Abrogation of postentry restriction of HIV-1-based lentiviral vector transduction in simian cells

ABSTRACT The Ref1 and Lv1 postentry restrictions in human and monkey cells have been analyzed for lentiviruses in the primate and ungulate groups, but no data exist for the third (feline) group. We compared feline immunodeficiency virus (FIV) to other restricted (human immunodeficiency virus type 1 [HIV-1], equine infectious anemia virus [EIAV]) and unrestricted (NB-tropic murine leukemia virus [NB-MLV]) retroviruses across wide ranges of viral inputs in cells from multiple primate and nonprimate species. We also characterized restrictions conferred to permissive feline and canine cells engineered to express rhesus and human TRIM5α proteins and performed RNA interference (RNAi) against endogenous TRIM5α. We find that expression of rhesus or human TRIM5α proteins in feline cells restricts FIV, impairing pseudotyped vector transduction and viral replication, but rhesus TRIM5α is more restricting than human TRIM5α. Notably, however, canine cells did not support restriction by human TRIM5α and supported minimal restriction by rhesus TRIM5α, suggesting that these proteins may not function autonomously or that a canine factor interferes. Stable RNAi knockdown of endogenous rhesus TRIM5α resulted in marked increases in FIV and HIV-1 infectivities while having no effect on NB-MLV. A panel of nonprimate cell lines varied widely in susceptibility to lentiviral vector transduction, but normalized FIV and HIV-1 vectors varied concordantly. In contrast, in human and monkey cells, relative restriction of FIV compared to HIV-1 varied from none to substantial, with the greatest relative infectivity deficit for FIV vectors observed in human T-cell lines. Endogenous and introduced TRIM5α restrictions of FIV could be titrated by coinfections with FIV, HIV-1, or EIAV virus-like particles. Arsenic trioxide had complex and TRIM5α-independent enhancing effects on lentiviral but not NB-MLV infection. Implications for human gene therapy are discussed.

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563. In Vivo Derivation of HIV-1 Resistant Transgenic T Cells by Lentiviral Vector Transduction of a Triple Combination of Anti-HIV Genes Anti-CCR5 Ribozyme, Tat-Rev siRNA and a TAR Decoy

Molecular Therapy ◽

10.1016/s1525-0016(16)44769-6 ◽

2007 ◽

Vol 15 ◽

pp. S217

Keyword(s):

T Cells ◽

Lentiviral Vector ◽

Triple Combination ◽

Anti Hiv ◽

Hiv 1 ◽

Vector Transduction

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384. Abrogation of Post-Entry Restriction of HIV-1 Based Lentiviral Vector Transduction in Simian Cells. Implications for Gene Therapy

Molecular Therapy ◽

10.1016/s1525-0016(16)40826-9 ◽

2003 ◽

Vol 7 (5) ◽

pp. S150

Keyword(s):

Gene Therapy ◽

Lentiviral Vector ◽

Post Entry ◽

Hiv 1 ◽

Vector Transduction

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Construction and identification of recombinant lentiviral vector containing HIV-1 Tat gene and its expression in 293T cells

Journal of Biomedical Research ◽

10.1016/s1674-8301(10)60009-7 ◽

2010 ◽

Vol 24 (1) ◽

pp. 58-63 ◽

Cited By ~ 6

Author(s):

Bingbing Wei ◽

Ninghan Feng ◽

Feng Zhou ◽

Chun Lu ◽

Jiantang Su ◽

...

Keyword(s):

Lentiviral Vector ◽

Hiv 1

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ZNF521 Represses Osteoblastic Differentiation in Human Adipose-Derived Stem Cells

International Journal of Molecular Sciences ◽

10.3390/ijms19124095 ◽

2018 ◽

Vol 19 (12) ◽

pp. 4095 ◽

Cited By ~ 8

Author(s):

Emanuela Chiarella ◽

Annamaria Aloisio ◽

Stefania Scicchitano ◽

Valeria Lucchino ◽

Ylenia Montalcini ◽

...

Keyword(s):

Stem Cells ◽

Zinc Finger Protein ◽

Lentiviral Vector ◽

Adipogenic Differentiation ◽

Bone Matrix ◽

Osteoblastic Differentiation ◽

Adipose Derived Stem Cells ◽

Matrix Mineralization ◽

Calcium Deposits ◽

Vector Transduction

Human adipose-derived stem cells (hADSCs) are multipotent mesenchymal cells that can differentiate into adipocytes, chondrocytes, and osteocytes. During osteoblastogenesis, the osteoprogenitor cells differentiate into mature osteoblasts and synthesize bone matrix components. Zinc finger protein 521 (ZNF521/Zfp521) is a transcription co-factor implicated in the regulation of hematopoietic, neural, and mesenchymal stem cells, where it has been shown to inhibit adipogenic differentiation. The present study is aimed at determining the effects of ZNF521 on the osteoblastic differentiation of hADSCs to clarify whether it can influence their osteogenic commitment. The enforced expression or silencing of ZNF521 in hADSCs was achieved by lentiviral vector transduction. Cells were cultured in a commercial osteogenic medium for up to 20 days. The ZNF521 enforced expression significantly reduced osteoblast development as assessed by the morphological and molecular criteria, resulting in reduced levels of collagen I, alkaline phosphatase, osterix, osteopontin, and calcium deposits. Conversely, ZNF521 silencing, in response to osteoblastic stimuli, induced a significant increase in early molecular markers of osteogenesis and, at later stages, a remarkable enhancement of matrix mineralization. Together with our previous findings, these results show that ZNF521 inhibits both adipocytic and osteoblastic maturation in hADSCs and suggest that its expression may contribute to maintaining the immature properties of hADSCs.

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434. Production of High-Quality TCR-Gene Modified T Cells Using siTCR Lentiviral Vector Transduction in Combination With Anti-CD3/CH296 Stimulation

Molecular Therapy ◽

10.1016/s1525-0016(16)35447-8 ◽

2014 ◽

Vol 22 ◽

pp. S166

Keyword(s):

T Cells ◽

Lentiviral Vector ◽

High Quality ◽

Vector Transduction

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Inhibition of HIV-1 by a Lentiviral Vector with a Novel Tat-Inducible Expression System and a Specific Tropism to the Target Cells

Human Gene Therapy ◽

10.1089/hum.2015.031 ◽

2015 ◽

Vol 26 (10) ◽

pp. 680-687 ◽

Cited By ~ 2

Author(s):

Touraj Farazmandfar ◽

Mohammad Reza Haghshenas ◽

Majid Shahbazi

Keyword(s):

Lentiviral Vector ◽

Expression System ◽

Inducible Expression ◽

Target Cells ◽

Inducible Expression System ◽

Hiv 1

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Immunogenicity, safety, and efficacy of sequential immunizations with an SIV-based IDLV expressing CH505 Envs

npj Vaccines ◽

10.1038/s41541-020-00252-w ◽

2020 ◽

Vol 5 (1) ◽

Author(s):

Maria Blasi ◽

Donatella Negri ◽

Kevin O. Saunders ◽

Erich J. Baker ◽

Hannah Stadtler ◽

...

Keyword(s):

Immune Responses ◽

Neutralizing Antibodies ◽

Lentiviral Vector ◽

Rhesus Macaques ◽

Infected Individual ◽

Neutralizing Antibody ◽

Antibody Responses ◽

Safety And Efficacy ◽

Env Protein ◽

Hiv 1

AbstractA preventative HIV-1 vaccine is an essential intervention needed to halt the HIV-1 pandemic. Neutralizing antibodies protect against HIV-1 infection in animal models, and thus an approach toward a protective HIV-1 vaccine is to induce broadly cross-reactive neutralizing antibodies (bnAbs). One strategy to achieve this goal is to define envelope (Env) evolution that drives bnAb development in infection and to recreate those events by vaccination. In this study, we report the immunogenicity, safety, and efficacy in rhesus macaques of an SIV-based integrase defective lentiviral vector (IDLV) expressing sequential gp140 Env immunogens derived from the CH505 HIV-1-infected individual who made the CH103 and CH235 bnAb lineages. Immunization with IDLV expressing sequential CH505 Envs induced higher magnitude and more durable binding and neutralizing antibody responses compared to protein or DNA +/− protein immunizations using the same sequential envelopes. Compared to monkeys immunized with a vector expressing Envs alone, those immunized with the combination of IDLV expressing Env and CH505 Env protein demonstrated improved durability of antibody responses at six months after the last immunization as well as lower peak viremia and better virus control following autologous SHIV-CH505 challenge. There was no evidence of vector mobilization or recombination in the immunized and challenged monkeys. Although the tested vaccines failed to induce bnAbs and to mediate significant protection following SHIV-challenge, our results show that IDLV proved safe and successful at inducing higher titer and more durable immune responses compared to other vaccine platforms.

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