HSV-2 triggers upregulation of MALAT1 in CD4+ T cells and promotes HIV latency reversal
Carl A. Pierce, Lip Nam Loh, Holly R. Steach, Natalia Cheshenko, Paula Preston-Hurlburt, Fengrui Zhang, Stephanie Stransky, Leah Kravets, Simone Sidoli, William Philbrick, Michel Nassar, Smita Krishnaswamy, Kevan C. Herold, Betsy C. Herold
Carl A. Pierce, Lip Nam Loh, Holly R. Steach, Natalia Cheshenko, Paula Preston-Hurlburt, Fengrui Zhang, Stephanie Stransky, Leah Kravets, Simone Sidoli, William Philbrick, Michel Nassar, Smita Krishnaswamy, Kevan C. Herold, Betsy C. Herold
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Research Article AIDS/HIV Virology

HSV-2 triggers upregulation of MALAT1 in CD4+ T cells and promotes HIV latency reversal

Abstract

Herpes simplex virus type 2 (HSV-2) coinfection is associated with increased HIV-1 viral loads and expanded tissue reservoirs, but the mechanisms are not well defined. HSV-2 recurrences result in an influx of activated CD4+ T cells to sites of viral replication and an increase in activated CD4+ T cells in peripheral blood. We hypothesized that HSV-2 induces changes in these cells that facilitate HIV-1 reactivation and replication and tested this hypothesis in human CD4+ T cells and 2D10 cells, a model of HIV-1 latency. HSV-2 promoted latency reversal in HSV-2–infected and bystander 2D10 cells. Bulk and single-cell RNA-Seq studies of activated primary human CD4+ T cells identified decreased expression of HIV-1 restriction factors and increased expression of transcripts including MALAT1 that could drive HIV replication in both the HSV-2–infected and bystander cells. Transfection of 2D10 cells with VP16, an HSV-2 protein that regulates transcription, significantly upregulated MALAT1 expression, decreased trimethylation of lysine 27 on histone H3 protein, and triggered HIV latency reversal. Knockout of MALAT1 from 2D10 cells abrogated the response to VP16 and reduced the response to HSV-2 infection. These results demonstrate that HSV-2 contributes to HIV-1 reactivation through diverse mechanisms, including upregulation of MALAT1 to release epigenetic silencing.

Authors

Carl A. Pierce, Lip Nam Loh, Holly R. Steach, Natalia Cheshenko, Paula Preston-Hurlburt, Fengrui Zhang, Stephanie Stransky, Leah Kravets, Simone Sidoli, William Philbrick, Michel Nassar, Smita Krishnaswamy, Kevan C. Herold, Betsy C. Herold

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Figure 7

HSV-2–induced reactivation is decreased in the absence of MALAT1.

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HSV-2–induced reactivation is decreased in the absence of MALAT1. (A) 2D...
(A) 2D10 cells were exposed to TNF (10 ng/mL), romidepsin (6.25 nM), or HSV-2(SD90) (MOI = 1 PFU/cell) or mock-treated, and MALAT1 expression was determined 24 hours after treatment. Results are expressed as log10 fold change (FC) relative to mock-treated cells (n = 3 independent experiments, **P < 0.01, ****P < 0.0001, compared with mock-treated cells by 1-way ANOVA). (B) 2D10 or ΔMALAT1 cells were exposed to HSV-2(SD90) (MOI = 1 PFU/cell) and HSV-2 ICP0 expression determined 24 hpi as log10 fold change relative to mock-infected cells. (C and D) 2D10 or ΔMALAT1 cells were infected with HSV-2(SD90) (MOI = 1 PFU/cell) (C) or treated with TNF or romidepsin (D), and HIV ltr expression was determined 24 hours after treatment. Results are presented as fold change in gene expression relative to mock-treated cells. ****P < 0.0001, unpaired t test comparing 2D10 vs. ΔMALAT1 cells. (E) 2D10 cells were infected with live or UV-inactivated HSV-2(G) (MOI = 1), treated with romidepsin (6.25 nM), or left untreated (UT), and after 24 hours of incubation, the cells were harvested, nuclei isolated, and HDAC activity measured using a colorimetric HDAC activity assay. (F) Primary CD4+ T cells were stimulated by CD3/CD28 cross-linking for 72 hours and infected with HSV-2(G) (MOI = 1), treated with romidepsin, or left untreated (UT). HDAC activity was assayed as in E. **P < 0.01, ****P < 0.0001 relative to untreated cells, 1-way ANOVA.