Induction of Specific Humoral and Cellular Immune Responses in a Mouse Model following Gene Fusion of HSP70C and Hantaan Virus Gn and S0.7 in an Adenoviral Vector

Abstract

Heat shock proteins (HSPs) display adjuvant functions when given as fusion proteins to enhance vaccination efficiency. To evaluate enhanced potency of Hantaan virus (HTNV) glycoprotein (GP) and nucleocapsid protein (NP) immunogenicity by heat shock protein 70 (HSP70), a recombinant adenovirus rAd-GnS0.7-pCAG-HSP70C expression vector was developed by genetically linking the HSP70 C-terminal gene (HSP70 359–610 aa, HSP70C) to the Gn and 0.7 kb fragment of the NP (aa1–274-S0.7). C57BL/6 mice were immunized with these recombinant adenoviral vectors. A series of immunological assays determined the immunogenicity of the recombinant adenoviral vectors. The results showed that rAd-GnS0.7-pCAG-HSP70C induced a stronger humoral and cellular immune response than other recombinant adenoviruses (rAd-GnS0.7-pCAG and rAd-GnS0.7) and the HFRS vaccine control. Animal protection experiments showed that rAd-GnS0.7-pCAG-HSP70C was effective at protecting C57BL/6 mice from HTNV infection. The results of the immunological experiments showed that HSP70C lead to enhanced vaccine potency, and suggested significant potential in the development of genetically engineered vaccines against HTNV.

Screening and Identification of an H-2Kb-Restricted CTL Epitope within the Glycoprotein of Hantaan Virus

“…The genome of HTNV consists of L, M, and S segments, which encode the viral RNA-dependent RNA polymerase, glycoprotein, and nucleoprotein, respectively (Cheng et al, 2014). HFRS is a severe, life-threatening illness characterized by fever, hemorrhage, and renal failure and is a serious public health threat in China (Hansen et al, 2015).…”

Construction and evaluation of DNA vaccine encoding Hantavirus glycoprotein N-terminal fused with lysosome-associated membrane protein

“…Other discoveries were also noted in this study. Expression of antigen matters in DNA vaccine to be efficacious [30,37,38]. Our results discovered an increasing intracellular Gn accumulation with pVAX-LAMP/Gn by the side of an identical mRNA transcription between two constructs transfection.…”

“…Our study referred to a novel infection model of HTNV, which was currently used for assessing vaccinal immune protection [30]. In addition with VL detection, practicability was confirmed and the result illustrated protection in vivo conferred by LAMP1 targeted Gn.…”

“…The cell microculture neutralization test using sandwich ELISA was performed as described earlier [30]. Monolayers of Vero E6 cells grown in RPMI-1640 medium supplemented with 10% FCS were plated at a density of 2 × 10 4 cells well −1 18-24 h before testing.…”

Incorporation of GM-CSF or CD40L Enhances the Immunogenicity of Hantaan Virus-Like Particles

“…HTNV strain 76–118 was provided by our library (Cheng et al, 2014). The pFastBac™ Dual expression system, which includes the pFastBac™ Dual vector, was purchased from Invitrogen (Carlsbad, CA, USA).…”