Schistosoma mansoni: two-dimensional gel electrophoretic analysis of antigens uniquely immunoreactive with protective rat serum.
Candidate vaccine antigens are defined by their differential immunoreactivity with antisera which are distinguishable by their capacity to confer passive resistance to infection. This "contrasting antisera" immunoassay has been successfully used in previous analyses of 4-week-old worm biosynthetically radiolabeled Schistosoma mansoni proteins to identify potentially protective antigens. Twice-infected Fischer (F-2x) and Wistar-Furth (W-2x) rat sera were the sources of protective and non-protective antibody, respectively. We have extended our original analysis by applying two-dimensional gel electrophoresis to resolve total and immunoreactive soluble proteins of the 4-week worms. Total proteins were characterized by silver staining and autoradiography. Radiolabeled protein antigens immunoprecipitated by F-2x and W-2x antisera were compared, and several were shown to be uniquely reactive with the protective immune serum. In a companion molecular approach to clone the candidate vaccine antigens, screening of a lambda gt11 adult S. mansoni cDNA expression library by the contrasting antisera assay has identified a clone (lambda 40) producing a fusion protein with epitopes uniquely reactive with F-2x. A rabbit antiserum to the lambda 40 fusion protein (anti-FP40) reacted with radiolabeled worm proteins in the 20-kDa size range. By 2D gel electrophoretic analysis, we can now demonstrate that anti-FP40 specifically immunoprecipitates most of the members of a multicomponent protein antigen subset 18-22 kDa in Mr, focusing over a pI range of 5.3-5.8, and recognized uniquely by F-2x.
Mark
H F
HF
Division of Biology and Medicine, Brown University, Providence, RI 02912.
el-Sherbeini
M
M
Goldberg
M
M
Suri
P K
PK
Sturley
S L
SL
Bostian
K A
KA
Knopf
P M
PM
D000818:Animals; D000909:Antibodies, Helminth / Q000276:immunology*; D000947:Antigens, Helminth / Q000032:analysis* / Q000276:immunology; D001345:Autoradiography; D015180:Electrophoresis, Gel, Two-Dimensional; D004591:Electrophoresis, Polyacrylamide Gel; D007106:Immune Sera / Q000276:immunology; D007525:Isoelectric Focusing; D011233:Precipitin Tests; D051381:Rats; D011916:Rats, Inbred F344; D011920:Rats, Inbred WF; D012550:Schistosoma mansoni / Q000276:immunology*
D000909:Antibodies, Helminth; D000947:Antigens, Helminth; D007106:Immune Sera