沙眼衣原体感染小鼠生殖道模型的初步研究

目的建立沙眼衣原体（C￡）小鼠生殖道感染模型。方法分别用10。、10‘、10。数量IFU的CtE型株阴道内接种雌性BALB／c小鼠,观察外阴炎症反应,检测排菌情况,PCR检测分泌物及对组织进行病理观察。结果接种后第5d,实验组小鼠即出现感染迹象;在接种后的5～20d均可在小鼠生殖道分离得到具感染性的c≠,但其持续排菌时期与接种的IFU数量成正比;且小鼠生殖道排菌量（IFU）与接种的IFU数量亦成正比,以10。剂量组为高;小鼠生殖道分泌物PCR扩增后电泳,可见与CtMOMP分子量相当的产物;小鼠生殖道组织切片观察证实,c￡E型株感染小鼠生殖道可产生生殖道炎症。结论成功建立了小鼠生殖道E型c￡感染模型;10。IFU是建立模型的适宜感染量。     

沙眼衣原体初次和再次感染小鼠生殖道的初步研究

目的 观察小鼠初次和再次生殖道感染沙眼衣原体后对病原体的清除情况,抗体产生规律及病理表现。方法 将小鼠生物型沙眼衣原体C. muridarum 1×10⁴ IFU阴道接种于C57B6背景雌性小鼠,分别取初次和再次感染后阴道拭子做沙眼衣原体培养,计算IFU,监测小鼠清除病原体情况;同时取小鼠尾静脉血,监测初次和再次感染后抗体产生情况;处死小鼠,检测子宫输卵管病理改变。结果 初次感染,小鼠生殖道感染病原体多,病程长,再次感染病原体明显减少,病程明显缩短;小鼠于初次感染2周后检测到抗体,之后迅速升高并维持在较高水平,再次感染后抗体维持高水平无明显改变;小鼠子宫输卵有明显病理改变,表现为炎症、官腔扩张积水,狭窄等。结论 成功建立沙眼衣原体初次再次感染小鼠生殖道模型,再次感染抗体水平无明显升高,但能有效控制感染,清除病原体,对炎症反应无明显改善。     

鼠衣原体质粒缺失株生物学特性及免疫保护性研究

目的以鼠衣原体(Chlamydia muridarum,CM)质粒缺失株CMUT3和CM972为对象研究质粒缺失对CM生物学特性的影响,评价质粒缺失株的免疫保护作用。方法碘染色观察衣原体包涵体中糖原聚集情况,间接免疫荧光法(IFA)和Western blot方法检测糖原合酶(GlgA)的表达,离心辅助感染试验检测衣原体对HeLa细胞的感染力。C3H/HeJ小鼠阴道感染CM,感染后60d内每隔3或7d取生殖道分泌物,检测包涵体形成单位(IFU);感染60d后处死小鼠,分离生殖道组织,观察输卵管水肿程度。CBA/J小鼠阴道感染CMUT3,感染后60d用CM菌株进行再次感染,观察质粒缺失菌株抗CM感染的免疫保护效果。结果 CMUT3和CM972菌株碘染色呈阴性;质粒影响GlgA的表达,质粒缺失后GlgA表达水平下降;离心因素能显著提高新分离质粒缺失株CMUT3对HeLa的感染力;C3H/HeJ小鼠下生殖道感染质粒缺失株CMUT3和CM972后不发生输卵管积水,而质粒缺失株在小鼠下生殖道的增殖与野生株比较差异不明显;CBA/J小鼠下生殖道免疫CMUT3后再感染CM,其单侧输卵管积水发生率为15%(2/13),而先行感染CM后再次感染CM的小鼠单侧或双侧输卵管积水发生率89%(8/9)。结论 CM质粒缺失株在小鼠生殖道中的致病力减弱,小鼠下生殖道免疫CMUT3后可抑制CM再感染所致的病理变化,CM质粒缺失株具有潜在的疫苗研究价值。     

肽聚糖识别蛋白1抗鼠衣原体生殖道感染的初步研究北大核心CSCD

目的探讨肽聚糖识别蛋白1(peptidoglycan recognition protein 1,PGLYRP-1)抗鼠衣原体(Chlamydia muridarum,Cm)生殖道感染作用,为衣原体感染性疾病的治疗提供实验依据。方法构建pET-28a(+)/PGLYRP-1原核表达重组体,IPTG诱导表达PGLYRP-1重组蛋白,重组蛋白经Ni^(2+)-NTA纯化后进行Western blot鉴定。将5×10^(5)包涵体形成单位(IFU)的Cm和不同剂量的PGLYRP-1(50、100、200μg)分别接种于BALB/c小鼠阴道后穹隆,PBS和Cm感染分别作为阴性和阳性对照。收集感染后不同时间(3、7、10、14、21、28、35d)小鼠生殖道分泌物进行衣原体包涵体计数;第21d处死小鼠,无菌分离小鼠脾脏,制备脾淋巴细胞,PGLYRP-1刺激后测定IFN-γ含量;第63d分离小鼠生殖道组织,观察组织病理学变化。结果PGLYRP-1可降低Cm在小鼠生殖道组织的定植并促进Cm的清除,Cm感染阳性对照组及50、100、200μg PGLYRP-1接种组小鼠生殖道Cm清除时间分别是第35、21、14、10d;IFN-γ含量分别为(959.6±104.6)、(1025.4±112.5)、(1537.8±118.6)、(2131.2±196.7)pg/ml,PBS阴性对照组为(224.0±23.5)pg/ml;83.3%的小鼠在Cm感染后第63d存在单侧或双侧输卵管积水现象,PGLYRP-1接种组有22.2%的小鼠存在类似情况。与阳性对照组相比,PGLYRP-1接种组小鼠生殖道组织炎症反应程度显著减轻,且与PGLYRP-1蛋白剂量呈正比(P<0.05)。结论PGLYRP-1具有抗Cm生殖道感染的作用,其作用机制可能与促进IFN-γ表达上调有关。     

沙眼衣原体在肠道定殖中的作用机制

生殖道沙眼衣原体(Chlamydia trachomatis,Ct)是一种最常见的细菌性传播病原体之一。最新研究表明68%的泌尿生殖道Ct核酸阳性的患者中，在其直肠中也能检测到Ct;由于小鼠生殖道感染鼠衣原体(Chlamydia muridarum,Cm)后与人类生殖道感染Ct产生相同的症状，Cm感染动物模型被广泛用于研究Ct的发病机制和免疫途径，研究发现Cm也可以长期定殖小鼠胃肠道。因此，Ct是否可以长期定殖于胃肠道及其作用机制引起众多学者的关注。本文拟对Ct生殖道感染可能播散到机体其他器官的传播途径、Ct主要致病因子及其作用机制作一综述。     

肽聚糖识别蛋白1抗鼠衣原体生殖道感染的初步研究

目的探讨肽聚糖识别蛋白1(peptidoglycan recognition protein 1,PGLYRP-1)抗鼠衣原体(Chlamydia muridarum,Cm)生殖道感染作用,为衣原体感染性疾病的治疗提供实验依据。方法构建pET-28a(+)/PGLYRP-1原核表达重组体,IPTG诱导表达PGLYRP-1重组蛋白,重组蛋白经Ni~(2+)-NTA纯化后进行Western blot鉴定。将5×10~5包涵体形成单位(IFU)的Cm和不同剂量的PGLYRP-1(50、100、200μg)分别接种于BALB/c小鼠阴道后穹隆,PBS和Cm感染分别作为阴性和阳性对照。收集感染后不同时间(3、7、10、14、21、28、35 d)小鼠生殖道分泌物进行衣原体包涵体计数;第21 d处死小鼠,无菌分离小鼠脾脏,制备脾淋巴细胞,PGLYRP-1刺激后测定IFN-γ含量;第63 d分离小鼠生殖道组织,观察组织病理学变化。结果 PGLYRP-1可降低Cm在小鼠生殖道组织的定植并促进Cm的清除,Cm感染阳性对照组及50、100、200μg PGLYRP-1接种组小鼠生殖道Cm清除时间分别是第35、21、14、10 d; IFN-γ含量分别为(959.6±104.6)、(1 025.4±112.5)、(1 537.8±118.6)、(2131.2±196.7)pg/ml, PBS阴性对照组为(224.0±23.5)pg/ml; 83.3%的小鼠在Cm感染后第63 d存在单侧或双侧输卵管积水现象,PGLYRP-1接种组有22.2%的小鼠存在类似情况。与阳性对照组相比,PGLYRP-1接种组小鼠生殖道组织炎症反应程度显著减轻,且与PGLYRP-1蛋白剂量呈正比(P0.05)。结论 PGLYRP-1具有抗Cm生殖道感染的作用,其作用机制可能与促进IFN-γ表达上调有关。     

衣原体感染

沙眼衣原体和解脲支原体感染与输卵管性不孕症关系的研究，不孕和生育不良结局与妇女沙眼衣原体感染，肺炎衣原体感染对C57BL／6J小鼠动脉粥样硬化形成的影响。[编按]     

间接血凝法检测孕妇血清、阴道分泌物沙眼衣原体抗体

沙眼衣原体可以引起多个部位的感染，泌尿生殖道感染是一种常见的性传播疾病，间接血凝法检测孕妇血清中的沙眼衣原体抗体能否反应孕妇泌尿生殖道的感染情况，是一个有待于解决的问题。孕妇沙眼衣原体感染对孕妇和新生儿健康的危害都比较严重，了解孕妇沙眼衣原体的感染情况可为制订预防措施提供理论依据。１　材料和方法１１　研究对象和标本采集　对４０２例于１９９５年３～１２月间每周２在山东医科大学附属医院妇产科进行产前检查的孕妇采上臂静脉血３ｍｌ，分离血清。１１４例于１９９５年４～６月间每周２就诊的孕妇采集阴道分泌物…     

衣原体感染

抗生素联用丙种球蛋白治疗泌尿生殖道沙眼衣原体感染178例疗效观察——韩武红等（湖北孝感市中心医院432100）；《中国男科学杂志》，2006，20（5）：40—41[目的：探索泌尿生殖道沙眼衣原体（CT）感染满意的治疗途径。方法：对178例泌尿生殖道沙眼衣原体患者进行随机分组治疗，A组91例只用抗生素治疗，B组87例用抗生素加丙种球蛋白治疗。结果：A组有效率84．6％，B组有效率94．3％，两组有效率有显著差别（P〈0．05）。结论：抗生素加丙种球蛋白治疗泌尿生殖道沙眼衣原体感染有一定的辅助治疗作用。]     

沙眼衣原体空气传播感染

沙眼衣原体是泌尿生殖道及结膜的病原体。通常认为直接接触或与感染的粘膜和分泌物接触后才会传播。本文报告3例沙眼衣原体感染,其中1例可能为空气传播.例1 29岁男性,发病开始为眼灼热及搔痒伴结膜分泌物。症状持续10个月。后因阴茎头红痒时才从咽部、结膜及尿道分离出沙眼衣原体。经用四环素及强力霉素治疗无效,改用利福平治疗。例2 27岁女性,为例1之妻子。在例1发病后10个月开始结膜灼热发红,随后阴道痒,性交困难及白带过多。从结膜、咽喉、尿道及宫颈找到沙眼衣原体。四环素及强力霉素治疗无效,采用利福平治疗。     

发热门诊呼吸道感染患者中的肺炎衣原体感染调查

目的 了解发热门诊呼吸道感染患者中的肺炎衣原体感染状况,为临床诊断和治疗提供依据。方法 采集2010年11月15日至12月15日在北京友谊医院发热门诊就诊,诊断为呼吸道感染患者的咽拭子标本92例,采用国际肺炎衣原体参考菌株TW-183作为阳性对照和优化PCR条件。用肺炎衣原体种特异性引物对其16S rRNA基因进行PCR检测,阳性产物进行基因测序,结果采用SPSS 10.0软件进行数据统计分析,计数资料比较用(²检验,P≤0.05为差异有统计学意义。结果 CpnA-CpnB PCR可检测到5 ×10^-1 IFU 的肺炎衣原体。92例呼吸道感染患者的咽拭子标本中PCR阳性者为30例,阳性率为32.6%。男性和女性、不同年龄组、以及上呼吸道和下呼吸道感染病例的阳性率间差异均无统计学意义(P>0.05)。结论 在发热门诊有呼吸道症状的病例中,肺炎衣原体感染率较高。临床医生应对肺炎衣原体感染加以重视。CpnA-CpnB PCR法是一灵敏快速的肺炎衣原体筛查方法。     

Experimental genital tract infection withChlamydia psittaci (GPIC agent) in male rats

Summary The course of experimental chlamydial infection of the male genital tract was studied. Inoculation of theChlamydia psittaci agent of guinea pig inclusion conjunctivitis (GPIC agent) into the vas deferens of rats resulted in chlamydial infection of the epididymis, testis and the prostate gland. The inflammatory response was most prominent at 14 days after infection. Chlamydiae were recovered from the epididymides and the prostate glands for up to 90 and 60 days post inoculation, respectively. Histopathological changes associated with chlamydial infection of the epididymis or prostate gland were characterized by intratubular and interstitial purulent inflammation. Chlamydia-specific IgM- and IgG-antibodies were found in sera of nearly all infected animals. Results of this study indicate that this animal model may be useful to study the pathogenesis, immune responses and sequelae of chlamydial infections of the male genital tract.     

IFN-γ抗鹦鹉热衣原体急性感染保护作用研究

目的探讨IFN-γ对鹦鹉热衣原体(Chlamydia psittaci,Cps)的抗感染作用,为进一步阐明机体抗衣原体免疫机制提供参考数据。方法不同浓度的重组人IFN-γ(5ng/mL、25ng/mL和50ng/mL)作用于感染Cps 6BC的HeLa细胞,48h后计数包涵体数量,并观察包涵体形态的改变。2×10~6 IFUs Cps 6BC滴鼻感染C57BL/6J小鼠,于感染前、后24h腹腔内注射10μg重组鼠IFN-γ,观察小鼠体重、活动状态、生存率等一般指标,分别于感染后5d和10d处死小鼠,取肝、肺组织进行HE染色检测其病理变化;并取感染后5d的肺组织,匀浆,计数其中Cps包涵体数量。结果感染48h后,Cps 6BC在5ng/mL、25ng/mL、50ng/mL重组人IFN-γ处理的HeLa细胞中包涵体数量低于对照组(包涵体数分别为(23.8±5.1)×10~6,(10±3.58)×10~6,(8.0±2.22)×10~6,(43.3±11.05)×10~6,衣原体包涵体形状不规则,体积变小。小鼠实验显示,腹腔注射IFN-γ可明显提高Cps 6BC感染小鼠生存率,并减轻急性临床表现及脏器病变。结论 IFN-γ可发挥早期抗Cps感染的保护作用。     

The effect of doxycycline treatment on the development of protective immunity in a murine model of chlamydial genital infection.

Chlamydia trachomatis is a major cause of sexually transmitted disease (STD) worldwide. Antibiotics are effective in treating infection; however, reinfection is common. This observation has led to the conclusion that infection fails to elicit a protective antichlamydial immune response. It was postulated that high reinfection rates might be due to early eradication of organisms from genital tissue after antibiotic intervention, which could negatively influence the development of naturally acquired protective immunity. This hypothesis was tested by use of a murine model of female genital infection. The findings show that doxycycline intervention of infection, although very effective in eradicating chlamydiae from genital tissue and preventing upper genital tract disease, significantly inhibits the development of protective immunity. If antibiotic intervention of human chlamydial genital infection has a similar effect on protective immunity, it could have important implications in the understanding of immunity to infection and future public health efforts to control chlamydial STD.     

Association between genital tract cytomegalovirus infection and bacterial vaginosis

Women with sexually transmitted diseases (STDs) and bacterial vaginosis (BV) have increased rates of cytomegalovirus (CMV) seroprevalence and CMV seroconversion. To characterize the association between genital tract CMV infection and BV, vaginal wash specimens from 52 women attending an STD clinic were analyzed. Significantly more women with BV shed CMV in the lower genital tract than did women without BV. In addition, most of the women who were shedding CMV were infected with >1 virus strain. These results suggest that local CMV replication and infection with multiple CMV strains is facilitated by the presence of BV.     

Genital Herpesvirus homonis infection in mice. II. Treatment with phosphonoacetic acid, adenine arabinoside, and adenine arabinoside 5'-monophosphate.

Genital infection of mice with Herpesvirus hominis type 2 provides an experimental model for screening potential antiviral chemotherapeutic agents before clinical trials in humans. Intravaginal treatment with phosphonoacetic acid (at a dose of 500 mg/kg in saline or as a 5% cream) initiated 3 hr after inoculation with H. hominis type 2 completely inhibited viral replication in the genital tract and prevented subsequent mortality. Although therapy initiated 24-72 hr after infection significantly reduced titers of virus in vaginal secretions from three- to 100-fold, most mice eventually died of encephalitis. Topical treatment with either adenine arabinoside or adenine arabinoside 5'-monophosphate at a dose of 500 mg/kg in saline or as a 10% cream failed to alter viral replication in the genital tract or to protect the mice from death due to encephalitis. Treatment by the intraperitoneal route with any of these three agents had no effect on local viral replication or final mortality.     

Murine Chlamydia trachomatis genital infection is unaltered by depletion of CD4+ T cells and diminished adaptive immunity

Chlamydia muridarum and Chlamydia trachomatis mouse models of genital infection have been used to study chlamydial immunity and vaccine development. To assess the protective role of CD4(+) T cells in resolving C. trachomatis and C. muridarum genital tract infections, we used the female mouse model and evaluated infection in the presence and absence of CD4(+) T cells. In contrast to C. muridarum infection, C. trachomatis infection was unaltered in the absence of CD4(+) T cells. Mice infected with C. trachomatis developed protective immunity to re-challenge, but unlike C. muridarum infection, optimum resistance required multiple infectious challenges, despite the generation of adaptive serum and local chlamydial specific immune responses. Thus, understanding the chlamydial pathogenic and host immunologic factors that result in a diminished protective role for CD4(+) T cells in C. trachomatis murine infection might lead to new insights important to human immunity and vaccine development.     

Quantitative Chlamydia trachomatis cultures: correlation of chlamydial inclusion-forming units with serovar, age, sex, and race

The number of inclusion-forming units (IFUs) observed in quantitative chlamydial cultures may be a surrogate for infectivity or transmissibility. Therefore, we conducted a cross-sectional study of 11,034 patients with Chlamydia trachomatis infection who presented to the Seattle-King County public health department clinics between 1988 and 1996, to determine relationships between the number of IFUs observed in culture and sex, age, race, and serovar class. Of the 11,034 cases of infection we studied, 6801 (62%) were cervical infections in women, and 4233 (38%) were urethral infections in men. The median count was 450 IFU for women and 72 IFU for men (P<.001). Overall, both men and women infected with B-class serovars had significantly higher IFU counts than did those infected with C-class serovars (P<.001). The median IFU count fell consistently with increasing age for both women (625 IFU for those <16 years old to 185 IFU for those >30 years old; P<.001) and men (210 IFU for those <16 years old to 40.5 IFU for those >30 years old; P<.001). We found, by use of multiple regression analysis, that sex, age, race, and serovar class remained independently related to IFU count, with counts being highest among young white women infected with B-class serovars.     

Experimental Chlamydia trachomatis salpingitis in mice: initial studies on the characterization of the leukocyte response to chlamydial infection

The murine biovar (mouse pneumonitis) of Chlamydia trachomatis was inoculated into the left oviduct of female Swiss Webster mice to establish acute salpingitis. Chlamydial inclusions were observed in secretory epithelial cells by both transmission electron microscopy and light microscopy using immunoperoxidase staining of deparaffinized sections. By days 5-8 after infection, a mixed polymorphonuclear and mononuclear cellular infiltrate was observed in the submucosa and mucosa. Epithelial cell deterioration occurred in the endosalpinx in areas of heavy mononuclear cellular infiltration. During the acute phase of the disease a cellular infiltrate consisting mainly of T cells was identified by staining frozen tissue sections with monoclonal antibodies to mouse lymphocyte antigens. Occasionally B lymphocytes were observed. Widespread deciliation of the mucosa was observed by scanning electron microscopy. No histopathologic or immunopathologic responses were observed in the control oviducts. These observations suggest an immunologic basis for the structural abnormalities seen in the infected oviducts.