Production of cytokine and chemokines by human mononuclear cells and whole blood cells after infection with Trypanosoma cruzi

Abstract

<jats:p>INTRODUCTION: The innate immune response is the first mechanism of protection against Trypanosoma cruzi, and the interaction of inflammatory cells with parasite molecules may activate this response and modulate the adaptive immune system. This study aimed to analyze the levels of cytokines and chemokines synthesized by the whole blood cells (WBC) and peripheral blood mononuclear cells (PBMC) of individuals seronegative for Chagas disease after interaction with live T. cruzi trypomastigotes. METHODS: IL-12, IL-10, TNF-α, TGF-β, CCL-5, CCL-2, CCL-3, and CXCL-9 were measured by ELISA. Nitrite was determined by the Griess method. RESULTS: IL-10 was produced at high levels by WBC compared with PBMC, even after incubation with live trypomastigotes. Production of TNF-α by both PBMC and WBC was significantly higher after stimulation with trypomastigotes. Only PBMC produced significantly higher levels of IL-12 after parasite stimulation. Stimulation of cultures with trypomastigotes induced an increase of CXCL-9 levels produced by WBC. Nitrite levels produced by PBMC increased after the addition of parasites to the culture. CONCLUSIONS: Surface molecules of T. cruzi may induce the production of cytokines and chemokines by cells of the innate immune system through the activation of specific receptors not evaluated in this experiment. The ability to induce IL-12 and TNF-α contributes to shift the adaptive response towards a Th1 profile.</jats:p>

Evaluation of Gene Expression of TGF-B and FOXP3 in Individuals Chronically Infected with Trypanosoma Cruzi

“…Few studies have investigated the role of soluble factors involved in the immune response against Chagas disease. Similar to the results reported here, studies such as Rezende-oliveira et al, [27] in which TGF-β levels were measured in the supernatant of mononuclear cells infected with T. cruzi, and Nougueira et al, [28], who analyzed TGF-β gene expression through real time PCR in patients with chronic Chagas disease, reported no statistically significant differences in TGF-β expression. This allows us to conclude that TGF-β synthesis assessed through qPCR after cultivation with CRA and FRA Rec-Ags may not be used as a prognostic marker of the evolution of the clinical forms of chronic Chagas disease.…”

Effects of Meglumine Antimoniate Treatment on Cytokine Production in a Patient with Mucosal Leishmaniasis and Chagas Diseases Co-Infection

“…In this way, our results seem to be promising by the fact that treatment decreased the percentage of circulating CD4 + CD25 + Foxp3 + T cells, concomitantly to the decreased ability of leukocytes to produce TNF-α, IL-6, and IL-10 in response to live parasites or their antigens. Live trypomastigotes of the same T. cruzi Y strain used in our study were able to induce TNF-α and IL-12 but not IL-10 production in PBMC of CD seronegative healthy individuals [12]. This suggests an important in vivo clonal expansion of parasite-specific IL-10-producing T cells.…”

“…After, TMB (tetramethylbenzidine) substrate solution (BD Pharmingen, San Diego, CA, USA) was used for assay revelation, which was read on a microplate Venous blood was collected in two different time periods: (1) at the moment of patient's admission, just after clinical evaluation and before any treatment regimen and (2) at the end of specific treatment for mucocutaneous leishmaniasis (40-day period of Glucantime ® regimen). In both periods, peripheral blood mononuclear cells (PBMC) were separated using Ficoll-Paque TM Plus gradient (GE Health Care, Uppsala, Sweden) and cultured in RPMI 1640 (GIBCO, Grand Island, NY, USA) in 8 different culture conditions: medium alone, 2 ug/mL Phytohaemagglutinin (Sigma, St. Louis, MO, USA), 3:1 T cruzi (Y strain) or 3:1 L. braziliensis (Lb2904 strain) live parasites, as well as 5 ug/mL T. cruzi and 5 ug/mL L. braziliensis soluble proteins, in a 5% CO 2 atmosphere at 37 °C for 24 h, as described elsewhere [11,12]. The supernatants were used for cytokine quantification, IL-6, IL-10, TNF-α and IL-12, by ELISA assay as previously described [11,13].…”

A Review on the Immunological Response against Trypanosoma cruzi

“…Innate immunity, consisting of phagocytes, especially macrophages, neutrophils and dendritic cells, constitutes the first line of defence that T. cruzi faces when invading a vertebrate host [ 21 , 22 , 23 ]. The role of these cells is to recognize pathogen-associated molecular patterns (PAMPs) and damage-associated molecular patterns (DAMPs) by membrane receptors such as toll-like receptors (TLRs).…”