Dataset Information

Proteic characterization of the IgM triplet involved in the diagnosis of congenital toxoplasmosis

ABSTRACT: Congenital toxoplasmosis is a fetal infection following the transplacental transmission of Toxoplasma gondii in a mother who seroconverted during pregnancy. Neonatal diagnosis has recently been improved through the identification by L’Ollivier et al. and Peyclit et al. of a congenital toxoplasmosis pathognomonic marker: the IgM triplet, corresponding to three bands of high molecular weight of 75, 90 and 100 kDa respectively found on the immunoblot pair profile mother-child. This is a new concept, as these three IgM bands do reflect an immunological response against proteins involved in mother-to-child transmission of Toxoplasma gondii. These proteins could be Toxoplasma gondii secreted effectors or not playing a role in the processes of invasion, modulation of the host cell immune system as well as in parasitic virulence. In this study, immunoproteomic techniques allowed us to identify 32 interesting protein spots on immunoblot, including 4 interesting spots specific to the IgM triplet. After protein identification of these spots by LC-MS/MS technique, we showed here that several Toxoplasma gondii proteins were good candidates for the IgM triplet. It turns out that each of these proteins is, directly or indirectly, involved in the process of cellular invasion and therefore probably in the process of transplacental invasion of Toxoplasma gondii. The identification of these proteins opens several fields of future diagnostic and therapeutic research that would improve management of congenital toxoplasmosis.

INSTRUMENT(S):

ORGANISM(S): Toxoplasma Gondii

TISSUE(S): Blood Serum

DISEASE(S): Congenital Toxoplasmosis

SUBMITTER: Hélène Diemer

LAB HEAD: Sarah Cianferani

PROVIDER: PXD060424 | Pride | 2025-12-08

REPOSITORIES: Pride

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Protein characterization of the IgM triplet involved in the diagnosis of congenital toxoplasmosis.

Carreno Maria M Villard Odile O Villena Isabelle I Peyclit Lucie L Diemer Helene H Schaeffer-Reiss Christine C L'Ollivier Coralie C

Parasite (Paris, France) 20251126

Congenital toxoplasmosis is a fetal infection resulting from the transplacental transmission of Toxoplasma gondii in mothers who seroconvert during pregnancy. Neonatal diagnosis has recently been improved through the identification by L'Ollivier et al. (2012) and Peyclit et al. (2023) of a pathognomonic marker for congenital toxoplasmosis: the IgM triplet, corresponding to three high molecular weight bands of 75, 90, and 100 kDa, respectively found on the mother-child immunoblot pair profile. Th ...[more]

PMID: 41296914

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Project description:The population structure of Toxoplasma gondii includes three highly prevalent clonal lineages, types I, II, and III, which differ greatly in virulence in the mouse model. Previous studies have implicated a family of serine threonine protein kinases found in rhoptries (ROPs) as important in mediating virulence differences between types I vs. III and II vs. III. Here, we explored the genetic basis of differences in virulence between the highly virulent type I lineage and moderately virulent type II based on a new genetic cross and linkage mapping. Genome-wide association revealed a single quantitative trait locus controls the > 4 log difference in lethality between these strains. Neither ROP16 nor ROP18, previously implicated in virulence differences in T. gondii, were found to contribute to differences between types I and II. Instead, the major virulence locus contained a cluster of pseudokinases denoted as rhoptry protein 5 (ROP5); this locus contains a tandem cluster of polymorphic alleles that differed in expression levels between strains. ROP5 alleles contained only part of the catalytic triad of canonical S/T kinases, and consistent with this they lack demonstrable kinase activity in vitro. Genetic disruption of the rop5 locus in the type I lineage lead to a > 5 log increase in the lethal dose, and surviving mice developed lasting immunity and were protected from an otherwise lethal challenge. These findings reveal that amplification of a polymorphic cluster of pseudokinases plays an important role in pathogenesis of toxoplasmosis in the mouse model. We used a new genotyping protocol based on hybridization of parental and recombinant progeny genomic DNA (gDNA) to Affymetrix gene arrays constructed for T. gondii (http://ancillary.toxodb.org/docs/Array-Tutorial.html). We identified 1,603 single feature polymorphisms (SFP) based on probes that successfully discriminated perfect match (type II) from mismatch (type I) hybridization across all clones and provided reliable SFP markers for this cross.

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