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 11 was screened with rabbit antisera to metacestodal T. solium and T. saginata crude extract. Primary clones (121) were identified, and after rescreening and lysogenization in Escherichia coli Y 1089, were tested in Western blot for reactivity with the same antisera. In addition, analyses were performed with rabbit antisera directed towards T. crassiceps and Echinococcus granulosus metacestode crude extract, sera from humans with neurocysticercosis (Mexico) and other important helminth diseases, mice and calves with experimental T. crassiceps and T. saginata infections and normal sera. Of those tested, 22 clones expressing beta-galactosidase fusion proteins (approximately 118-132 kDa) were reactive with IgG antibodies of cysticercotic patients and T. crassiceps infected mice. Of these clones, 11 were also sero-positive with calf-IgG antibodies against T. saginata larvae. None of the 22 clones reacted with IgG antibodies due to human cystic and alveolar echinococcosis, intestinal/hepatic or urinary schistosomiasis, African onchocerciasis or with sera from uninfected controls (man, rabbit, calf and mouse). Of these 22 clones, 15 have been subcloned into the plasmid vectors pGEX-2T (modified) and pT7T3 alpha 19. Expressed glutathione-S-transferase fusion proteins were again tested for sensitivity and specificity by Western blot, and concentrated by affinity chromatography. The nucleotide sequence of the cDNA inserts of 9 clones has been determined in pT7T3 alpha 19 and revealed identity in 4 and 5 clones, respectively.
Fischer
C
C
Fachbereich Biologie, Philipps-Universität Marburg, Germany.
Nosratian
R
R
Habtemichael
N
N
Tolle
R
R
Riemenschneider
V
V
Geyer
E
E
D000595:Amino Acid Sequence; D000818:Animals; D000947:Antigens, Helminth / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D001483:Base Sequence; D003551:Cysticercosis / Q000175:diagnosis*; D005260:Female; D008297:Male; D051379:Mice; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000276:immunology; D012698:Serologic Tests; D013621:Taenia / Q000276:immunology*
D000947:Antigens, Helminth; D011994:Recombinant Proteins