The effect of the tapeworm Hymenolepis nana on immunity to tuberculosis in mice.
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When Diphyllobothrium latum develops from larva to adult in a definitive host, it first sheds the entire larval 'body' before growth of an adult strobila starts. This process of shedding off the entire larval abothrial extremity, piece by piece, takes about 48 h. By this time the larva has usually reached the anterior third of the small intestine of the host. D. dendriticum and D. ditremum develop quite differently, although exhibiting similar anterior migrations. In these two species the larvae develop directly into adults without the larval 'body' first being shed. The implications of the observed differences in growth pattern between these three species of Diphyllobothrium to the classification of diphyllobothriid cestodes is discussed briefly.
In laboratory studies with 3 species of Calliphora more than half were shown to ingest eggs of Taenia hydatigena from the surface of dog faeces with a maximum of over 5000 eggs in 1 fly. Most eggs were voided within 48 h and between 38 and 48% of these eggs had lost their embryophores. In field studies carried out in the South Island of New Zealand, traps baited with dog faeces captured the following blowfly species in decreasing order of abundance: Hybopygia varia, Calliphora quadrimaculata, C. hortona and C. stygia. Peak numbers were trapped in January, February and March. Almost 25% of wild flies, caught after feeding for up to 3 min on dog faeces naturally contaminated with taeniid eggs, had eggs in their intestine. When administered to lambs all 4 species of fly transferred infection. Lambs grazed in winter on plots near kennels that had housed dogs with patent infections of T. hydatigena acquired higher worm burdens than those grazed further away. The burdens were greater downwind of the prevailing wind. After removal of the dogs, blowflies caught in the vicinity contained taeniid eggs but the contents of pitfall traps did not. Blowflies, and to a much lesser degree the insects and dust caught on sticky traps during the presence of the dogs, transferred infection when administered to naive lambs.
Infection with Hymenolepis microstoma significantly affected the lipid metabolism of young male Balb/C mice. Infection increased the rates of hepatic fatty acid and cholesterol synthesis and cholesterol synthesis by the gut. Decreases were recorded in testicular fatty acid synthesis and in the weights of testes and white epididymal adipose tissue. Plasma glucose decreased rapidly during infection. The observed changes in lipogenesis could not be attributed to changes in food intake or body temperature. The changes are discussed in relation to nutritional interactions between host and parasite and the possible effects on host hormone levels. The presence of newly synthesized fatty acid in H. microstoma is also reported.
Standard indirect immunocytochemical techniques have been interfaced with confocal scanning laser microscopy (for whole-mount preparations) and epifluorescence microscopy (for cryosections) to investigate the occurrence and distribution of serotoninergic and peptidergic nerve elements in adult H. diminuta. Serotonin (5-HT)-immunoreactivity (IR) was widespread throughout the worm, occurring in the paired cerebral ganglia, transverse commissure, the 10 longitudinal nerve cords and in a plethora of small nerve fibres of the peripheral nervous system. An abundance of serotoninergic nerve cell bodies was found in association with the lateral nerve cords. The genital atrium and accessory reproductive ducts were richly innervated with serotoninergic nerve fibres. Thirty-five antisera to 20 vertebrate regulatory peptides and 1 invertebrate peptide (FMRFamide) were used to screen the worm for neuropeptide IR. Immunostaining was obtained with antisera raised to pancreatic polypeptide (PP), peptide YY (PYY), neuropeptide Y (NPY), substance P (SP), peptide histidine isoleucine (PHI), xenopsin (XP) and FMRFamide. The most extensive pattern of IR occurred with antisera to PP and PYY, IR being evident in the cerebral ganglia, transverse commissure, longitudinal nerve cords and in small nerve fibres that ramified throughout the parenchyma. A series of bipolar nerve cell bodies between the median nerve cords displayed PP/PYY-IR. The distribution of FMRFamide-IR was reminiscent of the PP/PYY pattern but was less extensive. Comparison of the serotoninergic and peptidergic nervous systems has revealed general similarities and some distinct differences, especially with regard to the distribution of immunoreactive nerve cell bodies. Quantitative data are presented on the levels of PP-, SP-, PHI-, and gastrin-releasing peptide (GRP)-immunoreactivities demonstrable in acid-alcohol extracts of whole worms. The highest level of peptide IR determined was recorded for PP.
Cyclosporin A (CsA), administered in 5 daily subcutaneous doses of 50 mg/kg to MF1 mice immediately following infection with Hymenolepis diminuta enhanced parasite growth relative to controls. Drug administered at 24 h intervals for 10 days, and thereafter every 48 h to MF1 and CBA/Ca mice infected with H. diminuta, increased worm survival and growth, delayed host-mediated expulsion of the parasite and enabled some worms to develop to patency. Worm survival and weight both increased in a dose-dependent manner following daily CsA treatment of infected CBA/Ca and BALB/c mice (0-150 mg/kg CsA/day). Delay in parasite elimination was accompanied by increased frequency of worm-attachment in the anterior small intestine (MF1 mice given 5 daily doses of CsA [0-150 mg/kg] following infection); posteriad migration of worms was restricted in a dose-dependent manner. The data presented contrast markedly with the action of the same drug on H. microstoma in mice. Thus CsA treatment acts in opposing ways on two closely related parasites in the same host; this possibly reflects the mechanistic antagonism between immunosuppression and anthelmintic activity. This paper reports the first use of a specific T cell-suppressive drug on H. diminuta in the mouse, implicating the role of T cells in protective immunity to this parasite.
The free radical nitric oxide (NO), which is synthesized by nitric oxide synthase (NOS), has recently been discovered to function as a neuronal messenger. The presence of NOS was detected in the nervous system of adult Hymenolepis diminuta with NADPH-diaphorase (NADPH-d) histochemistry. The NADPH-d histochemical reaction is regarded as a selective marker for NOS in neuronal tissue. NADPH-d staining was observed in nerve fibres in the main and minor nerve cords and the transverse ring commissures, and in cell bodies in the brain commissure, along the main nerve cords, in the suckers and the rostellar sac. NADPH-d staining was also observed in the wall of the internal seminal vesicle and the genital atrium. The pattern of NADPH-d staining was compared with that of the 5-HT immunoreactive nervous elements. The NADPH-d staining reaction and the 5-HT immunoreactivity occur in separate sets of neurons. This is the first time the NADPH-d reaction has been demonstrated in the nervous system of a flatworm, indicating that NOS is present and that NO can be produced at this level of evolution.
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When incubated in vitro for 24 h, intact eggs, chemically shelled eggs (obtained by treating intact eggs with NaOCl), activated larvae (eggs in which the outer shell and inner envelope were removed), and oncospheres (activated larvae treated with papain to remove the embryophore) absorb and metabolize radioactive glucose. Intact eggs, which are covered by the impermeable shell, absorb only small amounts of exogenous radioactive glucose, while chemically shelled eggs, activated larvae, and oncospheres absorb much larger amounts. Only very small amounts of the exogenous glucose are incorporated into the ethanol-precipitable carbohydrate fraction (which would include glycogen) by any of the preparations of eggs/larvae. However, the glucose is incorporated into higher molecular weight end-products that are liberated into the incubation medium. There is a temporal shift in the ability of activated larvae and oncospheres to metabolize exogenous glucose. Activated larvae and oncospheres absorb but do not metabolize glucose during the first 8 h post-activation. Between 8 and 16 h post-activation, however, virtually all of the absorbed glucose is metabolized into higher molecular weight end-products that are liberated into the incubation media. This temporal shift suggests that activation of oncospheres and cysticercoid morphogenesis are accompanied by distinct changes in carbohydrate metabolism.
When incubated in vitro for 24 h, oncospheres of Hymenolepis diminuta absorb and metabolize radioactive glucose. Between 0 and 12 h post-activation, oncospheres absorb glucose, but glucose is neither metabolized into other carbohydrates nor incorporated into the ethanol-precipitable fraction (which would contain glycogen). Between 12 and 24 h post-activation glucose is incorporated into a number of higher molecular weight carbohydrates that are demonstrable in ethanol extracts of the larvae, as well as the incubation media. Furthermore, measurable amounts of radioactivity are incorporated into the ethanol-precipitable carbohydrate fraction of oncospheres. To determine if these temporal changes in carbohydrate metabolism occurred spontaneously following activation, oncospheres were pre-incubated for 12 h (0-12 h post-activation) in the absence or presence of glucose, and then transferred to media containing radioactive glucose for an additional 12 h (12-24 h post-activation). In these latter experiments, glucose absorption and metabolism between 12 and 24 h post-activation were virtually identical to glucose metabolism in oncospheres that were incubated in radioactive glucose for 0-12 h immediately following activation. Thus, these data do not support the hypothesis that the temporal shift in carbohydrate metabolism occurs spontaneously.
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The host-parasite relationship is exquisitely specific. In exploiting the host niche, a variety of helminth parasites have been shown to directly manipulate their hosts' immune responses. We assessed the ability of a whole-worm extract of Hymenolepis diminuta to modulate immune cell activation. Immune cells isolated from human blood or rodent spleens were activated with the T cell mitogen, concanavalin A (Con A) +/- H. diminuta extract and cytokine production (i.e. IL-2, -4, -10, -12) and proliferation assessed by ELISA and [3H]thymidine incorporation 24 and 72 h post-treatment, respectively. Co-treatment with the H. diminuta extract (100 microg protein/ml) virtually abolished Con A-induced immune cell proliferation, which was not due to increased apoptosis. Boiling of the worm extract reduced its anti-proliferative effect and fractionation indicated that a > 50 kDa component was predominantly responsible for the inhibition of Con A-induced immune cell proliferation. Cytokine determinations revealed that the H. diminuta extract significantly reduced Con A-stimulated IL-2 and IL-4, but enhanced the production of IFNy, IL-12 and IL-10. The increased IL-12 was due to an LPS contaminant in the extract and a helminth-derived 'IL-12'-like peptide that bound in the ELISA and Western blots. In contrast, a H. diminuta-derived factor directly stimulated IL-10 production by murine splenocytes, and contaminating LPS synergistically enhanced the production of IL-10. Thus, H. diminuta has the potential to block stimulated T cell proliferation and, by inhibiting IL-4 and promoting IL-10 production, may bias the immune environment towards one of immunoregulation and away from IL-4 dominated T helper 2 type events.
The complete Taenia asiatica mitochondrial genome was amplified by long extension polymerase chain reaction (long PCR) to yield overlapping fragments that were then completely sequenced. The whole mitochondrial genome was 13 703 bp long and contained 12 protein-encoding, 2 ribosomal RNA (small and large subunits), 22 transfer RNA genes and a short non-coding region. Thus, its gene contents are like those typically found in metazoan animal mitochondrial genomes (apart from the absence of atp8). All the genes were transcribed from the same strand. The 3' end 34 bp region of nad4L overlapped with the 5' end portion of nad4. The tRNA genes were 61-69 bp long, and the secondary structures of 18 tRNAs had typical clover-leaf shapes with paired DHU arms. However, trnC, trnS1, trnS2 and trnR had unpaired DHU arms that were 7-12 bp in length. The tRNAs that transferred serine lacked a DHU arm, as is also observed in a number of parasitic platyhelminths and metazoans. However, the trematode trnRs have paired DHU arms. The T. asiatica mtDNA non-coding region was like that in other cestodes since it was composed of a short non-coding region of 72 nucleotides and a long non-coding region of 176 nucleotides separated by a trnL1/, trnS2/, trnL2/, trnR/, nad5 gene cluster. The sequences of the cox1 genes between T. asiatica and T. saginata differ by 4.6%, while the T. asiatica cob gene differs by 4.1% and 12.9% from the cob genes of T. saginata and T. solium, respectively. In conclusion, the T. asiatica mitocondrial genome should provide a resource for comparative mitochondrial genomics and systematic studies of parasitic cestodes.
The equine intestinal cestode Anoplocephala perfoliata has been the subject of recent epidemiological and immunological studies because of its suspected association with intestinal disease in the horse. We have previously shown that the IgG(T) subtype antibody response to the 12/13 kDa component of the parasite excretory/secretory (E/S) antigen is positively correlated with parasite intensity. In this study, we utilize that correlation to examine the changes in natural infection intensity with age. Infection intensity based on IgG(T) responses showed a triphasic age-dependency pattern with peak mean worm burden in the 6 months-2 years age group, falling to a lower plateau level from 3 to 15 years, and rising again in older age groups. Anti-E/S total IgG was found to have a convex age-dependency curve, with maximal response in the 6 months-2 years old age group. IgG(a) showed a triphasic response similar to the age-intensity profile of IgG(T); IgG(c) showed steadily increasing levels of antibody with age. The IgG(b) age-dependency profile was intermediate between IgG(a) and IgG(c). Age-specific correlation coefficients between anti-12/13 kDa IgG(T) (as a measure of infection intensity) and IgG(a) and IgG(b) revealed statistically significant values for many age groups. The relative importance of exposure to infection and the development of acquired immunity as determinants of the observed age-intensity pattern is considered.