Construction Of HIV-1 Pseudovirus System For Broadly Activity Evaluation Of Neutralizing Antibodies

By the end of 2017,the number of people who have infected HIV is approaching 77.3 million or so,resulting in 35.4 million dying of AIDS-related diseases.There are currently about 36.9 million people living with HIV globally.The potential HIV infection,high mutagenicity and destruction of the immune system are the huge obstacles to the development of effective preventive and therapeutic vaccines.The research and development of HIV vaccine,the development of antiviral drugs and the evaluation and identification of neutralizing antibodies need an effective and standard evaluation system,and the cell evaluation method based on HIV pseudo virus is an important tool for HIV researchThe aim of this study was to construct a HIV Env pseudovirus system that can be used to evaluate the antiserum,neutralizing antibody potency and broad-spectrum of candidate vaccine,and to establish a pseudovirus two-plasmid system.It contains 10 global panel of HIV-1 Env reference strains and 8 global epidemic strains,involving 6 epidemic subtypes(A,B,C,D?DC and G)and 2 recombinant subtypes(CRF07 BC,CRF01_AE).The eukaryotic expression vector pVRC8400 with the universality of Env expression was determined by comparing the differences of the three expression vectors in the expression of recombinant Env and the packaging efficiency of pseudovirus.To optimize the transfection conditions,determine the content of key proteins and titer of virus,and determine the optimal conditions for the packaging of each strain.By comparing the differences in the titers of the three types of neutralizing antibodies and the evaluation of the titers of the three types of neutralizing antibodies and the pseudo viruses packaged in the plasmid system of HIV-1 epidemic strains 94UG114 and NL4-3 and the euvirus packaged by infectious molecular cloning system,it was proved that the pseudovirus constructed in this study can replace the euvirus.The pseudovirus disk was used as a tool to evaluate the neutralization potency and broad-spectrum of the broadly neutralization antibody 15A7 and gp140 cross-neutralization antibody obtained in the laboratory,providing an important evaluation tool for the follow-up research on HIV vaccine and the development of neutralization antibodyPseudovirus has the advantages of high safety,strong applicability and easy access,and it can replace the replicative living virus to evaluate various antibodies and drugs,which is of great significance for HIV-1 research.As an important tool for the evaluated of neutralizing antibodies,the pseudovirus panel does great favor to guiding vaccine design and tackling AIDS.