Single-domain Antibodies M36.4 And 10E8 Single-chain Variable Region Fragments For The Study Of AIDS Gene Therapy Strategies

HIV-1 broad spectrum neutralizing antibodies can target antigenic epitopes on HIV-1 envelope glycoprotein subunits and show extensive anti-HIV-1 activity.In our study,we used glycosyl-phosphatidylinositol(GPI)to express single chain variable region fragments(scFv)of 3BNC117,N6,PGT126,PGT128,35022 and 10E8,and single domain antibody m36.4 on the lipid raft region of HIV virus target cells.We found that all GPI-scFvs or GPI-m36.4 can be effectively expressed on the surface of transduced target cells and are targeted to the lipid raft site without affecting the expression of CD4,CCR5 and CXCR4.Transduced TZM.bl cells showed different degrees of resistance to different HIV-1 virus infections,while TZM.bl cells transduced by GPI-10E8 and GPI-m36.4 could achieve complete suppression of various subtypes of HIV-1 isolates.In addition,we also demonstrated that GPI-10E8 or GPI-m36.4 can effectively block the cell-to-cell transmission of HIV-1 and cell fusion mediated by multiple HIV-1 Envs.In particular,GPI-m36.4 could make cells completely resistant to it.Human T cell line expressing GPI-m36.4 and GPI-10E8 also showed resistance to HIV-1 infection,and during HIV-1 virus infection,they showed more obvious selective survival and expansion advantages than unmodified cells.In addition,we also found that GPI-10E8 and GPI-m36.4 can greatly affect the processing of HIV-1 Env glycoprotein,virus release and infectivity.In summary,our study identified that m36.4 and 10E8 can be used as membrane bound virus entry inhibitors by GPI anchor to research on gene therapy strategies for HIV-resistant cells.