Development of 5' LTR DNA methylation of latent HIV-1 provirus in cell line models and in long-term-infected individuals

Clinical Epigenetics
Katerina TrejbalovaJiri Hejnar

Abstract

Human immunodeficiency virus type 1 (HIV-1) latency represents the major barrier to virus eradication in infected individuals because cells harboring latent HIV-1 provirus are not affected by current antiretroviral therapy (ART). We previously demonstrated that DNA methylation of HIV-1 long terminal repeat (5' LTR) restricts HIV-1 reactivation and, together with chromatin conformation, represents an important mechanism of HIV-1 latency maintenance. Here, we explored the new issue of temporal development of DNA methylation in latent HIV-1 5' LTR. In the Jurkat CD4(+) T cell model of latency, we showed that the stimulation of host cells contributed to de novo DNA methylation of the latent HIV-1 5' LTR sequences. Consecutive stimulations of model CD4(+) T cell line with TNF-α and PMA or with SAHA contributed to the progressive accumulation of 5' LTR DNA methylation. Further, we showed that once established, the high DNA methylation level of the latent 5' LTR in the cell line model was a stable epigenetic mark. Finally, we explored the development of 5' LTR DNA methylation in the latent reservoir of HIV-1-infected individuals who were treated with ART. We detected low levels of 5' LTR DNA methylation in the resting CD4(+) T cells o...Continue Reading

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Citations

Oct 7, 2016·Folia Microbiologica·Zora MelkovaJosef Bodor
Mar 5, 2017·Virus Genes·Bizhan Romani, Elham Allahbakhshi
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Methods Mentioned

BETA
PMA
transfection
acetylation
FACS
flow cytometry
Assay
density gradient centrifugation
PCR
PCRs

Software Mentioned

CFX Manager
Quma ( Quantification tool for Methylation Analysis )
FlowJo
GraphPad Prism
R

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