Work

Next generation sequencing in a direct infection model reveals important parallels to in vivo reservoir dynamics

Year: 2019

Type: article

Abstract: Background: The crosstalk between intestinal epithelial cells (IEC) and Th17 cells is critical for mucosal homeostasis, with HIV causing major alterations in people living with HIV (PLWH).Mucosal Th17 cells are the first targets of infection and contribute to viral reservoir persistence during antiretroviral therapy (ART).Here, we investigated the effects of IL-17A, a Th17 hallmark cytokine, in regulating HIV dissemination/reactivation in a model of IEC:Th17 interaction in vitro.Methods: HIV trans infection was studied upon HT-29 IEC activation with IL-17A and/or TNF, exposure to transmitted founder HIV THRO , and co-culture with CD3/CD28-activated memory CD4+ T-cells from uninfected individuals.HIV replication and outgrowth were studied upon co-culture of cytokine-activated IEC with T cells exposed to HIV THRO in vitro and isolated from ART-treated PLWH, respectively.HIV-p24 levels were measured by FACS/ ELISA.RNA-sequencing was performed on cytokine-activated IEC before/after co-culture with CD3/CD28-activated T-cells from ART-treated PLWH.Type I IFN was measured using the HEK-blue 293T-cell-based assay.Results: The ability of IEC to trans infect T-cells was boosted by IL-17A in the presence/absence of TNF.Also, the highest levels of viral replication and outgrowth were observed when T-cells infected with HIV in vitro and isolated from ART-treated PLWH, respectively, were co-cultured with IL-17A-exposed IEC.Exposure of IEC to IL-17A coincided with decreased type I IFN levels in co-cultures.RNA sequencing revealed potential mechanisms for the IL-17A-mediated proviral effects, including decreased expression of transcripts linked to type I IFN production and response (the viral sensor DDX60, the IFN regulatory factor 7, STAT1) and genes relevant for HIV restriction at different steps of the viral cycle (viral entry: IFITM1; disassembly: TRIM5, transcription: IF2AK2, MX1, MX2; translation: IFIT1,3,5, PARP12; assembly: HERC6; release: ISG15, viperin, BST2) and viral RNA degradation (OAS2/3). Conclusions:In conclusion, IL-17A acts on IEC to create an environment prone to HIV dissemination during primary infection and viral reactivation/outgrowth during the chronic phase, mainly by impairing the type I IFN-mediated antiviral immunity.Thus, new therapeutic interventions aimed at restoring Th17 paucity during ART-treated HIV infection should consider the proviral features of IL-17A.PP 2.12

Source: Journal of Virus Eradication

DOI: 10.1016/s2055-6640(20)30126-6

Authors: U. O’Doherty, M. Pinzone, M.P. Bertuccio, D.J. Vanbelzen

Institutions: University of Pennsylvania, University of Messina, Northwestern University

Language: English

Topic: HIV Research and Treatment

Subfield: Virology

Field: Immunology and Microbiology

Domain: Life Sciences

SDG: Good health and well-being

Open Access status: gold

Estimated APC paid: $2,950