Complement-mediated killing of schistosomula of Schistosoma mansoni by rat eosinophils in vitro.

Eosinophils from the peritoneal cavity of normal rats, in the presence of fresh normal rat serum (NRS), adhered to schistosomula of Schistosoma mansoni in vitro and killed the majority of parasites within 18 h. The reaction differed from the previously described antibody-mediated eosinophil adherence to schistosomula which occurs in heat-inactivated immune rat serum (IRS) and where adherence is mediated through Fc receptors. Adherence of eosinophils in fresh NRS was shown to be due to the activation of complement at the schistosomular surface by the alternative pathway, and it was effected through C3 receptors. The ability of eosinophils to kill in Fc-mediated adherence. This enhancement of killer activity may be due to the generation by complement activation of eosinophil chemotactic factors which increase the concentration of cells at the target surface. It is suggested that eosinophil adherence mediated through complement activation could be the principla mechanism of destroying schistosomula in the host.

Iridium anomaly in the upper devonian of the canning basin, Western australia.

A moderate iridium anomaly, about 20 times the local background, has been found in Upper Devonian rocks in the Canning Basin. It occurs at or near the Frasnian-Famennian boundary, which is known to be associated with a major massextinction event of global extent. The anomaly occurs in an extremely condensed limestone sequence laid down under quiet deepwater conditions. Its occurrence suggests a causal link with some form of meteoroid impact. Moreover, carbon isotope data indicate that a large reduction in biomass could have occurred at this level. However, the anomaly coincides with a stromatolite bed containing the fossil cyanobacterium Frutexites; iridium, platinum, iron, manganese, cobalt, arsenic, antimony, and cerium are preferentially concentrated in filaments of this organism, with concentrations ranging from two to five times that of the matrix. It is possible that Frutexites extracted these elements directly from seawater, without the need for their derivation from an extraterrestrial source.

Analysis of total and surface membrane lipids of Schistosoma mansoni.

Lipids extracted from whole worm homogenates and tegumental outer membranes of guinea pig-derived 5-day, 2-, 3- and 6-week old schistosomes have been analysed by thin layer chromatography. Six-week hamster-derived parasites have been studied for comparative purposes. All homogenates contained neutral lipids, cholesterol and several phospholipids; phosphatidyl choline and phosphatidyl ethanolamine were major components. Phospholipid (P3) was absent from homogenates of 5-day worms but was present in older parasites. A single phospholipid (P4) which co-chromatographed with phosphatidyl glycerol was unique to hamster-derived parasites. One glycolipid (G1) was ubiquitous to all homogenates and co-chromatographed with the monogalactosyl ceramide marker. A second sugar-containing lipid (G2) was unique to 3-week worm homogenates, and was highly polar. It was resolved beneath the trigalactosyl ceramide marker. Tegumental membranes isolated from 6-week adults contained at least five glycolipids, four of which were also highly polar. Cholesterol and two dominant phospholipids occurred in the membranes of 2-, 3-, and 6-week worms. One phospholipid co-chromatographed with phosphatidyl choline; the other had an Rf value (relative band speed) equivalent to phosphatidyl ethanolamine. Membranes from liver stage parasites contained a further phospholipid which cochromatographed with sphingomyelin, and three additional, phosphate-staining lipids (P1, P3 and P6). Five sugar-containing lipids occurred in adult membranes only; four were highly polar, being resolved near the origin. Similar components were identified in extracts of host erythrocytes. The remaining membrane lipid appeared homologous to G1 identified in the whole worm homogenates. Important changes in lipid composition thus occur during schistosome growth and maturation in guinea pigs; moreover, worms derived from different rodents express different lipids.

Schistosoma mansoni: modulation of schistosomular lipid composition by serum.

Human serum and foetal calf serum have been compared in terms of their ability to modify the biochemical and immunological properties of the schistosomular surface. Artificially transformed schistosomula were incubated in the presence of serum for 24 h and then radioiodinated using the chloramine T method. With this method only lipids are labelled. Foetal calf serum produces a net loss of lipids from the schistosomula, particularly of mono- and diglycerides. Human serum however, promotes not only a loss of mono- and diglycerides, but also a substantial uptake of cholesterol and triglycerides. Schistosomula recovered from the lungs of mice could also be labelled and contained besides triglycerides, an increased amount of cholesterol esters. The modulation of surface lipids in worms cultured with human serum correlates with the observation that such schistosomula develop significantly greater protection against eosinophil-mediated cytotoxicity in vitro than do individuals incubated with foetal calf serum. On the other hand, schistosomula cultured in the presence of either human serum or foetal calf serum develop the same degree of protection against complement-dependent lethal antibody; this result indicates that resistance against complement-mediated damage may be independent of the uptake of cholesterol and/or triglycerides, and might involve only limited alterations in the surface configuration of the schistosomulum.

Serum-induced expression of a surface protein in schistosomula of Schistosoma mansoni: a possible receptor for lipid uptake.

When freshly transformed schistosomula of Schistosoma mansoni are incubated with human serum, a protein doublet (molecular weight approximately 45 kDa) is expressed on the surfaces of the parasites; parasites incubated in defined media fail to express the doublet proteins. A 30 min exposure to serum is sufficient to induce the expression of the doublet, and even when the parasites are separated from the serum by a dialysis membrane, the doublet is still expressed. The doublet proteins are shown to be synthesized by cercariae, but not by schistosomula; they can be extracted using detergents and immunoprecipitated by specific antisera. Following expression of the doublet, purified low density lipoproteins from human serum interact with the schistosomula, become adsorbed onto their surface membranes and are possibly internalized and degraded.

Trypanocidal and antitumour activity of platinum-metal and platinum-metal-drug dual-function complexes.

A number of antitumour platinum-metal complexes related to cis-platin showed trypanocidal activity against Trypanosoma rhodesiense in vitro but not in vivo. New platinum- and rhodium-metal complexes of diamidine and plenanthridinium trypanocides are described which showed higher therapeutic indices than the parent drugs, due to increased activity in the former drug type and decreased toxicity in the latter. Some evidence of potentiation of antitumour activity was noted in these drug complexes. At the ultrastructural level, complex-treated trypanosomes showed a number of nuclear effects and other lesions specifically attributable to platinum-metal action. Some of the lesions were similar to those induced by cis-platin in tumour cells.

The role of T cells in vaccine immunity in the murine model of schistosomiasis mansoni.

Naive CBA/Ca mice and mice vaccinated with gamma-irradiated cercariae of Schistosoma mansoni were challenged percutaneously with normal cercariae and depleted of L3T4+ T helper cells through the administration of a specific monoclonal antibody. Three regimes were utilized to target known phases of parasite migration. The in vivo depletion of L3T4+ cells resulted in a significant reduction in immunity (up to 65%) in vaccinated/challenged mice, provided the monoclonal antibody was targeted towards skin-resident schistosomula. When antibody was targeted towards lung phase challenge larvae, however, there was a significant reduction in worm recovery, but no correspondingly significant reduction in vaccine immunity. In contrast, the administration of monoclonal to naive mice, via all three treatment regimes, had no effect on the primary schistosome worm burden. Histopathological studies complemented these worm recovery data. Skin tissue biopsied from vaccinated/challenged mice treated with monoclonal to L3T4+ T cells rarely showed the inflammatory foci which normally characterize untreated vaccinated/challenged mice. This was true when antibody was given either before challenge, or just after challenge, and correlated with the recorded depression in vaccine immunity. Lung tissue collected from monoclonal-treated vaccinated/challenged mice (for all three treatment regimes) exhibited no changes in morphology compared to that from untreated vaccinated/challenged mice. This was not altogether surprising since in the NIMR vaccine mouse model, the lungs represent a poor site for challenge attrition and appear normal in morphology with the exception of a few, small inflammatory reactions. When the monoclonal was given to naive/infected mice, there was no change in the morphology of the pulmonary tissue, as compared to corresponding untreated cohorts. Immunohistochemical studies revealed that Thy-1+ cells dominated the subdermal inflammatory foci of vaccinated/challenged mice. Of the T cells identified, the T helper subset was the most common, with T suppressor cells being only weakly represented, and in some cases not at all. The proportion of macrophages (Mac-1+) varied between reactions.

Effect of praziquantel on the migration and survival of developmental stages of Schistosoma mansoni in mice.

Compressed organ autoradiography has been used to determine whether the anthelmintic drug praziquantel (Pzq) modifies the migration of isotopically labelled Schistosoma mansoni during the first 16 days of infection in CBA/Ca mice. The mice were treated with 500 mg kg-1 body weight of the drug on day 1 or day 6. Treatment caused a marked delay in parasite migration from the skin when the drug was administered intradermally at the site of infection on day 1; migration from the lungs was also delayed after such treatment. Pzq injected either intradermally on day 1 or intramuscularly on day 6 effectively reduced the number of parasites that finally arrived in the lungs and the livers by 41 and 47%, respectively. Intramuscular administration of the drug on day 1 had a negligible effect. Worm recoveries assessed on day 38 by perfusion of the hepatic portal system were greatly reduced when Pzq was administered on day 14. The worms proved less susceptible when the drug was administered on day 21 and were completely resistant following drug delivery on day 28. The influence of drug preparation and route of delivery on parasite migration and survival are discussed.

Surface antigens of male worms and microfilariae of Onchocerca gibsoni.

Living adult males and microfilariae of the cattle filarial parasite Onchocerca gibsoni were externally labelled with radioactive iodine using the iodogen and Bolton-Hunter procedures. Characterization of labelled surface proteins by sodium dodecyl sulphate (SDS)-polyacrylamide gel electrophoresis revealed clear cut differences in the two life cycle stages. In addition, the two radiolabelling procedures yielded some differences in the profiles of radiolabelled surface proteins for both adults and microfilariae. Immunoprecipitation analysis revealed a number of labelled antigens recognized by antibodies in human onchocerciasis serum pools, thereby demonstrating the usefulness of O. gibsoni as a model in Onchocerca volvulus vaccine studies. The reactivity of microfilarial antigens extended to antibodies from other human nematode infections, whereas male surface antigens, particularly those of low molecular weight, were Onchocerca specific. This indicates that O. gibsoni can provide a convenient source of specific diagnostic antigen.

Host antigens and parasite antigens of murine Schistosoma mansoni.

An indirect fluorescent antibody technique was used to detect mouse host antigens and parasite antigens on the surface of Schistosoma mansoni. A rabbit anti-mouse rbc antiserum to detect host antigens, and serum from mice immune to S. mansoni was used to detect parasite antigens. Schistosomula prepared after penetration of isolated mouse skin did not possess host antigens but bound antibody from immune serum (immune antibody); schistosomula recovered from the lungs of mice five days after infection possessed host antigens and failed to bind immune antibody. In contrast, schistosomula recovered from the skin of normal or immune mice three and 20 hours after cercarial penetration, adult worms, and cryostat sections of adult worms, were positive for host antigens but also bound immune antibody. Strong binding of immune antibody only occurred with the cryostat sections. Anti-schistosome antibody can therefore bind to schistosomes in the presence of host antigen. The lung forms of schistosomula however, may have different surface properties as there is no evidence that immune antibody binds to these forms.

Expression of cross-reactive surface antigens by microfilariae and adult worms of Brugia pahangi during infections in cats.

Microfilariae of Brugia pahangi were labelled with 125-Iodine using the reagent IODOGEN. Electron microscope autoradiographs of sections of iodinated microfilariae showed that the label was strictly confined to their sheath. Adult worms were also iodinated by the same procedure. Sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) analysis of detergent extracts of radio-labelled parasites revealed components of molecular weights 113, 81-71, 46 and 33 kDa in microfilariae, and of molecular weights 29, 20 and 16 kDa in adult worms. All but the 33 kDa component of microfilariae were immunoprecipitable with sera of infected cats and therefore antigenic. Antibodies to the 81-71 kDa and the 46 kDa microfilarial antigens were detected by immunoprecipitation before patency. Similarly, the 29 kDa antigen of adult worms was immunoprecipitable before the fourth moult. Therefore, during infection in cats, these antigens cross-react with epitopes present on earlier developmental stages.

Micro-organisms in filarial larvae (Nematoda).

Unusual bodies have been described in the hypodermal tissues of larval Dirofilaria immitis and Brugia pahangi. Ultrastructural evidence indicates that these bodies are probably Gram-negative micro-organisms. It appears that the presence of large numbers of these bodies in an early embryo may affect development adversely. Their importance at later stages of development of filariae is not known.

A freeze-fracture study of the tegumental membrane of Schistosoma mansoni (Platyhelminthes:Trematoda).

Two fracture faces in each half of the freeze-fractured tegumental membrane of adult Schistosoma mansoni indicate the presence of two trilaminate membranes. This result is compatible with the heptalaminate appearance of the tegumental membrane in ultrathin sections. Intramembranous particles are located mainly in the outermost leaflet of the outer membrane and in the cytoplasmic leaflet of the inner membrane. The tegumental membrane of the cercaria (infective larva) has a single fracture plane, which conforms with its trilaminate appearance in sections. Intramembranous particles are extremely numerous and are almost all located in the cytoplasmic leaflet.