Hepatitis C virus E2 protein encapsulation into poly D, L-lactic-co-glycolide microspheres could induce mice cytotoxic T-cell response

Abstract

Piyachat Roopngam,1,2 Kewei Liu,1,2 Lin Mei,2 Yi Zheng,2 Xianbing Zhu,1,2 Hsiang-I Tsai,1,2 Laiqiang Huang1,2 1School of Life Sciences, Tsinghua University, Beijing, People’s Republic of China; 2The Shenzhen Key Laboratory of Gene and Antibody Therapy, Center for Biotechnology and Biomedicine, State Key Laboratory of Health Sciences and Technology, Division of Life and Health Sciences, Graduate School at Shenzhen, Tsinghua University, Shenzhen, People’s Republic of China Abstract: Hepatitis C virus (HCV) is known to cause hepatitis and hepatocellular carcinoma. E2 envelope glycoprotein of HCV type (HCV-E2) has been reported to bind human host cells and is a major target for developing anti-HCV vaccines. However, the therapeutic vaccine for infected patients still needs further development. The vaccine aims to provide cytotoxic T-cells to eliminate infected cells and hepatocellular carcinoma. Currently, there is no effective HCV therapeutic vaccine because most chronically infected patients rarely generate cytotoxic T-cells, even though they have high levels of neutralizing antibodies. Therefore, the adjuvant must be applied to enhance the efficacy of the therapeutic vaccine. In this study, we constructed HCV1b-E2 recombinant protein, a truncated form of peptide, to combine with an effective vaccine adjuvant and delivery system by using poly D, L-lactic-co-glycolide (PLGA) microspheres. HCV1b-E2 protein was effectively encapsulated into PLGA microspheres (HCV1b-E2-PLGA) as a strategy to deliver an insoluble form of HCV1b-E2 protein. The size and shape of PLGA microspheres were generated properly to carry an insoluble form of viral peptide in vivo. The encapsulated viral protein was slowly and continuously released from PLGA microspheres, which indicated the property of the adjuvant. HCV1b-E2-PLGA can trigger a cell-mediated immune response by inducing an expression of mice CD8+ T-cells. Our results demonstrated that HCV1b-E2-PLGA-immunized mice have a significantly increased CD8+ T-cell number, whereas HCV1b-E2-immunized mice have a lower number of CD8+ T-cells. Moreover, HCV1b-E2-PLGA could induce a specific antibody to viral protein, and the immune cells could secrete IFN-γ, which is a significant cytokine for viral response. Thus, HCV1b-E2-PLGA is shown to have adjuvant property and efficacy in the murine model, which is a good strategy to develop HCV prophylactic and therapeutic vaccines. Keywords: hepatitis C virus, HCV, therapeutic vaccine, PLGA microspheres, HCV-E2 peptide, cytotoxic T-cells, hepatitis, hepatocellular carcinoma

Keywords

• Hepatitis C virus (HCV). Therapeutic vaccine. PLGA microsheres. HCV-E2 peptide. Cytotoxic T-cells. Hepatitis. Hepatocellular carcinoma.