title: Sublingually administered Bacillus subtilis cells expressing tetanus toxin C fragment induce protective systemic and mucosal antibodies against tetanus toxin in mice.
abstract: Sublingual (SL) immunization against infectious agents or bacterial toxins is not a common route for antigen delivery. However, in our continued search for a needle-free platform for vaccine administration, we evaluated the efficacy of SL immunization with Bacillus subtilis engineered to express tetanus toxin fragment C (TTFC). We compared the results obtained with those for intranasal (IN) immunization with the same vaccine, which we recently reported to induce complete protection in mice against a 2×LD100 challenge of tetanus toxin (Lee et al., Vaccine 28:6658-65). Groups of animals received 3-4 immunizations of 10(9)B. subtilis vegetative cells expressing TTFC given IN or SL. Other SL immunized groups received either purified recombinant TTFC (rTTFC) or B. subtilis placebo. A non-toxic mutant of Escherichia coli heat labile enterotoxin (mLT) was included as adjuvant in some of the studies. Mice inoculated by either IN or SL administration developed protective IgG antibodies against tetanus toxin challenge. Similar of higher IgA levels in saliva, vaginal wash and feces were detected in animals immunized SL with B. subtilis cells expressing TTFC compared with IN-immunized mice or mice immunized SL with rTTFC. SL immunization promoted a mixed Th1/Th2 response, based on cytokine analysis (IL-2, IL-4, IL-10 and INFγ). Antigen-stimulated tissues (lung, intestine, spleen and lymph nodes) revealed a dramatic increase in the density of MHC class II+ expressing cells compared to all other groups. The antibody response to TTFC was superior when the adjuvant mLT was excluded from IN and SL immunizations. However, SL administration of mLT induced strong systemic and mucosal antibody responses, indicating that successful use of this route of immunization is not specific to tetanus toxin. We conclude that SL immunization is a promising, effective, safe, non-invasive and convenient method for mucosal delivery of B. subtilis cells expressing tetanus vaccine and, potentially, other immunogens. SL immunization appears to induce both systemic and mucosal immune responses.
authors.affiliation: Division of Infectious Diseases, Tufts University Cummings School of Veterinary Medicine, 200 Westboro Road, North Grafton, MA 01536, USA.
authors.forename: J Hellen
authors.initials: JH
authors.lastname: Amuguni
authors.forename: Sangun
authors.initials: S
authors.lastname: Lee
authors.forename: Kathryn O
authors.initials: KO
authors.lastname: Kerstein
authors.forename: David W
authors.initials: DW
authors.lastname: Brown
authors.forename: Boris R
authors.initials: BR
authors.lastname: Belitsky
authors.forename: John E
authors.initials: JE
authors.lastname: Herrmann
authors.forename: Gerald T
authors.initials: GT
authors.lastname: Keusch
authors.forename: Abraham L
authors.initials: AL
authors.lastname: Sonenshein
authors.forename: Saul
authors.initials: S
authors.lastname: Tzipori
mesh_terms: D000276:Adjuvants, Immunologic / Q000008:administration & dosage; D000281:Administration, Intranasal; D000286:Administration, Sublingual; D000818:Animals; D000907:Antibodies, Bacterial / Q000097:blood*; D000992:Antitoxins / Q000097:blood*; D001412:Bacillus subtilis / Q000235:genetics / Q000276:immunology*; D001427:Bacterial Toxins / Q000008:administration & dosage; D016207:Cytokines / Q000378:metabolism; D004768:Enterotoxins / Q000008:administration & dosage; D029968:Escherichia coli Proteins / Q000008:administration & dosage; D005243:Feces / Q000737:chemistry; D005260:Female; D018928:Immunity, Mucosal; D007070:Immunoglobulin A / Q000032:analysis; D007074:Immunoglobulin G / Q000097:blood; D051379:Mice; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology; D012463:Saliva / Q000737:chemistry; D013744:Tetanus Toxin / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology*; D013745:Tetanus Toxoid / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology*; D014611:Vaccination / Q000379:methods; D014614:Vaccines, Synthetic / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology; D014621:Vagina / Q000737:chemistry
chemical_list: D000276:Adjuvants, Immunologic; D000907:Antibodies, Bacterial; D000992:Antitoxins; D001427:Bacterial Toxins; D016207:Cytokines; D004768:Enterotoxins; D029968:Escherichia coli Proteins; D007070:Immunoglobulin A; D007074:Immunoglobulin G; D011994:Recombinant Proteins; D013744:Tetanus Toxin; D013745:Tetanus Toxoid; D014614:Vaccines, Synthetic; C027538:heat-labile enterotoxin, E coli