Potent inhibition of HIV-1 by TRIM5-cyclophilin fusion proteins engineered from human components

Article Article scientifique Article Potent inhibition of HIV-1 by TRIM5-cyclophilin fusion proteins engineered from human components eng 2009 author Neagu Martha Raluca 677894 author Ziegler Patrick author Pertel Thomas 674561 author Strambio-De-Castillia Caterina author Grutter Christian author Martinetti Gladys author Mazzucchelli Luca author Grutter Markus author Manz Markus G. author Luban Jeremy 659379 3035-3047 10.1172/JCI39354 19741300 0021-9738 New World monkeys of the genus Aotus synthesize a fusion protein (AoT5Cyp) containing tripartite motif-containing 5 (TRIM5) and cyclophilin A (CypA) that potently blocks HIV-1 infection. We attempted to generate a human HIV-1 inhibitor modeled after AoT5Cyp, by fusing human CypA to human TRIM5 (hT5Cyp). Of 13 constructs, 3 showed substantial HIV-1-inhibitory activity when expressed in human cell lines. This activity required capsid binding by CypA and correlated with CypA linkage to the TRIM5a capsid-specificity determinant and the ability to form cytoplasmic bodies. CXCR4- and CCR5-tropic HIV-1 clones and primary isolates were inhibited from infecting multiple human macrophage and T cell lines and primary cells by hT5Cyp, as were HIV-2ROD, SIVAGMtan, FIVPET, and a circulating HIV-1 isolate previously reported to be AoT5Cyp resistant. The anti-HIV-1 activity of hT5Cyp was surprisingly more effective than that of the well-characterized rhesus TRIM5alpha, especially in T cells. hT5Cyp also blocked HIV-1 infection of primary CD4+ T cells and macrophages and conferred a survival advantage to these cells without disrupting their function. Extensive attempts to elicit HIV-1 resistance to hT5Cyp were unsuccessful. Finally, Rag2-/-gammac-/- mice were engrafted with human CD4+ T cells that had been transduced by optimized lentiviral vectors bearing hT5Cyp. Upon challenge with HIV-1, these mice showed decreased viremia and productive infection in lymphoid organs and preserved numbers of human CD4+ T cells. We conclude that hT5Cyp is an extraordinarily robust inhibitor of HIV-1 replication and a promising anti-HIV-1 gene therapy candidate. Adoptive Transfer Amino Acid Sequence Animals Anti-HIV Agents/*metabolism Aotidae CD4-Positive T-Lymphocytes/cytology/immunology Carrier Proteins/genetics/*metabolism Cell Line Cyclophilin A/genetics/*metabolism DNA-Binding Proteins/genetics/metabolism HIV-1/*metabolism Humans Mice Mice, Knockout Models, Molecular Molecular Sequence Data Protein Conformation Protein Engineering Recombinant Fusion Proteins/genetics/*metabolism Dewey 616 54 Département de microbiologie et médecine moléculaire 21B false 510 Réplication virale, pathogénèse et immunité 848 The Journal of clinical investigation 119 10 660b9608-30ab-41df-b109-53097ac1e089 Article Restricted file1.pdf 3237715 application/pdf restrictedAccess Faculté de médecine / Section de médecine fondamentale / Département de microbiologie et médecine moléculaire

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Potent inhibition of HIV-1 by TRIM5-cyclophilin fusion proteins engineered from human components	The Journal of clinical investigation			2009	620	0

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Neagu, Martha Raluca