Is nitric oxide involved in the tolerance of Calomys callosus as a reservoir host towards Trypanosoma cruzi infection?

Trypanosoma cruzi, the agent of Chagas disease, is known to cause enhanced nitric oxide (NO) production, which might be involved in host resistance. The inducible nitric-oxide-synthase (iNOS) is assumed to be responsible for the NO increase after several infections. We studied the potential role of NO in Calomys callosus, a natural reservoir of this protozoan parasite. The concentration of NO was determined in spleen and liver of animals infected with two different T. cruzi strains, BOL and BOL-SB. Furthermore, the iNOS mRNA expression was quantified in the same cell types. NO production was detectable in both tissues exhibiting only slight differences compared to non-infected controls. All measured NO values were significantly lower than those reported for a number of different mouse strains, which displayed extremely enhanced NO levels after T. cruzi infection. Surprisingly, iNOS mRNA expression was induced in infected C. callosus but without subsequent increase of NO levels, indicating a post-transcriptional regulation mechanism. In summary, our results, indicate that the tolerance of C. callosus to T. cruzi is only accompanied by non-toxic NO intracellular concentrations.

Six Trypanosoma cruzi strains characterized by specific gene expression patterns.

The intracellular parasite Trypanosoma cruzi, the causative agent of Chagas disease, is known to comprise heterogeneous populations. One possibility to explain the obviously distinct phenotypes of different T. cruzi strains is differential expression of particular genes. This could result in environmental adaptations of the parasite within host organs, leading to distinct clinical symptoms. With the aim of identifying differentially expressed genes, we examined different T. cruzi strains by suppression subtractive hybridization analysis. The isolated clones were sequenced and Blasted for sequence-homology with known T. cruzi genes. A stage-specific glycoprotein (82gp), an 85-kDa protein with homology to heat-shock proteins, a beta-tubulin gene, a hexosetransporter, a dehydrogenase/ prostaglandin F2alpha-synthase and a cathepsin B-like protease were identified. The expression of these genes was analyzed by RT-PCR. Diverse expression patterns were detected for different T. cruzi strains, but no specific correlation between the gene expression and the classification of groups could be found. We discuss the presumed importance of these T. cruzi gene expression patterns for future strategies of molecular therapy of Chagas disease. For pathological studies, other parameters such as distinct gene/antigen expression could also be of interest, because they probably likewise correlate with distinct phenotypes.

Molecular genetic characterization of different Trypanosoma cruzi strains and comparison of their development in Mus musculus and Calomys callosus.

Trypanosoma cruzi populations are characterized by diverse morphology, heterogeneous biological behavior, high genetic variability, and distinctly different clinical courses. The first objective of this work was to characterize different strains of T. cruzi with various molecular markers [simple-sequence-repeat PCR, randomly amplified polymorphic DNA (RAPD)-PCR, mini-exon genes]. All examined strains could be divided into two major lineages. Only one strain showed a different banding pattern in RAPD-PCR, which could be a further indication of the existence of a third lineage. The second aim was to examine the biological behavior of the different strains. Two animal models, Calomys callosus and Mus musculus, were infected. The results provide strong evidence that the biological behavior of the strains is not only lineage-specific. It appears that all factors, such as the infecting strain belonging to a certain lineage, the predominant morphological form of the isolate, and the immune response of the respective infected host, play an important role in the course of this infection.