Hepatitis C Virus Influences HIV-1 Viral Splicing in Coinfected Patients

Abstract

<jats:p>Coinfection with hepatitis C virus (HCV) influences HIV reservoir size. However, it is unknown whether this coinfection also induces a higher provirus transcription. Viral transcription is promoted by synergy between cellular factors such as NF-κB and the viral regulator Tat. The impact of HCV coinfection on HIV provirus transcription was analyzed in resting (r)CD4 T+ cells (CD3+CD4+CD25-CD69-HLADR-) and rCD4 T cells-depleted PBMCs (rCD4 T- PBMCs) from a multicenter cross-sectional study of 115 cART-treated HIV patients: 42 HIV+/HCV+ coinfected individuals (HIV+/HCV+), 34 HIV+ patients with HCV spontaneous clearance (HIV+/HCV−) and 39 HIV patients (HIV+). Viral transcription was assessed in total RNA through the quantification of unspliced, single spliced, and multiple spliced viral mRNAs by qPCR. Linear correlations between viral reservoir size and viral splicing were determined. A 3-fold increase of multiple spliced transcripts in rCD4 T+ cells of HIV+/HCV+ patients was found compared to HIV+ individuals (p &lt; 0.05). As Tat is synthesized by multiple splicing, the levels of Tat were also quantified in these patients. Significant differences in single and multiple spliced transcripts were also observed in rCD4 T- PBMCs. Levels of multiple spliced mRNAs were increased in rCD4 T+ cells isolated from HIV+/HCV+ subjects, which could indicate a higher Tat activity in these cells despite their resting state.</jats:p>

Dynamics of HIV Reservoir and HIV-1 Viral Splicing in HCV-Exposed Individuals after Elimination with DAAs or Spontaneous Clearance

“…Previously, our group described how HIV/HCV coinfection increases HIV reservoir size in rCD4+ T cells from HCV spontaneous clarifiers (HIV+/HCV-) and HCV chronic-infected individuals (HIV+/HCV+), in comparison with HIV-monoinfected individuals [19]. We also disclosed that HIV+/HCV+-coinfected subjects showed an increase in multiple-spliced transcripts compared to HIV-monoinfected PLWH [20]. Both findings suggest that the elimination of the HIV viral reservoir in HCV-exposed subjects may be hampered.…”

“…Viral-splicing forms were quantified in rCD4+ T cells and rCD4 T-PBMCs by q-RT-PCR, as described previously [20,23]. Briefly, 250 ng of extracted total RNA were used in a master mix containing 5× buffer, 25 mM ClMg, dNTPs, RNAsin, and retrotranscriptase.…”

Interferon-Regulated Expression of Cellular Splicing Factors Modulates Multiple Levels of HIV-1 Gene Expression and Replication

“…In both groups, we also observed higher ISG15 expression levels, confirming hnRNP A0 as an IFN-I-repressed cellular splicing factor. Interestingly, co-infections exacerbating the inflammatory state exhibited elevated expression and altered HIV-1 splicing, correlating with an augmented HIV-1 reservoir size in cART-treated PWH [ 175 , 176 ].…”

Annual Meeting of the Canadian Association for the Study of the Liver (CASL), the Canadian Network on Hepatitis C (CanHepC), the Canadian Association of Hepatology Nurses (CAHN), and the Canadian NASH Network 2022 Abstracts

“…Higher HIV DNA levels were observed in resting CD4 T cells from HCV/HIV co-infected individuals as compared to HIV mono-infected participants ( 2 ). Even following HCV cure with Directly Acting Antivirals (DAA), a significant rise in HIV DNA levels was reported in the peripheral blood of some HIV/HCV co-infected patients compared to their pre-treatment baseline ( 3 ). This is associated with superior expression of HIV transcripts in DAA-treated co-infected compared to HIV mono-infected subjects ( 4 ).…”