Methodological approach to the ex vivo expansion and detection of T. cruzi-specific T cells from chronic Chagas disease patients

Acevedo, Gonzalo Raúl; Longhi, Silvia Andrea; Bunying, Alcinette; Sabri, Nazila; Atienza, Augusto; Zago, M. Paola; Santos, Radleigh; Judkowski, Valeria; Pinilla, Clemencia; Gómez, Karina Andrea

doi:10.1371/journal.pone.0178380

Cited by 12 publications

(13 citation statements)

References 45 publications

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“…3b). Furthermore, as seen in previously reported results 26,33 , PBMC from asymptomatic and cardiac Chagas patients proliferated ( Fig. 3c; and see Supplementary material, Fig.…”

Section: Cellular Immune Response Triggered By C-txnpx In Patients Wisupporting

confidence: 88%

Evaluation of the immune response against Trypanosoma cruzi cytosolic tryparedoxin peroxidase in human natural infection

et al. 2018

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Trypanosoma cruzi, the aetiological agent of Chagas disease, has a highly efficient detoxification system to deal with the oxidative burst imposed by its host. One of the antioxidant enzymes involved is the cytosolic tryparedoxin peroxidase (c-TXNPx), which catalyses the reduction to hydrogen peroxide, small-chain organic hydroperoxides and peroxynitrite. This enzyme is present in all parasite stages, and its overexpression renders parasites more resistant to the oxidative defences of macrophages, favouring parasite survival. This work addressed the study of the specific humoral and cellular immune response triggered by c-TXNPx in human natural infection. Thus, sera and peripheral blood mononuclear cells (PBMC) were collected from chronically infected asymptomatic and cardiac patients, and non-infected individuals. Results showed that levels of IgG antibodies against c-TXNPx were low in sera from individuals across all groups. B-cell epitope prediction limited immunogenicity to a few, small regions on the c-TXNPx sequence. At a cellular level, PBMC from asymptomatic and cardiac patients proliferated and secreted interferon-γ after c-TXNPx stimulation, compared with mock control. However, only proliferation was higher in asymptomatic patients compared with cardiac and non-infected individuals. Furthermore, asymptomatic patients showed an enhanced frequency of CD19 CD69 cells upon exposure to c-TXNPx. Overall, our results show that c-TXNPx fails to induce a strong immune response in natural infection, being measurable only in those patients without any clinical symptoms. The low impact of c-TXNPx in the human immune response could be strategic for parasite survival, as it keeps this crucial antioxidant enzyme activity safe from the mechanisms of adaptive immune response.

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“…3b). Furthermore, as seen in previously reported results 26,33 , PBMC from asymptomatic and cardiac Chagas patients proliferated ( Fig. 3c; and see Supplementary material, Fig.…”

Section: Cellular Immune Response Triggered By C-txnpx In Patients Wisupporting

confidence: 88%

Evaluation of the immune response against Trypanosoma cruzi cytosolic tryparedoxin peroxidase in human natural infection

et al. 2018

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“…However, T cell proliferative response is hampered in Chagas disease patients, at least in vitro , upon exposure to a strong, non-specific mitogen, and decreased expression of receptors involved in T cell activation is observed. Furthermore, proliferation of T cells from non-infected donors is inhibited in the presence of T. cruzi antigens (117, 118). T cell response is particularly important for the maintenance of the typically low parasitaemia in the chronic phase of the disease (119), and if impaired (e.g., in VIH co-infection cases), it leads to rapid clinical onset (120, 121).…”

Section: Adaptive Immunitymentioning

confidence: 99%

The Unsolved Jigsaw Puzzle of the Immune Response in Chagas Disease

2018

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Trypanosoma cruzi interacts with the different arms of the innate and adaptive host's immune response in a very complex and flowery manner. The history of host-parasite co-evolution has provided this protozoan with means of resisting, escaping or subverting the mechanisms of immunity and establishing a chronic infection. Despite many decades of research on the subject, the infection remains incurable, and the factors that steer chronic Chagas disease from an asymptomatic state to clinical onset are still unclear. As the relationship between T. cruzi and the host immune system is intricate, so is the amount and diversity of scientific knowledge on the matter. Many of the mechanisms of immunity are fairly well understood, but unveiling the factors that lead each of these to success or failure, within the coordinated response as a whole, requires further research. The intention behind this Review is to compile the available information on the different aspects of the immune response, with an emphasis on those phenomena that have been studied and confirmed in the human host. For ease of comprehension, it has been subdivided in sections that cover the main humoral and cell-mediated components involved therein. However, we also intend to underline that these elements are not independent, but function intimately and concertedly. Here, we summarize years of investigation carried out to unravel the puzzling interplay between the host and the parasite.

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“…Although in this study TNF‐α was measured, the data is not shown 48. In the same investigation, TCR usage in two CD4+ T cell lines displayed predominance of TCR‐Vβ 5.2 and 5.1 families 48. Interestingly, we identified the same TCR‐Vβ families among TNF‐α producing CD4+ and CD8+ T cells in two chagasic patients (Fig.…”

Section: Discussionmentioning

confidence: 53%

“…In a recent study, following ex vivo expansion of PBMCs from chagasic patients, IFN‐γ and GM‐CSF were considered the best read out of antigen specificity among cytokines tested in CD4+ T cells clones. Although in this study TNF‐α was measured, the data is not shown 48. In the same investigation, TCR usage in two CD4+ T cell lines displayed predominance of TCR‐Vβ 5.2 and 5.1 families 48.…”

Section: Discussionmentioning

confidence: 70%

“…Detection of anti‐ T. cruzi specific T cells can be difficult in peripheral blood due to the heterogeneity of T cells population and the size of T. cruzi proteome with a high potential of HLA restricted T cells epitopes 7, 48. In a recent study, following ex vivo expansion of PBMCs from chagasic patients, IFN‐γ and GM‐CSF were considered the best read out of antigen specificity among cytokines tested in CD4+ T cells clones.…”

Section: Discussionmentioning

confidence: 99%

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T cells responding to Trypanosoma cruzi detected by membrane TNF‐α and CD154 in chagasic patients

Ripoll

Giraldo

Bolaños

et al. 2017

Immunity Inflam & Disease

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IntroductionChagas disease is a parasitic infection whose pathogenesis is related to parasite persistence and a dysfunctional cellular immune response. Variability in cytokine secretion among chronic Trypanosoma cruzi‐infected patients might preclude the identification of the pool of antigen specific T cells. The goal of this study was to determine the fraction of T cells responding to T. cruzi antigen measured by the expression of membrane TNF‐α and CD154.MethodsA total of 21 chagasic patients, 11 healthy and 5 non‐chagasic cardiomyopathy controls were analyzed. PBMCs were short‐term cultured in the presence of anti‐CD28, anti‐CD49d, anti‐TNF‐α, and TACE (TNF‐α converting enzyme) inhibitor either under T. cruzi‐lysate or polyclonal stimuli. Cells were stained with anti‐CD3, anti‐CD4, anti‐CD8, and anti‐CD154, and analyzed with flow cytometry.ResultsCD4+ and CD8+ T cells in chagasic patients displayed higher percentages of membrane‐bound TNF‐α+ and CD154+ compared with controls after T. cruzi‐antigen stimulation. Both markers displayed a positive correlation in the T cell subpopulations analyzed. Symptomatic chagasic patients were differentiated from asymptomatic patients based on the expression of CD154 and membrane TNF‐α in TCD4+ and TCD8+ compartments, respectively.ConclusionsThese results show that both markers could be useful for assessing the pool of antigen‐specific T cells in chronic chagasic patients.

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Methodological approach to the ex vivo expansion and detection of T. cruzi-specific T cells from chronic Chagas disease patients

Cited by 12 publications

References 45 publications

Evaluation of the immune response against Trypanosoma cruzi cytosolic tryparedoxin peroxidase in human natural infection

Evaluation of the immune response against Trypanosoma cruzi cytosolic tryparedoxin peroxidase in human natural infection

The Unsolved Jigsaw Puzzle of the Immune Response in Chagas Disease

T cells responding to Trypanosoma cruzi detected by membrane TNF‐α and CD154 in chagasic patients

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