title: Characterization of various recombinant antigens from Echinococcus multilocularis for use in the immunodiagnosis.
abstract: Using four clones isolated from Echinococcus multilocularis cDNA library with alveolar echinococcosis (AE) patient sera, various antigens were expressed as ThioHis tag-fused protein. Recombinant EmII/3 antigen was produced as the five fragments divided into the N-terminal (#5 and #5s), the central (#6 and #6s) and the C-terminal domain (#7). Immunoblot analysis revealed that the #7 showed significant reactivity whereas those of #5 and #5s were relatively low. The #6 and #6s also showed lower reactivity than that of #7, although the two minor bands of #6 reacted with every serum. These results suggested that an immunodominant region of EmII/3 locate within the C-terminal one third. The #8s recombinant antigen, Ser23-Glu176 of actin filament fragmenting protein (AFFP), apparently reacted with the AE patient sera, while the #1 antigen synthesized as a full-length antigen BI did not show such high reactivity. Thus, #7 and #8s antigens showed significant potential for use in immunodetection of AE. In addition, the specific antibodies against #7 and #8s reacted with specific antigens in crude extract of E. multilocularis cyst, indicating that these antigens retained antigenicity common to native EmII/3 and AFFP, respectively.
authors.affiliation: Hokkaido Institute of Public Health, Kita-Ku, Sapporo 060-0819, Japan. kouguchi@iph.pref.hokkaido.jp
authors.forename: Hirokazu
authors.initials: H
authors.lastname: Kouguchi
authors.forename: Tomohiro
authors.initials: T
authors.lastname: Suzuki
authors.forename: Kimiaki
authors.initials: K
authors.lastname: Yamano
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Honma
authors.forename: Yukiharu
authors.initials: Y
authors.lastname: Sawada
mesh_terms: D000818:Animals; D000941:Antigens / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification; D015153:Blotting, Western; D003955:Diagnostic Tests, Routine / Q000379:methods*; D004445:Echinococcosis, Pulmonary / Q000175:diagnosis* / Q000276:immunology*; D048210:Echinococcus multilocularis / Q000276:immunology*; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D006801:Humans; D007106:Immune Sera / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification
chemical_list: D000941:Antigens; D007106:Immune Sera; D011994:Recombinant Proteins