Dendritic cells play a vital important role in HIV-1infection and transmission, since they are the first target cell and the primary defence of HIV-1. On meeting the viruses, they can capture, process and present the cleaved antigen to T cells and finally activated the immune system, which relies on the properly differentiation and maturation of DCs. Howerver, when being confronted with HIV-1, DCs seem to be regulated into a disfunctional state for HIV-1 encods 6 accessory proteins which can somehow regulate the differentiation and muration of DCs, and these auxiliary proteins make HIV-1 superior over other lentivirus for their high pathogenicity. Functions of DCs can be downregulate by auxiliary proteins; especially antigen presenting function, this regulatory effect is also a key step for pathegenesis of HIV-1 and morbidity of AIDS. Thus it is vital improtant to known how the auxiliary proteins lead to the dysfunction of DCs, since it’s helpful to understand the mechanism of the interaction between the proteins and the whole immune system and the overal course of morbidity and it’s also important to the prevention and curation of the disease and development of new effective vaccine of HIV-1. Since the interaction between DCs and each auxiliary protein is not clear, it is significant to set a proper cell line model and research system to study the interplay. In this work, THP-1, a leukimia derived cell line which can be effectively infected by HIV-1 in vitro, was chosen to stimulate into DCs. It was at first evaluated as a DC progenitor using established and modified methods. Then six HIV-1 auxiliary protein genes were electroporated into THP-1, selected and sorted under the same conditions. Two stably expressed cell lines, THP-1-Rev and THP-1-Vpr, were successfully established from the six transfected cells with G418 selection and sorting by flow cytometry and detected by RT-PCR assay. While Nef and Tat could not be stably expressed by transfected cells, the cells was analysed by apoptosis detection using Annexin V kit, and the results support the hypothesis that the transfected cells themselves with the two genes were induced into apoptosis. These results has provided an experimental basis for the next research of affact of Rev and Vpr on DC differentiation and maturation and the it’s function as well as the underline mechanism. Next work will be focused on the pathgenic effects of auxiliary proteins of HIV-1. As vif and vpu were not effeciently expressed in THP-1, we here analysed the possible reasons. It was reported that Vif prteins may be cleaved into several peptides in the host cells, so it cannot be detected by Western Blot and flow cytometry. As is referred to Vpu, APOBEC3G may has something to do with the low tranfection efficiency based on our experiment result in RT-PCR of APOBEC3G in several different cell lines and the transfection assay with their comterparts, which developed a new field for the research on interaction between the auxiliary proteins and the host resitant factors, much more work will be done in our lab to focus on the new interaction of Vpu and HIV-1 resistant factors. This will provide a basis to better understand the mechanism of the pathogenesis and morbidity, and the resistance of host to HIV-1.
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