In vivo Serial Passaging of Human–Simian Immunodeficiency Virus Clones Identifies Characteristics for Persistent Viral Replication

We previously reported that a human immunodeficiency virus type 1 with a simian immunodeficiency virus vif substitution (HSIV-vifNL4-3) could replicate in pigtailed macaques (PTMs), demonstrating that Vif is a species-specific tropism factor of primate lentiviruses. However, infections did not resul...

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Autores principales:	Rajesh Thippeshappa, Patricia Polacino, Shaswath S. Chandrasekar, Khanghy Truong, Anisha Misra, Paula C. Aulicino, Shiu-Lok Hu, Deepak Kaushal, Jason T. Kimata
Formato:	article
Lenguaje:	EN
Publicado:	Frontiers Media S.A. 2021
Materias:	HIV-1 SIV HSIV-vif nonhuman primates pigtailed macaques animal model Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/89b431c456c245b6913d3f3f0bc88295
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spelling	oai:doaj.org-article:89b431c456c245b6913d3f3f0bc882952021-11-18T05:31:53ZIn vivo Serial Passaging of Human–Simian Immunodeficiency Virus Clones Identifies Characteristics for Persistent Viral Replication1664-302X10.3389/fmicb.2021.779460https://doaj.org/article/89b431c456c245b6913d3f3f0bc882952021-11-01T00:00:00Zhttps://www.frontiersin.org/articles/10.3389/fmicb.2021.779460/fullhttps://doaj.org/toc/1664-302XWe previously reported that a human immunodeficiency virus type 1 with a simian immunodeficiency virus vif substitution (HSIV-vifNL4-3) could replicate in pigtailed macaques (PTMs), demonstrating that Vif is a species-specific tropism factor of primate lentiviruses. However, infections did not result in high-peak viremia or setpoint plasma viral loads, as observed during simian immunodeficiency virus (SIV) infection of PTMs. Here, we characterized variants isolated from one of the original infected animals with CD4 depletion after nearly 4years of infection to identify determinants of increased replication fitness. In our studies, we found that the HSIV-vif clones did not express the HIV-1 Vpr protein due to interference from the vpx open reading frame (ORF) in singly spliced vpr mRNA. To examine whether these viral genes contribute to persistent viral replication, we generated infectious HSIV-vif clones expressing either the HIV-1 Vpr or SIV Vpx protein. And then to determine viral fitness determinants of HSIV-vif, we conducted three rounds of serial in vivo passaging in PTMs, starting with an initial inoculum containing a mixture of CXCR4-tropic [Vpr-HSIV-vifNL4-3 isolated at 196 (C/196) and 200 (C/200) weeks post-infection from a PTM with depressed CD4 counts] and CCR5-tropic HSIV (Vpr+ HSIV-vif derivatives based NL-AD8 and Bru-Yu2 and a Vpx expressing HSIV-vifYu2). Interestingly, all infected PTMs showed peak plasma viremia close to or above 105 copies/ml and persistent viral replication for more than 20weeks. Infectious molecular clones (IMCs) recovered from the passage 3 PTM (HSIV-P3 IMCs) included mutations required for HIV-1 Vpr expression and those mutations encoded by the CXCR4-tropic HSIV-vifNL4-3 isolate C/196. The data indicate that the viruses selected during long-term infection acquired HIV-1 Vpr expression, suggesting the importance of Vpr for in vivo pathogenesis. Further passaging of HSIV-P3 IMCs in vivo may generate pathogenic variants with higher replication capacity, which will be a valuable resource as challenge virus in vaccine and cure studies.Rajesh ThippeshappaPatricia PolacinoShaswath S. ChandrasekarKhanghy TruongAnisha MisraPaula C. AulicinoShiu-Lok HuShiu-Lok HuDeepak KaushalJason T. KimataFrontiers Media S.A.articleHIV-1SIVHSIV-vifnonhuman primatespigtailed macaquesanimal modelMicrobiologyQR1-502ENFrontiers in Microbiology, Vol 12 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	HIV-1 SIV HSIV-vif nonhuman primates pigtailed macaques animal model Microbiology QR1-502
spellingShingle	HIV-1 SIV HSIV-vif nonhuman primates pigtailed macaques animal model Microbiology QR1-502 Rajesh Thippeshappa Patricia Polacino Shaswath S. Chandrasekar Khanghy Truong Anisha Misra Paula C. Aulicino Shiu-Lok Hu Shiu-Lok Hu Deepak Kaushal Jason T. Kimata In vivo Serial Passaging of Human–Simian Immunodeficiency Virus Clones Identifies Characteristics for Persistent Viral Replication
description	We previously reported that a human immunodeficiency virus type 1 with a simian immunodeficiency virus vif substitution (HSIV-vifNL4-3) could replicate in pigtailed macaques (PTMs), demonstrating that Vif is a species-specific tropism factor of primate lentiviruses. However, infections did not result in high-peak viremia or setpoint plasma viral loads, as observed during simian immunodeficiency virus (SIV) infection of PTMs. Here, we characterized variants isolated from one of the original infected animals with CD4 depletion after nearly 4years of infection to identify determinants of increased replication fitness. In our studies, we found that the HSIV-vif clones did not express the HIV-1 Vpr protein due to interference from the vpx open reading frame (ORF) in singly spliced vpr mRNA. To examine whether these viral genes contribute to persistent viral replication, we generated infectious HSIV-vif clones expressing either the HIV-1 Vpr or SIV Vpx protein. And then to determine viral fitness determinants of HSIV-vif, we conducted three rounds of serial in vivo passaging in PTMs, starting with an initial inoculum containing a mixture of CXCR4-tropic [Vpr-HSIV-vifNL4-3 isolated at 196 (C/196) and 200 (C/200) weeks post-infection from a PTM with depressed CD4 counts] and CCR5-tropic HSIV (Vpr+ HSIV-vif derivatives based NL-AD8 and Bru-Yu2 and a Vpx expressing HSIV-vifYu2). Interestingly, all infected PTMs showed peak plasma viremia close to or above 105 copies/ml and persistent viral replication for more than 20weeks. Infectious molecular clones (IMCs) recovered from the passage 3 PTM (HSIV-P3 IMCs) included mutations required for HIV-1 Vpr expression and those mutations encoded by the CXCR4-tropic HSIV-vifNL4-3 isolate C/196. The data indicate that the viruses selected during long-term infection acquired HIV-1 Vpr expression, suggesting the importance of Vpr for in vivo pathogenesis. Further passaging of HSIV-P3 IMCs in vivo may generate pathogenic variants with higher replication capacity, which will be a valuable resource as challenge virus in vaccine and cure studies.
format	article
author	Rajesh Thippeshappa Patricia Polacino Shaswath S. Chandrasekar Khanghy Truong Anisha Misra Paula C. Aulicino Shiu-Lok Hu Shiu-Lok Hu Deepak Kaushal Jason T. Kimata
author_facet	Rajesh Thippeshappa Patricia Polacino Shaswath S. Chandrasekar Khanghy Truong Anisha Misra Paula C. Aulicino Shiu-Lok Hu Shiu-Lok Hu Deepak Kaushal Jason T. Kimata
author_sort	Rajesh Thippeshappa
title	In vivo Serial Passaging of Human–Simian Immunodeficiency Virus Clones Identifies Characteristics for Persistent Viral Replication
title_short	In vivo Serial Passaging of Human–Simian Immunodeficiency Virus Clones Identifies Characteristics for Persistent Viral Replication
title_full	In vivo Serial Passaging of Human–Simian Immunodeficiency Virus Clones Identifies Characteristics for Persistent Viral Replication
title_fullStr	In vivo Serial Passaging of Human–Simian Immunodeficiency Virus Clones Identifies Characteristics for Persistent Viral Replication
title_full_unstemmed	In vivo Serial Passaging of Human–Simian Immunodeficiency Virus Clones Identifies Characteristics for Persistent Viral Replication
title_sort	in vivo serial passaging of human–simian immunodeficiency virus clones identifies characteristics for persistent viral replication
publisher	Frontiers Media S.A.
publishDate	2021
url	https://doaj.org/article/89b431c456c245b6913d3f3f0bc88295
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In vivo Serial Passaging of Human–Simian Immunodeficiency Virus Clones Identifies Characteristics for Persistent Viral Replication

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