title: Immunization with a Double-Mutant (R192G/L211A) of the Heat-Labile Enterotoxin of Escherichia coli Offers Partial Protection against Campylobacter jejuni in an Adult Mouse Intestinal Colonization Model.
abstract: We have previously shown that antibodies to cholera toxin (CT) reacted with the major outer membrane proteins (MOMPs) from Campylobacter jejuni strains on Western blot. Further, oral immunization with CT significantly protected against challenge with C. jejuni in an adult mouse colonization model of infection. CT and the heat-labile enterotoxin (LT) of enterotoxigenic Escherichia coli are structurally and functionally related. LT and its mutants including the double-mutant LT (R192G/L211A) (dmLT), are powerful mucosal adjuvants. Unlike LT which is reactogenic, dmLT has been shown to be safe for human use. In the current study, we determined whether rabbit anti-dmLT antibodies reacted with MOMPs from C. jejuni strains and whether immunization with dmLT would afford protection against C. jejuni. On Western blot, the MOMPs from C. jejuni 48 (Penner serotype O:19), C. jejuni 75 (O:3) and C. jejuni 111 (O:1,44) were probed with rabbit antibodies to dmLT or LT-E112K (a non-toxic LT mutant), which showed a lack of reaction. Adult BALB/c mice were orally immunized with dmLT and orally challenged with C. jejuni 48 or 111. Protection from colonization with the challenge bacteria was studied by enumerating Campylobacter colonies in feces daily for 9 days. Vaccination produced robust serum and stool antibody responses to dmLT and no antibody responses to C. jejuni MOMP. Vaccinated mice showed reduced colonization and excretion of both challenge strains compared to control mice. However, the differences were not statistically significant. The protective efficacy of the dmLT vaccine varied from 9.1% to 54.5%. The lack of cross-reaction between the MOMP and dmLT suggests that protection is not mediated by cross-reacting antibodies, but may be due to activation of innate immunity. As dmLT is safe for humans, it could be incorporated into a C. jejuni vaccine to enhance its efficacy.
authors.affiliation: Departments of Microbiology, Faculty of Medicine, Kuwait University, Jabriya, Kuwait.
authors.forename: M John
authors.initials: MJ
authors.lastname: Albert
authors.affiliation: Departments of Microbiology, Faculty of Medicine, Kuwait University, Jabriya, Kuwait.
authors.forename: Shilpa
authors.initials: S
authors.lastname: Haridas
authors.affiliation: Departments of Microbiology, Faculty of Medicine, Kuwait University, Jabriya, Kuwait.
authors.forename: Mathew
authors.initials: M
authors.lastname: Ebenezer
authors.affiliation: Departments of Microbiology, Faculty of Medicine, Kuwait University, Jabriya, Kuwait.
authors.forename: Raj
authors.initials: R
authors.lastname: Raghupathy
authors.affiliation: Department of Biochemistry, Faculty of Medicine, Kuwait University, Jabriya, Kuwait.
authors.forename: Islam
authors.initials: I
authors.lastname: Khan
mesh_terms: D000276:Adjuvants, Immunologic / Q000494:pharmacology; D000818:Animals; D000907:Antibodies, Bacterial / Q000276:immunology; D001427:Bacterial Toxins / Q000276:immunology*; D001428:Bacterial Vaccines / Q000276:immunology; D016123:Campylobacter jejuni / Q000276:immunology*; D004195:Disease Models, Animal; D054307:Enterotoxigenic Escherichia coli / Q000276:immunology*; D004768:Enterotoxins / Q000276:immunology*; D029968:Escherichia coli Proteins / Q000276:immunology; D005243:Feces / Q000382:microbiology; D006358:Hot Temperature; D007114:Immunization / Q000379:methods; D007422:Intestines / Q000276:immunology* / Q000382:microbiology*; D051379:Mice; D008807:Mice, Inbred BALB C; D014611:Vaccination / Q000379:methods
chemical_list: D000276:Adjuvants, Immunologic; D000907:Antibodies, Bacterial; D001427:Bacterial Toxins; D001428:Bacterial Vaccines; D004768:Enterotoxins; D029968:Escherichia coli Proteins