M.J. Doenhoff

Immunological control of hepatotoxicity and parasite egg excretion in Schistosoma mansoni infections: stage specificity of the reactivity of immune serum in T-cell deprived mice

Within seven weeks of infection with 200 Schistosoma mansoni cercariae, T-cell deprived mice have been shown to suffer from extensive microvesicular damage to hepatocytes, and an inability to excrete parasite eggs at the same rate as comparably infected, immunologically intact controls. Administration of serum (CIS) from chronically infected, immunologically intact donors prevented the development of microvesicular cell damage and partially restored egg excretion rates in infected deprived mice. Serum pools obtained from mice injected either with intact S. mansoni eggs or with a homogenate of eggs emulsified in Freunds complete adjuvant (FCA) were as effective as CIS in preventing hepatotoxicity and restoring the rate of egg excretion in infected deprived recipients. The degree of protection of liver tissue afforded by immune sera could be monitored either by histopathological examination of liver sections or by estimation of serum transaminase concentrations, the results from both assays being generally in agreement. Sera from donor mice injected with cercarial or worm antigens in FCA were relatively inactive either in protecting against S. mansoni-induced liver damage or in reconstituting egg excretion rates in infected deprived mice. Serum from donor mice infected with 25 cercariae became hepato-protective between 49 and 53 days after infection of the donors, and the degree of hepatoprotective activity and egg excretion reconstituting capacity in the serum of 25 cercariae-infected donors was shown to increase between 8 and 16 weeks after infection. Increasing the size of infection of the serum donors to 100 cercariae gave only a marginal increase of hepatoprotective activity at 7 weeks when compared with serum donors infected with 25 cercariae for 7 weeks. Liver parenchymal cells of very heavily infected, immunologically intact mice were found to show microvesicular damage similar to that in livers of infected deprived mice, and administration of CIS to these normal mice was histopathologically protective. However, the elevated serum transaminase concentrations obtaining in the infected normal mice were not reduced to any extent by CIS. The results obtained from serum-reconstituted deprived mice are discussed in terms of the contribution they may make to a better understanding of the host-parasite relationship in immunologically intact mice.

Evidence for an immune-dependent action of praziquantel on Schistosoma mansoni in mice

Effective schistosomicidal action of praziquantel against Schistosoma mansoni infections in mice appears to be dependent to some extent on appropriate immunological stimulation. Indirect evidence consistent with this hypothesis was obtained by demonstrating a positive relationship between drug efficacy and both the intensity and the age of the parasitic infection. More directly, it has previously been shown that praziquantel kills fewer S. mansoni worms in immunosuppressed T cell-deprived mice than in immunologically intact controls; and we show here that infections 5 weeks old, against which the drug alone is sub-optimally active, are more effectively killed by a combination of drug and a rabbit antiserum raised against adult worm antigens.

Identification and partial purification of an antigen (ω1) from Schistosoma mansoni eggs which is putatively hepatotoxic in T-cell deprived mice.

T-cell deprived mice heavily infected with Schistosoma mansoni suffer from severe microvesicular damage to hepatocytes within seven weeks of infection. The damage can be prevented by administration of serum (CIS) obtained from mice chronically infected with S. mansoni or from mice immunized with intact or homogenized S. mansoni eggs. Reaction of serum samples from individual chronically infected mice in immunoelectrophoresis with S. mansoni egg homogenate has enabled the identification of at least 12 distinct immuno precipitation reactions. Precipitating antibody against one S. mansoni egg antigen, omega 1, has been detected in all mice with patent chronic infections, and anti-omega 1 antibody is the most concentrated of the precipitating anti-egg antibody species in pooled CIS. Pooled serum obtained from infected intact mice reacting predominantly against omega 1 was found partially to prevent the hepatotoxicity reaction on transfer to infected deprived mice. Serum samples from mice injected with egg homogenate fractionated either by preparative electrophoresis or by cation exchange chromatography, and containing antibodies reactive with antigen omega 1 in immunoprecipitation, were fully protective against liver cell damage induced by S. mansoni in deprived mice. Sera from mice immunized with other S. mansoni egg fractions, and which did not contain antibodies reactive with omega 1, were not hepatoprotective. Antigen omega 1 is compared and contrasted with other S. mansoni egg antigens that have been described.

Resistance against Schistosoma mansoni induced by highly irradiated infections: studies on species specificity of immunization and attempts to transfer resistance

Significant levels of resistance against Schistosoma mansoni challenge were developed by mice exposed to highly irradiated (20 krad.) cercariae of the homologous species (53-67%), whereas vaccination with S. bovis, S. haematobium or S. japonicum failed to confer significant levels of resistance (-5-12%), thus confirming the specificity of the immunizing procedure. Attempts to transfer resistance to naive recipients by injection of serum and of spleen or lymph node cells from donor mice vaccinated with highly irradiated cercariae were largely unsuccessful. However, significant levels of resistance could be transferred to mice by injection of serum from rabbits exposed to irradiated cercariae. Comparable levels of resistance were conferred by injection of serum at the time of challenge (34-69%) or 5-6 days later (31-56%). In contrast, sera from rabbits injected with soluble egg antigen or homogenized cercariae failed to confer protection upon recipient mice. Sera from vaccinated mice, vaccinated rabbits and antigen-injected rabbits all caused cell adherence to skin-transformed schistosomula but neither the level of adherence nor the serum titre correlated with the ability to confer protection to mice.

Seroepidemiology and serodiagnosis of schistosomiasis in Kenya using crude and purified egg antigens of Schistosoma mansoni in ELISA

The performance of antibody detection for the diagnosis of schistosomiasis has been evaluated in Kenya. Approximately 1500 blood samples from 3 areas with endemic schistosomiasis (Schistosoma mansoni only, S. haematobium only, and a mixed infection area), and from a non-endemic control area, were tested for their antibody reactivity in an enzyme-linked immunosorbent assay (ELISA). The results were compared with infection status determined by parasitological examination. Two test antigens were used: unfractionated S. mansoni egg homogenate (SEA), and CEF6, a previously described, partially purified fraction of SEA containing 2 cationic antigens. The antigens prepared from eggs of Kenya and Puerto Rico S. mansoni isolates gave very similar results. Bloods from patients with S. haematobium infection cross-reacted significantly with the two S. mansoni antigen preparations, but reactivity against CEF6 appeared more specifically indicative of S. mansoni infection. Of 254 blood samples from schoolchildren in the non-endemic area, 100% gave ELISA optical density readings at 492 nm (OD492) < 0.20 against SEA, and 98% were < 0.20 against CEF6. With 887 blood samples from subjects of all ages in the area endemic for S. mansoni alone, using an ELISA OD492 cut-off point of 0.20, SEA and CEF6 had sensitivities of 94% and 97% respectively, and specificities of 64% and 59% respectively. Increasing the OD492 cut-off value reduced the sensitivity and increased the specificity of both test antigens. Specificity of both antigens was poor with samples from 234 children in an area endemic for both S. mansoni and S. haematobium (< 20% for both antigens at an OD492 cut-off value of 0.20).(ABSTRACT TRUNCATED AT 250 WORDS)

Serodiagnosis of human Schistosoma mansoni infections: enhanced sensitivity and specificity in ELISA using a fraction containing S. mansoni egg antigens ω1 and α1

Abstract Six fractions of Schistosoma mansoni egg homogenate obtained by cation exchange chromatography were tested in an enzyme-linked immunosorbent assay (ELISA) for their efficacy in serodiagnosis of human S. mansoni infections. One cationic fraction (CEF6) containing egg antigens ω1 and α1 was found to be highly reactive with a S. mansoni positive reference serum pool. 94 of 95 sera from individuals infected with S. mansoni gave a positive reaction with CEF6, and no cross reactivity was obtained with this antigenic fraction and sera from donors infected with heterologous non-schistosome parasites and cases of avian cercarial dermatitis. Only 10 of 165 sera from patients with S. haematobium and three of 10 sera from patients with S. japonicum were positive with S. mansoni egg CEF6. In one group of patients from West Africa, examined both serologically and parasitologically at a six-monthly interval, the measurement of antibody to CEF6 provided a more sensitive diagnostic aid than stool examination.

The immune dependence of chemotherapy

In this review, Mike Doenhoff and colleagues discuss the immune dependency of chemotherapy and the consequences for drug resistance. They also consider the implications for the control of infections that are relatively unresponsive to drugs, such as opportunistic infections in immunosuppressed patients.

The stage-, strain- and species-specificity of a Schistosoma mansoni egg antigen fraction (CEF6) with serodiagnostic potential

The immunological properties of CEF6, a cationic fraction of Schistosoma mansoni egg homogenate (SEA) containing two antigens, omega 1 and alpha 1, have been further investigated in two assays, immunoelectrophoresis (IEP) and enzyme immunoassay (ELISA). Although precipitating antibodies to antigen omega 1 were amongst the first to appear in mice with patient infections, IgG reactivity against CEF6 in ELISA trailed somewhat behind that of IgG activity against unfractionated SEA. Specificity of the two antigens in CEF6 to the egg stage of the parasite life-cycle was demonstrated by the failure of immunizations with cercarial or worm antigens to induce antibodies which reacted against either alpha 1 or omega 1 in IEP, or against CEF6 in ELISA. Mice infected with S. mansoni strains derived from geographically distinct areas, including the Caribbean, South America and Africa, produced antibodies which were reactive against CEF6 prepared from eggs of a Puerto Rican S. mansoni strain that had been maintained in the laboratory for many years. Of the different precipitating anti-SEA antibody species induced by the various S. mansoni strains in mice, those reactive against antigen omega 1 appeared to be present in highest titre. Sera from mice chronically infected with S. japonicum and S. haematobium failed to precipitate CEF6 in IEP and were less reactive with CEF6 than with S. mansoni SEA in ELISA. However, similar degrees of ELISA reactivity against S. mansoni CEF6 and SEA were given by sera from mice infected with S. bovis. The results support the notion that the antigens in CEF6 may be useful in the serodiagnosis of schistosomiasis mansoni infections.

Does the immunopathology induced by schistosome eggs potentiate parasite survival

Infection with schistosomes is accompanied by inflammatory lesions resembling delayed hypersensitivity reactions, that are induced by eggs in host tissues. Here Michael Doenhoff and his colleagues demonstrate that T-cell-deprived mice excrete much fewer eggs than do intact control animals. From the results of serum and cell transfer experiments, they conclude that immune processes assist egg expulsion and thus contribute to the parasites success.

Factors affecting the acquisition of resistance against Schistosoma mansoni in the mouse. VIII. Failure of concurrent infections with Plasmodium chabaudi to affect resistance to reinfection with S. mansoni.

Infection of mice with Plasmodium chabaudi induced a state of relative unresponsiveness to a challenge of sheep erythrocytes in terms of serum antibody titres, but failed to affect the degree of acquired resistance to reinfection with Schistosoma mansoni irrespective of whether the primary or challenge schistosome infection was administered during the peak malaria parasitaemia.