title: Development of an animal model to assess the immunogenicity of single-dose tetanus and diphtheria vaccines based on controlled release from biodegradable polymer microspheres.
abstract: We have determined threshold doses of aluminium phosphate (AlPO4) adsorbed tetanus toxoid (TT) and diphtheria toxoid (DT) in mice and guinea pigs with a view to developing an animal model to assess the immunogenicity of controlled release vaccines. A dose was sought (threshold dose) which produces little antibody after primary injection and a moderate response after a booster injection, thus mimicking the adult human response to TT and DT vaccines adsorbed on to aluminium adjuvants. After the first injection, mice and guinea pigs showed a dose response for both tetanus toxin IgG antibodies and tetanus antitoxin over a wide range of doses of AlPO4-adsorbed TT (0.01 to 0.2 Lf). After the second and third injections, there was no clear dose response for doses between 0.05 and 0.2 Lf However, doses between 0.01 and 0.04 Lf still showed a dose response after the second injection. Dilution of AlPO4-adsorbed TT in saline just before injection did not alter immunogenicity after the first injection, but stronger booster responses were seen in mice to diluted versus undiluted vaccine after the second and third injections. The threshold dose of AlPO4-adsorbed TT for both mice and guinea pigs was 0.01 Lf given in 100 microliters. For AlPO4-adsorbed DT, three strains of mice (inbred, Balb/c and C57/B6, and outbred, CD-1) showed a dose response after the first injection for DT IgG antibodies and diphtheria antitoxin at 0.1 and 0.2 Lf doses. Outbred CD-1 mice showed a dose response after the second and third injections also, whereas inbred mice showed inconsistent dose responses after the second injection and none after the third injection. In contrast to AlPO4-adsorbed TT, mice injected with undiluted AlPO4-adsorbed DT elicited significantly higher antibodies than those injected with diluted formulations, particularly after the first injection. The threshold dose of AlPO4-adsorbed DT for mice was 0.1 Lf in a volume of 250 microliters. Lower doses did not produce consistent antibody responses in mice. We propose that a single dose of controlled release formulations at doses not greater than 1/10 of a single human dose when injected into mice and guinea pigs should produce an antibody response similar or higher than two to three threshold doses of AlPO4-adsorbed TT or DT.
authors.affiliation: Massachusetts Public Health Biologic Laboratories, Boston, USA.
authors.forename: R K
authors.initials: RK
authors.lastname: Gupta
authors.forename: P
authors.initials: P
authors.lastname: Griffin
authors.forename: R
authors.initials: R
authors.lastname: Rivera
authors.forename: G R
authors.initials: GR
authors.lastname: Siber
mesh_terms: D000276:Adjuvants, Immunologic; D000327:Adsorption; D000328:Adult; D017607:Aluminum Compounds / Q000276:immunology; D000818:Animals; D001672:Biocompatible Materials; D003692:Delayed-Action Preparations; D004168:Diphtheria Toxoid / Q000008:administration & dosage / Q000276:immunology*; D006168:Guinea Pigs; D006801:Humans; D019344:Lactic Acid; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D008863:Microspheres; D008954:Models, Biological; D010710:Phosphates / Q000276:immunology; D011100:Polyglycolic Acid; D000077182:Polylactic Acid-Polyglycolic Acid Copolymer; D011108:Polymers; D013745:Tetanus Toxoid / Q000008:administration & dosage / Q000276:immunology*
chemical_list: D000276:Adjuvants, Immunologic; D017607:Aluminum Compounds; D001672:Biocompatible Materials; D003692:Delayed-Action Preparations; D004168:Diphtheria Toxoid; D010710:Phosphates; D011108:Polymers; D013745:Tetanus Toxoid; D000077182:Polylactic Acid-Polyglycolic Acid Copolymer; D011100:Polyglycolic Acid; D019344:Lactic Acid; C012714:aluminum phosphate