Construction and immunological characterization of CD40L or GM-CSF incorporated Hantaan virus like particle

// Qikang Ying 1, * , Tiejun Ma 1, * , Linfeng Cheng 1, * , Xiaoxiao Zhang 1 , Agnieszka D. Truax 2 , Ruixue Ma 1 , Ziyu Liu 1 , Yingfeng Lei 1 , Liang Zhang 1 , Wei Ye 1 , Fanglin Zhang 1 , Zhikai Xu 1 , Lei Shang 3 , Rongrong Liu 1 , Fang Wang 1 , Xingan Wu 1 1 Department of Microbiology, School of Basic Medicine, Fourth Military Medical University, Xi'an, 710032, China 2 The Lineberger Comprehensive Cancer Center, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599-7295, USA 3 Department of Statistics, Fourth Military Medical University, Xi'an, 710032, China * These authors have contributed equally to this work Correspondence to: Xingan Wu, email: wuxingan@fmmu.edu.cn Fang Wang, email: wangf07@fmmu.edu.cn Rongrong Liu, email: rong4713@163.com Keywords: Hantaan virus, virus like particle, virus vaccine, CD40 ligand, granulocyte macrophage colony-stimulating factor Received: March 28, 2016 Accepted: July 10, 2016 Published: August 17, 2016 ABSTRACT Infection of Hantaan virus (HTNV) usually causes hemorrhagic fever with renal syndrome (HFRS). China has the worst epidemic incidence of HFRS as well as high fatality. Inactivated whole virus has been used for HFRS vaccination, however there are still problems such as safety concerns. CD40 ligand (CD40L) and granulocyte macrophage colony-stimulating factor (GM-CSF) are well-known immune stimulating molecules that can enhance antigen presenting, lymphocytes activation and maturation, incorporation of CD40L and GM-CSF to the surface of virus like particles (VLPs) can greatly improve the vaccination effect. We constructed eukaryotic vectors expressing HTNV M segment and S segment, as well as vectors expressing HTNV M segment with CD40L or GM-CSF, our results showed successful production of CD40L or GM-CSF incorporated HTNV VLPs. In vitro stimulation with CD40L or GM-CSF anchored HTNV VLP showed enhanced activation of macrophages and DCs. CD40L/GM-CSF incorporated VLP can induce higher level of HTNV specific antibody and neutralizing antibody in mice. Immunized mice splenocytes showed higher ability of secreting IFN-γ and IL-2, as well as enhancing CTL activity. These results suggest CD40L/GM-CSF incorporated VLP can serve as prospective vaccine candidate.