Apoptosis: a mechanism of immunoregulation during human Schistosomiasis mansoni.

People infected with schistosomes may present with a variety of clinical manifestations ranging from the relatively asymptomatic intestinal (INT) form to the hepatointestinal (HI) or hepatosplenic (HS) forms characterized by hepatomegaly and hepatosplenomegaly with severe portal hypertension, respectively. Flow cytometry analyses were used to evaluate the contribution of apoptosis in specific cell populations from schistosomiasis patients to the development of the different clinical forms of the disease. The results showed that cell death induced by combinations of specific antigen and cytokines corresponds with specific clinical presentations. It was shown that soluble egg antigen (SEA) increased the level of apoptosis only in T cells from INT patients. Stimulation with soluble lung worm antigen preparation (SLAP) did not induce significant differences in the levels of apoptosis in T cells from the patients with the different clinical forms of schistosomiasis. These results suggest for the first time that apoptosis plays an important role in the modulation of the anti-SEA response in INT patients.

Cytokines as determinants of resistance and pathology in human Schistosoma mansoni infection.

The role of different cytokines in the peripheral blood mononuclear cell (PBMC) proliferative response and in in vitro granuloma formation was evaluated in a cross-sectional study with patients with the different clinical forms and phases of Schistosoma mansoni infection, as well as a group of individuals "naturally" resistant to infection named normal endemic (NE). The blockage of IL-4 and IL-5 using anti-IL-4 and anti-IL-5 antibodies significantly reduced the PBMC proliferative response to soluble egg (SEA) and adult worm (SWAP) antigens in acute (ACT), chronic intestinal (INT) and hepatosplenic (HS) patients. Similar results were obtained in the in vitro granuloma formation. Blockage of IL-10 had no significant effect on either assay using PBMC from ACT or HS. In contrast, the addition of anti-IL-10 antibodies to PBMC cultures from INT patients significantly increased the proliferative response to SEA and SWAP as well as the in vitro granuloma formation. Interestingly, association of anti-IL-4 and anti-IL-10 antibodies did not increase the PBMC proliferative response of these patients, suggesting that IL-10 may act by modulating IL-4 and IL-5 secretion. Addition of recombinant IL-10 decreased the proliferative response to undetectable levels when PBMC from patients with the different clinical forms were used. Analysis of IFN-gamma in the supernatants showed that PBMC from INT patients secreted low levels of IFN-gamma upon antigenic stimulation. In contrast, PBMC from NE secreted high levels of IFN-gamma. These data suggest that IL-10 is an important cytokine in regulating the immune response and possibly controlling morbidity in human schistosomiasis mansoni, and that the production of IFN-gamma may be associated with resistance to infection.

Cytokines as determinants of resistance and pathology in human Schistosoma mansoni infection

The role of diferent cytokines in the peripheral blood mononuclear cell (PBMC) proliferative response and in in vitro granuloma formation was evaluated in a cross-sectional study with patients with the different clinical forms and phases of Schistosoma mansoni infection, as well as a group of individuals "naturally" resistant to infection named normal endemic (NE). The blockage of IL-4 and IL-5 using anti-IL-4 and anti-IL-5 antibodies significantly reduced the PBMC proliferative response to soluble egg (SEA) and adult worm (SWAP) antigens in acute (ACT), chronic intestinal (INT) and hepatosplenic (HS) patients. Similar esults were obtained in the in vitro granuloma formation. Blockage of IL-10 had no significant effect on either assay using PBMC from ACT or HS. In contrast, the addition of anti-IL-10 antibodies to PBMC cultures from INT patients significantly increased the proliferative response to SEA and SWAP as well as the in vitro granuloma formation. Interestingly, association of anti-IL-4 and anti-IL-10 antibodies did not increase the PBMC proliferative response of these patients, suggesting that IL-10 may act by modulating IL-4 and IL-5 secretion. Addition of recombinant IL-10 decreased the proliferative response to undetectable levels when PBMC from patients with the different clinical forms were used. Analysis of IFN-gamma in the supernatants showed that PBMC from INT patients secreted low levels of IFN-gamma upon antigenic stimulation. In contrast, PBMC from NE secreted high levels of IFN-gamma. These data suggest that IL-10 is an important cytokine in regulating the immune response and possibly controlling morbidity in human schistosomiasis mansoni, and that the production of IFN-gamma may be associated with resistance to infection.

Factors affecting high and low human IgE responses to schistosome worm antigens in an area of Brazil endemic for Schistosoma mansoni and hookworm.

We have previously examined the antibody isotype responses to schistosome worm and egg antigens in human populations living in areas of Kenya and the Philippines endemic for Schistosoma mansoni and S. japonicum, respectively. Here, we have analyzed antibody isotype responses to S. mansoni adult worm (AW) antigen and soluble egg antigen (SEA) in more than 500 Brazilian individuals, and found similar relationships with age and sex as in the Kenyan and Filipino populations. Isotype responses to AW antigen broadly increased with age whereas isotype responses to SEA decreased, and a higher proportion of males than females had detectable IgE against AW antigen. Most isotype responses to AW antigen and SEA correlated positively with intensity of infection with S. mansoni except AW antigen-specific IgG2, which correlated negatively. The overall prevalence of infection with S. mansoni in this area was relatively low at only 39.5%; hookworm prevalence was higher at 57.4%. The majority of those infected with S. mansoni were also infected with hookworm (76%). Those individuals with high IgE responses to AW antigen were matched for sex, age, and total IgG to AW antigen as closely as possible with individuals with low levels of AW antigen-specific IgE. The two groups were compared for factors potentially influential in IgE production. No difference was found between the high and low IgE responders for 1) intensity or prevalence of infection with S. mansoni, 2) relative exposure to S. mansoni, 3) number of previous treatments for schistosomiasis, or 4) prevalence of infection with hookworm, but differences were found in other isotype responses to S. mansoni. The high IgE responders had higher IgA and IgG4 against both AW antigen and SEA but lower IgG3 responses to AW antigen than the low IgE responders. The IgE responses to three S. mansoni antigens (paramyosin, Sm22.6, and a 12-kD AW antigen band) were detected in individuals with IgE against AW antigen only.

Evidence that cellular immune responses to soluble and membrane associated antigens are independently regulated during human schistosomiasis mansoni.

We have made a comparative analysis of human cellular and antibody responses to membrane associated adult worm antigens (Mb-A), soluble adult worm antigens (SWAP) and soluble egg antigens (SEA) derived from Schistosoma mansoni. Chronically infected patients with the intestinal (I) and hepatosplenic (HS) forms of the disease as well as non-infected putative immune 'endemic normals' (EN), were studied. We observed that the cellular responses, of individuals, to the two adult worm preparations, SWAP and Mb-A, may be distinct and can be related to the occurrence of resistance or pathology. The resistant group (EN) presented higher levels of both cellular proliferation, and IFN-gamma production, in response to Mb-A as compared with SWAP whereas HS individuals presented higher levels of cellular proliferation to SWAP as compared with Mb-A. Individuals with intestinal disease had similar levels of proliferation to both antigens. The response to SEA by all groups was generally similar, and not predictive of any clinical form. The specific antibody response to the three antigens were in general higher among infected patients than in resistant EN individuals. These results support the hypothesis that the response to adult worm antigens may be pivotal in determining both the development of resistance and severity of disease.

Comparison of antibody isotype responses to Schistosoma mansoni antigens by infected and putative resistant individuals living in an endemic area.

The isotypic patterns of antibodies against Schistosoma mansoni antigenic preparations from eggs (SEA), adult worms (SWAP) and cercariae (CERC) have been studied in sera from two groups of individuals living in an area endemic for S. mansoni. One of the groups was comprised of individuals diagnosed as having S. mansoni infections based on their patency, i.e. those passing eggs in their faeces (patent infections, PI). The other group has been consider 'putatively resistant' due to their residence in an endemic area, their documented exposure to positive transmission sites, and their repeated negativity upon stool examinations (endemic normals, EN). There are strong specific responses of IgG1, IgG4 and IgM, particularly to SEA and CERC, by both groups. The reactivities of all isotypes were lower to SWAP. The responses of IgG4, IgM and IgE anti-CERC in EN and PI are higher than those found in normal individuals from outside endemic areas. In general, EN individuals express a relative higher level of anti-STEG IgE as compared to IgG4. On the other hand the pool of sera from PI showed the opposite pattern of higher IgG4 as compared to IgE. Several correlations are seen between isotypic responses to SEA, SWAP and CERC based on comparisons to the anti-SWAP IgE responses of the individuals in the two groups. These comparisons indicate the presence of distinct immunologic differences between individuals in the PI and the EN groups.

Interferon-gamma production by peripheral blood mononuclear cells from residents of an area endemic for Schistosoma mansoni.

During human schistosomiasis host responses to antigens of various parasite life-cycle stages may contribute to whether the severe, hepatosplenic state develops or the patient remains relatively asymptomatic throughout infection, and may play a role in resistance. This study evaluated production of interferon gamma (IFN-gamma) in vitro by schistosome antigen-stimulated peripheral blood mononuclear cells (PBMCs) from asymptomatic patients, and by PBMCs from apparently uninfected, untreated persons living in areas endemic for Schistosoma mansoni ('endemic normals'). IFN-gamma production parallels PBMC proliferation in that schistosomal egg antigens stimulate patent patients' cells poorly, but strongly stimulate PBMCs from 'endemic normals'. This is proportionally true for antigens from adult worms and cercariae. Although asymptomatic patent patients' cells produced little or no IFN-gamma in response to the 3 schistosomal antigenic extracts, their PBMCs, and PBMCs from 'endemic normals', produced expected amounts of IFN-gamma when exposed to phytohaemagglutinin. This implies that persons with patent infections have schistosome antigen-specific defects in their ability to respond to IFN-gamma production that are not exhibited by putatively resistant 'endemic normals'.

Differential cellular reactivity to adult worm antigens of patients with different clinical forms of schistosomiasis mansoni.

Analysis of the proliferation in vitro of peripheral blood mononuclear cells and the production of interferon-gamma (IFN-gamma) in individuals infected with Schistosoma mansoni and showing different clinical forms of the disease, as well as normal putatively immune individuals from an endemic area, was undertaken using total and fractionated soluble adult worm antigens (SWAP). A higher frequency of detectable response to fractionated antigens in T cell Western blot assays was observed in individuals with the more severe forms of the disease. Analysis of variance showed that, in the Western blot assays, there was a statistically significant difference in the level of cellular proliferation to antigens with low molecular weight (less than 21 kDa) between hepatosplenic patients and those with intestinal and hepatointestinal forms of the disease. No correlation between cellular proliferation and IFN-gamma production was observed. Most of the normal individuals from an endemic area failed to show significant proliferative responses to SWAP T cell Western blot assays or to antigen immobilized on nitrocellulose; they did show significant proliferative responses to whole soluble SWAP with positive IFN-gamma production. The results are consistent with the hypothesis that variations in the cellular immune responses to SWAP influence both the development of pathology and resistance to infection in schistosomiasis mansoni.

Immunological profiles of patients from endemic areas infected with Schistosoma mansoni.

Crude extracts of eggs (SEA) adult worms (SWAP) or cercariae (Cerc) have been used to stimulate Peripheral Blood Mononuclear cells (PBMC) and have provided rather distinct profiles of responses in different types of patients. In general it is clear that patients with early infections respond strongly to SEA while response to SWAP are develop more slowly. As infection progresses into the more chronic phases, a general pattern is seen which leads to lower anti-SEA proliferative responses in the face of higher responses to SWAP and variable anti-cerc responsiveness. Cured not re-exposed patients express very high levels of anti-SEA proliferation. It has recently been seen that those individuals who live in endemic areas and have continued water contact, but are repeatedly stool-negative (who are presumed to have self-cured or be putatively resistant; endemic normals) are strongly responsive to antigenic extracts, particularly to SEA. Furthermore, our results show that endemic normal individuals have significantly higher IFN gamma production upon PBMC stimulation with schistosome antigens than infected individuals. With the emergence of more studies it is becoming apparent that both the intensity and the prevalence of a given area may influence or shape the general responsiveness of the population under study.

Human antibody responses against schistosomal antigens. I. Antibodies from patients with Ancylostoma, Ascaris lumbricoides or Schistosoma mansoni infections react with schistosome antigens.

Sera of patients with either hookworm infections or ascariasis, who are from areas nonendemic for Schistosoma mansoni, contain antibody reactivity against extracts of S. mansoni schistosomula. Such patients with hookworm, but not those with Ascaris lumbricoides, also express considerable antibody activity against intact schistosomula and schistosomular membrane preparations. Sera from patients with schistosomiasis mansoni also contain these activities. Because these three helminthic infections often occur in the same areas, and concomitantly in patients, anti-schistosomular reactivity should not be ascribed only to exposure to schistosomes. The demonstration and definition of these surface-specific cross-reactivities point out the problems of serodiagnosis, and the potential of interactive influences in the common occurrence of multiple helminthic infections in humans.