Bioinformatics and expression analysis of the Xeroderma Pigmentosum complementation group C (XPC) of Trypanosoma evansi in Trypanosoma cruzi cells

Nucleotide excision repair (NER) acts repairing damages in DNA, such as lesions caused by cisplatin. Xeroderma Pigmentosum complementation group C (XPC) protein is involved in recognition of global genome DNA damages during NER (GG-NER) and it has been studied in different organisms due to its importance in other cellular processes. In this work, we studied NER proteins in Trypanosoma cruzi and Trypanosoma evansi, parasites of humans and animals respectively. We performed three-dimensional models of XPC proteins from T. cruzi and T. evansi and observed few structural differences between these proteins. In our tests, insertion of XPC gene from T. evansi (TevXPC) in T. cruzi resulted in slower cell growth under normal conditions. After cisplatin treatment, T. cruzi overexpressing its own XPC gene (TcXPC) was able to recover cell division rates faster than T. cruzi expressing TevXPC gene. Based on these tests, it is suggested that TevXPC (being an exogenous protein in T. cruzi) interferes negatively in cellular processes where TcXPC (the endogenous protein) is involved. This probably occurred due interaction of TevXPC with some endogenous molecules or proteins from T.cruzi but incapacity of interaction with others. This reinforces the importance of correctly XPC functioning within the cell.

STING Signaling Drives Production of Innate Cytokines, Generation of CD8+ T Cells and Enhanced Protection Against Trypanosoma cruzi Infection

A variety of signaling pathways are involved in the induction of innate cytokines and CD8+ T cells, which are major players in protection against acute Trypanosoma cruzi infection. Previous data have demonstrated that a TBK-1/IRF3-dependent signaling pathway promotes IFN-β production in response to Trypanosoma cruzi, but the role for STING, a main interactor of these proteins, remained to be addressed. Here, we demonstrated that STING signaling is required for production of IFN-β, IL-6, and IL-12 in response to Trypanosoma cruzi infection and that STING absence negatively impacts activation of IRF-dependent pathways in response to the parasite. We reported no significant activation of IRF-dependent pathways and cytokine expression in RAW264.7 macrophages in response to heat-killed trypomastigotes. In addition, we showed that STING is essential for T. cruzi DNA-mediated induction of IFN-β, IL-6, and IL-12 gene expression in RAW264.7 macrophages. We demonstrated that STING-knockout mice have significantly higher parasitemia from days 5 to 8 of infection and higher heart parasitism at day 13 after infection. Although we observed similar heart inflammatory infiltrates at day 13 after infection, IFN-β, IL-12, CXCL9, IFN-γ, and perforin gene expression were lower in the absence of STING. We also showed an inverse correlation between parasite DNA and the expression of CXCL9, IFN-γ, and perforin genes in the hearts of infected animals at day 13 after infection. Finally, we reported that STING signaling is required for splenic IFN-β and IL-6 expression early after infection and that STING deficiency results in lower numbers of splenic parasite-specific IFN-γ and IFN-γ/perforin-producing CD8+ T cells, indicating a pivotal role for STING signaling in immunity to Trypanosoma cruzi.

The Colombian Strain of Trypanosoma cruzi Induces a Proinflammatory Profile, Neuronal Death, and Collagen Deposition in the Intestine of C57BL/6 Mice Both during the Acute and Early Chronic Phase

The objective of this study was to evaluate the histopathological changes caused by infection with the Colombian strain of Trypanosoma cruzi (T. cruzi) in the acute and chronic experimental phases. C57Bl/6 mice were infected with 1000 trypomastigote forms of the Colombian strain of T. cruzi. After 30 days (acute phase) and 90 days (early chronic phase) of infection, the animals were euthanized, and the colon was collected and divided into two parts: proximal and distal. The distal portion was used for histopathological analysis, whereas the proximal portion was used for quantification of pro- and anti-inflammatory cytokines. In addition, the weight of the animals and parasitemia were assessed. The infection induced gradual weight loss in the animals. In addition, the infection induced an increase in interferon gamma (IFNγ) and tumor necrosis factor-alpha (TNF-α) in the intestine in the acute phase, in which this increase continued until the early chronic phase. The same was observed in relation to the presence of intestinal inflammatory infiltrates. In relation to interleukin (IL)-10, there was an increase only in the early chronic phase. The Colombian strain infection was also able to induce neuronal loss in the myenteric plexus and deposition of the collagen fibers during the acute phase. The Colombian strain of T. cruzi is capable of causing histopathological changes in the intestine of infected mice, especially in inducing neuronal destructions. Thus, this strain can also be used to study the intestinal form of Chagas disease in experimental models.

An Iron Transporter Is Involved in Iron Homeostasis, Energy Metabolism, Oxidative Stress, and Metacyclogenesis in Trypanosoma cruzi

The parasite Trypanosoma cruzi causes Chagas’ disease; both heme and ionic Fe are required for its optimal growth, differentiation, and invasion. Fe is an essential cofactor in many metabolic pathways. Fe is also harmful due to catalyzing the formation of reactive O2 species; for this reason, all living systems develop mechanisms to control the uptake, metabolism, and storage of Fe. However, there is limited information available on Fe uptake by T. cruzi. Here, we identified a putative 39-kDa Fe transporter in T. cruzi genome, TcIT, homologous to the Fe transporter in Leishmania amazonensis and Arabidopsis thaliana. Epimastigotes grown in Fe-depleted medium have increased TcIT transcription compared with controls grown in regular medium. Intracellular Fe concentration in cells maintained in Fe-depleted medium is lower than in controls, and there is a lower O2 consumption. Epimastigotes overexpressing TcIT, which was encountered in the parasite plasma membrane, have high intracellular Fe content, high O2 consumption—especially in phosphorylating conditions, high intracellular ATP, very high H2O2 production, and stimulated transition to trypomastigotes. The investigation of the mechanisms of Fe transport at the cellular and molecular levels will assist in elucidating Fe metabolism in T. cruzi and the involvement of its transport in the differentiation from epimastigotes to trypomastigotes, virulence, and maintenance/progression of the infection.

Trypanosoma cruzi Parasite Load Modulates the Circadian Activity Pattern of Triatoma infestans

American trypanosomiasis is a disease caused by the flagellate protozoan Trypanosoma cruzi, which is transmitted mainly in endemic areas by blood-sucking triatomine vectors. Triatoma infestans is the most important vector in the southern cone of South America, exhibiting a nocturnal host-seeking behavior. It has been previously documented that the parasite produces changes in some triatomine species, but this is the first time that the behavior of a vector has been evaluated in relation to its parasite load. After comparing the movement events and distance traveled of infected and non-infected T. infestans, we evaluated the change produced by different T. cruzi parasite loads on its circadian locomotor activity. We observed differences between infected and non-infected triatomines, and a significant relation between the parasite load and the increase in locomotor activity of T. infestans, which was accentuated during the photophase. This could have direct implications on the transmission of T. cruzi, as the increased movement and distance traveled could enhance the contact of the vector with the host, while increasing the predation risk for the vector, which could both constitute a risk for vectorial and oral transmission to mammals.

Biological and Molecular Effects of Trypanosoma cruzi Residence in a LAMP-Deficient Intracellular Environment

Trypanosoma cruzi invades non-professional phagocytic cells by subverting their membrane repair process, which is dependent on membrane injury and cell signaling, intracellular calcium increase, and lysosome recruitment. Cells lacking lysosome-associated membrane proteins 1 and 2 (LAMP1 and LAMP2) are less permissive to parasite invasion but more prone to parasite intracellular multiplication. Several passages through a different intracellular environment can significantly change T. cruzi’s gene expression profile. Here, we evaluated whether one single passage through LAMP-deficient (KO) or wild-type (WT) fibroblasts, thus different intracellular environments, could influence T. cruzi Y strain trypomastigotes’ ability to invade L6 myoblasts and WT fibroblasts host cells. Parasites released from LAMP2 KO cells (TcY-L2−/−) showed higher invasion, calcium signaling, and membrane injury rates, for the assays in L6 myoblasts, when compared to those released from WT (TcY-WT) or LAMP1/2 KO cells (TcY-L1/2−/−). On the other hand, TcY-L1/2−/− showed higher invasion, calcium signaling, and cell membrane injury rates, for the assays in WT fibroblasts, compared to TcY-WT and TcY-L1/2−/−. Albeit TcY-WT presented an intermediary invasion and calcium signaling rates, compared to the others, in WT fibroblasts, they induced lower levels of injury, which reinforces that signals mediated by surface membrane protein interactions also have a significant contribution to trigger host cell calcium signals. These results clearly show that parasites released from WT or LAMP KO cells are distinct from each other. Additionally, these parasites’ ability to invade the cell may be distinct depending on which cell type they interact with. Since these alterations most likely would reflect differences among parasite surface molecules, we also evaluated their proteome. We identified few protein complexes, membrane, and secreted proteins regulated in our dataset. Among those are some members of MASP, mucins, trans-sialidases, and gp63 proteins family, which are known to play an important role during parasite infection and could correlate to TcY-WT, TcY-L1/2−/−, and TcY-L2−/− biological behavior.

Hyperglycemia associated with Chagas disease

Chagas disease and type 2 diabetes mellitus are the two main causes of Years Lived with Disability. The consequences and possible interaction between both illnesses have been scarcely studied. A population study of blood donors from the Centro Estatal de Transfusión Sanguinea (CETS) was carried out with a reagent related to the Chagas test. Detection was carried out with an enzyme-linked immunosorbent assay (ELISA) of anti- Trypanosoma cruzi antibodies and also, an HbA1C test was performed with 4 mL of whole blood. Descriptive statistics were used to analyze quantitative and qualitative data. The Chi-square test was applied to determine the existence of an association between hyperglycemia and Chagas disease. A total of 4,952 participants were screened. All of the participants (100%) were from Hidalgo State and did not have a chagoma as evidence of having been bitten or denied contact with a “kissing bug”. Of the total, 26 reacted to the first screening test to detect anti-Trypanosoma cruzi antibodies; of these, five were confirmed as positive on the second screening test. The seroprevalence of T. cruzi was 0.01%. The results do not show a clear association but the frequency of hyperglycemia in population with Chagas coincides with that reported by other authors. Considering the scarce clinical–epidemiological evidence between Chagas disease, obesity, and hyperglycemia, long-term follow up of both morbidities is an area of opportunity for clinical and epidemiological study of T. cruzi reactivity.

Detection of Trypansoma cruzi in Kissing Bugs (Hemiptera: Reduviidae: Triatominae) Collected Across Oklahoma

Trypanosoma cruzi is the causative agent of Chagas disease in humans and dogs in the Americas. Transmission predominantly occurs via the feces of infected kissing bugs (Hemiptera: family Reduviidae; subfamily Triatominae) contaminating bite site wounds or mucous membranes. To better understand Chagas disease entomologic risk in Oklahoma, kissing bugs collected from within the state were tested for T. cruzi DNA. Data including county of insect collection, species and instar, and specific locations where specimens were found were collated. Triatomines were also tested by PCR to potentially identify DNA of vertebrate species on which specimens had recently fed. In total, 110 kissing bugs from 22 counties were tested. All triatomines were identified as Triatoma sanguisuga nymphs or adults, with the exception of one possible T. lecticularia adult. Trypanosoma cruzi DNA was detected in 22 (20%) triatomines from 12 counties spanning the state. The majority of T. cruzi PCR positive kissing bugs were found inside homes or associated structures (i.e., garages, porches). Vertebrate DNA was identified in 27 (24.5%) triatomines, with human DNA detected in 25 (92.6%) of these specimens, and canine and raccoon DNA detected in one specimen each (3.7%). Two specimens tested positive for both T. cruzi and human DNA and one specimen tested positive for both T. cruzi and raccoon DNA. Results from this study indicate that kissing bugs carrying T. cruzi are widespread in Oklahoma, that positive kissing bugs infest homes and associated structures, and that human-vector, canine-vector, and wildlife-vector contact all occur within the state.

Actualización de la distribución geográfica de triatominos en el departamento del Putumayo

La enfermedad de Chagas es una zoonosis producida por la infección del Trypanosoma cruzi (T. cruzi) (1), cuya principal vía de transmisión es vectorial (2). Esta enfermedad se caracteriza por ser una infección crónica que puede ocasionar daños cardiacos, digestivos y neurológicos irreversibles (3). En el departamento del Putumayo, de acuerdo con los datos del Sistema de Vigilancia Epidemiológica (SIVIGILA), entre el año 2015 y el 2020, se han notificado 19 casos de Chagas crónico y 4 casos de Chagas agudos (4). Por este motivo resulta de gran interés compartir con los lectores de la revista MedUNAB la actualización de la distribución geográfica de los triatominos, vectores de la enfermedad de Chagas, y establecer el riesgo epidemiológico que representan para la población Putumayense, donde hay hallazgos de gran importancia porque se identifican especies en municipios y localidades donde antes no se conocían.