Neurocysticercosis (NCC) occurs following brain infection by larvae of the cestode Taenia solium. It is the leading cause of preventable epilepsy worldwide and therefore constitutes a critical health challenge with significant global relevance. Despite this, much is still unknown about many key pathogenic aspects of the disease, including how cerebral infection with T. solium results in the development of seizures. Over the past century, valuable mechanistic insights have been generated using both clinical studies and animal models. In this review, we critically assess model systems for investigating disease processes in NCC. We explore the respective strengths and weaknesses of each model and summarize how they have contributed to current knowledge of the disease. We call for the continued development of animal models of NCC, with a focus on novel strategies for understanding this debilitating but often neglected disorder.

Neurocysticercosis is a parasitic disease caused by encysted larvae of Taenia solium in the human central nervous system. Cysts mainly affect the cerebral hemispheres, although they can also be found in ventricles, basal cisterns, and subarachnoid spaces, and rarely in the cerebellum. Given the impossibility of studying the disease in human patients, Cardona et al. (1999) developed a mouse model of neurocysticercosis, using Mesocestoides corti, a closely related cestode. This allows us to study the parasite–host relationship and the mechanisms involved in the disease, in order to improve the therapy. In this murine model of neurocysticercosis, the location of tetrathyridia in parenchyma, ventricles and meninges has already been reported. The aim of this work is to report the cerebellum as a new location for M. corti tetrathyridia in the murine model of neurocysticercosis. A murine model that reproduces the human pathology is essential to evaluate the symptomatology and response to drug treatment in experimentally infected mice.

The histone chaperone SET/TAF-Iβ is implicated in processes of chromatin remodelling and gene expression regulation. It has been associated with the control of developmental processes, but little is known about its function in helminth parasites. In Mesocestoides corti, a partial cDNA sequence related to SET/TAF-Iβ was isolated in a screening for genes differentially expressed in larvae (tetrathyridia) and adult worms. Here, the full-length coding sequence of the M. corti SET/TAF-Iβ gene was analysed and the encoded protein (McSET/TAF) was compared with orthologous sequences, showing that McSET/TAF can be regarded as a SET/TAF-Iβ family member, with a typical nucleosome-assembly protein (NAP) domain and an acidic tail. The expression patterns of the McSET/TAF gene and protein were investigated during the strobilation process by RT-qPCR, using a set of five reference genes, and by immunoblot and immunofluorescence, using monospecific polyclonal antibodies. A gradual increase in McSET/TAF transcripts and McSET/TAF protein was observed upon development induction by trypsin, demonstrating McSET/TAF differential expression during strobilation. These results provided the first evidence for the involvement of a protein from the NAP family of epigenetic effectors in the regulation of cestode development.

Neurocysticercosis (NCC) is the most common parasitic disease of the central nervous system (CNS) caused by the larval form of the tapeworm Taenia solium. NCC has a long asymptomatic period with little or no inflammation, and the sequential progression to symptomatic NCC depends upon the intense inflammation associated with degeneration of larvae. The mechanisms involved in these progressive events are difficult to study in human patients. Thus it was necessary to develop an experimental model that replicated NCC. In this review, we describe studies of a murine model of NCC in terms of the release/secretion of parasite antigens, immune responses elicited within the CNS environment and subsequent pathogenesis. In particular, the kinetics of leukocyte subsets infiltrating into the brain are discussed in the context of disruption of the CNS barriers at distinct anatomical sites and the mechanisms contributing to these processes. In addition, production of various inflammatory mediators and the mechanisms involved in their induction by the Toll-like receptor signaling pathway are described. Overall, the knowledge gained from the mouse model of NCC has provided new insights for understanding the kinetics of events contributing to different stages of NCC and should aid in the formulation of more effective therapeutic approaches.

The influence of nonspecific immunomodulation on the course of experimental infection was examined in larval cestodosis (Mesocestoides corti) and ascaridosis (Ascaris suum) in mice. Immunosuppressive treatment (with azathioprine or hydrocortisone) resulted in a decrease of resistance in both models. The subsequent administration of T-activin to immunosuppressed mice led to the restoration of resistance to a level equal to that of untreated control mice. The administration of different immunomodulators partially protected mice against M. corti (T-activin. thymomodulin) or A. suum (T-activin. thymomodulin. thymosin fr.5. bursa-activin) infection. The protective effect of different treatments did not correlate with the level of specific antibody in the sera of infected mice. These results, which confirmed the decisive role of T-cell immunity in the resistance to the helminth infections, raise the possibility of the use of immunomodulators (thymic preparations) in the immunoprophylaxis of helminthoses.

The activities of selected enzymes of carbohydrate metabolism were measured in tetrathyridia of Mesocestoides corti and in adult females and males of Heterakis spumosa. When the species were compared, only lactate dehydrogenase and phosphoenolphyruvate carboxykinase activities were considerably higher in M. corti. Activities of other enzymes were higher in H. spumosa, with malate dehydrogenase activity being considerable so. In H. spumosa, enzyme activity was higher, and succinate dehydrogenase markedly so in males, when compared with females. Tetrathyridia aged 170 and 210 days show relatively stable malate and lactate dehydrogenase activities, and mice of ICR and BALB/c strains are suitable for the maintenance of tetrathyridia.

Many liver-invasive parasites cause extensive liver damage which may result in an impaired ability to catabolize endotoxin. The influence of endogenous endotoxin on the progress of liver-invasive parasitic diseases has been investigated in murine Mesocestoides corti infection. Invasion of liver tissue by tetrathyridia resulted in extensive parenchymal destruction with fibrosis. In association with this, undetoxified endotoxin, in potentially biologically active concentration, was found on peritoneal macrophages, 5 months post-M. corti infection. Host susceptibility was influenced by the Lps gene for responsiveness to lipopolysaccharide (LPS). The parasite burden of LPS-responsive (C3H/HeN) mice was significantly increased in the livers of these mice when compared to LPS-resistant (C3H/HeJ) mice. LPS reduced the ability of normal peritoneal macrophages to kill tetrathyridia, when co-cultured in vitro. LPS also abrogated the ability of recombinant interferon-γ (r.IFN-γ) to enhance macrophage larvicidal activity. These in vitro findings were confirmed in vivo. Daily intraperitoneal administration of LPS, at low concentration, caused a 4-fold increase in parasite burden in the liver, while r.IFN-γ at optimal concentration reduced parasite burden by 57%. Post-infection macrophages have previously been shown to be refractory to cytokine-activation for larval killing. In this report, we conclude that (1) this refractoriness may be due to the presence of undetoxified endotoxin on post-infection macrophages and (2) endotoxin may reduce host resistance by abrogating effector macrophage response to IFN-γ.

The genetic influences upon host variation in eosinophilia and resistance to helminth infection, and the relationship between these parameters, was investigated in 9 inbred and 3 hybrid strains of mice infected with Mesocestoides corti. Blood, bone marrow, spleen and peritoneal fluid eosinophilia were far higher in SJL mice than in any other inbred strain. SWR, NIH, C3H and BALB/c mice were high responders to M. corti whereas CBA and 3 congenic strains sharing the B10 background (C57BL/10, B10.S, B10.G) were low responders. Some of the genes for high eosinophil responsiveness appeared to be dominant, as F1 hybrids from high and low response parental strains were intermediate to high in response to infection. SJL and NIH strains were highly susceptible to infection with M. corti, larval burdens at 21 days after infection with 100 tetrathyridia being considerably higher (>1000) than all other strains. BALB/c (≃ 700 larvae) were designated susceptible, SWR (>400 larvae) were resistant and the BIO congenics (<400 larvae) were highly resistant. Genes influencing resistance also appeared to be dominant, as F1 hybrids between resistant and susceptible parental strains were intermediate to resistant on infection. The overall response patterns indicate a direct correlation between susceptibility to infection and high eosinophil responsiveness, but this relationship is not consistent in all strains.

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.

Peritoneal macrophages from Mesocestoides corti-infected mice showed a marked and progressive loss of ability to act as accessory cells for syngeneic Con A-stimulated mesenteric lymph node lymphocytes. The same effect on the macrophages could be induced by intraperitoneal injection of M. corti culture supernatant, despite a concurrent increase in numbers of peritoneal adhesive macrophages. The findings are used to compare and contrast the known immunomodulatory effects of M. corti and taeniid metacestodes, the latter differing chiefly in their potential for modifying T-cell as well as macrophage behaviour.

The regulatory role of tumour necrosis factor (TNF) was investigated in murine infection with tetrathyridia of Mesocestoides corti. Recombinant TNFα reduced macrophage larvicidal activity in vitro. M. corti primed mice for TNF release in response to bacterial lipopolysaccharide (LPS) in vivo. TNF activity was amplified 100-fold at 14 days post-infection (p.i.), with a further rise at day 28 p.i. Maximal inflammatory reaction was observed histologically in the liver at the height of TNF activity. Hepatic necrosis was located within inflammatory foci, but not within the vicinity of the parasite itself, suggesting that TNF may contribute to the pathogenesis of infection. Peritoneal cells from infected mice, when stimulated with tetrathyridia in vitro, showed a 4-fold increase in TNFα activity at day 14 p.i. However, when peritoneal cells were stimulated with LPS in vitro, a marked increase in TNFα secretion was observed at 2 months post-infection followed by a slow decline. It is suggested that impaired macrophage effector function, previously attributed to endogenous endotoxin, which gains access to peritoneal macrophages through an inability of the liver to detoxify endotoxin, may be mediated through TNFα.

Killing of metacestodes by normal or post-infection macrophages and the regulation of this activity by cytokines were studied in vitro. The protoscolecidal activity of normal macrophages against Echinococcus granulosus was inhibited by a product of naive T-enriched lymphocytes co-cultured with protoscoleces (PSC). By contrast, supernates from co-cultures of Mesocestoides corti tetrathyridia (MCT) and T-enriched or B-enriched normal lymphocytes increased killing of MCT by normal macrophages. Larvicidal activity (against both PSC and MCT) was enhanced by high concentrations of macrophage-activating factors produced by Con A-stimulated rat lymphocytes (Con A-LK), but was reduced by low concentrations of these factors.

Three strains of male and female mice, CFLP, BALB/c and CBA/ca, were infected i.p. with the tetrathyridia of Mesocestoides corti and examined in groups of 5 at 0, 7, 14, 21, 28 and 42 days post-infection. At post-mortem the numbers of parasites both loose in the peritoneal cavity and in the liver tissue were counted, the livers weighed fresh and sections of liver stained to examine the inflammatory response, encapsulation of the tetrathyridia and for eosinophils, neutrophils, mast cells and plasma cells. BALB/c mice had significantly more parasites loose in the peritoneal cavity than CFLP and CBA/ca mice. Infected livers of all three strains were significantly heavier than control livers; the heaviest livers were those of the CFLP followed by the CBA/ca mice which also showed the greatest rate of weight increase. The tetrathyridia from the peritoneal cavities of the CBA/ca and CFLP mice were covered with a ‘mucilage-like’ substance. Tetrathyridia within the host capsules of the CBA/ca mice contained host cells. There was no strain difference with regard to numbers of tetrathyridia in the liver but male mice harboured significantly more parasites than females. Differences in the numbers of cell types within the liver were detected between the strains but no one strain showed any consistent pattern. There was an overall increase in total white blood counts as well as an increase in the number of eosinophils, monocytes, neutrophils and lymphocytes during the course of the infection.

Fifty tetrathyridia of Mesocestoides corti showing various degrees of scolex fission during asexual multiplication were subjected to estimation of the total number of neurons in the central ganglia, using a ‘fractionator’ designed sampling scheme on every 10th Historesin section through the specimen. Neurons were stained with Cresyl fast violet and viewed by transmission light microscopy. Cells were only counted if a nucleolus was present. The animals were divided into groups according to their stage in the asexual reproduction process indicated by head shape and sucker number. Animals which had a narrow head with 4 suckers contained about 250 neurons. As their heads became wider, in preparation for fission, neurons were overproduced by up to 5-fold. As additional suckers were developed (8 in total), neurons were reduced to about 500. Finally, when the animals had 2 clearly discernible heads, with 4 suckers on each, the total number of neurons (per head) was again approximately 250. As the heads became wider again, neuron over-production recommenced. Exceptionally shaped animals with an X- or T-shaped single head were observed. These results suggest that there is a preparatory stage for neuronal regeneration, during which the basic population of neurons is about 250. This is followed by neuron over-production followed by a selective cell death of neurons which occurred during asexual multiplication. Another cycle for neuronal regeneration can occur before an animal completes its fission process.

The activation of peritoneal macrophage effector functions after therapy with free PZQ and PZQ incorporated in liposomes (lip.PZQ) was studied in the Mesocestoides corti–mouse model system. Each drug formulation was administered to an infected group of mice in 6 daily doses from day 14 p.i. Phagocytic activity of macrophages increased significantly after the administration of both drug formulations, more after lip.PZQ with an earlier peak observed for PZQ (day 3) than for lip.PZQ (day 6). Empty liposomes had no significant effect. The average counts of ingested particles in phagocytosing cells were significantly higher only after lip.PZQ administration. The pattern of changes in phagocytic activity correlated with the reduction of parasite numbers in the peritoneal cavity, with the highest observed on day 6 after therapy with lip.PZQ. Phagocytosis of lip.PZQ in vivo stimulated significantly the respiratory burst in peritoneal macrophages, with the highest concentration of superoxide anions recorded on day 1 after the last dose, whereas therapy with PZQ itself did not increase this process significantly. The capacity for the respiratory burst declined in all groups with progressing infection. It is proposed that the phagocytic activity of peritoneal macrophages after therapy was stimulated indirectly as a consequence of activation of the specific immune response. The larvicidal effect of lip.PZQ on the tetrathyridia in the peritoneal cavity was synergistic with the phagocytic activity and might be the result of double action of drug and superoxide anions generated during the respiratory burst stimulated by this drug formulation.