Replacement of pr gene with Japanese encephalitis virus pr using reverse genetics reduces antibody-dependent enhancement of dengue virus 2 infection

Abstract

Severe dengue is more likely found during secondary heterologous dengue virus (DENV) infection or primary infection of infants born to dengue-immune mothers and led to the hypothesis of antibody-dependent enhancement (ADE). It has been reported that pre-membrane (prM)-reactive antibodies do not efficiently neutralize DENV infection but instead potently promote ADE infection. Meanwhile, these enhancing anti-prM antibodies mainly react with the precursor (pr) peptide. To evaluate the effect of pr gene substitution on neutralization and ADE of DENV infection, a novel chimeric dengue virus (JEVpr/DENV2) was rationally constructed by replacing the DENV pr gene with Japanese encephalitis virus (JEV) pr gene, based on the full-length infectious complementary DNA (cDNA) clone of DENV2 ZS01/01. We found that chimeric JEVpr/DENV2 showed reduced virulence and good immunogenicity. In addition, anti-JEVpr/DENV2 sera showed broad cross-reactivity and efficient neutralizing activity with all four DENV serotypes and immature DENV2 (ImDENV2). Most importantly, compared with anti-DENV2 sera, anti-JEVpr/DENV2 sera showed significantly reduced enhancing activity of DENV infection in K562 cells. These results suggest that the ADE activities could be reduced by replacing the DENV pr gene with JEV pr gene. These findings may help us better understand the pathogenesis of DENV infection and provide a reference for the development of a vaccine against DENV.

Global Epidemiology of Dengue Outbreaks in 1990–2015: A Systematic Review and Meta-Analysis

“…Cross-reactive and non-neutralizing antibodies from the primary infection can also bind to the new DENV serotype and facilitate virus entry into susceptible cells. It was a phenomenon known as antibody-dependent enhancement of infection (ADE), considered as the most rational explanation for severe dengue (Simmons et al, 2007 ; Wang Y. et al, 2015 ). This can also explain the significantly association between secondary infection and DHF in our study (OR: 1.86, 95% CI: 1.46–2.37).…”

Antibody dependent enhancement: Unavoidable problems in vaccine development

“…The use of reverse genetics to construct chimeric JEVpr/DENV2 with the Japanese encephalitis virus pr instead of the pr gene showed reduced virulence and good immunogenicity. In addition, anti-JEVpr/DENV2 sera showed extensive cross-reactivity and effective neutralizing activity against all four DENV serotypes and immature DENV2 (ImDENV2), and the ADEV activity of DENV decreased ( Wang et al, 2015 ). Wang replaced DENV pr with JEV pr to construct a novel chimeric protein of JEV pr and DENV2 M peptide (JEVpr/DENV-M).…”

Antibody-Dependent Enhancement of Viral Infections

“…DENV NS1 has also been considered as a vaccine candidate as it avoids ADE (Hertz et al 2017). Chimeric DENV constructs with pr gene replaced with Japanese Encephalitis virus pr and DNA vaccines with substitutions or removal of enhancing epitopes have demonstrated reduction in ADE (Crill et al 2012;Tang et al 2015;Wang et al 2015). Another strategy for evasion of ADE is the development of vaccine candidates inducing protective CD4+ and CD8+ T-cell responses.…”