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CD1d-restricted invariant natural killer T (iNKT) cells represent a heterogeneous population of lipid-reactive T cells that are involved in many immune responses, mediated through T-cell receptor (TCR)–dependent and/or independent activation. Although numerous microbial lipid antigens (Ags) have been identified, several lines of evidence have suggested the existence of relevant Ags of endogenous origin. However, the identification of their precise nature as well as the molecular mechanisms involved in their generation are still highly controversial and ill defined. Here, we identified two mammalian gangliosides—namely monosialoganglioside GM3 and disialoganglioside GD3—as endogenous activators for mouse iNKT cells. These glycosphingolipids are found in Toll-like receptor-stimulated dendritic cells (DC) as several species varying in their N-acyl fatty chain composition. Interestingly, their ability to activate iNKT cells is highly dependent on the ceramide backbone structure. Thus, both synthetic GM3 and GD3 comprising a d18:1-C24:1 ceramide backbone were able to activate iNKT cells in a CD1d-dependent manner. GM3 and GD3 are not directly recognized by the iNKT TCR and required the Ag presenting cell intracellular machinery to reveal their antigenicity. We propose a new concept in which iNKT cells can rapidly respond to pre-existing self-molecules after stress-induced structural changes in CD1d-expressing cells. Moreover, these gangliosides conferred partial protection in the context of bacterial infection. Thus, this report identified new biologically relevant lipid self-Ags for iNKT cells., Although the existence of self-antigens for invariant Natural Killer T (iNKT) cells is widely accepted, their precise nature remains a matter of debate. This study shows that two mammalian ganglioside species activate iNKT cells in a CD1d-dependent manner., Author summary Invariant natural killer T (iNKT) cells are a population of unconventional T lymphocytes that activate rapidly during inflammation due to their innate-like features. They are unconventional since they do not react to peptidic antigens (Ags) presented by classical major histocompatibility complex (MHC) molecules; instead, they recognize lipid-based Ags in the context of the MHC class I-like molecule CD1d. While numerous Ags of microbial origins have been described, their endogenous Ags are far less understood and remain a matter of strong debate. Here, we report that engagement of an innate receptor on the Ag-presenting cells leads to modulation of their lipid metabolism. This results in an enrichment of particular glycosphingolipid species that differ in both the nonpolar tail and polar head structures. Among those, two species have the potential to activate iNKT cells in a CD1d-dependent manner after further intracellular modifications. Based on these data, we propose a concept that iNKT cells can rapidly respond to pre-existing self-molecules after stress-induced changes in CD1d-expressing cells. Given the presence of closely related molecules in some pathological conditions such as cancer, it will be interesting to evaluate the biological relevance of these Ags in disease states.

We have previously shown that a mAb that inhibits the enzymatic activity of the Schistosoma mansoni 28-kDa glutathione S-transferase (Sm28 GST) also reduces female worm fecundity and egg viability in vivo and in vitro. By peptidic epitope mapping and an activity reconstitution assay, the carboxyl terminus (CT) amino acid residues 190-211 and to a lesser extent the truncated amino terminus (NT) residues 10-43 of the enzyme were identified as mAb recognition sites. Sera from rats immunized with the NT (10-43) and CT (190-211) peptides showed a partial inhibitory effect on Sm28 GST activity in a late phase (6 to 7 wk) but not in an early phase (2 to 4 wk) after immunization. Passive transfer of Sm28 GST-inhibiting anti-N- and C-terminal sera, but not of the noninhibitory sera, protected the infected mice by reducing tissue egg deposition and the ability of eggs to hatch. In active immunization experiments, the CT peptide significantly decreased the worm burden (37 to 40%) in mice as did the rSm28 GST (28 to 52%). In terms of tissue egg deposition and egg-hatching ability, immunization with both the NT and CT peptides reproduced the reduction observed after immunization with rSm28 GST. A constant reduction in egg numbers was noted in the small intestines and the livers of the immunized mice. A clear reduction in the ability of intestinal or hepatic eggs to hatch was observed. The results are discussed in terms of the conformational participation of the NT and CT of Sm28 in the expression of GST activity.

The parasite Schistosoma mansoni infects its definitive mammalian host through an obligatory cutaneous penetration. In this work, we studied early immune response following migration of larvae through human skin, the first immunocompetent organ encountered by the parasite. For this purpose we used an experimental model of severe combined immunodeficient mice engrafted with human skin and injected with autologous PBL. Six days after percutaneous infection, we observed an infiltration of lymphocytes within the human skin, predominantly composed of CD4+ T cells. Moreover, among the cytokines potentially present in the infected skin, immunohistochemistry analysis revealed an in vivo expression of IL-7 in the epidermal layers and strikingly at the level of vascular endothelium. Using an in vitro coculture system, we showed that the S. mansoni larvae directly trigger IL-7 production by human dermal microvascular endothelial cells but not by keratinocytes. Finally, measurements of IL-7 concentrations in plasma of 187 S. mansoni-infected individuals showed that the youngest, which are also the most infected, displayed the highest IL-7 levels. Together, these findings describe dermal endothelial cells as a novel source of IL-7, a cytokine particularly important in schistosomiasis.

The role of adhesion molecules in antibody-dependent cell-mediated cytotoxicity (ADCC) of macrophages toward the extracellular parasite Schistosoma mansoni was investigated by using 1) a panel of blocking mAbs directed against adhesion molecules and 2) different soluble ligands as candidate inhibitors of ADCC. The results show that the beta2-integrin Mac-1 (CD11b/CD18), L-selectin (CD62-L), and the carbohydrate determinant sialyl Lewis(x) (sLe(x); sCD15) are required for macrophage effector function toward schistosomula targets. On the other hand, the parasite counter-receptors involved in ADCC were found to share common motifs with the mammalian selectin-carbohydrate families. One family of parasite receptor(s) involved in ADCC carries the Lewis(x) (Le(x); CD15) carbohydrate structure, whereas a second family of receptor(s) appears to display selectin-like properties with affinity for the sLe(x) tetrasaccharide. Immunostaining experiments confirmed that schistosomula express on their surface hostlike molecules recognized by anti-Le(x) (CD15) and by anti-human E-selectin (CD62-E) mAbs. The double receptor-ligand interaction between macrophages and parasite targets provides new insights into the biologic roles of selectins and Le(x)-related structures in immunity against helminthic parasites.