HBVsvp-Pulsed Dendritic Cell Immunotherapy Induces Th1 Polarization and Hepatitis B Virus-Specific Cytotoxic T Lymphocytes Production

Mohamed M S Farag; Reda A Suef; Ghada M Al-Toukhy; Mohamed A Selim; Mostafa A Elbahnasawy; Nahla El Sharkawy; Sameera Ezzat; Nashwa Shebl; Mohamed T M Mansour

doi:10.2147/IDR.S265681

HBVsvp-Pulsed Dendritic Cell Immunotherapy Induces Th1 Polarization and Hepatitis B Virus-Specific Cytotoxic T Lymphocytes Production

Infect Drug Resist. 2020 Aug 5:13:2699-2709. doi: 10.2147/IDR.S265681. eCollection 2020.

Authors

Mohamed M S Farag¹, Reda A Suef¹, Ghada M Al-Toukhy², Mohamed A Selim¹, Mostafa A Elbahnasawy¹, Nahla El Sharkawy³, Sameera Ezzat⁴, Nashwa Shebl⁵, Mohamed T M Mansour⁶

Affiliations

¹ Botany and Microbiology Department, Faculty of Science, Al-Azhar University, Cairo 11884, Egypt.
² Virology & Immunology Department, Children's Cancer Hospital, Cairo 57357, Egypt.
³ Clinical Pathology Department, National Cancer Institute, Cairo University and Children Cancer Hospital, Cairo 57357, Egypt.
⁴ Epidemiology & Preventive Medicine Department, National Liver Institute, Menoufia University, Al Minufya, Egypt.
⁵ Hepatology Department, National Liver Institute, Menoufia University, Al Minufya, Egypt.
⁶ Virology & Immunology Department, National Cancer Institute, Cairo University and Children Cancer Hospital, Cairo 57357, Egypt.

Abstract

Background: In chronic hepatitis B virus (CHB) patients, both dendritic cells (DCs) and T cells are functionally impaired and consequently the HBV-specific cellular immune responses are downregulated. The present study aims to investigate whether monocyte-derived DC (MoDCs)-pulsed-HBV subviral particles (HBVsvp) can polarize Th1 cells to induce HBV-specific cytotoxic T-lymphocytes (CTL) responses in CHB patients.

Methods and materials: To this end, the human hepatoma HepG2.2.15 cell line was used to produce HBVsvp as a culturing system, and HBVsvp were concentrated for highly virus titer using the polyethylene glycol protocol. Peripheral blood mononuclear cells (PBMCs), collected from CHB patients and healthy donors, were differentiated into MoDCs and T cells. PBMCs-derived MoDCs were first pulsed with HBVsvp and then cultured with PBMCs-derived T cells. MoDCs and/or T subsets cells were identified for phenotypic activation by FACS analysis. The cytokine secretion of IL-4, IL-12, and IFN-γ in the culture supernatants was detected.

Results: The MoDCs were restored for their activation upon pulsing with HBVsvp in vitro, as identified by significantly overexpression of both CD86 and HLA-DR, and overproduction of IL-4 and IL-12. Furthermore, MoDCs-pulsed-HBVsvp induced Th1 frequencies and activated HBV-specific CTL to produce significantly highest amount of IFN-γ. Enhanced HBV-specific CTL led to strong cytolytic capacity against HepG2.2.15.

Conclusion: Overall, our data suggest that in vitro activation of MoDCs with HBVsvp overcomes the functionally impaired DCs and T cells in CHB patients offering a promising tool for therapeutic or vaccine-based approaches against HBV.

Keywords: CHB; HBV-specific CTL; MoDCs-pulsed-HBVsvp; immunotherapy.