A Taenia solium glutathione-S-transferase fraction (SGSTF) was isolated from a metacestode crude extract by affinity chromatography on reduced glutathione (GSH)-sepharose. The purified fraction displayed a specific glutathione S-transferase (GST) activity of 2.8 micromol/min/mg and glutathione peroxidase selenium-independent activity of 0.22 micromol/min/mg. Enzymatic characterization of the fraction suggested that the activity was closer to the mammalian mu-class GSTs. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis, gel filtration, and enzyme activity analysis showed that the fraction was composed of a major band of Mr = 26 kd and that the active enzyme was dimeric. Immunohistochemical studies using specific antibodies against the major 26-kd band of the SGSTF indicated that GST protein was present in the tegument, parenchyma, protonephridial, and tegumentary cytons of the T. solium metacestode. Antibodies generated against the SGSTF tested in western blot showed cross-reactivity against GSTs purified from Taenia saginata, T. taeniaeformis, and T. crassiceps, but did not react with GSTs from Schistosoma mansoni, or mice, rabbit, and pig liver tissue. Furthermore, immunization of mice with SGSTF reduced the metacestode burden up to 74.2%. Our findings argue in favor of GST having an important role in the survival of T. solium in its hosts.
Taenia crassiceps cysticerci were disrupted through trypsinization to isolate cells which can be maintained in culture for up to 15 days. When injected intraperitoneally into susceptible BALB/cAnN mice, complete cysticerci were recovered in a number that is proportional to the quantity of injected cells. Thus, cysticerci contain cells which can reconstitute complete cysts, suggesting that individual cells play a role, independent to budding, during asexual multiplication of T. crassiceps cysticerci in the peritoneal cavity of mice. In contrast, injection of the cells into resistant C57BL/6J mice does not result in the recovery of complete cysts. These findings provide a new experimental model to identify resistance factors in the hosts, for the in vitro screening of anti-cysticerci drugs and for the genetic manipulation of cysticerci through recombinant DNA techniques.
Evaginated Taenia solium metacestodes dissected from infected pork meat were incubated in vitro in RPMI 1640 medium with tritiated thymidine, washed, and further incubated for various chase periods. Worms were fixed and embedded in Poly/Bed and sections were processed for autoradiography. Results showed that all longitudinal sections had a germinative region located 500-700 mm posterior to the apex of the scolex with tegumentary cytons arranged in staggered columns perpendicular to the tegument. After 6-hr pulse and 0-12-hr chase periods, a large number of labeled cells were found in the parenchyma and tegumentary wall, included were myocytons, calcareous corpuscle cells, flame cells, osmoregulatory channel cells, and, in the medullary parenchyma, labeled undifferentiated round cells with a large nucleus, prominent nucleolus, abundant ribosomes, and no cytoplasmic organelles. These undifferentiated cells were not labeled after 24-hr and 48-hr chase periods, an observation that strongly suggests these cells divide and migrate toward the tegument in a pattern similar to that described for other cestodes. The morphology and localization of these cells support the view that they are stem cells that give rise to the various cell types of the tegumentary wall. The results indicate that T. solium contains a germinative tissue similar to that described in other cestodes, in which stem cells proliferate continuously, differentiate, and migrate to the tegument, constituting the main process by which these worms develop from metacestode to the adult stage.
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