title: A new assay system detecting antibody production and delayed-type hypersensitivity responses to trinitrophenyl hapten in an individual mouse.
abstract: A new assay system detecting antibody production and delayed-type hypersensitivity (DTH) responses to trinitrophenyl hapten in an individual mouse (AS-DAD) was established. BALB/c mice were immunized intraperitoneally with varying amounts of 2,4,6-trinitrophenylated sheep red blood cells (TNP-SRBC) on day 0. Venous blood was collected on days 2, 4, 6, 8 and 10. Levels of anti-TNP IgM and IgG serum were assayed by enzyme-linked immunosorbent assay (ELISA). After series of bleeding the mice were challenged with 2,4,6-trinitrobenzene sulfonic acid (TNBS) solution in the footpad on day 14. Footpad swelling was measured 24 or 48 h after the challenge. Peak responses of the anti-TNP IgM and IgG production were detected 4 or 6 days after the immunization with 10(9) TNP-SRBC. Maximum DTH response was also observed with 10(9) TNP-SRBC 24 h after the challenge on day 14. The antibody and DTH responses were also induced in other normal inbred strains such as C3H/He and DBA/1 but not BALB/c nu/nu mice. To evaluate AS-DAD in immunopharmacological studies, various immunomodulating agents were examined in BALB/c mice by subcutaneous administration on days 0, 1, 2 and 3. Cyclosporin or cyclophosphamide at 100 mg/kg/day completely inhibited not only the anti-TNP IgM and IgG production but also the TNP-specific DTH response. Prednisolone at 0.5 mg/kg/day had no significant effect on the IgM and IgG production, whereas it inhibited the TNP-specific DTH response. Interestingly, histamine-added mouse gamma-globulin at 150 MG/kg/day clearly enhanced the anti-TNP IgM and IgG production, while it showed a suppressive effect on the TNP-specific DTH response. Levamisole at 5.0 mg/KG/day showed suppressive effects on the anti-TNP IgG production without affecting the IgM production and the DTH response. These results suggest that AS-DAD is useful for evaluating the immunopharmacological action of various agents.
authors.affiliation: Department of Haematology and Immunology, Nippon Zoki Pharmaceutical Company, Hyogo, Japan.
authors.forename: H
authors.initials: H
authors.lastname: Yoshii
authors.forename: Y
authors.initials: Y
authors.lastname: Fukata
authors.forename: K
authors.initials: K
authors.lastname: Yamamoto
authors.forename: H
authors.initials: H
authors.lastname: Yago
authors.forename: S
authors.initials: S
authors.lastname: Suehiro
authors.forename: Y
authors.initials: Y
authors.lastname: Yanagihara
authors.forename: H
authors.initials: H
authors.lastname: Okudaira
mesh_terms: D000818:Animals; D004797:Enzyme-Linked Immunosorbent Assay; D004912:Erythrocytes / Q000276:immunology; D006241:Haptens / Q000276:immunology*; D006968:Hypersensitivity, Delayed / Q000276:immunology*; D007118:Immunoassay / Q000379:methods*; D007074:Immunoglobulin G / Q000096:biosynthesis*; D007075:Immunoglobulin M / Q000096:biosynthesis*; D051379:Mice; D008807:Mice, Inbred BALB C; D008809:Mice, Inbred C3H; D008811:Mice, Inbred DBA; D008819:Mice, Nude; D014301:Trinitrobenzenes / Q000276:immunology*; D010132:p-Azobenzenearsonate / Q000276:immunology
chemical_list: D006241:Haptens; D007074:Immunoglobulin G; D007075:Immunoglobulin M; D014301:Trinitrobenzenes; D010132:p-Azobenzenearsonate