SCHISTOSOMA-MANSONI - 2-DIMENSIONAL GEL-ELECTROPHORETIC ANALYSIS OF ANTIGENS UNIQUELY IMMUNOREACTIVE WITH PROTECTIVE RAT SERUM

被引：5

作者：

MARK, HFL ^{[1
]}

ELSHERBEINI, M ^{[1
]}

GOLDBERG, M ^{[1
]}

SURI, PK ^{[1
]}

STURLEY, SL ^{[1
]}

BOSTIAN, KA ^{[1
]}

KNOPF, PM ^{[1
]}

机构：

[1] BROWN UNIV, DIV BIOL & MED, PROVIDENCE, RI 02912 USA

来源：

EXPERIMENTAL PARASITOLOGY | 1991年 / 72卷 / 03期

关键词：

CONTRASTING ANTIBODY IMMUNOASSAY; FISCHER RAT; HELMINTH ANTIGENS; PASSIVE IMMUNIZATION; PROTECTIVE SERUM; RECOMBINANT PROTEINS; SCHISTOSOMA-MANSONI; TREMATODE; 2-DIMENSIONAL GEL ELECTROPHORESIS; VACCINES; WISTAR-FURTH RAT; BOVINE SERUM ALBUMIN (BSA); FUSION PROTEIN (FP); ISOELECTRIC FOCUSING (IEF); ISOELECTRIC POINT (PI); KILODALTONS (KDA); MOLECULAR WEIGHT (MW); NORMAL FISCHER RAT SERUM (FNRS); NORMAL RABBIT SERUM (NRBS); PHOSPHATE-BUFFERED SALINE (PBS); PROTEIN-A CONJUGATED TO SEPHAROSE (PAS); POLYACRYLAMIDE GEL ELECTROPHORESIS IN SODIUM DODECYL SULFATE (SDS-PAGE); RELATIVE MW (MR); SOLUBLE FRACTION RELEASED FROM 4-WEEK WORMS BY FREEZE THAW + HOMOGENIZATION (S2; TRICHLOROACETIC ACID (TCA); TWICE-INFECTED FISCHER RAT SERUM (F-2X); TWICE-INFECTED WISTAR-FURTH RAT SERUM (W-2X); 2-DIMENSIONAL (2D);

D O I：

10.1016/0014-4894(91)90149-Q

中图分类号：

R38 [医学寄生虫学]; Q [生物科学];

学科分类号：

07 ; 0710 ; 09 ; 100103 ;

摘要：

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 γgt11 adult S. mansoni cDNA expression library by the contrasting antisera assay has identified a clone (γ40) producing a fusion protein with epitopes uniquely reactive with F-2x. A rabbit antiserum to the γ40 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. © 1991.

引用

页码：294 / 305

页数：12

共 32 条

[21] CONTROL OF PARASITIC INFECTIONS IN NONPERMISSIVE HOSTS - APPLICATIONS TO IMPROVEMENT OF VACCINE EFFICACY [J].