Immunosuppression in Experimental Chagas Disease Is Mediated by an Alteration of Bone Marrow Stromal Cell Function During the Acute Phase of Infection.

After infection with Trypanosoma cruzi, the etiologic agent of Chagas disease, immunosuppression, and apoptosis of mature lymphocytes contribute to the establishment of the parasite in the host and thereby to persistence and pathology in the chronic stage of infection. In a systemic mouse model of experimental Chagas disease, we have demonstrated a strong depletion of mature B cells in the spleen during the first 2 weeks of infection. Remarkably, the decrease in this cell population commenced already in the bone marrow from infected mice and was a concomitant of an increased apoptosis in pro- and pre-B cell populations. Pro- and pre-B cells in the bone marrow showed a significant reduction accompanied by a functional disturbance of bone marrow-derived stromal cells resulting in diminished levels of IL-7, an essential factor for the development of B cell precursors. Ex vivo, stromal cells isolated from the bone marrow of infected mice had a strikingly impaired capacity to maintain the development of pro- and pre-B cells obtained from uninfected animals. Together, the reduction of an active humoral immune response during acute Chagas disease suggests to be an initial immune evasion mechanism of the parasite to establish persistent infection. Therefore, prevention of B cell depletion by rescuing the stromal cells during this early phase, could give rise to new therapeutic approaches.

An insight into the transcriptome of the digestive tract of the bloodsucking bug, Rhodnius prolixus.

The bloodsucking hemipteran Rhodnius prolixus is a vector of Chagas' disease, which affects 7-8 million people today in Latin America. In contrast to other hematophagous insects, the triatomine gut is compartmentalized into three segments that perform different functions during blood digestion. Here we report analysis of transcriptomes for each of the segments using pyrosequencing technology. Comparison of transcript frequency in digestive libraries with a whole-body library was used to evaluate expression levels. All classes of digestive enzymes were highly expressed, with a predominance of cysteine and aspartic proteinases, the latter showing a significant expansion through gene duplication. Although no protein digestion is known to occur in the anterior midgut (AM), protease transcripts were found, suggesting secretion as pro-enzymes, being possibly activated in the posterior midgut (PM). As expected, genes related to cytoskeleton, protein synthesis apparatus, protein traffic, and secretion were abundantly transcribed. Despite the absence of a chitinous peritrophic membrane in hemipterans - which have instead a lipidic perimicrovillar membrane lining over midgut epithelia - several gut-specific peritrophin transcripts were found, suggesting that these proteins perform functions other than being a structural component of the peritrophic membrane. Among immunity-related transcripts, while lysozymes and lectins were the most highly expressed, several genes belonging to the Toll pathway - found at low levels in the gut of most insects - were identified, contrasting with a low abundance of transcripts from IMD and STAT pathways. Analysis of transcripts related to lipid metabolism indicates that lipids play multiple roles, being a major energy source, a substrate for perimicrovillar membrane formation, and a source for hydrocarbons possibly to produce the wax layer of the hindgut. Transcripts related to amino acid metabolism showed an unanticipated priority for degradation of tyrosine, phenylalanine, and tryptophan. Analysis of transcripts related to signaling pathways suggested a role for MAP kinases, GTPases, and LKBP1/AMP kinases related to control of cell shape and polarity, possibly in connection with regulation of cell survival, response of pathogens and nutrients. Together, our findings present a new view of the triatomine digestive apparatus and will help us understand trypanosome interaction and allow insights into hemipteran metabolic adaptations to a blood-based diet.

The innate immune system of mammals and insects.

Infectious agents threaten any organism. Therefore, mammals and insects have evolved a complex network of cells and humoral factors termed immune system able to control and eliminate pathogens. Immunity varies between different groups of animals but always contains an innate immune system that can act fast and often effectively against a wide range of distinct pathogens (i.e. viruses, bacteria, fungi, and eukaryotic parasites). In mammals and insects, the communication between and regulation of immune cells is carried out by cytokines which orchestrate the defense against the invaders. The major challenge to recognize and to fight pathogens is the same for any host. In insects and mammals, the pathogens are recognized as non-self by recognition of pathogen-associated molecular patterns. In addition, similar pathogen recognition receptors and signaling pathways activate the immune response in insects and mammals. The pathogens have to be opsonized and/or ingested and controlled/eliminated by antimicrobial peptides or small effector molecules (reactive oxygen and nitrogen intermediates). Interestingly, even invertebrates have evolved certain forms of adaptive immunity, i.e. specific immune priming, and in some invertebrates alternative splicing of pathogen recognition receptors allows for a more specific recognition of a wide variety of pathogens. This enhanced specificity of pattern recognition conveys a special form of memory to their invertebrate hosts. In this chapter, we also consider gut immunity of insects and compare it with the response in mammals.

The development of Blastocrithidia triatomae (Trypanosomatidae) in the reduviid bug Triatoma infestans (Insecta): influence of feeding.

The population density and composition of an established infection of Blastocrithidia triatomae in the intestinal tract of fifth instars of Triatoma infestans were compared in unfed bugs, at 4 h and up to 15 days after feeding, and also in feces and urine deposited in the first 4 h after feeding. In unfed bugs, about 1-2 million B. triatomae colonized the small intestine and rectum, mainly epimastigotes (85% and 80%, respectively). During excretion, the percentage of cysts increased within the first two drops (from 15% to 35%) and then decreased slowly, indicating a washing-out of these unattached stages. The initial reduction in the B. triatomae population lasted up to 6 days after feeding. By 15 days after feeding, the populations had strongly increased in the small intestine and rectum, to 22 million and 2 million flagellates, respectively, as had cysts, comprising some 50% of the total population in the rectum.

The development of Blastocrithidia triatomae (Trypanosomatidae) in the reduviid bug Triatoma infestans (Insecta): influence of starvation.

Fifth instars of Triatoma infestans with established Blastocrithidia triatomae infections were dissected after different periods of starvation. After a short starvation period of 30 days, 60% of the total population (2,700,000 flagellates) occurred in the small intestine. Within the following 3 months, the numbers of living flagellates there (epimastigotes, cysts) were reduced by about 70% and the percentage of dead mastigotes increased to 30% of the respective total population. Epimastigotes always dominated (about 90%), followed by cysts and only up to 3% spheromastigotes. These relations were only slightly changed by starvation. In the rectum, at 30-120 days after feeding, the total population of living epimastigotes was reduced by 90% and the percentage of those attached to the rectal wall decreased from 10% to <3%. During this period, the proportion of dead from all epimastigotes increased from 34% to >99%. In the rectum, the percentage of cysts from the total population of living parasites increased from 41% to 88% at 30-60 days after feeding and remained at this percentage and total numbers, showing that especially the early phase of starvation strongly induced the encystment of B. triatomae.