Fragment of Japanese encephalitis virus envelope protein produced in Escherichia coli protects mice from virus challenge.
A fragment from the N-terminal part (E(A)) and a fragment from the C-terminal part (E(B)) of the envelope (E) protein of Japanese encephalitis virus (JEV) was synthesized in Escherichia coli. These two fragments were overlapping with each other by nine amino acids, however, they were not cross-reacting with each other at the antisera level. Both E(A)and E(B)are antigenic by themselves when injected into mice, but when tested against sera from mice, rabbit, swine and human that had been immunized or naturally infected with JEV, E(B)acted as a better antigen than E(A)by ELISA assays. E(B)also proved to be a better immunogen in protection against lethal JEV infection than E(A). The protection appears to be correlated with the neutralizing titres of the anti-JEV sera. The response elicited by E(B)is a Th1 response and the antibody produced contained higher neutralizing titre than E(A)fragment. The major difference between E(A)and E(B)fragments is the solubility during expression in E. coli, while E(B)fragment is soluble, E(A)was isolated from the insoluble inclusion bodies. Therefore the antigenicity and immunogenicity expressed by the E(B)fragment may probably be due to its proper folding to assume a correctly assembled form during expression in E. coli, a quality that is important for a protein to qualify as a good vaccine candidate.
Chia
S C
SC
Institute of Biomedical Sciences, Academia Sinica, Taipei, Taiwan 11529, Republic of China.
Leung
P S
PS
Liao
C P
CP
Huang
J H
JH
Lee
S T
ST
D000328:Adult; D000818:Animals; D000914:Antibodies, Viral / Q000276:immunology; D000956:Antigens, Viral / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification; D002648:Child; D004664:Encephalitis Virus, Japanese / Q000235:genetics / Q000276:immunology; D004672:Encephalitis, Japanese / Q000097:blood / Q000276:immunology / Q000517:prevention & control*; D004926:Escherichia coli; D015870:Gene Expression; D006801:Humans; D007074:Immunoglobulin G / Q000276:immunology; D007132:Immunoglobulin Isotypes; D022321:Japanese Encephalitis Vaccines / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification; D008562:Membrane Glycoproteins / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification; D051379:Mice; D009500:Neutralization Tests; D010446:Peptide Fragments / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification; D011817:Rabbits; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification; D013552:Swine; D014611:Vaccination; D014614:Vaccines, Synthetic / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification; D014759:Viral Envelope Proteins / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification
D000914:Antibodies, Viral; D000956:Antigens, Viral; D007074:Immunoglobulin G; D007132:Immunoglobulin Isotypes; D022321:Japanese Encephalitis Vaccines; D008562:Membrane Glycoproteins; D010446:Peptide Fragments; D011993:Recombinant Fusion Proteins; D014614:Vaccines, Synthetic; D014759:Viral Envelope Proteins; C080628:glycoprotein E, Japanese encephalitis virus