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A Broadly Cross-protective Vaccine Presenting the Neighboring Epitopes within the VP1 GH Loop and VP2 EF Loop of Enterovirus 71.

Xu L, He D, Yang L, Li Z, Ye X, Yu H, zhao H, Li S, Yuan L, Qian H, Que Y, Shih JW, Zhu H, Li Y, Cheng T, Xia N - Sci Rep (2015)

Bottom Line: We demonstrate that anti-VP2 (aa141-155) sera bound authentic CA16 viral particles, whereas anti-VP1 (aa208-222) sera could not.Moreover, the anti-VP2 (aa141-155) antibodies inhibited the binding of human serum to virions, which demonstrated that the VP2 epitope is immunodominant between EV71 and CA16.These results illustrated that the chimeric VLP HBc-E1/2 is a promising candidate for a broad-spectrum HFMD vaccine, and also reveals mechanisms of protection by the neighboring linear epitopes of the VP1 GH and VP2 EF loops.

View Article: PubMed Central - PubMed

Affiliation: National Institute of Diagnostics and Vaccine Development in infectious diseases, School of Public Health, Xiamen University, Xiamen 361102, PR China.

ABSTRACT
Human enterovirus 71 (EV71) and coxsackievirus A16 (CA16) are the major etiological agents of hand, foot and mouth disease (HFMD) and are often associated with neurological complications. Currently, several vaccine types are being developed for EV71 and CA16. In this study, we constructed a bivalent chimeric virus-like particle (VLP) presenting the VP1 (aa208-222) and VP2 (aa141-155) epitopes of EV71 using hepatitis B virus core protein (HBc) as a carrier, designated HBc-E1/2. Immunization with the chimeric VLPs HBc-E1/2 induced higher IgG titers and neutralization titers against EV71 and CA16 in vitro than immunization with only one epitope incorporated into HBc. Importantly, passive immunization with the recombinant HBc-E2 particles protected neonatal mice against lethal EV71 and CA16 infections. We demonstrate that anti-VP2 (aa141-155) sera bound authentic CA16 viral particles, whereas anti-VP1 (aa208-222) sera could not. Moreover, the anti-VP2 (aa141-155) antibodies inhibited the binding of human serum to virions, which demonstrated that the VP2 epitope is immunodominant between EV71 and CA16. These results illustrated that the chimeric VLP HBc-E1/2 is a promising candidate for a broad-spectrum HFMD vaccine, and also reveals mechanisms of protection by the neighboring linear epitopes of the VP1 GH and VP2 EF loops.

No MeSH data available.


Related in: MedlinePlus

Capture ELISA and competitive ELISA test for MAbs binding to virus.(A,B) The number of RNA genome copies/ml determined by the capture ELISA and TaqMan real-time RT-PCR. The log10 values of the viral RNA copies/ml were calculated by interpolating the Ct values from the standard curve. These experiments were repeated three times and a representative result is shown. (C,D)The dilution of nMAb BB1A5 and H3B10 competed with human serum samples for specifically binding to EV71 virus 52-3 (C) or CA16-190 (D) coated on the well. A nonrelated monoclonal antibody was used as a negative or positive control, respectively.
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f5: Capture ELISA and competitive ELISA test for MAbs binding to virus.(A,B) The number of RNA genome copies/ml determined by the capture ELISA and TaqMan real-time RT-PCR. The log10 values of the viral RNA copies/ml were calculated by interpolating the Ct values from the standard curve. These experiments were repeated three times and a representative result is shown. (C,D)The dilution of nMAb BB1A5 and H3B10 competed with human serum samples for specifically binding to EV71 virus 52-3 (C) or CA16-190 (D) coated on the well. A nonrelated monoclonal antibody was used as a negative or positive control, respectively.

Mentions: The in-house developed VP1 (aa208-222)-specific monoclonal antibody (H3B10) and the VP2 (aa141-155)-specific nMAb (BB1A5) were tested using Capture ELISA (Fig. 5A,B). An unrelated antibody was used as a negative control for the assay. Serial dilutions of JS 52-3 with an initial concentration of 107 TCID50 were captured by H3B10 and BB1A5. A dilution of 102 TCID50 of the EV71 virus continued to generate a detectable signal in the real-time PCR detection assay (38.61 copies/mL and 61.79 copies/mL). Importantly, the VP2 nMAb BB1A5 also captured a viral CA16 copy number of 1073.18/mL at a dilution of 102 TCID50.


A Broadly Cross-protective Vaccine Presenting the Neighboring Epitopes within the VP1 GH Loop and VP2 EF Loop of Enterovirus 71.

Xu L, He D, Yang L, Li Z, Ye X, Yu H, zhao H, Li S, Yuan L, Qian H, Que Y, Shih JW, Zhu H, Li Y, Cheng T, Xia N - Sci Rep (2015)

Capture ELISA and competitive ELISA test for MAbs binding to virus.(A,B) The number of RNA genome copies/ml determined by the capture ELISA and TaqMan real-time RT-PCR. The log10 values of the viral RNA copies/ml were calculated by interpolating the Ct values from the standard curve. These experiments were repeated three times and a representative result is shown. (C,D)The dilution of nMAb BB1A5 and H3B10 competed with human serum samples for specifically binding to EV71 virus 52-3 (C) or CA16-190 (D) coated on the well. A nonrelated monoclonal antibody was used as a negative or positive control, respectively.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4525384&req=5

f5: Capture ELISA and competitive ELISA test for MAbs binding to virus.(A,B) The number of RNA genome copies/ml determined by the capture ELISA and TaqMan real-time RT-PCR. The log10 values of the viral RNA copies/ml were calculated by interpolating the Ct values from the standard curve. These experiments were repeated three times and a representative result is shown. (C,D)The dilution of nMAb BB1A5 and H3B10 competed with human serum samples for specifically binding to EV71 virus 52-3 (C) or CA16-190 (D) coated on the well. A nonrelated monoclonal antibody was used as a negative or positive control, respectively.
Mentions: The in-house developed VP1 (aa208-222)-specific monoclonal antibody (H3B10) and the VP2 (aa141-155)-specific nMAb (BB1A5) were tested using Capture ELISA (Fig. 5A,B). An unrelated antibody was used as a negative control for the assay. Serial dilutions of JS 52-3 with an initial concentration of 107 TCID50 were captured by H3B10 and BB1A5. A dilution of 102 TCID50 of the EV71 virus continued to generate a detectable signal in the real-time PCR detection assay (38.61 copies/mL and 61.79 copies/mL). Importantly, the VP2 nMAb BB1A5 also captured a viral CA16 copy number of 1073.18/mL at a dilution of 102 TCID50.

Bottom Line: We demonstrate that anti-VP2 (aa141-155) sera bound authentic CA16 viral particles, whereas anti-VP1 (aa208-222) sera could not.Moreover, the anti-VP2 (aa141-155) antibodies inhibited the binding of human serum to virions, which demonstrated that the VP2 epitope is immunodominant between EV71 and CA16.These results illustrated that the chimeric VLP HBc-E1/2 is a promising candidate for a broad-spectrum HFMD vaccine, and also reveals mechanisms of protection by the neighboring linear epitopes of the VP1 GH and VP2 EF loops.

View Article: PubMed Central - PubMed

Affiliation: National Institute of Diagnostics and Vaccine Development in infectious diseases, School of Public Health, Xiamen University, Xiamen 361102, PR China.

ABSTRACT
Human enterovirus 71 (EV71) and coxsackievirus A16 (CA16) are the major etiological agents of hand, foot and mouth disease (HFMD) and are often associated with neurological complications. Currently, several vaccine types are being developed for EV71 and CA16. In this study, we constructed a bivalent chimeric virus-like particle (VLP) presenting the VP1 (aa208-222) and VP2 (aa141-155) epitopes of EV71 using hepatitis B virus core protein (HBc) as a carrier, designated HBc-E1/2. Immunization with the chimeric VLPs HBc-E1/2 induced higher IgG titers and neutralization titers against EV71 and CA16 in vitro than immunization with only one epitope incorporated into HBc. Importantly, passive immunization with the recombinant HBc-E2 particles protected neonatal mice against lethal EV71 and CA16 infections. We demonstrate that anti-VP2 (aa141-155) sera bound authentic CA16 viral particles, whereas anti-VP1 (aa208-222) sera could not. Moreover, the anti-VP2 (aa141-155) antibodies inhibited the binding of human serum to virions, which demonstrated that the VP2 epitope is immunodominant between EV71 and CA16. These results illustrated that the chimeric VLP HBc-E1/2 is a promising candidate for a broad-spectrum HFMD vaccine, and also reveals mechanisms of protection by the neighboring linear epitopes of the VP1 GH and VP2 EF loops.

No MeSH data available.


Related in: MedlinePlus