In mammals, chronic diseases resulting from infectious agents have been associated with functional T cell response deficiency, a high frequency of terminally differentiated T cells, the presence of monofunctional Ag-specific T cells, and increased expression of inhibitory receptors. Similar to other chronic diseases, the progressive loss of certain functional activities during Trypanosoma cruzi infection might result in the inability to control replication of this parasite. To examine this hypothesis, we evaluated the differentiation and cell effector function of CD8⁺ T cells and characterized the expression of inhibitory receptors and the presence of the parasite in the bloodstream of chagasic patients. The results showed that patients at an advanced severe disease stage had a higher frequency of terminally differentiated CD8⁺ T cells than patients at an early stage of the disease. A monofunctional CD8⁺ T cell response was observed in patients at an advanced stage, whereas the coexpression of markers that perform three and four functions in response to parasite Ags was observed in patients at a less severe disease stage. The frequency of CD8⁺ T cells producing granzyme B and perforin and those expressing inhibitory receptors was higher in symptomatic patients than in asymptomatic patients. Taken together, these findings suggest that during the course of Chagas disease, CD8⁺ T cells undergo a gradual loss of function characterized by impaired cytokine production, the presence of advanced differentiation, and increased inhibitory receptor coexpression.

CD8+ T cells have been shown to play a crucial role in Trypanosoma cruzi infection. Memory CD8+ T cells can be categorised based on their distinct differentiation stages and functional activities as follows: stem cell memory (TSCM), central memory (TCM), transitional memory (TTM), effector memory (TEM) and terminal effector (TTE) cells. Currently, the immune mechanisms that control T. cruzi in the chronic phase of the infection are unknown. To characterise the CD8+ T cell subsets that could be participating in the control of T. cruzi infection, in this study, we compared total and T. cruzi-specific circulating CD8+ T cells with distinctive phenotypic and functional features in chronic chagasic patients (CCPs) with different degrees of cardiac dysfunction. We observed a decreased frequency of total TSCM along with an increased frequency of TTE in CCPs with severe disease. Antigen-specific TSCM cells were not detectable in CCPs with severe forms of the disease. A functional profile of CD8+ T cell subsets among CCPs revealed a high frequency of monofunctional CD8+ T cells in the most severe patients with IFN-γ+- or TNF-α+-producing cells. These findings suggest that CD8+ TSCM cells may be associated with the immune response to T. cruzi and outcome of Chagas disease, given that these cells may be involved in repopulating the T cell pool that controls infection. Chagas disease is caused by the intracellular parasite Trypanosoma cruzi. After the onset of acute infection, all individuals enter the chronic phase and approximately 70% of them never have symptoms. However, nearly 30% of infected individuals develop symptoms, mainly of heart disease, even decades after the initial infection. Currently, it is unclear how the immune response controls infection and prevents the development of heart disease in some infected people. We have characterised the memory CD8⁺ T cell subsets in chronic chagasic patients, including a newly described population of cells called memory stem cells. This T cell subset seems important for replenishing the other T cell populations. The findings in this manuscript show that chronic chagasic patients with severe disease have the following: a) a low frequency of memory stem cells, b) no antigen-specific memory stem cells, and c) CD8⁺ T cells with less effector function compared with asymptomatic patients. These results indicate that the lack of T cell population renewal and the decrease in cells with multiple effector functions may be associated with the clinical outcome of chronic Chagas disease.

CD4+/CD8+ double positive (DP) T cells have been described in healthy individuals as well as in patients with autoimmune and chronic infectious diseases. In chronic viral infections, this cell subset has effector memory phenotype and displays antigen specificity. No previous studies of double positive T cells in parasite infections have been carried out. Seventeen chronic chagasic patients (7 asymptomatic and 10 symptomatic) and 24 non-infected donors, including 12 healthy and 12 with non-chagasic cardiomyopathy donors were analyzed. Peripheral blood was stained for CD3, CD4, CD8, HLA-DR and CD38, and lymphocytes for intracellular perforin. Antigen specificity was assessed using HLA*A2 tetramers loaded with T. cruzi K1 or influenza virus epitopes. Surface expression of CD107 and intracellular IFN-γ production were determined in K1-specific DP T cells from 11 chagasic donors. Heart tissue from a chronic chagasic patient was stained for both CD8 and CD4 by immunochemistry. Chagasic patients showed higher frequencies of DP T cells (2.1%±0.9) compared with healthy (1.1%±0.5) and non-chagasic cardiomyopathy (1.2%±0.4) donors. DP T cells from Chagasic patients also expressed more HLA-DR, CD38 and perforin and had higher frequencies of T. cruzi K1-specific cells. IFN-γ production in K1-specific cells was higher in asymptomatic patients after polyclonal stimulation, while these cells tended to degranulate more in symptomatic donors. Immunochemistry revealed that double positive T cells infiltrate the cardiac tissue of a chagasic donor. Chagasic patients have higher percentages of circulating double positive T cells expressing activation markers, potential effector molecules and greater class I antigenic specificity against T. cruzi. Although K1 tetramer positive DP T cell produced little IFN-γ, they displayed degranulation activity that was increased in symptomatic patients. Moreover, K1-specific DP T cells can migrate to the heart tissue. Chagas disease, produced by the blood parasite Trypanosoma cruzi, is considered a public health problem in Central and South America. Non sterile immunity can be achieved after acute infection. Parasite persistence can induce tissue damage in nearly 20% to 30% of chronically infected individuals. Indeed, chagasic cardiomyopathy is one of the consequences of the chronic infection. Antigen persistence and dysfunctional cellular immune response have been implicated in T. cruzi pathogenesis. Here, a higher frequency of circulating CD4+/CD8+ double positive T cells in chronic chagasic patients is reported as compared with non infected donors, including those with a non-chagasic cardiomyopathy. This cell subset also expressed more activation markers and stored more intracellular perforin. We have previously reported that CD8+ T cells from T. cruzi infected donors recognized the HLA-A*0201 restricted K1-peptide derived from the KMP-11 protein. Here, double positive T cells displayed higher percentages of recognition for the K1 peptide than single CD8+ T cells. These cells produce little IFN-γ, but display degranulation activity that was increased in the symptomatic group. Finally, double positive T cells can be localized in the heart tissue from a chronic chagasic donor.

Chagas disease is a chronic infection caused by Trypanosoma cruzi, an intracellular protozoan parasite. Chronic chagasic patients (CCPs) have dysfunctional CD8⁺ T cells that are characterized by impaired cytokine production, high coexpression of inhibitory receptors, and advanced cellular differentiation. Most patients diagnosed in the chronic phase of Chagas disease already exhibit heart involvement, and there is no vaccination that protects against the disease. Antiparasitic treatment is controversial as to its indication for this stage of the disease. There is a lack of biological markers to evaluate the effectiveness of antiparasitic treatment, and little is known about the effect of the treatment on CD8⁺ T cells. Thus, the aim of the current study was to analyze the early effects of antiparasitic treatment on CD8⁺ T cells from CCPs with asymptomatic clinical forms of disease. To evaluate the CD8⁺ T cell subsets, expression of inhibitory receptors, and functionality of T cells in CCPs, PBMCs were isolated. The results showed that treatment of CCPs with the asymptomatic form of the disease induces an increase in the frequency of CD8⁺ central memory T cells and terminal effector T cells, a decrease in the coexpression of inhibitory receptors, an improved Ag-specific CD8⁺ T cell response exhibited by the individual production of IFN-γ or IL-2, and a multifunctional CD8⁺ T cell profile of up to four functions (IFN-γ⁺IL-2⁺Perforin⁺Granzyme B⁺). These findings suggest that, in CCPs, antiparasitic treatment improved the quality of Ag-specific CD8⁺ T cell responses associated with a decrease in inhibitory receptor coexpression, which could serve as biomarkers for monitoring the effectiveness of antiparasitic treatment.