Decreased CD4+CD25+CD127dim/- Regulatory T Cells and T Helper 17 Cell Responsiveness to Toll-Like Receptor 2 in Chronic Hepatitis C Patients with Daclatasvir Plus Asunaprevir Therapy

Viral Immunol. 2018 Oct;31(8):559-567. doi: 10.1089/vim.2018.0055. Epub 2018 Aug 1.


Direct-acting antivirals (DAAs) not only rapidly inhibited hepatitis C virus (HCV) replication but also modulated innate and adaptive immune response in chronic hepatitis C patients. However, the regulatory activity of DAAs to Toll-like receptor 2 (TLR2) stimulation on CD4+CD25+CD127dim/- regulatory T cells (Tregs) and T helper (Th) 17 cells was not completely understood. In the present study, a total of 23 patients with chronic HCV genotype 1b infection were enrolled, and blood samples were collected at baseline (treatment naive), end of therapy (EOT), and 12 weeks after EOT (SVR12) with daclatasvir plus asunaprevir therapy. TLR2 expression on Tregs and Th17 cells was measured by flow cytometry. Cellular proliferation, cytokine production, and suppressive activity were also tested in purified CD4+CD25+CD127dim/- Tregs in response to the stimulation of Pam3Csk4, an agonist of TLR2. Inhibition of HCV RNA by daclatasvir and asunaprevir did not affect either percentage of Tregs/Th17 cells or TLR2 expression on Tregs/Th17 cells. Pam3Csk4 stimulation also did not influence either cellular proliferation or Tregs/Th17 proportion at each time point. Stimulation with Pam3Csk4 only enhanced the suppressive function and interleukin (IL)-35 production by Tregs purified from baseline, but not those from EOT or SVR12. Similarly, Pam3Csk4 stimulation only elevated Th17 cell frequency of CD4+ T cells from baseline, but not those from EOT or SVR12. Moreover, daclatasvir and asunaprevir therapy did not promote TLR2-induced shift of Tregs toward Th17-like phenotype and function. These data suggested that daclatasvir plus asunaprevir therapy resulted in the decreased responsiveness of Tregs/Th17 cells to TLR2 stimulation in chronic hepatitis C patients, which might provide a novel mechanism underlying DAA-induced immunoregulation.

Keywords: T helper 17 cells; antiviral; chronic hepatitis C; regulatory T cells; toll-like receptor 2.

MeSH terms

  • Adolescent
  • Adult
  • Antiviral Agents / administration & dosage
  • Antiviral Agents / therapeutic use*
  • CD4-Positive T-Lymphocytes / immunology*
  • Carbamates
  • Female
  • Hepacivirus / drug effects
  • Hepatitis C, Chronic / drug therapy
  • Hepatitis C, Chronic / immunology*
  • Hepatitis C, Chronic / virology
  • Humans
  • Imidazoles / administration & dosage
  • Imidazoles / therapeutic use
  • Interleukin-2 Receptor alpha Subunit / immunology
  • Interleukin-7 Receptor alpha Subunit / immunology
  • Isoquinolines / administration & dosage
  • Isoquinolines / therapeutic use
  • Liver Cirrhosis / diagnosis
  • Liver Cirrhosis / diagnostic imaging
  • Liver Cirrhosis / virology
  • Male
  • Middle Aged
  • Pyrrolidines
  • Sulfonamides / administration & dosage
  • Sulfonamides / therapeutic use
  • T-Lymphocytes, Regulatory / immunology*
  • Th17 Cells / immunology*
  • Toll-Like Receptor 2 / immunology*
  • Tomography, X-Ray Computed
  • Valine / analogs & derivatives
  • Young Adult


  • Antiviral Agents
  • Carbamates
  • Imidazoles
  • Interleukin-2 Receptor alpha Subunit
  • Interleukin-7 Receptor alpha Subunit
  • Isoquinolines
  • Pyrrolidines
  • Sulfonamides
  • Toll-Like Receptor 2
  • Valine
  • daclatasvir
  • asunaprevir