V3-Independent Competitive Resistance of a Dual-X4 HIV-1 to the CXCR4 Inhibitor AMD3100

Maeda, Yosuke; Terasawa, Hiromi; Nakano, Yoshiaki; Monde, Kazuaki; Yusa, Kosuke; Oka, Shinichi; Takiguchi, Masafumi; Harada, Shinji

doi:10.1371/journal.pone.0089515

Cited by 2 publications

(2 citation statements)

References 55 publications

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“…Pseudotyped HIV-1 production and determination of coreceptor usage. For coreceptor usage by HIV-1 that carries a V3 region from plasma vRNA, an Env expression vector to carry the V3 loop from the plasma vRNA was constructed as previously described (38,66,(75)(76)(77). The V3-spanning region that carries AflII and NheI sites was amplified from the first PCR product for the V3 sequencing as mentioned above, using forward primer 5=-GCACCTTAAGAAATCTGTAGAAATCAATTG-3= (812.7AF1) and reverse primer 5=-GCTAGCTACCTGTTTTAAAGCTTTATACC-3= (812.7NR1) (underlines denote the AflII and NheI site, respectively) (75).…”

Section: Methodsmentioning

confidence: 99%

“…The AflII-NheI-carrying fragment of the V3 region was then introduced into the AflII-NheI cloning site of the pCXN-FLan vector as previously described (39,75). A luciferase reporter HIV-1 pseudotyped with Env carrying the V3 region or full-length Env was prepared by transfecting 293T cells with pNL-LucΔBglII and each Env expression vector (25,38,(75)(76)(77). The virus-containing culture supernatant was recovered and filtered through a 0.45-m filter and then stored at Ϫ80°C until use.…”

Section: Methodsmentioning

confidence: 99%

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Existence of Replication-Competent Minor Variants with Different Coreceptor Usage in Plasma from HIV-1-Infected Individuals

Maeda

Takemura

Chikata

et al. 2020

J Virol

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Cell entry by HIV-1 is mediated by its principal receptor, CD4, and a coreceptor, either CCR5 or CXCR4, with viral envelope glycoprotein gp120. Generally, CCR5-using HIV-1 variants, called R5, predominate over most of the course of infection, while CXCR4-using HIV-1 variants (variants that utilize both CCR5 and CXCR4 [R5X4, or dual] or CXCR4 alone [X4]) emerge at late-stage infection in half of HIV-1-infected individuals and are associated with disease progression. Although X4 variants also appear during acute-phase infection in some cases, these variants apparently fall to undetectable levels thereafter. In this study, replication-competent X4 variants were isolated from plasma of drug treatment-naive individuals infected with HIV-1 strain CRF01_AE, which dominantly carries viral RNA (vRNA) of R5 variants. Next-generation sequencing (NGS) confirmed that sequences of X4 variants were indeed present in plasma vRNA from these individuals as a minor population. On the other hand, in one individual with a mixed infection in which X4 variants were dominant, only R5 replication-competent variants were isolated from plasma. These results indicate the existence of replication-competent variants with different coreceptor usage as minor populations. IMPORTANCE The coreceptor switch of HIV-1 from R5 to CXCR4-using variants (R5X4 or X4) has been observed in about half of HIV-1-infected individuals at late-stage infection with loss of CD4 cell count and disease progression. However, the mechanisms that underlie the emergence of CXCR4-using variants at this stage are unclear. In the present study, CXCR4-using X4 variants were isolated from plasma samples of HIV-1-infected individuals that dominantly carried vRNA of R5 variants. The sequences of the X4 variants were detected as a minor population using next-generation sequencing. Taken together, CXCR4-using variants at late-stage infection are likely to emerge when replication-competent CXCR4-using variants are maintained as a minor population during the course of infection. The present study may support the hypothesis that R5-to-X4 switching is mediated by the expansion of preexisting X4 variants in some cases.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Existence of Replication-Competent Minor Variants with Different Coreceptor Usage in Plasma from HIV-1-Infected Individuals

Maeda

Takemura

Chikata

et al. 2020

J Virol

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Separate Cellular Localizations of Human T-Lymphotropic Virus 1 (HTLV-1) Env and Glucose Transporter Type 1 (GLUT1) Are Required for HTLV-1 Env-Mediated Fusion and Infection

Maeda

Terasawa

Tanaka

et al. 2015

J Virol

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Interaction of the envelope glycoprotein (Env) of human T-lymphotropic virus 1 (HTLV-1) with the glucose transporter type 1 (GLUT1) expressed in target cells is essential for viral entry. This study found that the expression level of GLUT1 in virus-producing 293T cells was inversely correlated with HTLV-1 Env-mediated fusion activity and infectivity. Chimeric studies between GLUT1 and GLUT3 indicated that the extracellular loop 6 (ECL6) of GLUT1 is important for the inhibition of cell-cell fusion mediated by Env. When GLUT1 was translocated into the plasma membrane from intracellular storage sites by bafilomycin A1 (BFLA1) treatment in 293T cells, HTLV-1 Env-mediated cell fusion and infection also were inhibited without the overexpression of GLUT1, indicating that the localization of GLUT1 in intracellular compartments rather than in the plasma membrane is crucial for the fusion activity of HTLV-1 Env. Immunoprecipitation and laser scanning confocal microscopic analyses indicated that under normal conditions, HTLV-1 Env and GLUT1 do not colocalize or interact. BFLA1 treatment induced this colocalization and interaction, indicating that GLUT1 normally accumulates in intracellular compartments separate from that of Env. Western blot analyses of FLAGtagged HTLV-1 Env in virus-producing cells and the incorporation of HTLV-1 Env in virus-like particles (VLPs) indicate that the processing of Env is inhibited by either overexpression of GLUT1 or BFLA1 treatment in virus-producing 293T cells. This inhibition probably is due to the interaction of the Env with GLUT1 in intracellular compartments. Taken together, separate intracellular localizations of GLUT1 and HTLV-1 Env are required for the fusion activity and infectivity of HTLV-1 Env. IMPORTANCEThe deltaretrovirus HTLV-1 is a causative agent of adult T-cell leukemia (ATL) and HTLV-1-associated myelopathy/tropical spastic paraparesis (HAM/TSP). Although HTLV-1 is a complex retrovirus that has accessory genes, no HTLV-1 gene product has yet been shown to regulate its receptor GLUT1 in virus-producing cells. In this study, we found that a large amount of GLUT1 or translocation of GLUT1 to the plasma membrane from intracellular compartments in virus-producing cells enhances the colocalization and interaction of GLUT1 with HTLV-1 Env, leading to the inhibition of cell fusion activity and infectivity. The results of our study suggest that GLUT1 normally accumulates in separate intracellular compartments from Env, which is indeed required for the proper processing of Env. Human T-lymphotropic virus 1 (HTLV-1) is a complex deltaretrovirus and a causative agent of adult T-cell leukemia (ATL) (62-64) and HTLV-1-associated myelopathy/tropical spastic paraparesis (HAM/TSP) (1, 2). The envelope glycoprotein (Env) of HTLV-1 is synthesized in virus-infected cells as a polyprotein precursor (gp62), which subsequently is cleaved by cellular proteinase(s) localized in the Golgi apparatus into two proteins, surface glycoprotein (gp46; SU) and transmembrane glycoprotein (gp21; TM...

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V3-Independent Competitive Resistance of a Dual-X4 HIV-1 to the CXCR4 Inhibitor AMD3100

Cited by 2 publications

References 55 publications

Existence of Replication-Competent Minor Variants with Different Coreceptor Usage in Plasma from HIV-1-Infected Individuals

Existence of Replication-Competent Minor Variants with Different Coreceptor Usage in Plasma from HIV-1-Infected Individuals

Separate Cellular Localizations of Human T-Lymphotropic Virus 1 (HTLV-1) Env and Glucose Transporter Type 1 (GLUT1) Are Required for HTLV-1 Env-Mediated Fusion and Infection

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