摘要
探讨登革病毒对人树突状细胞(DC)的感染性。人外周新鲜血常规分离单核细胞,经细胞因子GMCSF、IL4诱导培养成DC,通过形态学特征、细胞表型和淋巴细胞刺激能力鉴定。用登革病毒2型(DV2)感染DC,于作用后6h、24h、48h、72h、96h分别收集上清液和细胞,甲基纤维素微量空斑试验测定病毒滴度,间接免疫荧光法检测细胞上病毒抗原表达,透射电镜观察病毒在细胞内的定位。病毒感染后6h即可在培养上清中测出病毒,病毒滴度在48h达到高峰,以后逐渐下降。间接免疫荧光法证明感染的DC胞浆及胞膜上携带病毒抗原。透射电镜下在病毒感染48h后DC胞浆内可见大量病毒颗粒。树突状细胞是登革病毒感染的靶细胞,病毒可感染DC并产生大量病毒颗粒,可能在其发病机制中起重要作用。
To study Dengue virus (DV) infection of human dendritic cells(DC). Monocyte isolated from healthy human peripheral blood were incubated in medium with GM-CSF and IL-4 for more than 7 days. DCs were then collected and identified by transmission electron microscope, immunohistochemistry and lymphocytes stimulatory ability. Dengue virus type ΙΙ (DV-2) were infected with human dendritic cells(DC) in vitro,culture supernatants and cells were collected by different time postinfection (6h,12h,24h,48h,72h).Viral titers were evaluated by microplaque forming assay on C6/36 monolayer cells;DV antigen in human dendritic cells were demonstrated by an indirect immunofluorescent assay(IFA).Localization of DV in DC was observed under a transmission electron microscope. The viruses were detected in the culture supernatants as early as 6h after infection; the highest viral titers were obtained at 48h,and then declined to very low titers at 96h. DV-2 antigen was detected in infected DC by IFA. After infection for 48h, DV particles were obvious in cystic vesicle, vacuoles . Human dendritic cells are targets of dengue virus infection. DV could efficiently infect DC and produce virus particles, DC possibly plays a role in the pathogenesis of DV infection.
出处
《微生物学报》
CAS
CSCD
北大核心
2005年第4期598-600,i003,共4页
Acta Microbiologica Sinica
基金
国家"863计划"(2003AA219060)
广东省医学科研基金(A2005340)~~
关键词
登革病毒
人树突状细胞
感染
Dengue virus, Human dendritic cells, Infection
