ABSTRACT
In the present study, we characterized the phagocytic capacity, cytokine profile along with the FCγ-R and TLR expression in leukocytes from Chagas disease patients (indeterminate-IND and cardiac-CARD) before and one-year after Bz-treatment (INDT and CARDT). A down-regulation of IL-17, IFN-γ and IL-10 synthesis by neutrophils was observed in CARDT. The Bz-treatment did not impact on the expression of phagocytosis-related surface molecules or monocyte-derived cytokine profile in INDT. Although CARDT showed unaltered monocyte-phagocytic capacity, up-regulated expression of Fcγ-RI/III and TLR-4 may be related to their ability to produce IL-10 and TGF-ß. Down-regulation of lymphocyte-derived cytokine was observed in INDT whereas up-regulated cytokine profile was observed for lymphocytes in CARDT. Analysis of cytokine network revealed that IND displayed a multifaceted cytokine response characterized by strong connecting axes involving pro-inflammatory/regulatory phagocytes and lymphocytes. On the other hand, CARD presented a modest cytokine network. The Bz-treatment leads to distinct cytokine network: decreasing the links in INDT, with a pivotal role of IL-10(+) monocytes and expanding the connections in CARDT. Our findings highlighted that the Bz-treatment contributes to an overall immunomodulation in INDT and induces a broad change of immunological response in CARDT, eliciting an intricate phenotypic/functional network compatible with beneficial and protective immunological events.
Subject(s)
Chagas Disease/drug therapy , Neutrophils/drug effects , Nitroimidazoles/administration & dosage , Trypanocidal Agents/administration & dosage , Adolescent , Adult , Chagas Disease/immunology , Controlled Before-After Studies , Cytokines/metabolism , Female , Host-Pathogen Interactions/drug effects , Humans , Immunomodulation , Inflammation Mediators/metabolism , Male , Middle Aged , Neutrophils/immunology , Receptors, IgG/genetics , Receptors, IgG/metabolism , Toll-Like Receptor 4/genetics , Toll-Like Receptor 4/metabolism , Young AdultABSTRACT
The distinct ability of phagocytes to present antigens, produce cytokines and provide co-stimulatory signals may contribute to the severity of the outcome of Chagas disease. In this paper, we evaluate the phenotypic features of phagocytes along with the cytokine signature of circulating T-cells from Chagas disease patients with indeterminate (IND) and cardiac (CARD) clinical forms of the disease. Our data demonstrated that neutrophils from IND patients displayed an impaired ability to produce cytokines. A lower Trypanosoma cruzi phagocytic index and higher nitric oxide levels were characteristics of monocytes from IND. The impaired phagocytic capacity did not reflect on the levels of anti-T. cruzi IgG, but was detectable in the downregulation of Fc-γR, TLR and CR1 molecules. The monocyte-derived cytokine signature demonstrated that a down-regulated synthesis of IL-12 and a modulatory state were evidenced by a positive correlation between IL-12 and IL-10 with a lower synthesis of TNF-α. The down-regulation of MHC-II and CD86 in monocytes supports the occurrence of particularities in the APC-activation-arm in IND, and may be involved in the T-cell pro-inflammatory pattern counterbalanced by a potent IL-10 response. Our findings support the hypothesis that differential phenotypic features of monocytes from IND may be committed to the induction of a distinct immune response related to low morbidity in chronic Chagas disease.
Subject(s)
Chagas Disease/immunology , Cytokines/biosynthesis , Monocytes/immunology , Neutrophils/immunology , Phagocytosis/immunology , Antibodies, Protozoan/blood , Antibodies, Protozoan/immunology , Antigen-Presenting Cells/immunology , Antigen-Presenting Cells/metabolism , Antigens, Protozoan/immunology , B7-2 Antigen/metabolism , Cells, Cultured , Chagas Disease/metabolism , Humans , Immunoglobulin G/blood , Immunoglobulin G/immunology , Immunomodulation , Monocytes/metabolism , Neutrophils/metabolism , Nitric Oxide/biosynthesis , Receptors, Complement 3b/metabolism , Receptors, IgG/metabolism , T-Lymphocytes/immunology , T-Lymphocytes/metabolism , Toll-Like Receptors/metabolism , Trypanosoma cruzi/immunologyABSTRACT
Flow cytometry has been proposed as an alternative method for direct determination of intracellular NO by using the 4,5-diaminofluorescein-diacetate (DAF-2DA) as a fluorescent probe. In the present study, the protocol for intracellular NO determination in peripheral blood monocytes and neutrophils of by flow cytometry was optimized and applied to monitor chronic graft nephropathy patients. The optimize method consists to incubate plasma-free whole blood samples with DAF-2DA at 2.0 microM for 180 min at 37 degrees C to determine the percentage of DAF-2T+ monocytes and neutrophils. Distinct intracellular NO profiles in monocytes and neutrophils from chronic graft nephropathy patients as compared to the healthy individuals. Although the pre-incubation with LPS was able to trigger higher percentages of DAF-2T+ monocytes and neutrophils in both groups, our data demonstrated that LPS had a greater impact on monocytes as compared to neutrophils, selectively in the group of healthy individuals. Moreover, our findings demonstrated that LPS had lower impact on monocytes from chronic graft nephropathy as compared to healthy individuals. Supplementary analysis revealed that the LPS impact tends to be resorted in those patients with longer post-transplant time, as demonstrated by a significant positive correlation index. Furthermore, our results demonstrated that AG had lower inhibitory impact on neutrophils as compared to monocytes, selectively in the group of chronic graft nephropathy patients. Taken together, this study showed a new approach to monitor the immunological status of patients with chronic graft nephropathy opening new perspectives of research regarding the monocyte and neutrophil functions in patient undergoing immunosuppressive therapy.
