Monthly Archives: December 2012

Neutralization activity. Part 2

DNA vaccine has been demonstrated as a promising vaccination strategy for various viral infections. Previous studies have shown good immunogenicity and protection efficacy of hantavirus DNA vaccine. Hooper et al. demonstrated that DNA vaccination with a plasmid containing a cDNA representing the Seoul virus (SEOV) M segment elicited neutralizing antibody responses in mice and hamsters. […]

Neutralization activity

Neutralizing antibodies, which conferring protective immunity induced by DNA vaccine plasmids against hantavirus were evaluated by microneutralization assays. Pre-immune sera from all the groups exhibited no neutralizing activity. In contrast, immune sera collected 21 days after the first immunization with 100 μg of HTNV DNA vaccine plasmids in the presence or absence of CpG motifs […]

Expression and identification of eCTLA4-N/GP fusion proteins

The expressing of eCTLA4-N fusion protein was firstly determined in prokaryotic system. Supernatant and cell lysates of E.coli DH5α transformed with pET30a/eCTLA-S or pET30a/S plasmid were analyzed by SDS-PAGE (Figure 1A). Neither eCTLA4-N fusion protein nor HTNV N was expressed into supernatant (Figure 1A, lane 1 and lane 3). Instead, eCTLA4-N fusion protein was shown […]

Serologic assays

HTNV N-specific IgG antibodies in mice sera were determined, by enzyme-linked immunosorbent assay (ELISA) in [26,27]. 96-well microtiter plates (Costar, USA) coated with 100 μl of purified recombinant N protein of HTNV strain A9 at a concentration of 1 μg/ml as described previously. Hantaan virus glycoproteins specific IgG antibodies were evaluated by IFA using insect […]

Cells and viruses

The 293T, Vero E6 cells and Baby hamster kidney cell (BHK) cells were purchased from ATCC (ATCC number: CRL-1586) and cultured in Dulbecco modified Eagle medium (DMEM) supplemented with 10% heat-inactivated fetal calf serum, 100 U of penicillin, and 100 μg of streptomycin per ml at 37 with 5% CO2. HTNV strain 84Fli, isolated from […]

Fusion with extracellular domain of cytotoxic T-lymphocyte-associated-antigen 4 leads to enhancement of immunogenicity of Hantaan virus DNA vaccines in C57BL/6 mice

Background Hantaan virus (HTNV) is the causative agent of the most severe form of a rodent-borne disease known as hemorrhagic fever with renal syndrome (HFRS). A safe and effective HTNV vaccine is needed. Vaccination with DNA constructs expressing fused antigen with bioactive factors, has shown promising improvement of immunogenicity for viral agents in animal models, […]