Journal of Vaccines & Vaccination

LLO as a genetic adjuvant enhances hepatitis C virus NS3 DNA vaccine immunogenicity

12^th Asia Pacific Global Summit and Expo on Vaccines & Vaccination

November 24-25, 2016 Melbourne, Australia

Mohammad Reza Aghasadeghi, Mohammad Hassan Pouriayevali, Taravat Bamdad, Seyed Mehdi Sadat, Farzaneh Sabahi, Seyed Davar Siadat and Sepehr Soleimani

Pasteur Institute of Iran, Iran
Tarbiat Modares University, Iran

Posters & Accepted Abstracts: J Vaccines Vaccin

Abstract:

Introduction: Hepatitis C virus (HCV) chronic infection is a worldwide health problem which affects almost 3% of the world�??s population. To develop HCV vaccine, induction of potent T-cell response is consider to that target the immunogenic and conserved region. Listeriolysin O (LLO) of Listeria monocytogenes can be as an adjuvant to initiate T-cell immune responses. Herein, we cloned and expression fusion LLO-NS3 in mammalian cell line then evaluated T-cell immune response of recombinant DNA vaccine in mice model. Methods: cDNA corresponding to partial immunogenic length of NS3 (rNS3, aa 1191-1380) was constructed by RT-PCR on RNA purified from HCV patients. rNS3 was cloned on pcDNA3.1-LLO and then recombinant plasmid evaluated by sequencing. Finally the plasmid was transfected to HEK293T cell line and fusion protein expression was confirmed by Western blotting. Mice were immunized three doses of the recombinant vector or pcDNA3.1-NS3 (as control group) and pcDNA3.1 (as negative control group) in 3-week intervals and their immune responses were evaluated using cytotoxic T-lymphocyte (CTL) activity by lactate dehydrogenase releasing method. Results: Proper expression of the recombinant protein in the expected size (around 70 kDa) was confirmed using Western blotting. The immunization results indicated that cytotoxicity T-cell responses of vaccinated mice were significantly increased compared to control group (p<0.05). Discussion: pcDNA3.1-LLO-NS3 demonstrated strong T-cell immunogenicity in a murine model. Our primary results demonstrated that truncated region of NS3 as immunogenic truncated region and LLO as genetic adjuvant can induce cell immune responses by activating the Th1 pathway.

Biography :

Email: mr_sadeqi@yahoo.com