Pathological changes in mice infected with tetrathyridia of Mesocestoides corti.

Long-term studies in mice experimentally infected with the tetrathyridia of Mesocestoides corti have revealed disseminated lesions in the liver, lung, kidney, epididymis, and testicle.     

Circulating antigens in infections of mice by tetrathyridia of Mesocestoides corti Hoeppli, 1925

Two circulating antigens were detected in the serum of ICR/Timco female mice infected intraperitonealy with tetrathyridia of the cestode Mesocestoides corti Hoeppli, 1925. One circulating antigen appeared by day 2 postinfection (p.i.) and remained in all mice until at least 90 days p.i. A second antigen appeared in the serum on day 14 p.i. and disappeared from all mice by day 28 p.i. Infected mouse serum also contained antibodies against one secretory/excretory antigen and two antigens in crude homogenate, as judged by double diffusion in two dimensions (Ouchterlony). Immune deposits were observed in the kidney tissue of Rockland mice by transmission electron microscopy, and their identity as products of tetrathyridia was confirmed by immunofluorescence. Further studies showed that the main antibody subclass associated with the mesangial immune deposits was 7S gamma l, and that other subclasses of IgG and IgM were not involved. Antigen was found in the proximal renal tubules of infected mice, as demonstrated by fluorescein-labeled IgG fraction of rabbit antitetrathyridia secretory/excretory antigen antisera. The presence of tetrathyridia antigen in the urine of infected mice was confirmed using the Ouchterlony technique.     

Maternal transmission of asexually proliferative Mesocestoides corti tetrathyridia (Cestoda) in mice

Asexually proliferative Mesocestoides corti tetrathyridia were studied to test the hypothesis of in utero transmission in mice and define more clearly the path of transmammary transmission. In utero transmission was not observed in 132 fetuses (22 litters) taken by caesarean section from infected mothers. However, 19 of these mothers had tetrathyridia in their mammary glands at the time of operation, nine had worms in the uterine lumen, and one had a single worm in the maternal blood space of a placenta. No tetrathyridia were found in amniotic cavities. No infection was found in 32 young (7 litters) examined immediately after birth to infected mothers, but before nursing. No infection was found in 30 young (5 litters) removed from infected mothers before nursing and raised by uninfected fosters. Of 29 uninfected young (5 litters) allowed to nurse on infected mothers, 18 became infected. Whole mounts and sections of infected mammary glands showed proliferating tetrathyridia free in larger milk ducts and free and encapsulated in mammary parenchyma. These data suggest that maternal transmission of M. corti tetrathyridia in mice occurs primarily or perhaps exclusively by the transmammary route.     

Factors influencing the establishment of Mesocestoides corti in mice following oral inoculation of tetrathyridia

Factors which influence the establishment of tetrathyridia of Mesocestoides corti in mice following inoculation per os were examined. Only a proportion of the tetrathyridia penetrate the gut wall and gain access to the peritoneal cavity and liver, and most of these penetrate through the wall of the small intestine. It appears that tetrathyridia must attach to the intestinal mucosa and commence penetration immediately or they pass into the large intestine and are voided. Establishment was not influenced by strain, sex or age of host. However, the temperature at which tetrathyridia were maintained before inoculation influence their ability to penetrate the intestinal wall. Additionally it appears that tetrathyridia have to undergo a morphological change before or during, this penetration phase.     

Mesocestoides corti: cation concentration in calcareous corpuscles of tetrathyridia grown in vitro

The tetrathyridia larval stage of the cestode Mesocestoides corti has been maintained for 5 days in vitro in the presence of Cr(III), Cu(II), Ga(III), In(III) T1(I), VO(II), Zn(II), and Zr (IV) cations. With the exception of the VO(II) culture, all tetrathyridia concentrated these cations in the calcareous corpuscles. Vanadium, as the vanadyl ion, appeared to have a toxic effect on the organisms and no vanadium was detected in these corpuscles by emission spectrographic analysis. Incorporation of the other ions has been shown by emission spectrographic analysis and, in the case of Ga, In, and T1, compounds containing these ions were identified by X-ray powder diffraction. The identification by emission spectrography of cations whose compounds were not identifiable by X-ray diffraction has led to the speculation that these ions may have been incorporated into the organic portion of the calcareous corpuscles rather than the inorganic portion.     

Distribution of [3H]cholesterol-labelled liposomes with or without praziquantel in mice infected with Mesocestoides corti (Cestoda) tetrathyridia

The anthelmintic drug praziquantel (PZQ) has a short half-life in the circulation, necessitating repeated daily administration of PZQ for the therapy of larval stages of cestodes. The effect of incorporation of PZQ into multilamellar liposomes on their biodistribution in Mesocestoides corti (syn. M. vogae) infected mice has been examined using [3H]cholesterol as a liposomal marker. Incorporation of PZQ significantly increased the average size of liposomes with 70.3% of [3H]lip.PZQ particles up to 1.9 microm, whereas higher portion of [3H]liposomes (66.3% of total) were of smaller (up to 1.3 microm). Both liposome preparations were given intraperitoneally to avoid rapid sequestration in the liver. There were significant differences between [3H]liposomes and [3H]lip.PZQ-associated radioactivity in peritoneal adherent cells, liver- and peritoneal larvae, liver, spleen and lymph nodes within 16 days of examination. The highest uptake (about 2-fold more [3H]lip.PZQ than [3H]liposomes from the total dose) was found in peritoneal cells on day 1 post therapy (p.t.) followed by a rapid decline. The kinetic of decline in these cells recovered on day 1 p.t. was studied also in vitro. Disappearance of the marker due to the breakdown of liposomes and efflux of lipids and PZQ from cells was slower for [3H]lip.PZQ in comparison with drug-free liposomes and was not completed after 4 days-incubation. Significantly increased levels of radioactivity, more in [3H]liposomes treated groups, were recorded in the liver- and peritoneal larvae between days 8-16 p.t. indicating re-utilization of cholesterol by the larvae. The data suggest that incorporation of PZQ into liposomes contributes to the enlargement of liposome average size and slows down their degradation in phagocytosing cells. In this respect, these cells could serve as the secondary circulating depots for PZQ releasing it slowly to the circulation.     

Anaerobic and aerobic energy metabolism in the larvae (tetrathyridia) of Mesocestoides corti

The tetrathyridia of Mesocestoides corti produce lactate, succinate, acetate, and CO₂ as major carbon-containing end products during in vitro incubation with glucose as the substrate. Differences in the rate of glucose consumption and lactate production under anaerobic or aerobic conditions were observed, but their significance could not be determined. However, succinate production was greatly decreased in the presence of oxygen.The relative activities and intracellular distribution of various enzymes involved in energy-supplying metabolism of the larvae appear to conform to the pathways observed in other parasitic helminths known to produce lactate, succinate, and volatile fatty acids as metabolic end products. Some common features found in this respect are the relatively low pyruvate kinase activity, the presence of a highly active cytoplasmic phosphoenolpyruvate carboxylase and the capability of mitochondrial membrane bound fumarate reductase to reduce fumarate by means of NADH. Although a stimulatory effect of fructose-1,6-diphosphate on the reaction velocity of pyruvate kinase occurred, the absolute activity of this enzyme is very low.Nearly all the enzymes required for Krebs cycle activity are available in the tetrathyridia. Under the assay conditions employed by us, only NAD-dependent isocitrate dehydrogenase could not be demonstrated. The small amounts of ¹⁴CO₂ liberated from 6-¹⁴C-glucose suggest that the cycle in its classical form probably only functions at a very low rate. The incorporation of ¹⁴C from labeled glucose into glycogen indicates the presence of enzymes capable of glycogenesis. The incorporation rate was found to be higher in the presence of oxygen than under anaerobic conditions. On account of the very low NAD-linked glycerol-3-phosphate dehydrogenase activity the glycerolphosphate cycle may be of minor importance for the tetrathyridia.As a result of these studies a scheme for the main carbohydrate dissimilating pathways in the tetrathyridia is proposed and the significance of oxygen with respect to energy-supplying metabolism is discussed.     

Effect of sex hormones on the growth and multiplication of tetrathyridia of Mesocestoides corti (Cestoda: Cyclophyllidea) in mice

The number of tetrathyridia in the peritoneal cavities of mice increases exponentially with time. Thirty days post infection more larvae are in the cavities than in the livers. After that the increase of intraperitoneal populations continues, whereas the number of tetrathyridia in the livers remains more or less constant.Exogenous testosterone propionate, 10 $\text{μg}\text{g}$, twice a week, for 5 weeks, increases significantly the total volumes of tetrathyridial populations in the peritoneal cavities, whereas oestradiol benzoate, 5 $\text{μg}\text{g}$ or 10 $\text{μg}\text{g}$, also for 5 weeks, accelerates the rate of growth and multiplication of coelomic tetrathyridia to a much lesser extent, but increases significantly the infection of the livers.     

Environmental temperature and the growth of Mesocestoides corti populations in mice

The volumes of tetrathyridial populations in the peritoneal cavities of mice of both sexes kept at 5 ± 1dgC for 20 days were significantly larger than those in control mice kept at 21 ± 1dgC; the volumes of these populations in mice kept at 35 ± 1dgC were significantly lower in males but not in females. The liver weights in different groups were proportional to the intensity of liver infection: for both sexes livers were heaviest at 5 ± 1dgC and lightest at 35 ± 1dgC.     

Cortisone and the growth of populations of Mesocestoides tetrathyridia in mice.

Volumes and weights of populations of Mesocestoides tetrathyridia increased faster in SEC mice treated with cortisone, than in the controls. Cortisone did not alter the sex resistance of the host and the populations always grew faster in male than in female hosts. Larvae of the populations from cortisone treated mice were smaller than those from control mice, and those from males smaller than those from females.The average size of the larvae decreased as the population size increaed. Fast growing populations contained a higher percentage of larvae with 2 suckers. These forms may serve as indicators of accelerated division of larvae.     

The isotype potential of B cells present in BALB/c mice chronically infected with Mesocestoides corti

Infection of BALB/c mice with Mesocestoides corti results in a chronic infection with a pronounced splenomegaly and hypergammaglobulinemia. A prominent feature of this infection is that the vast majority of serum immunoglobulin produced is restricted to IgG1 and IgM. As much as 30-fold increases in serum IgG1 levels have been noted. To ascertain whether, as a result of infection, the resident B cell pool is committed to IgG1, B cells from infected animals were tested for their ability to produce various isotypes after stimulation. In one series of experiments, B cells from normal and infected animals were used as donor cells in the splenic fragment assay. The results show that the frequency of 2,4-dinitrophenyl-specific and phosphorylcholine-specific B cells remains unaltered in infected animals compared to controls. Importantly, the hapten-specific B cell clones induced were found to express multiple isotypes. These results demonstrate that the nonactivated B cell pool in spleens of infected mice is not committed to IgG1 and IgM production.     

The influence of Mesocestoides corti on subsequent Angiostrongylus cantonensis infections in mice.

The influence of Mesocestoides corti on subsequent Angiostrongylus cantonensis infection in mice (C57BL/6 and BALB/c) was assessed. Both strains of mice infected with M. corti demonstrated a peak blood eosinophilia at around 3 weeks post-infection (p.i.). C57BL/6 and BALB/c mice primarily infected with M. corti were given A. cantonensis infection 18 days later, but pre-existing M. corti infection did not affect the recovery of intracranial worms of A. cantonensis at day 21 p.i. BALB/c mice with mixed parasite infections showed low morbidity and mortality as compared with mice singly infected with A. cantonensis and some mice demonstrated a pulmonary migration of intracranial worms. In C57BL/6 mice, intracranial worms were killed and thus all mice survived. C57BL/6 mice with mixed parasite infections failed to resist A. cantonensis reinfection. The blastogenic responses of spleen cells against A. cantonensis antigen were lower in BALB/c than in C57BL/6 mice and mixed parasite infections also resulted in less blastogenic responses against both concanavalin A and A. cantonensis antigen than monoinfection. The recovery of M. corti biomass was significantly higher in mice with mixed parasite infections than mice with monoinfection with M. corti. These data suggest a distinct difference in response to A. cantonensis infection between C57BL/6 and BALB/c mice, and the induction of immunosuppression in both mouse strains following M. corti infection. Blood eosinophilia provoked by M. corti infection is not directly associated with the killing of worms in subsequent A. cantonensis infection.