Formalin Inactivation of Japanese Encephalitis Virus Vaccine Alters the Antigenicity and Immunogenicity of a Neutralization Epitope in Envelope Protein Domain III.
Formalin-inactivated Japanese encephalitis virus (JEV) vaccines are widely available, but the effects of formalin inactivation on the antigenic structure of JEV and the profile of antibodies elicited after vaccination are not well understood. We used a panel of monoclonal antibodies (MAbs) to map the antigenic structure of live JEV virus, untreated control virus (UCV), formalin-inactivated commercial vaccine (FICV), and formalin-inactivated virus (FIV). The binding activity of T16 MAb against Nakayama-derived FICV and several strains of FIV was significantly lower compared to live virus and UCV. T16 MAb, a weakly neutralizing JEV serocomplex antibody, was found to inhibit JEV infection at the post-attachment step. The T16 epitope was mapped to amino acids 329, 331, and 389 within domain III (EDIII) of the envelope (E) glycoprotein. When we explored the effect of formalin inactivation on the immunogenicity of JEV, we found that Nakayama-derived FICV, FIV, and UCV all exhibited similar immunogenicity in a mouse model, inducing anti-JEV and anti-EDII 101/106/107 epitope-specific antibodies. However, the EDIII 329/331/389 epitope-specific IgG antibody and neutralizing antibody titers were significantly lower for FICV-immunized and FIV-immunized mouse serum than for UCV-immunized. Formalin inactivation seems to alter the antigenic structure of the E protein, which may reduce the potency of commercially available JEV vaccines. Virus inactivation by H2O2, but not by UV or by short-duration and higher temperature formalin treatment, is able to maintain the antigenic structure of the JEV E protein. Thus, an alternative inactivation method, such as H2O2, which is able to maintain the integrity of the E protein may be essential to improving the potency of inactivated JEV vaccines.
Graduate Institute of Microbiology and Public Health, National Chung Hsing University, Taichung, Taiwan.
Yi-Chin
YC
Fan
Department of Periodontology, School of Dentistry, National Defense Medical Center and Tri-Service General Hospital, Taipei, Taiwan.
Hsien-Chung
HC
Chiu
College of Medicine, Tzu-Chi University, Hualien, Taiwan.
Li-Kuang
LK
Chen
Arboviral Diseases Branch, Center for Disease Control and Prevention, Fort Collins, Colorado, United States of America.
Gwong-Jen J
GJ
Chang
Graduate Institute of Microbiology and Public Health, National Chung Hsing University, Taichung, Taiwan.
Shyan-Song
SS
Chiou
D000818:Animals; D057134:Antibodies, Neutralizing / Q000097:blood*; D000914:Antibodies, Viral / Q000097:blood; D000956:Antigens, Viral / Q000276:immunology; D004202:Disinfectants / Q000494:pharmacology*; D004664:Encephalitis Virus, Japanese / Q000187:drug effects*; D000939:Epitopes / Q000276:immunology*; D005557:Formaldehyde / Q000494:pharmacology*; D022321:Japanese Encephalitis Vaccines / Q000276:immunology*; D008562:Membrane Glycoproteins / Q000276:immunology; D008807:Mice, Inbred BALB C; D015164:Vaccines, Inactivated / Q000276:immunology; D014759:Viral Envelope Proteins / Q000276:immunology; D038301:Virus Inactivation
D057134:Antibodies, Neutralizing; D000914:Antibodies, Viral; D000956:Antigens, Viral; D004202:Disinfectants; D000939:Epitopes; D022321:Japanese Encephalitis Vaccines; D008562:Membrane Glycoproteins; D015164:Vaccines, Inactivated; D014759:Viral Envelope Proteins; C080628:glycoprotein E, Japanese encephalitis virus; D005557:Formaldehyde