title: In vivo induction of interleukin-12 mRNA expression after oral immunization with Salmonella dublin or the B subunit of Escherichia coli heat-labile enterotoxin.
abstract: Mice orally immunized with Salmonella dublin EL23, a nonreverting, aromatic-dependent, histidine-requiring mutant transformed with a plasmid which carries a gene that codes for production of the B subunit of the heat-labile toxin (LT-B) of enterotoxigenic Escherichia coli, or with purified LT-B alone were compared for their ability to initiate expression of interleukin-12 (IL-12) mRNAs at mucosal sites. At 6 or 20 h following oral immunization, the Peyer's patches and mesenteric lymph nodes were removed, and polyadenylated mRNA was prepared from each tissue. Constitutive expression of an mRNA encoding the p35 subunit of IL-12 was observed in control as well as immunized mice. Conversely, expression of an mRNA encoding the p40 subunit of IL-12 was not detected in control animals but was dramatically upregulated in immunized mice. By using semiquantitative reverse transcription-PCR (RT-PCR) followed by competitive RT-PCR, differences in the magnitude of IL-12 p40 mRNA expression were quantified. Six hours after oral immunization with the Salmonella construct, mice had 12.1- and 8.4-fold increases in expression of IL-12 p40 mRNA in the Peyer's patches and mesenteric lymph nodes, respectively, compared with control mice receiving only saline. By 20 h, the pattern of increased mRNA expression was reversed, showing 2.5- and 17.6-fold increases in the Peyer's patches and mesenteric lymph nodes, respectively. Oral immunization with LT-B alone also stimulated IL-12 p40 mRNA expression, but to a lesser extent. The constitutive expression of IL-12 p35 mRNA at these mucosal sites coupled with a rapid and dramatic induction of IL-12 p40 mRNA following immunization with wild-type or attenuated strains of S. dublin is consistent with other investigations which support a role for IL-12 in modulating cell-mediated immune responses against intracellular pathogens.
authors.affiliation: Department of Microbiology and Immunology, Tulane University Medical Center, New Orleans, Louisiana 70112.
authors.forename: K L
authors.initials: KL
authors.lastname: Bost
authors.forename: J D
authors.initials: JD
authors.lastname: Clements
mesh_terms: D000284:Administration, Oral; D000818:Animals; D000942:Antigens, Bacterial / Q000276:immunology; D001427:Bacterial Toxins / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology*; D001428:Bacterial Vaccines / Q000008:administration & dosage / Q000276:immunology*; D001483:Base Sequence; D004768:Enterotoxins / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology*; D029968:Escherichia coli Proteins; D005260:Female; D018664:Interleukin-12 / Q000096:biosynthesis* / Q000235:genetics; D008198:Lymph Nodes / Q000276:immunology; D008221:Lymphoid Tissue / Q000276:immunology*; D008643:Mesentery / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D010581:Peyer's Patches / Q000276:immunology; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000096:biosynthesis; D011994:Recombinant Proteins / Q000276:immunology; D012475:Salmonella / Q000276:immunology*
chemical_list: D000942:Antigens, Bacterial; D001427:Bacterial Toxins; D001428:Bacterial Vaccines; D004768:Enterotoxins; D029968:Escherichia coli Proteins; D012333:RNA, Messenger; D011994:Recombinant Proteins; D018664:Interleukin-12; C027538:heat-labile enterotoxin, E coli