Sophia Lafitte

Identification of a Novel Antigen of Schistosoma mansoni Shared with Plasmodium falciparum and Evaluation of Different Cross-Reactive Antibody Subclasses Induced by Human Schistosomiasis and Malaria

ABSTRACT Plasmodium falciparum and Schistosoma mansoni are often found in human coinfections, and cross-reactive antibodies to different components of the two parasites have been detected. In this work, we identified a cross-reactive S. mansoni gene product, referred to as SmLRR, that seems to belong to the leucine-rich repeat protein family. Comparative analysis of SmLRR revealed 57% similarity with a putative gene product encoded in the P. falciparum genome. Antibodies to SmLRR were found in experimental infections and in both S. mansoni- and P. falciparum-infected individuals. Correlative analysis of human anti-SmLRR responses in Kenya and Uganda suggested that malaria and schistosomiasis drive the immunoglobulin G3 (IgG3) and IgG4 isotypes, respectively, against SmLRR, suggesting that there is differential regulation of cross-reactive isotypes depending on the infection. In addition, the levels of anti-SmLRR IgG4, but not the levels of IgG3, correlated positively with the intensity of S. mansoni infection.

The age-related resistance of rats to Plasmodium berghei infection is associated with differential cellular and humoral immune responses

In this study, we investigated how the age of rats would affect the course of infection of and the immune response to Plasmodium berghei. Both young (4-week-old) and adult rats (8-week-old) can be infected with P. berghei ANKA strain, with significantly higher levels of infected red blood cells in young rats. While 100% of young rats succumbed to infection, adult rats were able to clear blood parasites and no mortality was observed. Analysis of cellular distribution and circulating cytokines demonstrated the persistence of CD4+/CD25+ T cells and high expression of circulating interleukin-10 (IL-10) during the progression of infection in young-susceptible rats, whereas high levels of CD8+ T cells and natural killer T cells are detected in adult-resistant rats. Analysis of antibody isotypes showed that adult rats produced significantly higher levels of interferon-gamma (IFN-gamma)-dependent IgG2c antibodies than young rats during infection. Further evaluation of the role of IL-10, IFN-gamma and of immune cells showed that only the adoptive transfer of spleen cells from adult-resistant rats was able to convert susceptibility of young-susceptible rats to a resistant phenotype. These observations suggest that cell-mediated mechanisms are crucial for the control of a primary infection with P. berghei in young rats.

Definition and mapping of epitopes recognized by specific monoclonal antibodies to Schistosoma bovis 28 kDa glutathione S-transferase: relation with anti-egg viability immunity

Monoclonal antibodies to the 28kDa glutathione S‐transferase of Schistosoma bovis have been constructed in mice and used to characterize the epitope(s) potentially implied in the induction of anti‐fecundity and anti‐egg viability immune responses. Among the MoAbs produced three were particularly studied: Sb4‐50 (IgG2a) and Sb4‐56 (IgG1) which inhibited Sb28GST activity and Sb4‐10 (IgG1) which did not. The use of overlappting peptides covering the entire amino acid sequence of Sb28GST, allowed us to define the linear epitopes recognized by these anti‐Sb28GST MoAbs. Amino acid residues 202‐211 were recognized by both MoAbs Sb4‐50 and Sb4‐56 and MoAb Sb4‐10 recognized amino acid residues 58‐67. Their capacity to inhibit GST activity suggested binding to the active site or to neighbouring regions, which include the C‐terminal domain (a.a. 190‐211) of the protein. When passively transfered into BALB/c mice MoAbs induced a significant reduction in egg hatching and an increase in immature eggs. Effects on worm burdens were, however, variable and no clear‐cut association between the inhibition of enzyme activity and anti‐fecundity or anti‐viability activities was recorded. Our data indicate that beside the anti‐fecundity and anti‐viability immunity related to the impairment of GST activity, immune response to epitopes located in other regions of the molecule also contribute to the reduction of egg viability.

Egg‐hatching inhibition in mice immunized with recombinant Schistosoma bovis 28 kDa glutathione S‐transferase

The capacity of a recombinant glutathione S‐transferase from Schistosoma bovis (rSb 28GST) to protect BALB/c mice against homologous and heterologous infections with, respectively, S. bovis or Schistosoma mansoni has been studied. Two injections of the rSb 28GST and an intravenous boost resulted in a marked specific IgG response on the day of experimental challenge with S. bovis or S. mansoni cercariae. Immunization of BALB/c mice led to a reduction in egg maturation and egg viability after infection with S. bovis or S. mansoni. Adult worm recoveries after an S. bovis challenge infection and tissue egg densities (intestine and liver) in S. mansoni challenge infection were also reduced in the immunized groups, but these differences were not statistically significant. No association between in vitro inhibition of GST enzymatic activity induced by immunized mouse sera and worm burden reduction was recorded. The analysis of the immune response, on the day of perfusion, showed the production of immunoglobulin (Ig)G1, IgG2a and IgG2b specific antibodies and the production of interleukin (IL)‐4 and IL‐5 by spleen cells after rSb 28GST stimulation. These data suggest that rSb 28GST immunization induces a moderate effect upon egg maturation and egg hatching, suggesting the involvement of similar mechanisms of action and common, but not exclusive, targets during S. bovis and S. mansoni infections. As a consequence, immunization with rSb 28GST may prove useful in affecting the pathology and transmission of African schistosomes.

Murine Schistosoma bovis infection: analysis of parasitic and immune parameters

Humoral and cellular responses to Schistosoma bovis antigens have been evaluated over a period of 11 weeks in mice exposed to S. bovis cercariae and data analysed in the context of the parasitic parameters (worm and egg loads) recorded at days 30, 60 and 80 of the ongoing infection. Results revealed a decrease of worm burden, particularly marked for female worms, between day 60 and day 80 of infection suggesting a higher susceptibility of female schistosomes to attrition mechanisms. The B‐cell response, studied by measuring the production of different isotypes, was directed against different stage specific antigens, with a predominance of IgG1 antibodies associated with a significant increase of IgA and IgE antibodies after egg deposition. The T‐cell response, assessed after in vitro stimulation of splenocytes, showed a predominant production of Th‐2 cytokines (IL‐4, IL‐5 and IL‐10) occurring after egg laying. Interestingly in contrast to S. mansoni infection the Th‐2 polarization did not seem to be exclusively triggered by egg‐associated antigens since significant amounts of IL‐10 were produced after stimulation with adult worm antigen preparation (SWAP) before the beginning of egg deposition.

Gene profiling analysis reveals the contribution of CD24 and P2Y6R to the susceptibility of young rats to Plasmodium berghei infection

Our previous studies have shown that Plasmodium berghei infection induces distinct clinical, parasitological and immunological states in young susceptible rats versus adult resistant rats. This susceptibility was mainly found to be related to inadequate cellular responses. In this study we first identified the altered genes in young susceptible rats. Unexpectedly, transcriptome analysis did not reveal any alteration of effector cytokines or their receptors. At day 13 p.i., six transcripts corresponding to faim3, mesothelin, gas3 (PMP22), gas7, CD24 and P2Y6R were significantly decreased in young infected rats when compared with adult infected rats. Because CD24 and P2Y6R participate in cellular immune responses, we next evaluated their role in the course of infection. Adoptive transfer experiments showed a transient but robust participation of CD24+ cells in the control of parasitaemia. The role of P2Y6R was investigated via its specific ability to be activated by Uridine di‐Phosphate (UDP). Young rats treated with UDP partially restored the expression of P2Y6R, controlled parasitaemia and survived thereafter. In conclusion, this study contributes to the discovery of novel biomarkers in young susceptible rats and suggests that the decrease in their expression could be among the reasons for the development of severe pathology in malaria.