Although the normal intermediate hosts of Hymenolepis microstoma are insects, oncospheres invade the intestinal tissue of the definitive hosts, mice, within 30 min after oral egg inoculation.

The indirect immunofluorescent technique was used to determine the occurrence of IgA, IgM and IgG1 immunoglobulin-containing cells in local intestinal mucosal immune responses to Hymenolepis citelli, H. diminuta and H. microstoma infections in mice. In the intestinal lamina propria of H. citelli and H. diminuta infected mice there was no increase in the mean numbers of immunoglobulin-containing cells when compared with uninfected control mice, but there was in H. microstoma infected mice. The numbers of IgG1 – positive cells in both infected and uninfected mice were very small relative to IgA and IgM immunocytes. The distribution of immunocytes in the lamina propria of infected and uninfected mice was essentially similar and the localization of isotypes in duodenal sections showed no immunoglobulins in the villous epithelial cells. There was also no marked difference between primary and secondary infections indicating that immunoglobulin-containing cells play no major role in functional immunity against hymenolepid infections in the mouse. The presence of IgA and IgM was also demonstrated on the tegument of the tapeworms, although the distribution was patchy and more abundant on H. microstoma than on H. diminuta or H. citelli. The time of appearance of both isotypes was latest on H. citelli.

Patent, but not prepatent, Schistosoma mansoni infections in mice enhanced the expulsion of a superimposed infection with Hymenolepis diminuta. An antagonistic effect was also directed against a superimposed H. microstoma infection in mice harbouring patent S. mansoni infections.

The in vivo activity of two cyclosporins, cyclosporin A (CsA) and a non-immunosuppressive derivative of dihydrocylosporin A (DHCsA-d) against three tapeworms, Hymenolepis microstoma, H. diminuta and Mesocestoides corti, has been assessed. CsA reversibly reduced the dry weight of H. microstoma in the mouse, briefly delayed oviposition and had a statistically significant effect on worm numbers recovered. Oral and subcutaneous treatments of both CsA and DHCsA-d were effective in reducing worm weight; juvenile worms were most susceptible but worms of all ages responded to drug by a dramatic reduction in weight from which they recovered. Multiple courses of CsA were no more active than single courses of treatment but dose response suggested that a threshold level of drug was necessary to evoke activity. By contrast, H. diminuta in the rat was completely unaffected by CsA but no explanation for the differences in drug response by these two closely related helminths is forthcoming. Mesocestoides corti responded reversibly to CsA in the mouse by a reduction in asexual proliferation of both liver and peritoneal cavity tetrathyridia. The data presented argue in favour of a range of anti-parasitic activities by cyclosporins but the details of the various putative modes of action remain to be defined.

Cyclosporin A (CsA) acts as a powerful immunosuppressant through its binding to the cytosolic isomerase, cyclophilin (CyP), forming a complex which inhibits the phosphatase activity of calcineurin. The drug is also selectively anti-parasitic but its mode of action remains unknown. The mouse tapeworm, Hymenolepis microstoma is sensitive to CsA, but the rat tapeworm, H. diminuta is not susceptible either in rats, mice or in vitro. Using these two tapeworm models, the uptake and binding of CsA were examined in relation to parasite cyclophilins. Uptake and compartmentalization of the drug were markedly different in the two species: H. microstoma takes up more drug than does H. diminuta and sequesters more drug into intracellular compartments. Characterization of cyclophilins using both CsA binding and isomerase activity assays reveals that H. microstoma possesses two cyclophilin isoforms (Mr 17700 and 21400) with isomerase activity that is inhibited by CsA. Using identical assays, we have been unable to demonstrate CsA-binding proteins or CsA-sensitive isomerase activity in H. diminuta. These data suggest that the anthelmintic action of CsA relates in some way to the presence and function of parasite cyclophilins.

Cyclosporin A (CsA) induced significant changes in parasite morphology when administered to mice infected with Hymenolepis microstoma. Gross morphological damage consisted of proglottis swelling and the formation of protuberances from the worm surface, visible with a low-power dissecting microscope, occurring most frequently in the posterior third of the strobila. Gross morphology and ultrastructure were examined further using scanning and transmission electron microscopy. Swollen proglottides exhibited areas covered in small pits and fissures (diameter approximately 1–2 μm) but it was not possible to establish the significance of this damage. The brush-border and distal cytoplasm appeared largely intact although some evidence of swelling of the basal membrane invaginations and possible fluid accumulation was seen in drug-treated TEM sections. The apparent oedematous condition of many of the proglottides from drug-treated mice may indicate that CsA treatment mediates permeability changes in the worm surface membrane but the mechanisms by which this may occur remain to be elucidated. The effects of CsA on the morphology of H. microstoma correlate with the previously described anthelmintic activity of the drug against this parasite where CsA treatment dramatically reduces worm growth, retards migration into the bile duct and limits parasite survival.

Antineoplastic cyclophosphamide (CY) significantly reduced the survival and the weight of Hymenolepis microstoma in mice. Corticosteroid dexamethasone (DEX), on the other hand, did not influence the worm survival and the worms in DEX-treated mice became significantly heavier. Both immunosuppressants abolished H. microstoma-induced bile duct hypertrophy and submucous enteritis and lowered the number of mast cells (MC) in these regions. CY appeared to be more effective in depleting the bile duct MC whereas DEX seemed to affect more the duodenal MC population.

The transport of [14C]glucose by Hymenolepis microstoma in vitro following in vivo treatment with cyclosporin A (CsA) was determined over a range of concentrations. For untreated (control) worms glucose uptake showed saturation kinetics with a small diffusion component. Estimates of the maximum velocity of glucose uptake (Vmax) and the affinity of substrate for the glucose transporter (Kt) revealed that untreated 8-day-old worms had a Vmax twice that of 15-day-old worms and that younger worms had a lower Kt. An inverse relationship was demonstrated between log10 worm weight and the rate of uptake of [14C]glucose, reflecting the relatively greater number of glucose transporters due to the larger surface area: volume ratio of smaller worms. Treatment of H. microstoma with CsA in vivo significantly increased the diffusion component of glucose uptake in vitro. Parasites from drug-treated mice had a significantly lower Vmax for glucose uptake than size-matched controls. The affinity of glucose for its transporter in CsA-treated worms (Kt) was not significantly different from size-matched controls. Both juvenile and adult worms underwent transient depletion in total glycogen content after CsA treatment in vivo. The data confirm that CsA treatment in vivo disrupts the functional integrity of the worm tegument, one facet of which is impaired acquisition of glucose.

Cyclosporin A (CsA) is a widely investigated experimental anti-parasitic drug whose mode of action remains unresolved. The immunosuppressive action of CsA depends on the drug binding to the intracellular receptor cyclophilin (CyP). This study investigates whether complexing of CsA with parasite CyP is equally essential for its anthelmintic action. The correlation between initial cyclosporin-induced damage in vitro and drug binding to parasite CyP has been examined. CsA and the analogues B-5-49, CsH and CsA-acetate all induced similar damage to the tapeworm Hymenolepis microstoma in vitro in incubations between 2h and 4 days. The initial foci of drug damage were the parasite surface and mitochondria in the syncytium. In a competitive binding assay only B-5-49 displaced [H3]-CsA from either crude parasite cytosolic CyP or affinity-purified CyP while CsH and CsA-acetate had no effect. These data suggest strongly that cyclosporins act on the surfaces of helminth parasites but that drug action does not involve complex formation with CyP. An alternative drug-binding site must therefore be identified which may lead to the rational design of novel anthelmintic drugs.

Three hymenolepidid tapeworms, Hymenolepis diminuta, H. nana and H. microstoma, are commonly maintained in laboratory rodents and used in many experimental model systems of tapeworm infections. We examined partial sequences from the mitochondrial cytochrome c oxidase subunit 1 (CO1) gene and nuclear ribosomal internal transcribed spacer 2 (ITS2) sequences to infer phylogenetic relationships of the 3 hymenolepidid species. Parts of the CO1 gene and ITS2 were amplified by PCR and sequenced directly. The CO1 gene sequence obtained was the same in length (391 bp) among all specimens. In the case of ITS2, however, several insertions and deletions were detected (671–741 bp) not only among species but also between an American isolate and a Japanese isolate of H. diminuta. Percentage nucleotide differences between H. diminuta and H. microstoma, or H. diminuta and H. nana were 16·6–18·2% for the CO1 gene and 21·3–22·9% for ITS2. The differences in both sequences between H. microstoma and H. nana were about 14%. Phylogenetic trees inferred from both of the nucleotide sequences showed similar topology, and suggest that H. diminuta may have diverged from the common ancestral line the earliest, and that H. nana is closer to H. microstoma than to H. diminuta.