title: Incomplete APOBEC3G/F Neutralization by HIV-1 Vif Mutants Facilitates the Genetic Evolution from CCR5 to CXCR4 Usage.
abstract: Incomplete APOBEC3G/F neutralization by a defective HIV-1Vif protein can promote genetic diversification by inducing G-to-A mutations in the HIV-1 genome. The HIV-1 Env V3 loop, critical for coreceptor usage, contains several putative APOBEC3G/F target sites. Here, we determined if APOBEC3G/F, in the presence of Vif-defective HIV-1 virus, can induce G-to-A mutations at V3 positions critical to modulation of CXCR4 usage. Peripheral blood mononuclear cells (PBMC) and monocyte-derived macrophages (MDM) from 2 HIV-1-negative donors were infected with CCR5-using 81.A-VifWT virus (i.e., with wild-type [WT] Vif protein), 81.A-VifE45G, or 81.A-VifK22E (known to incompletely/partially neutralize APOBEC3G/F). The rate of G-toA mutations was zero or extremely low in 81.A-VifWT- and 81.A-VifE45G-infected PBMC from both donors. Conversely, G-to-A enrichment was detected in 81.A-VifK22E-infected PBMC (prevalence ranging from 2.18% at 7 days postinfection [dpi] to 3.07% at 21 dpi in donor 1 and from 10.49% at 7 dpi to 8.69% at 21 dpi in donor 2). A similar scenario was found in MDM. G-to-A mutations occurred at 8 V3 positions, resulting in nonsynonymous amino acid substitutions. Of them, G24E and E25K strongly correlated with phenotypically/genotypically defined CXCR4-using viruses (P = 0.04 and 5.5e-7, respectively) and increased the CXCR4 N-terminal binding affinity for V3 (WT, -40.1 kcal/mol; G24E, -510 kcal/mol; E25K, -522 kcal/mol). The analysis of paired V3 and Vif DNA sequences from 84 HIV-1-infected patients showed that the presence of a Vif-defective virus correlated with CXCR4 usage in proviral DNA (P = 0.04). In conclusion, incomplete APOBEC3G/F neutralization by a single Vif amino acid substitution seeds a CXCR4-using proviral reservoir. This can have implications for the success of CCR5 antagonist-based therapy, as well as for the risk of disease progression.
authors.affiliation: University of Rome "Tor Vergata," Department of Experimental Medicine and Surgery, Rome, Italy.
authors.forename: Claudia
authors.initials: C
authors.lastname: Alteri
authors.affiliation: University of Rome "Tor Vergata," Department of Experimental Medicine and Surgery, Rome, Italy.
authors.forename: Matteo
authors.initials: M
authors.lastname: Surdo
authors.affiliation: University of Rome "Tor Vergata," Department of Experimental Medicine and Surgery, Rome, Italy.
authors.forename: Maria Concetta
authors.initials: MC
authors.lastname: Bellocchi
authors.affiliation: University of Rome "Tor Vergata," Department of Experimental Medicine and Surgery, Rome, Italy.
authors.forename: Patrizia
authors.initials: P
authors.lastname: Saccomandi
authors.affiliation: INMI L. Spallanzani, Rome, Italy.
authors.forename: Fabio
authors.initials: F
authors.lastname: Continenza
authors.affiliation: University of Rome "Tor Vergata," Department of Experimental Medicine and Surgery, Rome, Italy.
authors.forename: Daniele
authors.initials: D
authors.lastname: Armenia
authors.affiliation: Università Magna Graecia di Catanzaro, Dipartimento di Scienze della Salute, Campus Universitario, Catanzaro, Italy.
authors.forename: Lucia
authors.initials: L
authors.lastname: Parrotta
authors.affiliation: University of Rome "Tor Vergata," Department of Experimental Medicine and Surgery, Rome, Italy.
authors.forename: Luca
authors.initials: L
authors.lastname: Carioti
authors.affiliation: Università Magna Graecia di Catanzaro, Dipartimento di Scienze della Salute, Campus Universitario, Catanzaro, Italy.
authors.forename: Giosuè
authors.initials: G
authors.lastname: Costa
authors.affiliation: Department of Virology, Hospital "Pitie Salpietrere," Paris, France.
authors.forename: Slim
authors.initials: S
authors.lastname: Fourati
authors.affiliation: University of Rome "Tor Vergata," Department of Experimental Medicine and Surgery, Rome, Italy.
authors.forename: Fabiola
authors.initials: F
authors.lastname: Di Santo
authors.affiliation: University of Rome "Tor Vergata," Department of Experimental Medicine and Surgery, Rome, Italy.
authors.forename: Rossana
authors.initials: R
authors.lastname: Scutari
authors.affiliation: University of Rome "Tor Vergata," Department of Experimental Medicine and Surgery, Rome, Italy.
authors.forename: Silvia
authors.initials: S
authors.lastname: Barbaliscia
authors.affiliation: INMI L. Spallanzani, Rome, Italy.
authors.forename: Valentina
authors.initials: V
authors.lastname: Fedele
authors.affiliation: INMI L. Spallanzani, Rome, Italy.
authors.forename: Stefania
authors.initials: S
authors.lastname: Carta
authors.affiliation: University of Rome "Tor Vergata," Department of Experimental Medicine and Surgery, Rome, Italy.
authors.forename: Emanuela
authors.initials: E
authors.lastname: Balestra
authors.affiliation: Università Magna Graecia di Catanzaro, Dipartimento di Scienze della Salute, Campus Universitario, Catanzaro, Italy.
authors.forename: Stefano
authors.initials: S
authors.lastname: Alcaro
authors.affiliation: Department of Virology, Hospital "Pitie Salpietrere," Paris, France.
authors.forename: Anne Genevieve
authors.initials: AG
authors.lastname: Marcelin
authors.affiliation: Department of Virology, Hospital "Pitie Salpietrere," Paris, France.
authors.forename: Vincent
authors.initials: V
authors.lastname: Calvez
authors.affiliation: University of Rome "Tor Vergata," Department of Experimental Medicine and Surgery, Rome, Italy.
authors.forename: Francesca
authors.initials: F
authors.lastname: Ceccherini-Silberstein
authors.affiliation: Università Magna Graecia di Catanzaro, Dipartimento di Scienze della Salute, Campus Universitario, Catanzaro, Italy.
authors.forename: Anna
authors.initials: A
authors.lastname: Artese
authors.affiliation: University of Rome "Tor Vergata," Department of Experimental Medicine and Surgery, Rome, Italy INMI L. Spallanzani, Rome, Italy valentina.svicher@uniroma2.it cf.perno@uniroma2.it.
authors.forename: Carlo Federico
authors.initials: CF
authors.lastname: Perno
authors.affiliation: University of Rome "Tor Vergata," Department of Experimental Medicine and Surgery, Rome, Italy valentina.svicher@uniroma2.it cf.perno@uniroma2.it.
authors.forename: Valentina
authors.initials: V
authors.lastname: Svicher
mesh_terms: D000071478:APOBEC Deaminases; D000595:Amino Acid Sequence; D019943:Amino Acid Substitution / Q000235:genetics; D001483:Base Sequence; D002460:Cell Line; D003564:Cytidine Deaminase; D043525:Cytosine Deaminase / Q000235:genetics*; D019143:Evolution, Molecular; D057809:HEK293 Cells; D015658:HIV Infections / Q000235:genetics* / Q000821:virology; D015497:HIV-1 / Q000235:genetics*; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000821:virology; D009154:Mutation / Q000235:genetics*; D019713:Receptors, CCR5 / Q000235:genetics*; D019718:Receptors, CXCR4 / Q000235:genetics*
chemical_list: C584933:CCR5 protein, human; C527276:CXCR4 protein, human; D019713:Receptors, CCR5; D019718:Receptors, CXCR4; D043525:Cytosine Deaminase; D000071478:APOBEC Deaminases; C502889:APOBEC3 proteins, human; D003564:Cytidine Deaminase