title: Immunization of mice with the non-toxic HC50 domain of botulinum neurotoxin presented by rabies virus particles induces a strong immune response affording protection against high-dose botulinum neurotoxin challenge.
abstract: We previously showed that rabies virus (RABV) virions are excellent vehicles for antigen presentation. Here, a reverse genetic approach was applied to generate recombinant RABV that express a chimeric protein composed of the heavy chain carboxyterminal half (HC50) of botulinum neurotoxin type A (BoNT/A) and RABV glycoprotein (G). To promote surface expression and incorporation of HC50/A into RABV virions, the RABV glycoprotein (G) ER translocation sequence, various fragments of RABV ectodomain (ED) and cytoplasmic domain were fused to HC50/A. The HC50/A chimeric proteins were expressed on the surface of cells infected with all of the recombinant RABVs, however, the highest level of surface expression was detected by utilizing 30 amino acids of the RABV G ED (HV50/A-E30). Our results also indicated that this chimeric protein was effectively incorporated into RABV virions. Immunization of mice with inactivated RABV-HC50/A-E30 virions induced a robust anti-HC50/A IgG antibody response that efficiently neutralized circulating BoNT/A in vivo, and protected mice against 1000 fold the lethal dose of BoNT/A.
authors.affiliation: Department of Microbiology and Immunology, Jefferson Medical College, Thomas Jefferson University, Philadelphia, PA, United States.
authors.forename: Waleed
authors.initials: W
authors.lastname: Mustafa
authors.forename: Fetweh H
authors.initials: FH
authors.lastname: Al-Saleem
authors.forename: Zidoon
authors.initials: Z
authors.lastname: Nasser
authors.forename: Rebecca M
authors.initials: RM
authors.lastname: Olson
authors.forename: Jeffrey A
authors.initials: JA
authors.lastname: Mattis
authors.forename: Lance L
authors.initials: LL
authors.lastname: Simpson
authors.forename: Matthias J
authors.initials: MJ
authors.lastname: Schnell
mesh_terms: D000818:Animals; D000907:Antibodies, Bacterial / Q000097:blood*; D057134:Antibodies, Neutralizing / Q000097:blood; D000956:Antigens, Viral / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D001428:Bacterial Vaccines / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology*; D019274:Botulinum Toxins, Type A / Q000737:chemistry* / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D001906:Botulism / Q000276:immunology / Q000517:prevention & control*; D005260:Female; D005822:Genetic Vectors / Q000235:genetics / Q000276:immunology; D006023:Glycoproteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D007114:Immunization; D007074:Immunoglobulin G / Q000097:blood; D051379:Mice; D011820:Rabies virus / Q000235:genetics* / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D014614:Vaccines, Synthetic / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology; D014759:Viral Envelope Proteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D014771:Virion / Q000235:genetics* / Q000378:metabolism
chemical_list: D000907:Antibodies, Bacterial; D057134:Antibodies, Neutralizing; D000956:Antigens, Viral; D001428:Bacterial Vaccines; D006023:Glycoproteins; D007074:Immunoglobulin G; D011993:Recombinant Fusion Proteins; D014614:Vaccines, Synthetic; D014759:Viral Envelope Proteins; C064951:glycoprotein G, Rabies virus; D019274:Botulinum Toxins, Type A