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

Jiang, Duyin; Sun, Yuanjie; Cheng, Long; Zhang, Ge-Fei; Dong, Chen; Jin, Boquan; Song, Chaojun; Ma, Ying; Zhang, Fanglin; Yang, Kun

doi:10.1016/j.vaccine.2015.05.007

Cited by 26 publications

(19 citation statements)

References 46 publications

(47 reference statements)

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“…The improved plaque formation test is dependent on the low pH-induced cytopathic effects of hantavirus but is time-consuming and has low reproducibility. The most widely adopted approach to test hantavirus titers (especially for HTNV) is the TCID50 (50% tissue culture infective dose) calculation using ELISA as previously reported by our group (Xu et al, 2002; Cheng et al, 2014; Jiang et al, 2015; Ye et al, 2015a,b; Ying et al, 2016); however, virus propagation in Vero E6 cells takes at least 10 days. All the reported detective measurements have insurmountably objective drawbacks, such as high demanding experimental conditions for qRT-PCR and expensive apparatus and labware for FCM, which limits their applicability (Wan et al, 2010).…”

Section: Introductionmentioning

confidence: 99%

In-Cell Western Assays to Evaluate Hantaan Virus Replication as a Novel Approach to Screen Antiviral Molecules and Detect Neutralizing Antibody Titers

Chen

et al. 2017

Front. Cell. Infect. Microbiol.

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Hantaviruses encompass rodent-borne zoonotic pathogens that cause severe hemorrhagic fever disease with high mortality rates in humans. Detection of infectious virus titer lays a solid foundation for virology and immunology researches. Canonical methods to assess viral titers rely on visible cytopathic effects (CPE), but Hantaan virus (HTNV, the prototype hantavirus) maintains a relatively sluggish life cycle and does not produce CPE in cell culture. Here, an in-cell Western (ICW) assay was utilized to rapidly measure the expression of viral proteins in infected cells and to establish a novel approach to detect viral titers. Compared with classical approaches, the ICW assay is accurate and time- and cost-effective. Furthermore, the ICW assay provided a high-throughput platform to screen and identify antiviral molecules. Potential antiviral roles of several DExD/H box helicase family members were investigated using the ICW assay, and the results indicated that DDX21 and DDX60 reinforced IFN responses and exerted anti-hantaviral effects, whereas DDX50 probably promoted HTNV replication. Additionally, the ICW assay was also applied to assess NAb titers in patients and vaccine recipients. Patients with prompt production of NAbs tended to have favorable disease outcomes. Modest NAb titers were found in vaccinees, indicating that current vaccines still require improvements as they cannot prime host humoral immunity with high efficiency. Taken together, our results indicate that the use of the ICW assay to evaluate non-CPE Hantaan virus titer demonstrates a significant improvement over current infectivity approaches and a novel technique to screen antiviral molecules and detect NAb efficacies.

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Section: Introductionmentioning

confidence: 99%

In-Cell Western Assays to Evaluate Hantaan Virus Replication as a Novel Approach to Screen Antiviral Molecules and Detect Neutralizing Antibody Titers

Chen

et al. 2017

Front. Cell. Infect. Microbiol.

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“…15 A candidate ANDV vaccine is likely to be available for use in human trials in the near future. [16][17][18][19] A Hantaan virus (HTNV) inactivated vaccine has been in use in China since 1990. DNA vaccines carrying plasmids with M segments from pathogenic New and Old World hantaviruses are in development.…”

Section: Introductionmentioning

confidence: 99%

Knowledge, attitudes, and practices regarding hantavirus disease and acceptance of a vaccine trial in rural communities of southern Chile

Valdivieso

González

Nájera

et al. 2017

Human Vaccines & Immunotherapeutics

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Andes hantavirus cardiopulmonary syndrome, transmitted by Oligoryzomys longicaudatus, has no approved treatment, a case fatality rate of 35%, and documented person-to-person transmission. An Andes vaccine, highly needed for prevention, is in development. We aimed to evaluate knowledge, attitudes and practices (KAP) regarding hantavirus disease and willingness to participate in a future Andes vaccine trials through a cross sectional face-to-face oral survey of a randomly selected adult sample from 2 rural communes in southern Chile. Human subjects approval was obtained from our institutional IRBs, and participants signed informed consent. We enrolled 319 subjects from Corral and 321 from Curarrehue; 98% had heard about hantavirus disease and its reservoir but only half knew about transmission, symptoms and prevention. Participants fear the disease but are only partially aware of their own risk. One third of participants reported presence of rodents inside their homes. Despite moderate confidence in their health system, most subjects perceived vaccines as beneficial, and 93% would accept an approved hantavirus vaccine. Half would agree to participate in a vaccine trial and 29% would allow their children to participate. Motivations to participate were mainly altruistic, while risk perception was the main reason for declining. Knowledge about hantavirus disease and prevention practices require reinforcement, and a vaccine trial seems feasible in these populations.

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“…However, these inactivated vaccines have many shortcomings, including poor immunogenicity, which hinders the production of neutralizing antibodies or cell-mediated immunity (Song et al, 1992), and the concern of safety when using inactivated vaccines, which may contain some infectious particles. Furthermore, there is no study reporting that it could establish long-term memory immunity (Jiang et al, 2015). Thus, it is necessary to develop an alternative immunotherapy strategy that can induce robust humoral immune response to produce therapeutically effective antiviral antibodies and T cell-mediated autoimmune responses.…”

Section: Introductionmentioning

confidence: 99%

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

Cheng

Ying

et al. 2016

Front. Cell. Infect. Microbiol.

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The cytotoxic T lymphocyte (CTL) response plays a key role in controlling viral infection, but only a few epitopes within the HTNV glycoprotein (GP) that are recognized by CTLs have been reported. In this study, we identified one murine HTNV GP-derived H2-Kb-restricted CTL epitope in C57BL/6 mice, which could be used to design preclinical studies of vaccines for HTNV infection. First, 15 8-mer peptides were selected from the HTNV GP amino acid sequence based on a percentile rank of <=1% by IEDB which is the most comprehensive collection of epitope prediction and analysis tool. A lower percentile rank indicates higher affinity and higher immune response. In the case of the consensus method, we also evaluated the binding score of peptide-binding affinity by the BIMAS software to confirm that all peptides were able to bind H2-Kb. Second, one novel GP-derived CTL epitope, GP6 aa456-aa463 (ITSLFSLL), was identified in the splenocytes of HTNV-infected mice using the IFN-γ ELISPOT assay. Third, a single peptide vaccine was administered to C57BL/6 mice to evaluate the immunogenic potential of the identified peptides. ELISPOT and cell-mediated cytotoxicity assays showed that this peptide vaccine induced a strong IFN-γ response and potent cytotoxicity in immunized mice. Last, we demonstrated that the peptide-vaccinated mice had partial protection from challenge with HTNV. In conclusion, we identified an H2-Kb-restricted CTL epitope with involvement in the host immune response to HTNV infection.

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

Cited by 26 publications

References 46 publications

In-Cell Western Assays to Evaluate Hantaan Virus Replication as a Novel Approach to Screen Antiviral Molecules and Detect Neutralizing Antibody Titers

In-Cell Western Assays to Evaluate Hantaan Virus Replication as a Novel Approach to Screen Antiviral Molecules and Detect Neutralizing Antibody Titers

Knowledge, attitudes, and practices regarding hantavirus disease and acceptance of a vaccine trial in rural communities of southern Chile

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

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