Navegando por Assunto "Citocinas"

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Análise da proteção de citocinas após exposição celular in vitro com os antígenos ML2478 e ML0840 do Mycobacterium leprae
(Universidade Federal do Pará, 2019-09) MESSIAS, Ana Caroline Cunha; SALGADO, Claudio Guedes; http://lattes.cnpq.br/2310734509396125; http://orcid.org/0000-0003-3961-7764
Diagnosis of oligosymptomatic leprosy cases may enable interventions to be performed before the onset of physical disabilities. However, because the diagnosis is still essentially clinical and the disease progresses slowly, there is difficulty in recognizing these cases, since the lesions are discreets and with subtle changes in sensitivity. Most of the time patients are diagnosed when they already have obvious clinical characteristics and/or physical disabilities. Thus, is necessary to develop laboratory tools that help in the early diagnosis of the disease. The cell immunity assay Whole Blood Assay (WBA) is a low-cost, easy-to-perform technique that provides conditions for antigen screening and is favored in areas where leprosy is endemic and may facilitate incorporation of a test into sites with less access to sophisticated laboratories. The objective of this study was to evaluate the cellular immune response after in vitro exposure of peripheral blood to Mycobacterium leprae antigens ML2478 and ML0840. Eighty-seven individuals were selected for quantitation the cytokines of Interferon-γ (IFN-γ), Interleukin (IL)-10, IL-17 and Transforming Growth Factor-β1 after exposure with specific M. leprae antigens by WBA for 24 hours. A total of 47 leprosy cases were evaluated distributed in: 6 tuberculoid and 14 borderline tuberculoid, 13 borderline lepromatous leprosy, 6 lepromatous leprosy; and 8 schoolchildren diagnosed with leprosy during the group active search strategy (oligosymptomatic cases in the clinical forms: 1 primary neural, 1 undetermined, 6 borderline tuberculoid). The remaining 47 individuals corresponded to 20 contacts, 13 healthy schoolchildren and 7 individuals with other skin diseases. The analysis of cytokines suggests the balance between IFN-γ and IL-10 may indicate individuals who are progressing to the Th2 pole. IL-17 and TGF-β1 may be used to follow-up individuals with similar response to leprosy cases. The production of IFN-γ, IL-10, IL-17 and TGF-β1 cytokines by stimulation with proteins ML2478 and ML0840 did not differ between healthy students and case students. And the cytokine IL-17 demonstrated higher production in cases attended at URE than in case students and individuals in control groups.
Acesso aberto (Open Access)
Análise genômica comparativa e os polimorfismos nos genes TNFA, IFNG IL6 e IL10 associados à expressão de citocinas na infecção por Plasmodium vivax no município do Itaituba, Estado do Pará
(Universidade Federal do Pará, 2018-07-19) PIMENTA, Tamirys Simão; MACHADO, Ricardo Luiz Dantas; http://lattes.cnpq.br/0307356330748427; OLIVEIRA, Edivaldo Herculano Correa de; http://lattes.cnpq.br/0094007714707651
In endemic areas of Asia, Oceania, Central and South America and in the horn of Africa P. vivax malaria is a major cause of morbidity with 35 million cases annually. In Brazil, the Amazon region concentrates almost all cases and infections registered countrywide, with more than three hundred thousand cases per year. Several evidences suggest that an exacerbated inflammatory response associated to density parasite is likely to aggravate the malaria symptoms. We assessed the haematological and immunological aspects, genetic alterations related to CNVs that could lead to phenotypic alterations, conferring resistance or susceptibility to malaria, as well the presence of polymorphisms in cytokine genes and their association with the infection in patients living in a gold-mining area in a gold-mining in the Brazilian Amazon Region, establishing patterns of immune response characteristic of primary malaria, recurrent malaria and endemic control. Six SNPs (TNFA-308G/A, IFNG+874T/A, IL6-174G/C, IL10-1082G/A, -819C/T, -592C/A) in four genes were determined; blood cell count was conducted on automatic analyzer; plasmatic cytokines IL-6, IL-10, TNF-α and IFN-γ were quantified by flow cytometry and density parasite was estimated by thick blood films with confirmation by nested-PCR; the CNV was estimated by aCGH and association between copy number and phenotypes (parasite load, mean number of clinical infections of malaria and gender) was assessed. The statistical analyzes were performed by Graph-pad prism 6.0 and Bioestat 5.0. No significant association was found between SNPs and malaria infection; cytokine levels were higher in malaria group when compared to endemic control; production of IL-10 was higher in the presence of GCC/GCC haplotype; IFN-γ levels were correlated with previous malaria episodes; malaria patients showed lower platelet numbers, reduction on white blood cells count and an increased monocyte percentage; significant increase in the IL-6 and IL-10 plasmatic levels in both malaria groups; the primary malaria patients displayed the highest significant plasmatic IFN-γ levels; recurrent malaria patients displayed the highest significant plasmatic TNF-α; malaria infection demonstrated correlation between parasite density and TNF-α, IL-6 and IL-10 levels; a total of 112 amplified genes and 12 deleted genes were observed and the CNVs found did not include any gene related to receptors or vivax malaria resistance factors. There were no statistically significant correlations between the clinical and pathological data (parasite load, mean number of clinical infections of malaria and gender) and the presence of CNVs in the patients studied. This study provides additional data on Plasmodium-host immune response and describes the quantitative changes in the human genome in P. vivax infection in an endemic area of garimpo.
Acesso aberto (Open Access)
Analysis of polymorphisms in the interleukin 18 gene promotor (-137 g/c and -607 c/a) in patients infected with hepatitis c virus from the brazilian amazon
(Universidade Federal do Pará, 2015-09) SANTOS, Kemper Nunes dos; ALMEIDA, Marcella Kelly Costa de; FECURY, Amanda Alves; COSTA, Carlos Araújo da; MARTINS, Luisa Caricio
Background - The hepatitis C virus has been recognized as the leading cause of chronic liver disease in the world. Host genetic factors have been implicated in the persistence of hepatitis C virus infection. Single nucleotide polymorphisms at positions -607 C/A (rs1946518) and -137 G/C (rs187238) in the IL-18 gene promoter have been suggested to be associated with delayed hepatitis C virus clearance and persistence of the disease. Objective - Identify these polymorphisms in a population infected with hepatitis C virus from the Brazilian Amazon region. Methods - In a cross-sectional analytical study conducted in Belém, Pará, Brazil, 304 patients infected with hepatitis C virus were divided into two groups: group A, patients with persistent infection; group B, patients with spontaneous clearance. The control group consisted of 376 volunteers not infected with hepatitis C virus. Samples were analyzed by RT-PCR for the detection of viral RNA and by RFLP-PCR to evaluate the presence of the -137 G/C and -607 C/A IL-18 gene promoter polymorphisms. Results - Comparison of polymorphism allele frequencies between the patient and control groups showed a higher frequency of allele C at position -607 among patients (P=0.02). When the association between the polymorphisms and viral infection was analyzed, patients carrying genotype C/A at position -607 were found to be at higher risk of persistent hepatitis C virus infection (P=0.03). Conclusion - the present results suggest a possible role of the -607 IL-18 gene promoter polymorphism in the pathogenesis of hepatitis C virus infection.
Acesso aberto (Open Access)
Associação entre marcadores da resposta inflamatória e a imunopatogênese de agentes infecciosos de natureza viral (Vírus da dengue, HTLV-1 e HTLV-2) e bacteriana (Chlamydia trachomatis e Chlamydia pneumoniae)
(Universidade Federal do Pará, 2010-11-29) FEITOSA, Rosimar Neris Martins; ISHAK, Ricardo; http://lattes.cnpq.br/5621101706909450
The genetic basis of diseases is frequently studied aiming the polymorphisms of cytocine genes. The present study investigated markers of the inflammatory response associated to the course of infection and disease caused by viruses and bacteria. Serum levels (measured by an ELISA assay) and the polymorphisms (using PCR, RFLP and allele specific PCR) of TNF-α (-308), TNF-β (+252), IFN-γ (+874) and C reactive protein were measured among persons with febrile disease, infected by dengue virus (n=80), not infected by DV (100), a group of HTLV infected (30 symptomatic and 47 asymptomatic), a group with coronary disease (58 seroreactive to Chlamydia and 31 with negative serology) and a control group (99 persons with no reaction to DV, HTLV and Chlamydia). No group showed association with demographic informations. Dengue virus 3 (66.2%) and HTLV-1 (90% symptomatic and 76.6% asymptomatic persons) were the most frequent agents found among their groups. The majority of those with coronary disease (65.1%) presented antibodies to Chlamydia (39.6% to C. trachomatis and C. pneumoniae, 58.6% solely to C. trachomatis and 1.7% to C. pneumoniae). Statistically significant levels of differences were found among: (i) serum levels of TNF-β, IFN-γ and PrtCR of positive and negative dengue and control groups (p< 0,01); (ii) serum levels of TNF-α, TNF-β and IFN-γ of HTLV (including its types) and control groups; (iii) serum levels of TNF-α, TNF-β, IFN-γ and PrtCR among patients with coronary disease, serum reactive to Chlamydia, and the control group; (iv) the presence of antibodies to C. trachomatis and C. pneumoniae and the control group comparing TNF-β, IFN-γ and PrtCR. Genotypic frequency distributions were statistically significant for the polymorphisms: (i) of TNF-α (p=0,0494) and IFN-γ (p= 0,0008) genes among positive, negative and control dengue groups and to IFN-γ (p= 0,0007) among groups DEN 1, DEN 2, DEN 3 and controls; (ii) of IFN-γ gene (p= 0,0023) among the group of patients with coronary disease and sero reactivity to C. trachomatis e C. pneumoniae, as well as to the mono reactants in the comparison between the positivity to C. trachomatis and the control group.
Acesso aberto (Open Access)
Bacterial vaginosis in pregnant adolescents: proinflammatory cytokine and bacterial sialidase profile: cross-sectional study
(Universidade Federal do Pará, 2015-12) FERREIRA, Carolina Sanitá Tafner; MARCONI, Camila; PARADA, Cristina Maria Garcia de Lima; DUARTE, Marli Teresinha Cassamassimo; GONÇALVES, Ana Paula Oliveira; RUDGE, Marilza Vieira Cunha; SILVA, Márcia Guimarães da
Bacterial vaginosis occurs frequently in pregnancy and increases susceptibility to sexually transmitted infections (STI). Considering that adolescents are disproportionally affected by STI, the aim of this study was to evaluate the cervicovaginal levels of interleukin (IL)-1 beta, IL-6, IL-8 and bacterial sialidase in pregnant adolescents with bacterial vaginosis. DESIGN AND SETTING: Cross-sectional study at mother and child referral units in Belém, Pará, Brazil. METHODS: Vaginal samples from 168 pregnant adolescents enrolled were tested for trichomoniasis and candidiasis. Their vaginal microbiota was classified according to the Nugent criteria (1991) as normal, intermediate or bacterial vaginosis. Cervical infection due to Chlamydia trachomatisand Neisseria gonorrhoeae was also assessed. Cytokine and sialidase levels were measured, respectively, using enzyme-linked immunosorbent assays and MUAN conversion in cervicovaginal lavages. Forty-eight adolescents (28.6%) were excluded because they tested positive for some of the infections investigated. The remaining 120 adolescents were grouped according to vaginal flora type: normal (n = 68) or bacterial vaginosis (n = 52). Their cytokine and sialidase levels were compared between the groups using the Mann-Whitney test (P < 0.05). RESULTS: The pregnant adolescents with bacterial vaginosis had higher levels of IL-1 beta, IL-6 and IL-8 (P < 0.05). Sialidase was solely detected in 35 adolescents (67.2%) with bacterial vaginosis. CONCLUSIONS: Not only IL-1 beta and sialidase levels, but also IL-6 and IL-8 levels are higher in pregnant adolescents with bacterial vaginosis, thus indicating that this condition elicits a more pronounced inflammatory response in this population, which potentially increases vulnerability to STI acquisition.
Acesso aberto (Open Access)
Caracterização da resposta imune citocínica na infecção humana pelo vírus oropouche e sua relação com o padrão de soroconversão e a presença de sintomas
(Universidade Federal do Pará, 2011-12-19) OLIVEIRA, Euzébio de; VASCONCELOS, Pedro Fernando da Costa; http://lattes.cnpq.br/0973550817356564
This thesis is the first global study that researches and analyzes the immune response of cytokine in human infections by Orthobunyavirus Oropuche virus. The study used 320 samples of human serum. Sixty were from the Blood Bank (negative control) and 260 were obtained from two outbreaks of the Oropouche virus in the State of Pará and Amapá (Brazil). The latter was divided into 8 subgroups for better data accuracy. The collected samples were analyzed for clinical data/symptoms with serologic testing by titration of antibodies by the hemagglutination inhibition (IgM/IgG) and the detection cytokines plasma levels by flow cytometry. This allowed for the technical description of cytokine. The data obtained allowed for the observation of the characteristics and the behavior of the cytokines signatures expressed by patients by the presence or not of the virus. This also allowed for the observation of changes to serum through the presence of specific symptoms such as fever, chills, headache and dizziness. This led to the following conclusions a) there is a pattern in the synthesis of pro-inflammatory and regulatory cytokines; b) there is a balance in the profile of the immune response between pro-inflammatory cytokines (Th1) and modulators (Th17); c) an infection by the Oropouche virus alters the production of cytokines in individuals; d) the results also show that whem comparing individuals no responders with early responders, there was an increase of IL-1β and decreased IL- 12; no responders with late responders, there was a decrease of IL-8, and increased IFN-α, IL-23 and IL-17; No responders occurred early responders compared with the increase IL-4 and IFN-g; However, when compared early responders and late responders, decreased IFN-α and IL-6; early responders generally showed a decrease in IL-10 and late responders showed an increase in IL-5; e) The results also show the expression of IL-5 in patients who showed symptoms specific for Oropouche infection (fever, chills, headache and dizziness), suggesting this signal to be directly associated with pathogenesis of the virus; f) there is a need to complement this research with more studies such as those related to the expression of chemokines.
Acesso aberto (Open Access)
Comprometimento pulmonar na malária: associação com fatores epidemiológicos, imunológicos e variantes do Plasmodium vivax
(Universidade Federal do Pará, 2014-11-20) OHNISHI, Maria Deise de Oliveira; LIBONATI, Rosana Maria Feio; http://lattes.cnpq.br/3818175484709618
Malaria is a disease caused by protozoan parasites of the genus Plasmodium and transmitted to humans by mosquitoes of the genus Anophelis. In 2011, the Brazilian Amazon reported 266.348 cases. In Brazil, the most prevalent species is Plasmodium vivax (80%). It´s considered responsible for benign forms of malaria, reports have shown severe forms of pulmonary malaria and deaths. The pathophysiological mechanisms are not well understood.Thus, we analyzed 247 patients with vivax malaria recruited from the Clinical Trials Program in Malaria Institute Evandro Chagas (IEC) Belém / PA) and malaria diagnosis service of the Secretariat of Health of the city of Goianésia / Pará between April / 2011 to October / 2013 to evaluate clinical, epidemiological, parasitological, radiological, genetic and immune (variants of P. vivax) parameters, associating them to the pulmonary involvement in vivax malaria. The project was approved by the Research Ethics Committee of the Instituto Evandro Chagas.Hematological parameters (blood count, transaminases, bilirubin), parasite density were analyzed as were measured at D0 and D14 serum levels of cytokines (TNF-, IL-10 and INFγ), relating to the pulmonary manifestations. D0 was determined prevalence of P. vivax variants (VK210 and VK247 and P. vivax-like) and established association of these variants with patients who had an abnormal chest X-ray and / or spirometry. The study was longitudinal, prospective analytical type involving clinical, immunological, hematological, and radiological aspects of respiratory function. Patients with pulmonary involvement confirmed by clinical and / or radiological and / or functional findings constituted the study group, and patients without such findings, the control group. Participants: those with vivax malaria diagnosed by thick blood, confirmed by PCR, 15-60 years old, both genders, with no chronic diseases, no use of corticosteroids at diagnosis and who signed the consent form. Project approved by the CEP of the IEC. Observed: 69.2% were males; mean age 35 years old; 64.4% lived in Belém; 79.8% contracted malaria in the state of Pará; First infection 42.5%; mean duration of disease 7.5 ± 5.9 days; average of 6979.25 ± 8692.22 parasitaemia parasites / mm3; 46.8% were overweight or obese I; smoking and previous lung disease were not risk factors for worsening of the disease in this sample; 92.3% had headache from malaria triad; among respiratory symptoms, cough was the most frequent (53.4%); hepatomegaly and splenomegaly was more frequent among naïve malaria; Thrombocytopenia was significant in D0; radiological abnormalities in 9.7% of cases; changes in X-ray were related to malaria shortness of breath and cough had statistical significance in those with changes in the X-ray and / or spirometry; IL-10 INFγ and cytokines were significantly higher compared D0 to D14, unlike that serum TNF levels were low in D0. VK210 P. vivax variant was more frequent (69.1%); there was no significant correlation in the appearance of changes in RX in this sample and / or spirometry related to any of the P. vivax subspecies. Among the cytokines, INFγ was significantly elevated in D0 associated to variants in the following situations: (VK210, VK247, VK210 / 247). IL-10 also significantly increased in D0 only when the subspecies was VK210. Lung malaria is a reality still underestimated, especially less prominent pulmonary manifestations, being necessary to investigate potential mechanisms involved linked to the host and the parasite, emphasizing the importance of early diagnosis, avoiding unfavorable outcome.
Acesso aberto (Open Access)
Detecção in situ de citocinas de perfil TH1 e TH2 por imunoistoquímica e relação com a atividade de macrófagos nas formas polares da doença de Hansen
(Universidade Federal do Pará, 2009) SILVA, Dayse Danielle de Oliveira; QUARESMA, Juarez Antônio Simões; http://lattes.cnpq.br/3350166863853054
Leprosy is an infectious disease that has a slow and progressive evolution, with high incidence and prevalence in the states that compose the Legal Amazon. Considering the number of cases, the Pará state plays a prominent role in the national context. In order to evaluate the cytokine profile and the macrophages activity in the polar forms of leprosy and correlate with their histopathological aspects, a transversal study was performed, with a sample of 29 patients who had one of the polar forms of leprosy, namely tuberculoid or virchowian, from who skin biopsies of leprosy lesions were taken. It could be observed, through the analysis of this immunohistochemical material, that the studied groups showed no significant variation in the levels of IL-10, TGF-β, CD68, iNOS and Lysozyme. The levels of IL-4 were not expressive in the analyzed tissues and the levels of IFN-γ were higher in patients in the MHV group. In the virchowian pole, all cytokines tended towards a negative correlation with the macrophages activity. In the tuberculoid pole, the level of IL-10 showed a negative correlation with the levels of iNOS, IFN-γ and lysozyme, indicating a macrophage activity inhibition for this cytokine. It can be concluded that there is no difference in cytokine expression of TH1 and TH2 profile in polar forms of Hansen's disease and cytokines have a tendency to negative correlation with the activity of macrophages in MHV pole and a tendency to positive correlation in the pole MHT. Further research is needed to clarify why some populations have more susceptibility to leprosy than others. A similar study with a larger number of individuals may have a more significant outcome.
Acesso aberto (Open Access)
Estudo da neuropatologia induzida pelo vírus Marabá em modelo murino
(Universidade Federal do Pará, 2014-02-28) FARIAS, Alexandre Maia de; DINIZ, Cristovam Wanderley Picanço; http://lattes.cnpq.br/2014918752636286; DINIZ JUNIOR, José Antônio Picanço; http://lattes.cnpq.br/3850460442622655
The Marabá virus (Be AR 411459) is an Vesiculovirus (VSV), member of the Rhabdoviridae family, isolated in 1983 from a pool of sandflies captured in Marabá - PA by Section Arbovirus the Evandro Chagas Institute. In literature There are few studies on experimental neuropathology Marabá virus induced, despite 30 years of isolation. A single study, however, revealed that viral infection in newborn mice causes necrosis and pyknosis in neurons in several regions of the central nervous system (CNS). The objective of this study was to investigate the distribution of Marabá virus in the CNS, reactive microglia and astrocytes, histopathologic features, and the expression of cytokines and nitric oxide (NO), in Marabá virus-induced encephalitis in BALB / c mice. Thus was performed processing of samples for histopathological examination; Immunohistochemistry for observation of microglia, astrocytes and viral antigen; tests for quantification of cytokines and NO, and statistical analyzes. The results showed that infected animals (Ai) 3 days after inoculation (d.p.i.) with discrete labeling of viral antigen, as well as the activation of microglia and astrocytes in the CNS. Moreover , in Ai 6 d.p.i. marking the viral antigen was observed in almost all brain regions, with intense microglial activation in these locations, although was less astrogliosis. Edema, necrosis and apoptosis of neurons were mainly observed in the olfactory bulb, septum and frontal cortex of Ai 6 d.p.i. Quantification of IL - 12p40, IL- 10, IL- 6, TNF- α, INF- ү, MCP-1 and NO, showed significant increases in Ai 6 d.p.i., when compared with controls animals and Ai 3 d.p.i. On the other hand, TGF-β, important immunosuppressant, has not been significant in all groups and evaluated times (3 and 6 d.p.i.). These results indicate that Marabá virus can infect various CNS regions of BALB /c mouse adult 6 d.p.i., producing anatomical and pathological changes and strong inflammatory immune response that can be lethal to the animal.
Acesso aberto (Open Access)
Estudo morfológico e imunológico da encefalite induzida pelo vírus juruaçá em modelo murino
(Universidade Federal do Pará, 2013-10-08) FERREIRA, Natalie Chaves; DINIZ JUNIOR, José Antônio Picanço; http://lattes.cnpq.br/1897655177251738
Many studies have been conducted to understand the neuropathogenesis of viral encephalitis from experimental work, however, no experimental studies have been devoted to understanding the neuropathogenesis of members of the Picornaviridae family isolated from bats in the Amazon region. The Juruaçá virus, one of these agents, partially characterized as a member of the Picornaviridae family by Araujo et al. (2006), caused lesions in the brain of neonatal mice with reactive gliosis presence, although not cause cytopathic effect (CPE) in primary cultures of central nervous system (CNS) cells, suggesting that this viral agent is responsible for the death of animals due to an intense immune response. The aim of this study was to investigate the immune response in the CNS and cellular changes caused by Juruaçá virus in newborn albino mice of strain BALB/c from histopathological analysis, microglial activation, and expression of cytokines, nitric oxide (NO) and reactive oxygen species (ROS). Thus, we performed sample processing for histopathology, immunosorbent assay, immunohistochemical and immunofluorescence assays, tests to quantify NO and superoxide radicals, and statistical analysis. Our results demonstrated that the Juruaçá virus induces lesions throughout the brain, with greater intensity in the cortical parenchyma. Immunohistochemical tests showed the presence of viral antigens and reactive microglias distributed throughout the brain and anterior spinal cord. Microglias with amoeboid shape, demonstrating intense activation, were observed in the cerebral cortex, olfactory bulb, anterior olfactory nucleus, midbrain and forebrain near the lateral ventricle. The production of anti-inflammatory cytokines (IL-10 and IL-4) decreased over time, whereas pro-inflammatory cytokines (IL -12, IL- 6, IL- 1β, TNF-α and IFN-γ) increased significantly from the 8th day. Assays for ROS detection showed great superoxide radicals production from the 4th day, as NO production was always lower in the infected animals. Probably, activation of glial cells, especially microglias, and subsequent production of proinflammatory cytokines and ROS promoted a devastating action on the cells of the CNS, which coincides with the intensification of clinical signs. In accordance with what has been explained above, became evident that our results indicate that the Juruaçá virus is responsible for a imprint inflammatory disease that leads to death 100% of infected neonates mice.
Acesso aberto (Open Access)
IL-2, IL-5, TNF-α and IFN-γ mRNA expression in epidermal keratinocytes of systemic lupus erythematosus skin lesions
(2011) CARNEIRO, José Ronaldo Matos; FUZII, Hellen Thais; KAYSER, Cristiane; ALBERTO, Fernando Lopes; SOARES, Fernando Augusto; SATO, Emília Inoue; ANDRADE, Luis Eduardo Coelho
OBJECTIVE: To analyze cytokine gene expression in keratinocytes from patients with systemic lupus erythematosus (SLE). INTRODUCTION: Keratinocytes represent 95% of epidermal cells and can secrete several cytokines. METHODS: Keratinocytes were obtained by laser microdissection from 21 patients with SLE (10 discoid and 11 acute lesions) at involved and uninvolved sites. All patients were receiving a low/moderate prednisone dose and 18 were receiving chloroquine diphosphate. IL-2, IL-5, TNF-α and IFN-γ gene expression was evaluated by real-time PCR and expressed as the ratio (R) to a pool of skin samples from 12 healthy volunteers. RESULTS: Heterogeneity in cytokine gene expression was found among patients with SLE. Eighteen of 38 valid SLE samples (47%) presented overexpression (R>1) of at least one cytokine. Lesional skin samples tended to show higher cytokine expression than samples from uninvolved skin (p = 0.06). IL-5 and IFN-γ were the most commonly overexpressed cytokines. Samples with cytokine overexpression corresponded to more extensive and severe lesions. Prednisone dose did not differ between samples without cytokine overexpression (15.71±3.45 mg/day) and those with overexpressed cytokines (12.68±5.41 mg/day) (p = 0.216). Samples from all patients not receiving diphosphate chloroquine had at least one overexpressed cytokine. CONCLUSIONS: The heterogeneous keratinocyte cytokine gene expression reflects the complex immunological and inflammatory background in SLE. Patients with severe/extensive skin lesions showed a higher frequency of cytokine gene overexpression. Increased IFN-γ and IL-5 expression suggests that Th1 and Th2 cells are involved in SLE skin inflammation. The possibility that prednisone and antimalarial drugs may have contributed to low cytokine gene expression in some samples cannot be ruled out.
Acesso aberto (Open Access)
Implicações do perfil citocínico TH22 nas formas polares da hanseníase
(Universidade Federal do Pará, 2016-08-29) SILVEIRA, Edvaldo Lima; QUARESMA, Juarez Antônio Simões; http://lattes.cnpq.br/3350166863853054
BACKGROUND: Leprosy is a chronic granulomatous disease caused by Mycobacterium leprae. Among the immunopathological aspects of leprosy is known that the defense is done by the cellular immune response, able to phagocyte and destroy the bacilli, mediated by cytokines and mediators from oxidation. The long-standing concept of a Th1-Th2 dichotomy in leprosy, with predominant Th1 in tuberculoid lesions and Th2 predominant in virchowian pacients, has recently been challenged. Furthermore, the Th22 response was identified as modulating Th1-Th2 in inflammatory skin diseases, but their roles in leprosy have not yet been elucidated. OBJECTIVE: Evaluate the tissue expression of cytokines involved in Th22 response in the polar forms of leprosy. METHOD: Patients with dermato-immunological diagnosis of leprosy were included and selected 31 patients, 16 with the tuberculoid (TT) form and 15 with lepromatous (LL). Immunohistochemistry for tissue immunostaining with antibodies against IL-13, IL-22, TNF-α and FGF-b was based on the method involving the formation of biotin-streptavidin peroxidase complex. Quantitation of the immunostaining was taken randomly from 05 fields viewed at 400x magnification microscope. In univariate analysis, frequencies, measures of central tendency and dispersion were obtained and for investigation of the hypothesis were applied the Mann-Whitney test and the Pearson correlation, considering a significance level of 5% (p ≤ 0.05). RESULTS: Regarding the immunostaining for IL-22 can be observed statistical difference among the groups and in the LL pole the average was 241.3 ± 44.63 cells/mm2, while in the TT form the mean was 90.39 ± 30.18 cells/mm2 (p <0.0001). Engaging the presence of IL-13, LL pole average occurrence was 85.76 ± 19.99 cells/mm2. In the TT pole the mean was 57.20 ± 14.73 cells/mm2 (p = 0.0002). Regarding the immunostaining for FGF-b, in the LL lesions, the mean incidence was 228.9 ± 45.13 cells/mm2, while in the TT form the mean was 47.80 ± 14.29 cells/mm2 (p <0,0001). For TNF-α, quantitative analysis was statistically significant in TT form where the average of the cells expressing the cytokine was 99.74 ± 30.14 cells/mm2 when compared to the LL form where the results were 62.08 ± 13.67 cells/ mm2 (p = 0.0008). CONCLUSION: The Th-22 response, mediated by IL-22, has fundamental importance in the pathogenesis of leprosy, relating directly to the clinical form of the disease and other cytokines.
Acesso aberto (Open Access)
Imunopatogenia da interação entre acrófagos e/ou células de Langerhans e Lacazia loboi
(Universidade Federal do Pará, 2014-04-24) YAMANO, Suellen Sirleide Pereira; SALGADO, Claudio Guedes; http://lattes.cnpq.br/2310734509396125
Lobomycosis is a chronic, granulomatous, fungal infection of the skin and subcutaneous caused by the fungus Lacazia loboi. This mycosis occurs predominantly in Amazon region and affect all populations groups. Histologically, there is a chronic inflammatory reaction characterized by the intense fibroplastia and histiocytosis, abundant number of macrophages, multinucleated giant cells and presence of large numbers of yeast. Macrophages are phagocytic cells that participate in the recognition and response to pathogens through phagocytosis, antigen presentation to T lymphocytes and cytokine production. Langerhans cells (LC) are a group of dendritic cells (DC) derived from bone marrow, which are located mainly in the suprabasal layer of the epidermis. Studies on hostfungal interaction in Jorge Lobo's disease are scarce. Therefore, this study constitutes an important step towards a better understanding of L. loboi biology and pathogenesis, enabling in vitro culture of this organism endemic in Amazon region and study of their antigenic properties, as well as new therapeutic approaches. The objective of this study was to analyze the in vitro interaction between non-activated peritoneal macrophages and/or Langerhans cells isolated from BALB/c mice and L. loboi, calculating infection, phagocytosis and cellular fusion rates; also measuring the production of TNF, IL-4, IL-6, IL-10 and IL-12. Results showed that L. loboi is phagocytosed by macrophages but not by LC. Phagocytosis and infection rates in the interaction between macrophages and L. loboi was similar to interaction between macrophages, Langerhans cells and L. loboi at all times analyzed. The mean number of yeast cell per macrophage was almost equal among interactions and over time, with mean variation from 1.2 to 1.6 yeast/macrophages. No giant cells formation in coculture studied. No significant difference occurred in IL-4, Il-2 and IL-10 production between interactions studied. TNF levels decrease in interaction between macrophages and L. loboi while adding LC induces increased production of TNF-α, especially after 48 hours. LC negatively modulate IL-6 production by macrophage and L. loboi also inhibits this production by macrophages alone or cocultured with LC. L. loboi stimulated significantly IL-12 production by macrophages co-cultured with LC, but not for LC alone or macrophages.
Acesso aberto (Open Access)
Investigação de polimorfismos no gene TNF em pacientes com hepatotoxicidade induzida por medicações antituberculosas no norte do Brasil
(Universidade Federal do Pará, 2015-08-27) VALENTE, Sonia Lopes; SANTOS, Ney Pereira Carneiro dos; http://lattes.cnpq.br/1290427033107137; SORTICA, Vinicius de Albuquerque; http://lattes.cnpq.br/2046482071071824
Tuberculosis still remains a serious public health problem worldwide. The hepatotoxicity induced by anti-tuberculosis drugs causes a large number of hospitalizations and may be fatal if treatment is not interrupted. The hepatitis induced by anti-tuberculosis drugs are not yet fully understood and clinical studies suggests that immunological mechanisms are involved in its pathogenesis. The cytokine TNF-α is a major mediator of inflammatory and immune changes in the levels of this cytokine may be related to pathogenesis of drug-induced hepatitis. These changes observed may be related to polymorphisms in the TNF gene. The knowledge of which polymorphisms in the TNF gene are involved in the risk of developing hepatotoxicity anti-tuberculosis drugs will permit the use of these molecular markers to improve the therapeutic management of these patients. This study investigated the influence of polymorphisms -308C>T (rs1800629), -1031C>T (rs1799964), -238A>G (rs361525) and -857C>T (rs1799724) in the TNF gene with drug-induced hepatotoxicity. The study included 68 patients with tuberculosis who had hepatotoxicity of the basic regimen consisting of rifampicin, isoniazid, pyrazinamide and ethambutol (2RHZE/4R) and 191 patients without adverse therapy effects. The polymorphisms were determined by real-time PCR with TaqMan probes. Comparing the frequency of genotypes between cases and controls, a significant difference in the distribution of genotypes of the SNP -1031C>T was identified (p = 0.003). The frequency of homozygous -1031CC was higher in the case group (8.8%) than in the control group (1.6%). The -1031CC homozygous patients had an increased risk for the development of hepatotoxicity when compared to homozygous -1031TT or the T allele carriers (OR = 8.632, p = 0.014, OR = 11.355, p = 0.004). We concluded that -1031C>T SNP was significantly associated with susceptibility to induced hepatitis anti-tuberculosis drugs in the north population of Brazil.
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)
Polimorfismos de citocinas (TNF-A, IL-10 e IL-17) no câncer gástrico
(Universidade Federal do Pará, 2016-02-02) OLIVEIRA, Gabriela Almeida de; SANTOS, Ândrea Kely Campos Ribeiro dos; http://lattes.cnpq.br/3899534338451625
Gastric cancer (GC) is the second most common malignancy among men and the third in women, and therefore, an important public health problem in northern Brazil. The investigation of genetic factors related to immunological characteristics can aid the understanding of carcinogenesis in CG. The objective of the present work was investigate polymorphisms present in interleukin genes IL17G-197A, IL 17FA7488G, TNFαG-308A, IL10G-1082A, IL10C-819T e IL10C-592A, on samples of patients with gastric cancer and healthy patients without cancer. Case group was composed of 100 patients diagnosed with CG, met in the Hospital HUJBB (Pará, Brazil). Control group was composed of 100 individuals without cancer, unrelated, of the same population. The genetic material was extracted from 5 mL of peripheral blood with the DNA commercial kit from Roche, followed by quantification with the NanoDrop 1000 spectrophotometer. Analysis of the molecular polymorphisms was performed by real-time PCR with Taqman® probes. Measures of ancestry were investigated using a panel of 48 autosomal ancestry informative markers (AIMs). The proportions of ancestry of European, African and Amerindian were estimated using the software STRUCTURE v. 2.3.3. It was observed that the ethnic composition of the case group was 27% African, 42% European and 31% of Amerindian, while in the control group 21% African, 52% of European and 27% of Amerindian. In relation to the set of markers of interleukin IL-10 (IL10G-1082A, IL10C-819T, IL10C-592A), when the genotypic and haplotypic patterns were compared, it was noted that the haplotype distribution related to high expression (GCC/GCC, GCA, GCC/GCC/GTC, GCA/GCA, GCA/GTA) was more frequent in the patients with gastric cancer (P = 1,15e-11; OR = 2.630; IC 95% = 2.116-3.271). Among the individuals with the genotype related to the high production of IL-10, it was observed that the control group had more European contribution in their ancestry (P = 1e-06) while the group of patients with CG had more African contribution in their ancestry (P = 1.4e-5). Patients who presented TNF-α AA and TNF-α AG genotypes for TNF-α gene mutation presented a higher risk for development of cancer (P<0.001; OR 10.375; IC 95% 3.149-34.061). It is concluded that patients with a distribution of haplotypic markers of interleukin IL-10 (IL10G-1082A, IL10C-819T, IL10C-592A) related to a higher expression and higher contribution of African ancestry have a high risk of developing gastric cancer.
Acesso aberto (Open Access)
Replicação viral, padrão de expressão de citocinas e cinética de morte celular em cultura primária de células neurais infectadas pelo vírus rocio
(Universidade Federal do Pará, 2015-07-13) SOUTO, Adriano da Paixão; FRANCO, Edna Cristina Santos; http://lattes.cnpq.br/5939607544965550
Arboviruses belonging to the genus Flavivirus (Flaviviridae family) are responsible for considerable morbidity and mortality, may cause severe cases of encephalitis, hemorrhagic fever, hepatitis and febrile disease in vertebrates, including humans. Rocio flavivirus (ROCV) is very important for public health in Brazil, as it represents a serious threat of occurrence of sudden encephalitis outbreaks. The immunopathology of encephalitis caused by flavivirus is not yet fully understood and many aspects involved in modulation of immune responses in the CNS need to be unraveled. Thus, the aim of this study was to analyze the pattern of gene expression of cytokines in primary culture of neural cells when exposed to infection with Rocio. Mixed primary cultures (neuron/glia), obtained from the brains of BALB/C lineage isogenic newborns mice were inoculated with ROCV (MOI equal to 2) on the seventh day of culture and confirmation of infection was made by immunocytochemistry (anti-VROC). Viral replication was quantified in infected primary cultures, depending on the infectious period, by viral titer method. Total cellular RNA was extracted and used for detection of cytokines by real time RT-qPCR techniques. The study showed that primary cultures of neural cells is a good experimental model for the study of CNS infections by Rocio flavivirus. ROCV efficiently infected the primary cultured neural cells leading to cytopathic changes from the 2nd day a.i., when it detected the higher viral titer. The ROCV-infected primary culture survived up to 7 days a.i. and quantification of viral load showed a viral replication kinetics compatible with cell death kinetics of culture infected by ROCV. During the first five days a.i. of the primary culture, there was reduction of IFN-α gene expression, IFN-β, IFN-γ and IL-1β, there was no change in TNF-α expression and there was an increase in TGF-β expression in the 1st, 3rd and 5th day a.i.. The findings of this study suggest that ROCV has the ability to downregulate modulators cytokines of antiviral and inflammatory immune responses and, in this experimental model, immunoregulation exerted by TGF-β predominates over the modulation of inflammatory and antiviral immune responses, larger studies are needed to elucidate immunopathology of CNS infection by ROCV.
Acesso aberto (Open Access)
Severe visceral leishmaniasis in children: the relationship between cytokine patterns and clinical features
(Universidade Federal do Pará, 2013-12) GAMA, Mônica Elinor Alves; GOMES, Claudia Maria de Castro; SILVEIRA, Fernando Tobias; AURENTI, Márcia Dalastra; GONÇALVES, Eloisa da Graça do Rosario; SILVA, Antônio Rafael da; CORBETT, Carlos Eduardo Pereira
Introduction: The relationship between severe clinical manifestations of visceral leishmaniasis (VL) and immune response profi les has not yet been clarifi ed, despite numerous studies on the subject. This study aimed to investigate the relationship between cytokine profi les and the presence of immunological markers associated with clinical manifestations and, particularly, signs of severity, as defi ned in a protocol drafted by the Ministry of Health (Brazil). Methods: We conducted a prospective, descriptive study between May 2008 and December 2009. This study was based on an assessment of all pediatric patients with VL who were observed in a reference hospital in Maranhão. Results: Among 27 children, 55.5% presented with more than one sign of severity or warning sign. Patients without signs of severity or warning signs and patients with only one warning sign had the highest interferon-gamma (IFN-γ) levels, although their interleukin 10 (IL-10) levels were also elevated. In contrast, patients with the features of severe disease had the lowest IFN-γ levels. Three patients who presented with more than two signs of severe disease died; these patients had undetectable interleukin 2 (IL-2) and IFN-γ levels and low IL-10 levels, which varied between 0 and 36.8pg/mL. Conclusions: Our results showed that disease severity was associated with low IFN-γ levels and elevated IL-10 levels. However, further studies with larger samples are needed to better characterize the relationship between disease severity and cytokine levels, with the aim of identifying immunological markers of active-disease severity.