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Análise proteômica de cultura macrófagica tratada com ácido kójico
(Universidade Federal do Pará, 2023-07) ALMEIDA, Heyder Coutinho; SANTOS, Agenor Valadares; http://lattes.cnpq.br/9530734927662735; https://orcid.org/0000-0002-2690-2841
Kojic acid (AK) is a natural fungal secondary metabolite, produced by some species of Aspergillus, Penicillium and Acetobacter, through the direct biotransformation of carbon substrates such as glucose, sucralose, ethanol, among others. AK was originally isolated in Japan, in 1907, from Aspergillus oryzae mycelia, it is used in the cosmetic industry in the form of gels and soap and is used to obtain the effect of whitening the skin by inhibiting the enzyme tyrosinase, reducing the production of formation of melanin, it also blocks the formation of pigmentation and evens out the skin tone. It is also applied as a food additive for the prevention of enzymatic browning, as it is a metal chelator and a potent antioxidant. Although AK has numerous biological functions, only recently studies on in vitro proliferation and cytotoxicity have been reported, but still limitedly regarding its effect on immune cells. Macrophages are among the most important defense cells that specifically recognize and respond to foreign bodies, apoptosis cells and pathogens. Through the activation process, there is an increased proliferation of macrophages, which undergo various morphological changes, such as an increase in dissemination and adhesion abilities, phagocytosis activity, ROS generation, antigen presentation and cytokine production. According to the classification of biological functions, our results revealed that in both Kojic acid concentrations, the 14 expressed proteins have a predicted function related to cell cycle and redox processes.
Acesso aberto (Open Access)
Estudo da ação imunomodulatória do ácido kójico sobre as células mononucleares da medula óssea de camundongos
(Universidade Federal do Pará, 2015-11-03) ALMEIDA, Caroline Martins; SILVA, Edilene Oliveira da; http://lattes.cnpq.br/7410116802190343
Bone marrow is soft and sponge-like material found inside bones that contains hematopoietic cells responsible for development and proliferation of peripheral blood cells. The monocytes proliferation generated in the bone marrow and the differentiation of this cells in macrophages plays a key role in the immune response. In this context, the research for drugs that enhance the innate immune response is needed to restore the homeostasis and the immune response. Kojic Acid (KA) is a secondary metabolite synthesized by some species of fungi from Aspergillus genera and has several applications (food additive, cosmetics, antitumor agent and macrophage activator). Thus, the aim of this study is to evaluate the immunomodulatory effects of kojic acid (KA) in the bone marrow cells of mice. These cells were obtained by flushing femurs, and maintained in cultures treated with KA at the concentration of 100 μg/mL for 24-96 hours of culture. It was observed by optical microscopy that KA promoted increased cell adhesion, spreading ability and high number of cytoplasmatic projections and vacuoles in cytoplasm in the mononuclear cells from bone marrow treated with AK. To confirm these results, Akt signaling pathway was analyzed by western blot. KA seems to be able to activate the Akt signaling pathway that have a critical regulatory role in cellular development and differentiation. In addition, we detected by cytometer analysis, increase in the F4/80 and in the CD11b expression, and decrease in CD11c when cells were treated for 96 hours, showing that KA induce the differentiation of bone marrow cells in macrophages, but not in dendritic cells. The Analysis of the microbicidal response revealed that KA also potentiated phagocytosis and increased the production superoxide anion, But not promoted increases of nitric oxide production. Furthermore, no cytotoxic effects were observed in cells treated with KA when compared to the untreated bone marrow cells. Thus, KA acts as an immunomodulatory and is able to induce bone marrow monocyte-macrophage differentiation process.
Acesso aberto (Open Access)
Estudo da função tímica em portadores de HTLV-1 com PET/MAH
(Universidade Federal do Pará, 2017) GOMES, Jéssica Antonia Nunes; FUZII, Hellen Thais; http://lattes.cnpq.br/0026958665547973
In HTLV-1 infeccions 90% of carriers remain asymptomatic, 2-3% develop Adult T-cell leukemia/lymphoma (ATL) and 0.25-4% develop HTLV-associated myelopathy/tropical spastic paraparesis (HAM/TSP). In HAM/TSP, specific CD8 + T cell infiltrates are found in the marrow that destroy infected CD4 + T cells, leading to a chronic activated immune response. Recently emigrated T cells (or naïve T cells) have excised circles by the rearrangement of T cell receptor (TREC) genes that do not double in cell proliferation, being a good indicator to quantify the number of naïve T cells and thus evaluate the thymic function. This study aimed to verify the thymic function of patients with HTLV-1 infection by quantifying the number of TREC in peripheral blood mononuclear cells. This is a cross-sectional, analytical study of 39 patients over 18 years of age, divided into two groups: HAM/TSP (PET) and without HAM/TSP (NPET). We performed a clinical and physiotherapeutic evaluation, blood collection, lymphomononuclear cell separation, DNA and RNA extraction, absolute TREC curve, DNA and RNA quantification, TREC particle detection and quantification, cDNA synthesis, cytokine IL-7 and statistical analysis with the Mann-Whitney tests and Spearman's correlation, with p ≤ 0.05 as significance level. Of the 39 patients studied, two were excluded from the study because they presented autoimmune disease. Regarding the comparison between groups of TREC quantification: there was a difference between the PET and NPET groups (p = 0.01), in patients with age ≤59 years between the PET and NPET groups (p = 0.04), in the (p = 0.003) and the group with a wheelchair and without a wheelchair (p = 0.05). As for the comparison of IL-7 gene expression between groups: in the NPET group there was a difference between the group ≤59 years and the ≥60 years (p = 0.02), in the female there was a difference between the PET and NPET groups (p = 0.04). Thymic function was impaired in patients with HTLV-1 with HAM/TSP compared to those without HAM/TSP, as there was a loss in naïve T cell production in this population, shown by the differences between variables in both PET and NPET groups With respect to the quantification of TREC. Although the importance of this compromise in the triggering and / or evolution of HAM/TSP is not yet clear, it is inferred that the reduction of naïve T cell production can alter the immunological response in these patients, directly affecting their clinical picture.
Acesso aberto (Open Access)
Indução de escleróticas in vitro e análise da resposta imune dos pacientes de cromoblastomicose em tratamento com itraconazol
(Universidade Federal do Pará, 2009-06-26) SILVA, Moisés Batista da; SALGADO, Claudio Guedes; http://lattes.cnpq.br/2310734509396125
Chromoblastomycosis (CBM) is a chronic fungal disease witch affects the skin, characterized for slowing development of polymorphic skin, that present infiltrated inflammatory granulomatous in the presence of sclerotics cells, characteristic of this illness. One of the objectives of this study was to evaluate the induction of scleroticts cells for natural mediums, with biomasses of Bactris gasipaes and Theobroma grandiflorum, whose respective species had induced in vitro similar sclerotics cells to those found in tissue of patients, in 10 and 2 days, respectively, what it made possible the production of a powder medium inductor, already donated to other groups that study the CBM. Another objective was to evaluate the histopathology of the CBM in the patients, before and during the use of itraconazole (ITZ). For this, the technique of ELISA for the cytokines was used TNF-α, circulating IL-4 and IL-10, and the immunohistochemestry of biópsias in different times of treatment - that it allowed to analyze the quantitative and qualitative alterations of the cellular types during 12 months of the treatment with ITZ in the 200 dose of mg/dia - with antibodies anti- CD20, anti-CD8 and anti-CD68. How much the cytokines, the circulating IL-10 did not show significant change, while IL-4 and TNF-α had presented an increase of the levels throughout 12 months of treatment. In relation to the immunophenotyping, it had a significant reduction in the inflammatory process and the cellular infiltrated during 3 and 6 months of the treatment, whereas only to the 12 months had the significant regression of the number of sclerotics cells. The immunophenotyping disclosed that the macrophages are mainly located in the areas central areas of granuloma, whereas cells TCD8+ are in the periphery and cells TCD20+, which were found throughout the tissues, with a significant increase after 6 months of the treatment, returning to the initial levels after one year. The cytotoxic macrophages and lymphocytes were having presented a significant increase after 12 months of treatment with ITZ. These results demonstrate that the formation of granuloma in the CBM is similar to those observed in other granulomatous infectious disease, and that the presence of IL-4 and IL-10 can be related with the persistence of fungi in the injuries and with the difficulty of cure observed in these patients.
Acesso aberto (Open Access)
Leishmania (L.) amazonensis inibe a maturação e a função ativadora das células de Langerhans da pele tratadas com TNF-α e anti-CD40 in vitro
(Universidade Federal do Pará, 2014-06-27) CAMPELO, Simone Rodrigues; SALGADO, Claudio Guedes; http://lattes.cnpq.br/2310734509396125
Leishmania amazonensis is one of the agents in a wide spectrum of clinical forms of cutaneous leishmaniasis. In general, the resistance against leishmaniasis depends on the development of an efficient immune response, however many studies have demonstrated that specific cytokines or combinations of cytokines may be factors of resistance or susceptibility to infection by L. amazonensis. Recent studies suggest the involvement of Langerhans cells (LCs) in the anti-Leishmania response, but the mechanisms involved in this interaction are still poorly understood. In this study, we investigated the role of TNF-α and anti-CD40 in L. amazonensis interaction with LCs in vitro, showing de profile of cytokines produced and the expression of surface molecules, besides verifing their abilities to activate the production of IFN-γ e IL-4 by lymph node cells. Methods: Fresh immature LCs, highly purified from BALB/c mouse skin, were incubated with L. amazonensis promastigotes, TNF-α and/or anti- CD40 mAb. After 24 h, LCs were co-cultured with lymph nodes cells of BALB/c mice for additional 72h. Culture supernatants were tested for IL-6, IL-12p70, IFN-γ and IL-4 by ELISA, while surface molecules were analyzed by FACS. Results: The levels of IL-6 and IL- 12p70 produced by LCs were significantly reduced after interaction with L. amazonensis, even after treatment of LCs with TNF-α or anti-CD40. Regarding surface molecules, there was no difference in the expression of CD207 in both groups, but the presence of L. amazonensis promoted a significant reduction in the expression of CD40 on LCs treated with TNF-α or anti-CD40, and increased expression CD86 in all groups. Lymph node cells showed a decreased production of IFN-γ in the presence of L. amazonensis and no change in IL-4. When co-cultured with LCs previously stimulated with L. amazonensis, the production of IFN-γ was also reduced, even in the presence of TNF-α and/or anti-CD40. No significant changes were observed in IL-4 by lymph cells co-cultured under the same experimental conditions. Conclusion: L. (L.) amazonensis exert an immunomodulatory effect on the immune response mediated by LCs by: 1) inhibiting the production of IL-6 and IL-12p70; 2) decreasing CD40 expression and; 3) preventing the activation of IFN-γ production by lymph node cells co-cultured with LCs, even after treatment with TNF-α and anti-CD40 antibody.
Acesso aberto (Open Access)
Modulação imunológica in vitro de células de Langerhans e macrófagos por drogas utilizadas no manejo de reações hansênicas
(Universidade Federal do Pará, 2008-04-03) CAMPELO, Simone Rodrigues; SALGADO, Claudio Guedes; http://lattes.cnpq.br/2310734509396125
Langerhans cells (LCs) are localized in the epidermis and performs a key role in the induction of immune response and immunologic tolerance. Macrophages are phagocytic cells that act as first line of defense of the organism, and they are involved in the granuloma formation in patients with leprosy. The immunopathogeny of the cellular response in the reactional states is yet little studied, however, several evidences suggest that the drugs prednisone, thalidomide, cyclosporine and amitriptyline, used in the control of leprosy reactions, perform their effects by the modulation of different immunocompetent cells functions. The objective of the present study was to analyze in vitro action of prednisone, thalidomide, cyclosporine and amitriptyline on the cytokine production by LCs and macrophages of BALB/c mice. LCs were isolated, purified and cultivated from the epidermis by the panning technique and macrophages were isolated by the peritoneal cavity of BALB/c mice. After 36 h of treatment with the drugs, the levels of TNF-, IL-12 and IL-10 were measured by ELISA. Prednisone, thalidomide, cyclosporine and amitriptyline inhibited TNF- produced by LCs, in both concentrations, however no important alterations in IL-12 production were detected. TNF- and IL-12 production by macrophages was also decreased after treatment, but IL-10 levels were not modified for none of the drugs tested. Our results show that these drugs can modulate the immune response by the regulation of pro-inflammatory cytokines TNF- and IL-12 by purified epidermal LCs and peritoneal macrophages, indicating that they constitute an important target for the drugs used in treatment of leprosy reactional states.
Acesso aberto (Open Access)
Polarização de resposta dos macrófagos e suas possíveis implicações na imunopatogênese da Doença de Hansen
(Universidade Federal do Pará, 2016) SOUSA, Jorge Rodrigues de; QUARESMA, Juarez Antônio Simões; http://lattes.cnpq.br/3350166863853054
Leprosy is a disease that causes changes in the tissue and in the course of the immune response. Macrophages are one of the main cells which participating in the response against M. leprae. These cells undergo a differentiation process, modifying their behavior and polarizing the response between the classical pathway, composed of M1 macrophages that produce pro-inflammatory mediators and the alternative pathway known as restorative pathway having as representative M2 macrophages and their subtypes that produce anti-inflammatory cytokines. In Hansen's disease, there are few studies that discuss the ways of polarization in the spectrum of the disease. In order to understand the behavior of the cell in the polar form soft he disease, this study investigated the ways of macrophage response polarization and its possible implications for Hansen's disease. We used 33 blocks with skin fragments of patients with confirmed diagnosis for the disease according to the Ridley and Jopling criteria. The distribution of cases: 17 were tuberculoid and 16 lepromatous. The immunohistochemical method was used for the investigation of the markers CD68, CD163, arginase 1, iNOS, STAT3, FGF b, TGF-β and cytokines, IL-10, IL-13, IL-22 in skin lesions. The results showed a statistically significant difference among the groups and the expression of CD68, CD163, enzymes, growth factors, cytokines and STAT3 were higher in lepromatous forms when compared tuberculoid form. In the correlation analysis, it was observed the synergism between the response of anti-inflammatory cytokines, growth factors, arginase 1 and CD163 in lepromatous form. In the context of iNOS response in susceptible pole of the disease, the study points to a new approach involving iNOS, STAT3 and IL-22 behavior and this new relationship in Hansen's disease, a statistically significant positive correlation was observed between the markers in the lepromatous form. This study concludes: the markers that compose the response of M1 and M2 macrophages are an alternative for a better understanding of innate immune response, in polar forms of the disease increasing the role of cytokines, enzymes, and receptors in the microbicidal response, repair and suppression in the lepromatous form of the disease.