女性下生殖道HPV感染和HPV相关的宫颈肿瘤

子宫颈病变是女性最常见的疾患之一.在女性癌瘤中,宫颈癌的发病率仅次于乳腺癌.人乳头状瘤病毒(HPV)感染在宫颈肿瘤的发病机制中起着重要的作用,许多学者关注HPV疫苗的预防和治疗.在女性生殖道HPV传播及继发性感染是局部性的,因此,针对这种局部性传播疫苗的有效性最好能用局部免疫的参数来评价.     

杭州地区性病门诊男性就诊者HPV感染的流行病学调查

目的 了解杭州地区性病（STD）门诊男性就诊者中人乳头瘤病毒（HPV）感染的流行情况。方法 利用HPV黏膜型通用引物MY09／11。PCR方法检测STD门诊年龄在18—70岁男性就诊者阴茎拭子中的HPV DNA，并对阳性PCR产物用限制性长度片段多态性方法和直接测序法确定病毒基因型。结果 在采集的375例阴茎拭子中，81．3％（305例）产生足够的DNA可用于PCR扩增。HPVDNA的阳性率为13．8％（42／305），其中低危型HPV的阳性率为8．5％（26／305），高危型HPV的阳性率为4．3％（13/305），混合型感染的阳性率为1．0％（3/305）。居住地在城市、受教育程度越低、性伴侣的人数越多HPV DNA的检出率越高（P〈0．05）。结论 HPV在男性高危人群的感染并不少见，HPV DNA的检出与就诊者的居住地、教育程度及性伴侣人数有相关性。对男性高危人群HPV的感染应引起重视，积极采取预防和控制措施。     

2016~2018年苏州市育龄妇女HPV感染因素分析

目的 调查苏州市育龄妇女人乳头状瘤病毒（HPV）感染情况及其影响因素。方法 以整群抽样方式选取2016年7月~2018年12月苏州市育龄妇女4275例为研究对象,进行HPV和TCT检查,并填写调查问卷,了解HPV感染情况。结果 4275例孕妇中存在HPV感染者964例（22.55%）;HPV发生与年龄、职业、在苏时间、文化程度、每周性生活频率、性伴侣数、避孕方式、性传播疾病史、CIN、吸烟等因素有关,差异有统计学意义（P＜0.05）;Logistic回归分析显示:文化程度、性生活频率、避孕方式、吸烟、年龄、职业是苏州市育龄妇女HPV发生的独立危险因素（P＜0.05）。结论 苏州市育龄妇女HPV感染率较高,文化程度、性生活频率、避孕方式、吸烟、年龄、职业是其发生的独立危险因素,需加强对育龄期女性HPV感染知识宣教,增强认知,定期体检,早发现、早治疗。     

多重HPV感染与宫颈病变的临床分析

目的探讨多重人乳头状瘤病毒（HPV）感染与宫颈病变之间的关系。方法采用核酸分子快速导流杂交基因芯片技术,对2008年7月至2010年7月在东莞市太平人民医院就诊且有病理确诊的332例女性患者（实验组）及100例正常妇女（对照组）进行HPV基因分型检测,比较不同宫颈病变类型与HPV多重感染的关系。结果 332例宫颈病变中HPV感染率为78.61%（261/332）,多重感染率为58.13%（168/289）,其中以二重感染为主,最常见的二重感染类型为HPV16、58及HPV52、58,以HPV16型感染多见;多重HPV感染比例随病变级别增加逐渐上升,由对照组的17.86%升高到宫颈鳞癌组的100%,各病变组与对照组相比差异有统计学意义（P〈0.05）。结论 HPV多重感染与宫颈病变的发生有关,可作为宫颈病变早诊早治的有效指标之一。     

人乳腺癌HPV感染的超微结构和DNA分子原位杂交研究

目的：对乳腺癌的HPV感染进行形态学定位研究。方法：对46例乳腺癌切除标本及7例良性病变标本进行透射电子显微镜以及HPV DNA分子原位杂交检测。结果：透射电镜下，14例（30．40％）乳腺癌细胞核内有HPV样病毒颗粒，颗粒直径30－50nm，或弥漫散布，或聚集成团呈假结晶状排列。颗粒圆形或略不规则，含有HPV样颗粒的癌细胞核异型性较明显。7例良性病变细胞核内未见到HPV样颗粒。HPV DNA分子原位杂交显示，21例（45．65％）乳腺癌细胞核内HPV31／33 DNA阳性，1例软肉瘤样化生性癌细胞核同时有HPV31／33及HPV16／18阳性；1例纤维腺癌（14．29％）也显示HPV31／33 DNA阳性（X^2=2．7431，P＜0．05）。电镜下85％病毒样颗粒阳性病例中可检测到HPV DNA存在，两种检测结果符合率达73．91％。结论：乳腺肿瘤内存在未组装的、裸型病毒样颗粒。该研究对阐明乳腺癌的发病原因以及对乳腺癌的预防和治疗均有重要意义。     

北京市北苑地区女性HPV感染现状及基因型分布研究

目的 调查我院辖区范围内即北京市北苑地区成年女性人乳头状瘤病毒(HPV)感染现状及亚型分布情况.方法 采用核酸分子快速导流杂交基因芯片分型技术对本院辖区范围内(北苑地区)来我院妇科门诊就诊的14582例女性患者采集宫颈脱落细胞标本进行HPV及21种亚型检测.结果 ①HPV感染阳性检出2419例,感染阳性率16.59％(2419/14582),在HPV感染阳性人群中,以单一高危型HPV感染为主,单一感染HPV阳性1951例,占HPV阳性感染人群80.65％(1951/2419);多重感染以二重感染为主,多重感染HPV阳性468例,二重感染HPV阳性361例,占HPV阳性感染人群14.92％(361/2419),以高危型感染以为主.②各HPV基因亚型检出总和为3035项次,高危型HPV感染占91.73％(2784/3035),排在前五位由高到低依次是HPV16(17.36％,527/3035)、HPV52(11.89％,361/3035)、HPV58(11.80％,358/3035)、HPV53(8.27％,251/3035)、CP8304(7.05％,214/3035),以HPV16型为主;低危型HPV感染占8.27％(251/3035),由高到低依次是HPVl1 (3.95％,120/3035)、HPV6(3.57％,108/3035)、HPV44(0.43％,13/3035)、HPV42 (0.16％,5/3035)、HPV43 (0.16％,5/3035),以HPV11为主.③按年龄段划分,≤20岁年龄组阳性率最高,阳性率31.30％ (36/115);其次是≥61岁年龄组,阳性率21.05％ (64/304).结论 本院辖区范围内(北苑地区)成年女性HPV感染阳性率16.59 ％(2419/14582),以单一高危型HPV感染为主,高危型HPV16、HPV52、HPV58、HPV53、CP8304是本院辖区范围内成年女性HPV感染的主要亚型.     

浙江沿海地区人群TTV和HPV混合感染研究

目的了解浙江沿海地区人群TTV与HPV混合感染状况,探讨TTV传播途径。方法建立巢式聚合酶链反应方法（nPCR）,对健康体检和患宫颈疾病妇女150例宫颈病变细胞学标本及其平行97例血清标本进行TTVDNA及TTV病毒滴度的nPCR检测;应用导流杂交方法检测95例宫颈病变细胞学标本的HPV基因型。结果TTVDNA在55例健康妇女宫颈细胞标本中检出率52．7％（29／55）,与其平行42例血清样本中检出率50．0％（21／42）。在患有疾病妇女宫颈细胞中TTVDNA检出率（74．7％）高于健康体检对照组（P=0．005）。TTVDNA在患者血清样本中检出率51％（28／55）。在宫颈细胞及其平行血清中均检测出TTV基因亚型G1b。TTV病毒滴度在宫颈细胞中高于在其平行血清10～1000倍。HPV在患者组中检出率98．9％（94／95）,在健康组中检出率27．3％（15／55）。HPV基因型是高危型HPV16、18、33和低危型HPV6。HPV阳性标本TTVDNA检测率明显高于HPV阴性标本（P=0．02）。结论TTV在宫颈细胞中具有高检出率,在宫颈细胞中TTV病毒滴度高于其平行血清。TTV与HPV随性传播感染人群,并在女性生殖道内繁殖。TTV与HPV协同作用有待研究。     

抗HPV16E6核酶对宫颈癌细胞免疫学特性影响的研究

目的 研究特异性抗体HPV16E6核酶的转染对宫颈癌细胞免疫学特性的影响。方法 以脂质体法将抗HPV16E6-ribozyme、空载体质粒分别导入CaSKi细胞,命名为CaSKi、CaSKi-P细胞。流式细胞仪分别检测3种细胞HLA－1、HL2－2、B7－1和B7－2基因的表达,分析CaSKi细胞转染酶后免疫学特性的变化。诱导制备NK、LAK、CD3AK和肿瘤细胞共激活杀伤细胞,检测各种免疫细胞对CaSKi-R、CaSKi-P、CaSKi细胞的杀伤效应。结果 CaSKi和CaSKi-p细胞中HLA－2、B7－1、B7－2表达都很低,两者差异无显著性。CaSKi-R中HLA－2、B7－1、B7－2表达率均明显升高。3种细胞中HLA－1表达率都很高。NK、LAK、CD3AK细胞对CaSKi-R细胞伤率显著高于CaSKi细胞,CaSKi、CaSKi-R分别与rIL－2共激活杀伤细胞（称CASKI和CASKI－R）的样伤活性无明显差别,对CaSKi-R的杀伤活性高于CaSKi细胞。而各种免疫细胞对CaSKi-P细胞的杀伤率与CaSKi细胞差异无显著性。结论 抗HPV16E6－nrbozyme的导入能使宫颈癌CaSKi细胞易于被机体免疫系统识别杀伤,难于免疫逃避,但不能增强其免疫原性。     

1448例妇科患者HPV感染型别分布及临床分析

目的分析女性HPV感染的阳性率和各年龄段的关系及常见亚型的分布情况,探讨女性HPVDNA基因分型技术检测在宫颈癌防治方面的应用价值。方法采用HPV基因分型技术对1448例妇科门诊就诊者进行HPV分型检测。结果1448例样本中,检出HPV阳性者531例,总阳性率为36．7％,其中高危型HPV468例,占感染率88％,低危型HPV63例,占感染率12％,混合感染HPV122例,占感染率23％,HPV基因型单一感染为77．2％,二重感染为17．9％,三重感染为3．6％,四重感染为1．5％。高危型中未检出HPV35、73、83、MM4型,低危型中未检出HPV43、44型。患者其亚型感染率由高到低依次为HPV58、16、52、33、18、31、53、68、66、51、39、56和59型,低危型依次为HPV11、6、42型。不同年龄组中HPV感染高峰在50～59岁和60～69岁这两个年龄段,感染率分别为78．9％和73．7％。结论HPV感染型别普遍化,各年龄段感染的阳性率差异有统计学意义（P〈0．05）,25岁前后及〉50岁的妇女更应予以重视,对阳性患者要加强定期随访,HPV基因分型及多种亚型的检测有利于宫颈癌的早期预警和早期治疗。     

斑点分子杂交检测HPV感染与宫颈癌的关系

采用ＤＮＡ－ＤＮＡ分子杂交技术，对经病理组织学确诊的慢性宫颈炎，宫颈癌和正常宫颈的宫颈活检组织中ＨＰＶ６，ＨＰＶ１１，ＨＰＶ１６，ＨＰＶ１８型ＤＮＡ进行同源序列检测，结果表明ＨＰＶ６，ＨＰＶ１１，ＨＰＶ１６，ＨＰＶ１８型ＤＮＡ的检出率在正常宫颈均为０；在慢性宫颈炎分别为１６．０９％，１２．６４％，１１．４９％，３．４５％；在宫颈癌组分别为３．９６％，１．９８％，４６．５３％，７．９２％。在宫颈癌组     

In situ immunological characterization of Langerhans cells with monoclonal antibodies: Comparison with other dendritic cells in skin and lymph nodes

Summary The antigenic properties of epidermal Langerhans cells (LC) were determined and compared with those of non-lymphoid dendritic dermal cells (DDC), interdigitating reticulum cells (IRC), dendritic reticulum cells (DRC), and histiocytic reticulum cells (HRC) by examination of serial and double immunoenzymatic and -fluorescence stained frozen sections of skin and lymph node biopsies.All of these cell types expressed leucocyte common antigen. LC, DDC, and IRC demonstrated similar antigenic phenotypes (HLA-DR⁺, Leu3⁺, OKT6^+/–, anti-C3 receptor^–, R4/23^–, Ig-complex^–, M02^–), whereas the antigenic properties on DRC (HLA-DR^–, Leu3^–, OKT6^–, anti-C3 receptor⁺, R4/23⁺, Ig-complex⁺, M02^–) and HRC (HLA-DR^+/–, Leu3^–, OKT6^–, anti-C3 receptor⁺ R4/23^–, Ig-complex⁺, MO2⁺) were markedly different.These data suggest that LC, DDC, and IRC are closely interrelated cell types, and support the concept that DRC and HRC are unique cell types which do not appear to be related to LC, DDC, or IRC. The lack of labelling of LC with monoclonal anti-C3b receptor antibody, and polyclonal antiserum recognizing C3b, C3bi, and C3d receptors strongly indicate that the EAC-rosetting of LC previously described is not due to the presence of C3 receptors on these cells. Alternatively, LC may express C3 receptor molecules different from those previously identified (C3b, C3bi, and C3d).     

Langerhans cells and viral immunity

Langerhans cells (LC) are a unique dendritic cell subset that are located in mucosal stratified squamous epithelium and skin epidermis. Their location is ideally suited for their function as antigen presenting cells that capture invading viruses and induce anti-viral immunity. However, it is becoming evident that the interaction between LC and viruses can result in different responses, depending on the virus and the receptors involved. Here we will discuss the recent data on the similarities and differences in roles of LC in viral immunity to and infection with HIV, herpes simplex and varicella-zoster virus. Although all three viruses interact with LC during initial infection, the effects can be quite different, reflecting differences in biology and pathogenesis.     

Murine epidermal Langerhans cells and splenic dendritic cells present tumor-associated antigens to primed T cells

We examined the ability of epidermal Langerhans cells and splenic dendritic cells to present tumor-associated antigens (Ag) to immune T cells. Methylcholanthrene (MCA)-induced subcutaneous fibrosarcomas derived from C57BL/6 mice were used as tumor models. Our data demonstrate that both murine Langerhans cells and splenic dendritic cells have the capacity to present tumor-associated Ag to primed T cells. We found that variously treated tumor preparations (irradiated viable tumor cells, irradiated frozen-stored tumor cells, mitomycin C-treated viable tumor cells, and snap freeze-thawed tumor cell lysates) can be utilized for tumor Ag-pulsing. Primed CD4⁺ T cells demonstrated in vitro specificity towards their respective tumors and did not cross-react to other syngeneic MCA-induced or non-MCA-induced tumors. The T cell proliferative response critically depended on the presence of immune CD4⁺ T cells. We discuss the implications of these findings for the adoptive immunotherapy of cancer using immune CD4⁺ T cells.     

人乳头瘤病毒11型L1/E7嵌合DNA疫苗的构建及其诱导的免疫效应

目的 构建人乳头瘤病毒11型L1/E7嵌合DNA疫苗pcDNA3 L1-E7并研究其在小鼠中诱导的免疫效应.方法 用分子克隆技术构建重组真核表达质粒peDNA3 L1-E7.重组质粒DNA股四头肌注射免疫小鼠,用ELISA方法 检测L1、E7特异性抗体和脾细胞分泌的IL-2、γ-INF;MTT法检测脾淋巴细胞增殖反应.结果 成功构建pcDNA3 L1-E7.免疫小鼠后,重组质粒可诱导机体产生特异性脾淋巴细胞增殖及IL-2、γ-INF分泌增加,并诱导机体产生HPV 11-E7 IgG和HPV 11-L1 IgG抗体.结论 嵌合DNA疫苗pcDNA3 L1-E7能诱导小鼠产生特异性的细胞免疫和体液免疫反应.     

Effects of TNF-alpha inhibitors on the number of epidermal Langerhans cells in uninvolved skin of psoriatic patients: A pilot study

Only limited data are available on the effects of TNF-alpha inhibitors on dendritic cells. However, TNF-alpha plays a central role in the biology of dendritic cells, both with regard to their maturity process and mobilization to secondary lymphoid organs. In particular, the effects of TNF-alpha inhibitors on Langerhans cells in healthy skin have never been investigated. In this pilot study, we aimed to assess the change of the density of Langerhans cells within the normal, not photo-exposed, skin of 17 psoriatic patients, before and after 16 weeks of treatment with TNF-alpha inhibitors. Most of the patients (88%) showed an increase or a similar density of Langerhans cells after 16 weeks of therapy with TNF-alpha inhibitors compared with baseline values. Only 2 patients (12%) showed a reduction of these cells following therapy with TNF-alpha inhibitors.     

热处理淋巴瘤细胞增强树突状细胞介导的抗瘤效应研究

目的:探讨淋巴瘤细胞热处理后作为抗原,冲击树突状细胞而引发的抗淋巴瘤免疫效应。方法:用健康人外周血分离出单核细胞,体外培养树突状细胞(Dendritic cells,DCs),将淋巴瘤细胞株热处理(42℃,2小时)培养24小时后负载于DCs,在流式细胞仪上检测DCs的免疫表型变化;负载抗原后的DCs与淋巴细胞混合反应,以MTT法评价细胞毒性及在流式细胞仪上检测淋巴细胞的免疫表型;ELISPOT法检测细胞内因子IFNγ-的释放。结果:在两实验组中,DCs负载了热处理的淋巴瘤抗原后,细胞表面的共刺激分子和MHCⅡ类分子表达水平较对照组明显增加(P0.05),而两组之间差别无显著性(P0.05);经热处理的肿瘤细胞负载于DCs后,与淋巴细胞混合后IFNγ-的释放量明显增加;混合淋巴细胞反应后,细胞毒性实验(MTT法)和流式细胞仪检测结果均显示两实验组的杀瘤作用强于对照组(P0.05),两组之间无显著差别(P0.05)。结论:用热处理的淋巴瘤细胞作为肿瘤抗原冲击DC,能够增强抗淋巴瘤效应。     

Extremely high Langerhans cell infiltration contributes to the favourable prognosis of HPV-infected squamous cell carcinoma and adenocarcinoma of the lung

AIMS: The infiltration of Langerhans cells in adenocarcinomas and squamous cell carcinomas of the lung was examined in relation to prognostic implications and human papillomavirus (HPV) infection. METHODS AND RESULTS: Samples from 62 adenocarcinoma and 59 squamous cell carcinoma patients in 1995-97, the prognosis of which had been followed up, were used. The Langerhans cells were demonstrated immunohistochemically using anti S100a and CD1 antibodies. Human papillomavirus (HPV) infection was examined by polymerase chain reaction (PCR) and nonisotopic in-situ hybridization (NISH) methods. Statistical analysis was carried out using the Kaplan-Meier method (Wilcoxon analysis) and multiple regression analysis. HPV infection was demonstrated in 12 cases (19.4%) of adenocarcinoma. The HPV-infected adenocarcinomas had abundant faintly eosinophilic cytoplasm, and were immunohistochemically positive for the surfactant apoprotein A. In the 59 cases of squamous cell carcinomas 19 were of the well differentiated form, and 29 and 11 were moderately and poorly differentiated cases, respectively. HPV was detected in 29 cases (49.2%) (13 well and 16 moderately differentiated cases). In all HPV-infected adenocarcinoma and squamous cell carcinoma cases, extremely large numbers of Langerhans cells (more than 100 per high-power field) were demonstrated in the tumour nests. In contrast, in the non-HPV-infected adenocarcinomas and squamous cell carcinomas, only a few (less than about 10 per high-power field) Langerhans cells were observed. The squamous cell carcinoma cases with high Langerhans cell infiltration, which were also infected with HPV, showed a significantly good prognosis (P = 0.007). The adenocarcinoma cases with high Langerhans cell infiltration tended to have a better prognosis than the cases with low Langerhans cell infiltration, but the difference was not statistically significant. The low number of highly infiltrated cases was insufficient for an adequate statistical analysis. Furthermore, there was no significant correlation between either Langerhans cell infiltration and smoking, or HPV infection and smoking, in either squamous cell carcinoma or adenocarcinoma cases. CONCLUSIONS: It was considered that the extremely high Langerhans cell infiltration in the tumours was caused by HPV infection. The extremely large number of Langerhans cells in the tumours contributes to the favourable prognosis for HPV-infected lung cancer.     

Roles of lymphoid cells in the differentiation of Langerhans dendritic cells in mice

Langerhans dendritic cells are antigen presenting cells (APC) that reside within the epidermis and are capable of stimulating naive T cells. Reciprocally, lymphocytes may play a role in Langerhans cells (LC) differentiation. Our results show that the differentiation of skin LC is unaffected in the absence of lymphocytes and/or signaling through the common cytokine receptor gamma chain (gammac) required for IL-2, IL-4, IL-7, IL-9, IL-15 and IL-21 signaling. Migration of LC and other dendritic cells (DC) from the skin to the draining lymph nodes (LNs) after FITC skin sensitization, is unaffected in the absence of lymphocytes or CD40. FITC+ LC/DC sorted from the LNs of lymphoid deficient or control mice stimulated naive T cells with similar efficiency. However, while the absence of lymphocytes did not appear to affect the phenotype or number of emigrating LN DC/LC, their persistence in the LN appears to depend on alphabeta T cells. Thus, DC are strikingly reduced in numbers in the peripheral LNs of T-cell deficient mice. Finally, CD8alpha expression on skin emigrants was low and dependent on the presence of CD8+ lymphocytes, while spleen CD8+ DC were present in the absence of lymphocytes. We conclude that the presence of T cells is not required for the differentiation and migration of resident skin DC but is critical for the maintenance of DC and LC migrating into the LNs.     

Langerhans cells are more efficiently transduced than dermal dendritic cells by adenovirus vectors expressing either group C or group B fibre protein: implications for mucosal vaccines

Vaccines against viruses need to target dendritic cells (DC) and stimulate mucosal immunity. Most vaccine studies have focussed on monocyte-derived or dermal DC (dDC) but recent evidence suggests that Langerhans cells (LC) may stimulate mucosal immunity more effectively. New chimeric adenovirus vectors expressing fibre protein from group B adenoviruses (rAd5/11), which utilise CD46 rather than the Coxsackie adenovirus receptor (CAR), have been developed as vaccines to improve transduction and overcome problems of pre-existing vector immunity. Transduction of LC and dDC by rAd5/11 and standard rAd5 expressing green fluorescent protein (GFP) showed that both DC types were more efficiently transduced by rAd5/11 than by rAd5. Although expression of CD46 and the integrins alphavbeta3 and alphavbeta5, which recognise the adenovirus penton base and mediate virus internalisation, was similar in LC and dDC, LC expressed higher levels of GFP. Transduction by electroporation of plasmid also resulted in higher GFP expression in LC, suggesting differences between the two DC populations at a post-entry stage. Transduction with either vector did not induce maturation of LC or dDC and did not affect their ability to stimulate T cells. These findings suggest that vaccine strategies that target LC with adenovirus vectors may be worthy of exploration.     

IgE on Langerhans cells in the skin of patients with atopic dermatitis and birch allergy

Epidermal cell suspensions from the skin of seven patients with atopic dermatitis (AD) and seven healthy non-atopic controls were investigated for the presence of surface HLA-DR and CD1 antigen, and IgE using indirect and double-staining immunofluorescence techniques. Fifty-seven percent of all CDI + and 68% of all HLA-DR + cells from the patients demonstrated IgE on their surface, indicating that Langerhans cells (Lc) in AD may be a heterogeneous population with regard to surface characteristics. No IgE + cells were found in the epidermal cell suspensions from the healthy non-atopic controls. An attempt lo demonstrate birch pollen antigen on the surface of Lc from the same patients all strongly allergic to birch pollen, using indirect immunofluorescence techniques, was unsuccessful, also after in vitro incubation of the Lc with high concentration of the birch antigen.