生殖道衣原体、支原体感染与盆腔炎症的相关性分析

目的:探讨女性生殖道衣原体、支原体感染发生与盆腔炎症的相关性,并为相应人群提出相应的预防和治疗措施。方法:对我院2012年1月到2013年5月诊治的盆腔炎患者280例及60名健康妇女进行了衣原体、支原体的培养,采用试剂盒进行检查,并进行药敏实验,分析比较两组生殖道衣原体和支原体感染情况。结果:盆腔炎症组中衣原体感染的检出率为58.5%,支原体感染率为26.2%,衣原体合并支原体感染率为12.4%。健康妇女组衣原体感染、支原体感染及衣原体合并衣原体感染的检出率分别为:9.1%,6.2%和4.9%。两组的差异有统计学意义(P0.05)。在盆腔炎症患者组中,30岁人群中单纯衣原体感染和单纯支原体感染检出率分别为51.3%和26.4%,均要明显高于30岁以上的人群,差异有统计学意义(P0.05)。结论:盆腔炎症与生殖道衣原体,支原体感染有密切的相关性,盆腔炎的发病可能与生殖道衣原体、支原体感染有关,针对临床上盆腔炎患者应密切关注生殖道支原体、衣原体的感染问题。     

GM-CSF transgene-based adjuvant allows the establishment of protective mucosal immunity following vaccination with inactivated Chlamydia trachomatis

Cellular and humoral immune responses induced following murine Chlamydia trachomatis infection confer almost sterile protection against homologous reinfection. On the other hand, immunization with inactivated organism induces little protective immunity in this model system. The underlying mechanism(s) that determines such divergent outcome remains unclear, but elucidating the mechanism will probably be important for chlamydial vaccine development. One of the distinct differences between the two forms of immunization is that chlamydia replication in epithelial cells causes the secretion of a variety of proinflammatory cytokines and chemokines, such as GM-CSF, that may mobilize and mature dendritic cells and thereby enhance the induction of protective immunity. Using a murine model of C. trachomatis mouse pneumonitis lung infection and intrapulmonary adenoviral GM-CSF transfection, we demonstrate that the expression of GM-CSF in the airway compartment significantly enhanced systemic Th1 cellular and local IgA immune responses following immunization with inactivated organisms. Importantly, immunized mice had significantly reduced growth of chlamydia and exhibited less severe pulmonary inflammation following challenge infection. The site of GM-CSF transfection proved important, since mice immunized with inactivated organisms after GM-CSF gene transfer by the i.p. route exhibited little protection against pulmonary challenge, although i.p. immunization generated significant levels of systemic Th1 immune responses. The obvious difference between i.p. and intrapulmonary immunization was the absence of lung IgA responses following i.p. vaccination. In aggregate, the findings demonstrate that the local cytokine environment is critical to the induction of protective immunity following chlamydial vaccination and that GM-CSF may be a useful adjuvant for a chlamydial vaccine.     

血清E型沙眼衣原体的Real-time PCR定量分析

目的沙眼衣原体感染是最常见的性传播疾病,本文拟建立一种准确快速、标准化的感染动物组织衣原体载量检测体系。方法体外扩增感染用沙眼衣原体血清E型,克隆衣原体特异基因OMP1基因片段作为标准品,用Real time PCR法测定衣原体基因组拷贝数进行衣原体定量。结果 Real time PCR在OMP1基因片段200至2×108拷贝检测结果成线性,在模板中加入小鼠基因组未出现非特异扩增,同时未影响扩增效率。结论针对衣原体特异基因OMP1的实时定量PCR方法可以较为灵敏的特异的定量检测感染动物样本中的衣原体。     

Control of Trachoma from Achham District,Nepal: A Cross-Sectional Study from the Nepal National Trachoma Program

Background

The WHO seeks to control trachoma as a public health problem in endemic areas. Achham District in western Nepal was found to have TF (trachoma follicular) above 20% in a 2006 government survey, triggering 3 annual mass drug administrations finishing in 2010. Here we assess the level of control that has been achieved using surveillance for clinical disease, ocular chlamydia trachomatis infection, and serology for antibodies against chlamydia trachomatis protein antigens.

Methods

We conducted a cross-sectional survey of children aged 1–9 years in communities in Achham District in early 2014 including clinical examination validated with photographs, conjunctival samples for Chlamydia trachomatis (Amplicor PCR), and serological testing for antibodies against chlamydia trachomatis protein antigens pgp3 and CT694 using the Luminex platform.

Findings

In 24 randomly selected communities, the prevalence of trachoma (TF and/or TI) in 1–9 year olds was 3/1124 (0.3%, 95% CI 0.1 to 0.8%), and the prevalence of ocular chlamydia trachomatis infection was 0/1124 (0%, 95% CI 0 to 0.3%). In 18 communities selected because they had the highest prevalence of trachoma in a previous survey, the prevalence of TF and/or TI was 7/716 (1.0%, 95% CI 0.4 to 2.0%) and the prevalence of ocular chlamydia trachomatis infection was 0/716 (0%, 95% CI 0 to 0.5%). In 3 communities selected for serological testing, the prevalence of trachoma was 0/68 (0%, 95% CI 0 to 5.3%), the prevalence of ocular chlamydia trachomatis infection was 0/68 (0%, 95% CI 0 to 0.5%), the prevalence of antibodies against chlamydia trachomatis protein antigen pgp3 was 1/68 (1.5%, 95% CI 0.04% to 7.9%), and the prevalence of antibodies against chlamydia trachomatis protein antigen CT694 was 0/68 (0%, 95% CI 0 to 5.3%).

Conclusion/Significance

This previously highly endemic district in Nepal has little evidence of recent clinical disease, chlamydia trachomatis infection, or serological evidence of trachoma, suggesting that epidemiological control has been achieved.     

Specific prophylaxis of influenza with inactivated split vaccine Vaxigrip

Influenza remains a serious health problem, annually causing epidemics embracing up to 10% of the population of the world, and at the periods of pandemics this number may rise 4- to 6-fold. In the overwhelming majority of the countries the prophylaxis of influenza is carried out at present out with the use of inactivated vaccines. One of such vaccines is the highly purified split vaccine Vaxigrip (Aventis-Pasteur, France), permitted for use in Russia since 1992. The article contains the review of the data of literature, as well as the materials provided by the authors, which indicate that the preparation has low reactogenicity (also for children starting from the age of 6 months) and high reactogenic properties, leading to antibody formation at protective levels with respect to all three components of the vaccine. The vaccine has been found to ensure pronounced prophylactic efficacy for 70-90% of vaccinees and a decrease in influenza morbidity even in case of using the preparation a week before the onset of the epidemic. This shows the advantage of Vaxigrip in comparison with other inactivated vaccines.     

抵抗女性生殖道沙眼衣原体感染的免疫新策略

沙眼衣原体是引起沙眼和泌尿生殖道感染的主要病原体。据世界卫生组织2015年统计,全球每年约有1.3亿沙眼衣原体感染新发病例。研究表明CD4^+Th1型细胞免疫应答在抵抗沙眼衣原体感染中发挥着重要作用。因此,研究者依照抗沙眼衣原体感染的免疫应答特点,构建出许多候选疫苗,但都没有成功地应用于临床。近年研究发现,生殖道黏膜组织不仅存在体液免疫和细胞免疫,还驻留着一些引人注目的免疫细胞,提示增强黏膜免疫可作为预防沙眼衣原体感染的潜在途径,是抵抗生殖道沙眼衣原体感染的免疫新策略。本文全面概述了黏膜免疫与女性生殖道沙眼衣原体感染的研究进展,并为今后研制沙眼衣原体疫苗提供一些建议。     

阿奇霉素对沙眼衣原体感染后生殖道黏膜免疫反应的影响

目的 研究阿奇霉素治疗生殖道沙眼衣原体感染过程中对局部黏膜免疫反应的影响,为其临床应用提供新的实验依据.方法 构建小鼠生殖道沙眼衣原体感染模型,随机分为生理盐水组和阿奇霉素组.阿奇霉素组一次性给予阿奇霉素（80mg/kg）,生理盐水组给予等量生理盐水.给药当天、给药第7天、给药第14天和给药第21天,阴道拭子取宫颈脱落细胞,分离沙眼衣原体.给药21天,处死动物.收集血清,ELISA测定血清IL-6和TNF-α水平,同时进行阴道、宫颈黏膜常规HE染色和肥大细胞甲苯胺蓝染色、树突状细胞免疫组织化学分析.结果 1.阿奇霉素组沙眼衣原体感染率明显低于生理盐水组,且未出现上行感染（P〈0.05）.2.阿奇霉素组生殖道黏膜内树突状细胞数量增加（P〈0.05）,肥大细胞数量无明显变化（P〈0.05）.3.阿奇霉素组血清内IL-6和TNF-α的水平,均高于对照组和生理盐水组（P〈0.05）.结论 阿奇霉素除了有效清除生殖道沙眼衣原体感染,亦可以调节生殖道黏膜的免疫反应,减轻免疫病理损伤,使沙眼衣原体感染有较好的预后.     

Comparative study of the reactogenicity and immunogenicity of new Soviet and commercial vaccines for preventing Japanese encephalitis

In this article the results of testing Soviet vaccines for the prophylaxis of Japanese encephalitis, cell-culture inactivated adsorbed liquid vaccine and brain-tissue purified dried vaccine, are generalized and the comparison of their reactogenicity and immunogenicity characteristics with those of a similar commercial preparation manufactured by Toshiba Kagaku Kogyo Co., Ltd. (Japan) is made. Cell-culture inactivated liquid vaccine has proved to be the optimal preparation: it produces the most prolonged and intensive immunity and, when introduced by two methods, shows low reactogenicity.     

Chlamydia trachomatis secretion of an immunodominant hypothetical protein (CT795) into host cell cytoplasm

The Chlamydia-specific hypothetical protein CT795 was dominantly recognized by human antisera produced during C. trachomatis infection but not by animal antisera raised against dead chlamydia organisms. The immundominant region recognized by the human antibodies was mapped to the N-terminal fragment T22-S69. The endogenous CT795 was detected in the cytoplasm of host cells during C. trachomatis infection and was highly enriched in the host cytosolic fraction but absent in the purified chlamydia organisms, suggesting that CT795 is synthesized and secreted into host cell cytoplasm without incorporation into the organisms. All C. trachomatis serovars tested secreted CT795. A predicted signal peptide of CT795 directed the mature PhoA to cross Escherichia coli inner membranes. The secretion of CT795 in Chlamydia-infected cells was inhibited by a C(16) compound targeting signal peptidase I, but not by a C(1) compound known to block the type III secretion pathway. These results suggest that CT795, like CPAF (a Chlamydia-secreted virulence factor), is secreted into the host cell cytoplasm via a sec-dependent mechanism and not by a type III secretion pathway. The above characterizations of CT795 have provided important information for further understanding the potential roles of CT795 in C. trachomatis pathogenesis.     

沙眼衣原体GlgA蛋白表达和分泌的调控机制

【背景】沙眼衣原体(Chlamydia trachomatis,Ct)的分泌蛋白在Ct与宿主细胞的相互作用、感染发育周期及致病过程中发挥着至关重要的作用。GlgA蛋白是课题组前期研究发现的一种新的Ct分泌蛋白,其表达和分泌的具体机制及作用还不清楚。【目的】寻找调控CtGlgA蛋白表达和分泌的分子机制,为后续Ct致病机制研究提供实验基础和新思路。【方法】采用Signal P 4.1软件对GlgA蛋白N端进行信号肽预测分析,并用细菌分泌蛋白特异性阻断剂C16和C1化合物分别或同时处理Ct感染的He La细胞,观察阻断Ⅱ型、Ⅲ型分泌途径对GlgA蛋白分泌的影响;经新生霉素处理、噬斑筛选及穿梭质粒转染技术,构建Ct质粒缺失株和缺失互补株,并鉴定质粒编码基因在两种菌株的缺失及表达情况;间接免疫荧光法观察质粒缺失对GlgA表达和分泌的影响。【结果】GlgA蛋白N端无信号肽序列,细菌Ⅱ型、Ⅲ型分泌途径特异性阻断剂C16和C1化合物不能阻断GlgA的胞浆分泌;Ct质粒缺失株CTD1的质粒编码基因pgp7丢失,且质粒编码蛋白Pgp3及基因组编码蛋白GlgA的表达和分泌现象均消失;Ct缺失互补株CTD1-pGFP::SW2重新获得pgp7基因,并恢复Pgp3蛋白和GlgA的表达和分泌。【结论】初步证实Ct糖原合酶GlgA蛋白的表达和分泌不依赖细菌Ⅱ型和Ⅲ型分泌途径,而且与衣原体质粒密切相关。     

Modern features of the epidemiology and prophylaxis of influenza

Modern concepts concerning influenza pandemics and epidemics in different countries of the world are presented. The influenza epidemics of the last decade in different countries of the world and their specific features linked with the "drifting" variability of influenza virus have been analyzed. Information on influenza morbidity during the last 30 years is given; on the basis of this information the role of vaccinal prophylaxis and mainly the mass vaccination of school children and students, is shown. The results of the efficacy of such vaccines as live influenza vaccine, American split vaccine, Russian live recombinant vaccine and Grippovac (1995-1996), as well as new-generation vaccine Grippol (1998), are presented. The prospects of the combined use of specific and unspecific prophylaxis have been determined.     

Patterns of immunoenhancement and suppression induced by Chlamydia trachomatis in vivo and in vitro

We investigated the cycle of immune enhancement and suppression seen in mice infected with Chlamydia trachomatis by using in vivo and in vitro model systems. BALB/c mice injected intravenously with chlamydia reveal a three- to seven-fold increase in numbers of plaque-forming cells producing antibodies against sheep red blood cells (SRBC), when immunized with SRBC 0 to 5 days after chlamydia infection. When mice are injected with SRBC 10 to 15 days after initial chlamydia infection, the specific anti-SRBC plaque-forming cell response is suppressed two- to three-fold. In vitro, low numbers (2 to 5 X 10(6) bacteria/ml) of chlamydia stimulate potent proliferative responses by B lymphocytes while high numbers (25 X 10(6) bacteria/ml) of bacteria generate strong, general T suppressor activity. This model has important implications for regulation of immune responses that arise at different times during chlamydial infections, as well as for the potential effectiveness of chlamydial vaccines.     

Chlamydia trachomatis as a cause of abortion

The possible role of C. trachomatis as a causative agent of abortion in humans was investigated using the tissue culture method for the isolation of chlamydia. Two cases of spontaneous abortion and another seven of recurrent abortion due to C. trachomatis were positively identified. Among those with recurrent abortion were four patients with history of second trimester abortion.     

Both species of chlamydia and two biovars of Chlamydia trachomatis stimulate mouse B lymphocytes

We have investigated the ability of both species of chlamydiae (C. trachomatis and C. psittaci), two major biovars of C. trachomatis (lymphogranuloma venereum and trachoma), and the two developmental forms of chlamydia (reticulate and elementary bodies) to stimulate murine spleen lymphocytes. All of these forms of the bacteria induce potent proliferation and differentiation to plaque-forming cells by B lymphocytes in vitro. Chlamydiae induce a broad antibody response, suggesting that stimulation is polyclonal in nature. Although all chlamydiae possess a lipopolysaccharide (LPS) genus-specific molecule similar to LPS found on Re mutant enterobacteria, polyclonal B cell stimulation is likely caused by molecules other than LPS, since i) polymyxin B failed to inhibit chlamydia-induced immunostimulation and ii) C3H/HeJ mice (LPS nonresponders) produced normal numbers of PFC after culture with chlamydia (but not LPS). Thus, a cross-species moiety that is not LPS is responsible for polyclonal stimulation by chlamydia. Because these bacteria can exist in latent forms in an animal, and all forms are immunostimulatory, the question of whether these bacteria can alter immune responses if released during other infections or immunizations has been raised.     

Localization of TLR2 and MyD88 to Chlamydia trachomatis inclusions. Evidence for signaling by intracellular TLR2 during infection with an obligate intracellular pathogen

Chlamydia trachomatis is an obligate intracellular gram-negative pathogen and the etiologic agent of significant ocular and genital tract diseases. Chlamydiae primarily infect epithelial cells, and the inflammatory response of these cells to the infection directs both the innate and adaptive immune response. This study focused on determining the cellular immune receptors involved in the early events following infection with the L2 serovar of C. trachomatis.We found that dominant negative MyD88 inhibited interleukin-8 (IL-8) secretion during a productive infection with chlamydia. Furthermore, expression of Toll-like receptor (TLR)-2 was required for IL-8 secretion from infected cells, whereas the effect of TLR4/MD-2 expression was minimal. Cell activation was dependent on infection with live, replicating bacteria, because infection with UV-irradiated bacteria and treatment of infected cells with chloramphenicol, but not ampicillin, abrogated the induction of IL-8 secretion. Finally, we show that both TLR2 and MyD88 co-localize with the intracellular chlamydial inclusion, suggesting that TLR2 is actively engaged in signaling from this intracellular location. These data support the role of TLR2 in the host response to infection with C. trachomatis. Our data further demonstrate that TLR2 and the adaptor MyD88 are specifically recruited to the bacterial or inclusion membrane during a productive infection with chlamydia and provide the first evidence that intracellular TLR2 is responsible for signal transduction during infection with an intracellular bacterium.     

Chlamydia trachomatis Mip-like protein

A 27 kDa Chlamydia trachomatis Mip-like protein with homology of a 175-amino-acid C-terminal fragment to the surface-exposed Legionella pneumophila mip-gene product has previously been described. In this paper the entire chlamydia Mip-like sequence of C. trachomatis serovar L2 (lymphogranuloma venereum (LGV) biovar) is presented. The sequence shows high similarity to the legionella Mip protein and its C-terminal region, like that of the legionella Mip, has high amino acid similarity to eukaryotic and prokaryotic FK506-binding proteins. The chlamydial mip-like gene was detected by polymerase chain reaction (PCR) in other C. trachomatis serovars and by sequencing of the mip-like genes of serovars B and E (trachoma biovar) was shown to be highly conserved within the two major biovars of C. trachomatis. Monoclonal and polyclonal antibodies raised against the recombinant Mip-like protein failed to demonstrate surface-exposed epitopes on infectious elementary bodies or reproductive reticulate body forms either by immunofluorescence or immuno-gold electron microscopy. However, a complement-dependent inhibition of up to 91% of infectivity for cell cultures was observed with antibodies to the N-terminal fragment of the Mip-like protein suggesting that antibody-accessible epitopes are present on infectious EBs.     

Controlling schistosomiasis by vaccination: A realistic option?

Schistosomiasis, a snail-transmitted trematode infection affecting some 200 million people in 74 countries, continues to defy control efforts in vast stretches of endemic areas. Although tools are available, the logistics and cost of their proper application prevent the implementation of sustained control in most parts of the world. Chemotherapy remains the cornerstone of intervention, but rapid reinfection demands frequent retreatment. The development of at least partial immunity would make it logical to combine drug therapy with vaccination, but vaccination is not yet a real option. However, the advance in immunology and molecular biology of the past few years has substantially increased the odds for effective immune prophylaxis. A range of promising candidate vaccine anti-gens is currently undergoing independent confirmatory testing with scaled-up production and human trials as the logical next step. In this communication, Robert Bergquist deals with the international effort, co-ordinated by WHO/TDR, to generate a schistosomiasis vaccine.     

冰片提取物对沙眼衣原体感染HeLa细胞CT703及CT259表达的影响

目的:探究冰片提取物对沙眼衣原体感染后的He La细胞模型CT703和CT259表达的影响。方法:将成功建立的40例感染L2血清型沙眼衣原体的人宫颈癌上皮(He La)细胞模型随机分成A、B两组,分别添加冰片提取物和等剂量生理盐水。观察感染后He La细胞模型的包涵体数目及大小、RNA抽提结果完整性以及CT70与CT259的表达变化。结果:染色后的40例受沙眼衣原体感染的He La细胞模型体内均发现包涵体,在同一时间点B组细胞内包涵体比A组大,且数目比A组多(P0.05);两组提取的总RNA的OD值均在1.8~2.0之间,通过RNA凝胶电泳结果可清楚发现28S、18S及5S条带;同一时间点CT259和CT703扩增产物的平均灰度值比较,A组感染后He La细胞模型样本基因表达量低于B组,差异具有统计学意义(P0.05)。结论:冰片提取物能够有效降低经沙眼衣原体感染的He La细胞模型中CT703与CT259基因表达量。     

Isolation and initial characterization of a series of Chlamydia trachomatis isolates selected for hydroxyurea resistance by a stepwise procedure.

Chlamydiae are obligate intracellular bacteria that are dependent on eukaryotic host cells for ribonucleoside triphosphates but not deoxyribonucleotide triphosphates. Ribonucleotide reductase is the only enzyme known to catalyze the direct conversion of a ribonucleotide to a deoxyribonucleotide. Hydroxyurea inhibits ribonucleotide reductase by inactivating the tyrosine free radical present in the small subunit of the enzyme. In this report, we show that Chlamydia trachomatis growth is inhibited by hydroxyurea in both wild-type mouse L cells and hydroxyurea-resistant mouse L cells. Hydroxyurea was used as a selective agent in culture to isolate, by a stepwise procedure, a series of C. trachomatis isolates with increasing levels of resistance to the cytotoxic effects of the drug. One of the drug-resistant C. trachomatis isolates (L2HR-10.0) was studied in more detail. L2HR-10.0 retained its drug resistance phenotype even after passage in the absence of hydroxyurea for 10 growth cycles. In addition, L2HR-10.0 was cross resistant to guanazole, another inhibitor of ribonucleotide reductase. Results obtained from hydroxyurea inhibition studies using various host cell-parasite combinations indicated that inhibition of host cell and C. trachomatis DNA synthesis by hydroxyurea can occur but need not occur simultaneously. Crude extract prepared from highly purified C. trachomatis reticulate bodies was capable of reducing CDP to dCDP. The CDP reductase activity was not inhibited by monoclonal antibodies to the large and small subunits of mammalian ribonucleotide reductase, suggesting that the activity is chlamydia specific. The CDP reductase activity was inhibited by hydroxyurea. Crude extract prepared from drug-resistant L2HR-10.0 reticulate bodies contained an elevation in ribonucleotide reductase activity. In total, our results indicate that C. trachomatis obtains the precursors for DNA synthesis as ribonucleotides with subsequent conversion to deoxyribonucleotides catalyzed by a chlamydia-specific ribonucleotide reductase.     

Serodiagnosis of chlamydiosis: "Chlamy-IgG-DS-Tr"-- the first domestic immunoenzyme recombinant test-system for determination of anti-Chlamydia trachomatis class G antibodies]

The first Native test-system for determination of class G antibodies to Chlamydia trachomatis with chlamydia recombinant antigen was elaborated Test-system efficacy was demonstrated in clinical trials. The sensibility, specific activity and suitability of the "Chlamy-IgG-DS-Tr" were the same as for import analogous.