A New Indicator Cell Line to Monitor Human Foamy Virus Infection and Stability in vitro

Li, Zhi; Yang, Ping; Liu, Hui; Li, Wenxin

doi:10.1159/000063232

Cited by 8 publications

(3 citation statements)

References 16 publications

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“…The PFV Tas protein can activate the LTR promoter, resulting in the expression of EGFP. Therefore, the viral titer was assessed by the indicator cells with EGFP fluorescence; the procedure has been described previously [ 29 ].…”

Section: Methodsmentioning

confidence: 99%

The DACH1 Gene Transcriptional Activation and Protein Degradation Mediated by Transactivator Tas of Prototype Foamy Virus

Ma,

Wei,

Song

et al. 2023

Viruses

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Foamy viruses are members of the Retroviridae family’s Spumaretrovirinae subfamily. They induce cell vacuolation and exhibit a foamy pathogenic impact after infecting cells. DACH1 (dachshund family transcription factor 1) is a crucial cytokine linked to tumor development, and is associated with the growth of many different malignant tumor cells. Additionally, DACH1 suppresses pancreatic cell proliferation and is involved in diabetes insulin signaling. Prototype foamy viruses (PFVs) were used for the investigation of the regulatory mechanism of FVs on cellular DACH1 expression. The results show that DACH1 expression in PFV-infected cells was inconsistent at both the transcriptional and protein levels. At the transcriptional level, DACH1 was significantly activated by PFV transactivator Tas, and dual-luciferase reporter gene tests, EMSA, and ChIP assays found a Tas response element of 21 nucleotides in the DACH1 promoter. PFV and Tas did not boost the levels of DACH1 protein in a manner consistent with the high levels of DACH1 transcription expression. It was noted that Tas increased the expression of the Ser/Thr protein phosphatase PPM1E, causing PPM1E-mediated post-translational SUMOylation alterations of DACH1 to prompt DACH1 to degrade. The reason for DACH1 protein degradation is that DACH1 inhibits PFV replication. To sum up, these findings show that PFV upregulated the transcription of DACH1, while urging its protein into PPM1E-mediated SUMOylation, to eliminate the adverse effect of DACH1 overexpression of host cells on viral replication and promote virus survival.

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Section: Methodsmentioning

confidence: 99%

The DACH1 Gene Transcriptional Activation and Protein Degradation Mediated by Transactivator Tas of Prototype Foamy Virus

Ma,

Wei,

Song

et al. 2023

Viruses

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“…Other methods, such as RT-PCR, which has been reported for the detection of prototype foamy viruses [11,12] , does not give the direct quantitation on viral loading during infection. Recently, a new method called the green fluorescent protein (GFP)-based indicator cell line was established for the titration of prototype foamy virus (PFV) and feline foamy virus (FeFV) [13,14] . The indicator cell line was constructed by the stable integration of the GFP gene under the control of the foamy virus LTR promoter.…”

Section: Introductionmentioning

confidence: 99%

Detection and analysis of bovine foamy virus infection by an indicator cell line

Qiao

Xuan

et al. 2007

Acta Pharmacologica Sinica

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Aim: To determine the infectivity and replication strategy of bovine foamy virus (BFV) in different cultured cells using the BFV indicator cell line (BICL) system. Methods: BFV infection was induced by the co‐culture method or the transient transfection of the infectious BFV plasmid [pCMV (cytomegalovirus) ‐ BFV] clone. The infectivity of BFV was monitored by the percentage of green fluorescent protein‐positive cells in the BICL. The effect of reverse transcriptase inhibitor zidovudine (AZT) on BFV replication was also evaluated in the BICL. Results: The titer of BFV in fetal bovine lung cells was 4‐5‐folds more than that in either 293T or HeLa (Cells from Henrietta lacks) cells using the co‐culture method, and in the meantime was significantly higher than that produced by the infectious clone pCMV‐BFV in the same cells. AZT had only a minor effect on viral titers when added to cells prior to the virus infection. In contrast, viral titers reduced sharply to the level of the negative control when the virus was produced from cells in the presence of AZT. Conclusions: BICL can be used for the titration of the BFV viral infection in non‐cytopathic condition. In addition, we provide important evidence to show that reverse transcription is essential for BFV replication at a late step of viral infection.

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“…Since the vector inoculum was not removed and since FVs are stable in cell culture medium, 32 low amounts of input vector were still detectable in the first and second passages in some of the experiments described here. After the 5th (#8) and 10th (#2) passages of the SIN vector-transduced cells, FFV infectivities greater than 10 2 FFU/ml were clearly detectable (Figure 2a).…”

Section: Introductionmentioning

confidence: 99%

Kinetics and characteristics of replication-competent revertants derived from self-inactivating foamy virus vectors

Bastone

Löchelt

2004

Gene Ther

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In this study, self-inactivating (SIN) retroviral vectors based on feline foamy virus (FFV) were constructed and analysed. The FFV SIN vectors were devoid of the core FFV long terminal repeat promoter plus upstream sequences but contained all structural and regulatory genes. This design allowed sensitive detection of replication-competent revertants (RCRs). The FFV SIN vectors efficiently transduced the green fluorescence protein into recipient cells. However, RCRs appeared after serial passages of transduced cells. In all RCR clones analysed, parts of the heterologous cytomegalovirus immediate early promoter, originally driving expression of the FFV vector genome, were taken up to restore the deleted SIN promoter function required for replication competence. The RCRs were strongly reduced in replication capacity compared with the parental replication-competent vectors containing the FFV promoter. In all RCR genomes analysed, the uptake of the heterologous promoter was accompanied by deletion of almost the complete marker gene. Although the RCRs described in this study may not have the capacity to spread in humans and animals, they may pose a theoretical risk, for instance during transduction of haematopoietic stem cells. Thus, FV-based SIN vectors require additional genetic modifications in order to avoid RCRs.

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A New Indicator Cell Line to Monitor Human Foamy Virus Infection and Stability in vitro

Cited by 8 publications

References 16 publications

The DACH1 Gene Transcriptional Activation and Protein Degradation Mediated by Transactivator Tas of Prototype Foamy Virus

The DACH1 Gene Transcriptional Activation and Protein Degradation Mediated by Transactivator Tas of Prototype Foamy Virus

Detection and analysis of bovine foamy virus infection by an indicator cell line

Kinetics and characteristics of replication-competent revertants derived from self-inactivating foamy virus vectors

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