Characterization of various recombinant antigens from Echinococcus multilocularis for use in the immunodiagnosis.
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.
Kouguchi
Hirokazu
H
Hokkaido Institute of Public Health, Kita-Ku, Sapporo 060-0819, Japan. kouguchi@iph.pref.hokkaido.jp
Suzuki
Tomohiro
T
Yamano
Kimiaki
K
Honma
Hiroshi
H
Sawada
Yukiharu
Y
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
D000941:Antigens; D007106:Immune Sera; D011994:Recombinant Proteins