Bordetella spp. were probed with antibodies to Ptl proteins for the presence of these transport proteins. Neither Bordetella parapertussis nor Bordetella bronchiseptica contained detectable levels of PtlE or PtlF. This lack of detectable Ptl protein may provide an explanation for previous observations which indicated that introduction of the genes encoding pertussis toxin subunits from B. pertussis into other Bordetella spp. results in production of the toxin but not secretion of the toxin.