PREPARATION AND SEQUENCE-ANALYSIS OF TAENIA-CRASSICEPS METACESTODE RECOMBINANT ANTIGENS WITH POTENTIAL FOR SPECIFIC IMMUNODIAGNOSIS OF HUMAN CEREBRAL CYSTICERCOSIS

A Taenia crassiceps metacestode cDNA expression library in lambda gt 1 1 was screened with rabbit antisera to metacestodal T. solium and T. s aginata crude extract. Primary clones (121) were identified, and after rescreening and lysogenization in Escherichia coli Y1089, were tested in Western blot for reactivity with the same antisera. In addition, a nalyses were performed with rabbit antisera directed towards T. crassi ceps and Echinococcus granulosus metacestode crude extract, sera from humans with neurocysticercosis (Mexico) and other important helminth d iseases, mice and calves with experimental T. crassiceps and T. sagina ta infections and normal sera. Of those tested, 22 clones expressing b eta-galactosidase fusion proteins (approximately 118-132 kDa) were rea ctive with IgG antibodies of cysticercotic patients and T. crassiceps infected mice. Of these clones, 11 were also sere-positive with calf-I gG antibodies against T. saginata larvae. None of the 22 clones reacte d with IgG antibodies due to human cystic and alveolar echinococ cosis , intestinal/hepatic or urinary schistosomiasis, African onchocerciasi s or with sera from uninfected controls (man, rabbit, calf and mouse). Of these 22 clones, 15 have been subcloned into the plasmid vectors p GEX-2T (modified) and pT7T3 alpha 19. Expressed glutathione-S-transfer ase fusion proteins were again tested for sensitivity and specificity by Western blot, and concentrated by affinity chromatography. The nucl eotide sequence of the cDNA inserts of 9 clones has been determined in pT7T3 alpha 19 and revealed identity in 4 and 5 clones, respectively.