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1.
J Immunol ; 197(5): 1905-13, 2016 09 01.
Article in English | MEDLINE | ID: mdl-27474073

ABSTRACT

The chronic course of lepromatous leprosy may be interrupted by acute inflammatory episodes known as erythema nodosum leprosum (ENL). Despite its being a major cause of peripheral nerve damage in leprosy patients, the immunopathogenesis of ENL remains ill-defined. Recognized by distinct families of germline-encoded pattern recognition receptors, endogenous and pathogen-derived nucleic acids are highly immunostimulatory molecules that play a major role in the host defense against infections, autoimmunity, and autoinflammation. The aim of this work was to investigate whether DNA sensing via TLR-9 constitutes a major inflammatory pathway during ENL. Flow cytometry and immunohistochemistry analysis showed significantly higher TLR-9 expression in ENL when compared with nonreactional lepromatous patients, both locally in the skin lesions and in circulating mononuclear cells. The levels of endogenous and pathogen-derived TLR-9 ligands in the circulation of ENL patients were also higher. Furthermore, PBMCs isolated from the ENL patients secreted higher levels of TNF, IL-6, and IL-1ß in response to a TLR-9 agonist than those of the nonreactional patients and healthy individuals. Finally, E6446, a TLR-9 synthetic antagonist, was able to significantly inhibit the secretion of proinflammatory cytokines by ENL PBMCs in response to Mycobacterium leprae lysate. Our data strongly indicate that DNA sensing via TLR-9 constitutes a major innate immunity pathway involved in the pathogenesis and evolution of ENL. Thus, the use of TLR-9 antagonists emerges as a potential alternative to more effectively treat ENL aiming to prevent the development of nerve injuries and deformities in leprosy.


Subject(s)
DNA/metabolism , Erythema Nodosum/immunology , Immunity, Innate , Leprosy, Lepromatous/immunology , Signal Transduction , Toll-Like Receptor 9/metabolism , Adult , Aged , Aged, 80 and over , Erythema Nodosum/microbiology , Female , Flow Cytometry , Humans , Leprosy, Lepromatous/microbiology , Leukocytes, Mononuclear/immunology , Leukocytes, Mononuclear/microbiology , Male , Middle Aged , Mycobacterium leprae/chemistry , Mycobacterium leprae/immunology , Toll-Like Receptor 9/immunology , Young Adult
2.
Eur J Immunol ; 42(11): 2925-36, 2012 Nov.
Article in English | MEDLINE | ID: mdl-22851198

ABSTRACT

Lepromatous macrophages possess a regulatory phenotype that contributes to the immunosuppression observed in leprosy. CD163, a scavenger receptor that recognizes hemoglobin-haptoglobin complexes, is expressed at higher levels in lepromatous cells, although its functional role in leprosy is not yet established. We herein demonstrate that human lepromatous lesions are microenvironments rich in IDO⁺CD163⁺. Cells isolated from these lesions were CD68⁺IDO⁺CD163⁺ while higher levels of sCD163 in lepromatous sera positively correlated with IL-10 levels and IDO activity. Different Myco-bacterium leprae (ML) concentrations in healthy monocytes likewise revealed a positive correlation between increased concentrations of the mycobacteria and IDO, CD209, and CD163 expression. The regulatory phenotype in ML-stimulated monocytes was accompanied by increased TNF, IL-10, and TGF-ß levels whereas IL-10 blockade reduced ML-induced CD163 expression. The CD163 blockade reduced ML uptake in human monocytes. ML uptake was higher in HEK293 cells transfected with the cDNA for CD163 than in untransfected cells. Simultaneously, increased CD163 expression in lepromatous cells seemed to be dependent on ML uptake, and contributed to augmented iron storage in lepromatous macrophages. Altogether, these results suggest that ML-induced CD163 expression modulates the host cell phenotype to create a favorable environment for myco-bacterial entry and survival.


Subject(s)
Antigens, CD/immunology , Antigens, Differentiation, Myelomonocytic/immunology , Leprosy, Lepromatous/immunology , Leprosy, Lepromatous/microbiology , Macrophages/immunology , Mycobacterium leprae/immunology , Receptors, Cell Surface/immunology , Antigens, CD/genetics , Antigens, Differentiation, Myelomonocytic/genetics , Biopsy , Flow Cytometry , Humans , Indoleamine-Pyrrole 2,3,-Dioxygenase/immunology , Interleukin-10/immunology , Leprosy, Lepromatous/pathology , Macrophages/microbiology , RNA, Messenger/chemistry , RNA, Messenger/genetics , Receptors, Cell Surface/genetics , Reverse Transcriptase Polymerase Chain Reaction
3.
J Immunol ; 187(5): 2548-58, 2011 Sep 01.
Article in English | MEDLINE | ID: mdl-21813774

ABSTRACT

The mechanisms responsible for nerve injury in leprosy need further elucidation. We recently demonstrated that the foamy phenotype of Mycobacterium leprae-infected Schwann cells (SCs) observed in nerves of multibacillary patients results from the capacity of M. leprae to induce and recruit lipid droplets (LDs; also known as lipid bodies) to bacterial-containing phagosomes. In this study, we analyzed the parameters that govern LD biogenesis by M. leprae in SCs and how this contributes to the innate immune response elicited by M. leprae. Our observations indicated that LD formation requires the uptake of live bacteria and depends on host cell cytoskeleton rearrangement and vesicular trafficking. TLR6 deletion, but not TLR2, completely abolished the induction of LDs by M. leprae, as well as inhibited the bacterial uptake in SCs. M. leprae-induced LD biogenesis correlated with increased PGE(2) and IL-10 secretion, as well as reduced IL-12 and NO production in M. leprae-infected SCs. Analysis of nerves from lepromatous leprosy patients showed colocalization of M. leprae, LDs, and cyclooxygenase-2 in SCs, indicating that LDs are sites for PGE(2) synthesis in vivo. LD biogenesis Inhibition by the fatty acid synthase inhibitor C-75 abolished the effect of M. leprae on SC production of immunoinflammatory mediators and enhanced the mycobacterial-killing ability of SCs. Altogether, our data indicated a critical role for TLR6-dependent signaling in M. leprae-SC interactions, favoring phagocytosis and subsequent signaling for induction of LD biogenesis in infected cells. Moreover, our observations reinforced the role of LDs favoring mycobacterial survival and persistence in the nerve. These findings give further support to a critical role for LDs in M. leprae pathogenesis in the nerve.


Subject(s)
Leprosy/pathology , Schwann Cells/microbiology , Schwann Cells/pathology , Toll-Like Receptor 6/immunology , Animals , Humans , Immunohistochemistry , Inclusion Bodies/immunology , Inclusion Bodies/metabolism , Inclusion Bodies/pathology , Inflammation/immunology , Inflammation/microbiology , Inflammation/pathology , Leprosy/immunology , Lipid Metabolism/physiology , Lipids/immunology , Mice , Mice, Inbred C57BL , Microscopy, Confocal , Mycobacterium leprae/immunology , Schwann Cells/immunology , Toll-Like Receptor 6/metabolism
4.
Neurobiol Aging ; 113: 108-117, 2022 05.
Article in English | MEDLINE | ID: mdl-35325812

ABSTRACT

Aged and photoaged skin exhibit fine wrinkles that are signs of epidermal inflammation and degeneration. It has been shown that healthy elderly skin expresses amyloidogenic proteins, including α-Synuclein, which are known to oligomerize and trigger inflammation and neurodegeneration. However, little is known about their putative role in skin physiology and sensitivity. To unravel this possible role, we investigated the impact of oligomeric α-Synuclein (Oα-Syn) in 2D and 3D keratinocyte human models. Exogenous Oα-Syn caused degeneration of reconstructed human epidermis (RHE) by diminishing proliferation and thickness of the stratum basale. Oα-Syn also increased NF-kB nuclear translocation in keratinocytes and triggered inflammation in the RHE, by increasing expression of interleukin-1ß and tumor necrosis factor-alpha, and the release of tumor necrosis factor-alpha in a time-dependent manner. Dexamethasone and an IL-1ß inhibitor partially diminished RHE degeneration caused by Oα-Syn. These findings suggest that Oα-Syn induces epidermal inflammation and decreases keratinocyte proliferation, and therefore might contribute to epidermal degeneration observed in human skin aging.


Subject(s)
Tumor Necrosis Factor-alpha , alpha-Synuclein , Aged , Epidermis/metabolism , Epidermis/pathology , Humans , Inflammation/metabolism , Keratinocytes/metabolism , Tumor Necrosis Factor-alpha/metabolism , alpha-Synuclein/metabolism
5.
PLoS One ; 16(11): e0259804, 2021.
Article in English | MEDLINE | ID: mdl-34797866

ABSTRACT

The diagnosis of pure neural leprosy is more challenging because patients share characteristics with other common pathologies, such as ulnar compression, which should be taken into consideration for differential diagnosis. In this study, we identify ulnar nerve conduction characteristics to aid in the differential diagnosis of ulnar neuropathy (UN) in leprosy and that of non-leprosy etiology. In addition, we include putative markers to better understand the inflammatory process that may occur in the nerve. Data were extracted from a database of people affected by leprosy (leprosy group) diagnosed with UN at leprosy diagnosis. A non-leprosy group of patients diagnosed with mechanical neuropathy (compressive, traumatic) was also included. Both groups were submitted to clinical, neurological, neurophysiological and immunological studies. Nerve enlargement and sensory impairment were significantly higher in leprosy patients than in patients with compressive UN. Bilateral impairment was significantly higher in the leprosy group than in the non-leprosy group. Leprosy reactions were associated to focal demyelinating lesions at the elbow and to temporal dispersion (TD). Clinical signs such as sensory impairment, nerve enlargement and bilateral ulnar nerve injury associated with eletrodiagnostic criteria such as demyelinating finds, specifically temporal dispersion, could be tools to help us decided on the best conduct in patients with elbow ulnar neuropathy and specifically decide if we should perform a nerve biopsy for diagnosis of pure neural leprosy.


Subject(s)
Leprosy/diagnosis , Leprosy/metabolism , Ulnar Neuropathies/diagnosis , Adolescent , Adult , Aged , Biomarkers , Brazil/epidemiology , Cross-Sectional Studies , Data Management , Databases, Factual , Diagnosis, Differential , Elbow Joint , Female , Humans , Leprosy, Tuberculoid , Male , Middle Aged , Neural Conduction , Ulnar Nerve/metabolism , Ulnar Neuropathies/physiopathology
6.
J Leukoc Biol ; 110(1): 167-176, 2021 07.
Article in English | MEDLINE | ID: mdl-33040382

ABSTRACT

The enzyme IDO-1 is involved in the first stage of tryptophan catabolism and has been described in both microbicidal and tolerogenic microenvironments. Previous data from our group have shown that IDO-1 is differentially regulated in the distinctive clinical forms of leprosy. The present study aims to investigate the mechanisms associated with IDO-1 expression and activity in human monocyte-derived dendritic cells (mDCs) after stimulation with irradiated Mycobacterium leprae and its fractions. M. leprae and its fractions induced the expression and activity of IDO-1 in human mDCs. Among the stimuli studied, irradiated M. leprae and its membrane fraction (MLMA) induced the production of proinflammatory cytokines TNF and IL-6 whereas irradiated M. leprae and its cytosol fraction (MLSA) induced an increase in IL-10. We investigated if TLR2 activation was necessary for IDO-1 induction in mDCs. We observed that in cultures treated with a neutralizing anti-TLR2 antibody, there was a decrease in IDO-1 activity and expression induced by M. leprae and MLMA. The same effect was observed when we used a MyD88 inhibitor. Our data demonstrate that coculture of mDCs with autologous lymphocytes induced an increase in regulatory T (Treg) cell frequency in MLSA-stimulated cultures, showing that M. leprae constituents may play opposite roles that may possibly be related to the dubious effect of IDO-1 in the different clinical forms of disease. Our data show that M. leprae and its fractions are able to differentially modulate the activity and functionality of IDO-1 in mDCs by a pathway that involves TLR2, suggesting that this enzyme may play an important role in leprosy immunopathogenesis.


Subject(s)
Dendritic Cells/immunology , Dendritic Cells/metabolism , Immune Tolerance , Indoleamine-Pyrrole 2,3,-Dioxygenase/metabolism , Leprosy/etiology , Leprosy/metabolism , Mycobacterium leprae/immunology , Toll-Like Receptor 2/metabolism , Biomarkers , Flow Cytometry , Humans , Leprosy/pathology , Lymphocytes/immunology , Lymphocytes/metabolism , Monocytes/immunology , Monocytes/metabolism , Tumor Necrosis Factor-alpha/metabolism
7.
Cell Microbiol ; 10(6): 1274-85, 2008 Jun.
Article in English | MEDLINE | ID: mdl-18284419

ABSTRACT

The effects of capsular polysaccharides, galactoxylomannan (GalXM) and glucuronoxylomannan (GXM), from acapsular (GXM negative) and encapsulate strains of Cryptococcus neoformans were investigated in RAW 264.7 and peritoneal macrophages. Here, we demonstrate that GalXM and GXM induced different cytokines profiles in RAW 264.7 macrophages. GalXM induced production of TNF-alpha, NO and iNOS expression, while GXM predominantly induced TGF-beta secretion. Both GalXM and GXM induced early morphological changes identified as autophagy and late macrophages apoptosis mediated by Fas/FasL interaction, a previously unidentified mechanism of virulence. GalXM was more potent than GXM at induction of Fas/FasL expression and apoptosis on macrophages in vitro and in vivo. These findings uncover a mechanism by which capsular polysaccharides from C. neoformans might compromise host immune responses.


Subject(s)
Bacterial Capsules/chemistry , Cryptococcus neoformans/chemistry , Fas Ligand Protein/metabolism , Macrophages/drug effects , Macrophages/physiology , Polysaccharides, Bacterial/pharmacology , Polysaccharides/pharmacology , Animals , Apoptosis , Cells, Cultured , Cryptococcosis/immunology , Mice , Mice, Inbred BALB C , Nitric Oxide/metabolism , Nitric Oxide Synthase Type II/metabolism , Polysaccharides/isolation & purification , Polysaccharides, Bacterial/isolation & purification , Transforming Growth Factor beta/metabolism , Tumor Necrosis Factor-alpha/metabolism
9.
PLoS One ; 9(10): e109672, 2014.
Article in English | MEDLINE | ID: mdl-25340550

ABSTRACT

Previous results demonstrate that the hybrid synthetic pterocarpanquinone LQB-118 presents antileishmanial activity against Leishmania amazonensis in a mouse model. The aim of the present study was to use a hamster model to investigate whether LQB-118 presents antileishmanial activity against Leishmania (Viannia) braziliensis, which is the major Leishmania species related to American tegumentary leishmaniasis. The in vitro antileishmanial activity of LQB-118 on L. braziliensis was tested on the promastigote and intracellular amastigote forms. The cell death induced by LQB-118 in the L. braziliensis promastigotes was analyzed using an annexin V-FITC/PI kit, the oxidative stress was evaluated by 2',7'-dichlorodihydrofluorescein diacetate (H2DCFDA) and the ATP content by luminescence. In situ labeling of DNA fragments by terminal deoxyribonucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) was used to investigate apoptosis in the intracellular amastigotes. L. braziliensis-infected hamsters were treated from the seventh day of infection with LQB-118 administered intralesionally (26 µg/kg/day, three times a week) or orally (4,3 mg/kg/day, five times a week) for eight weeks. LQB-118 was active against the L. braziliensis promastigotes and intracellular amastigotes, producing IC50 (50% inhibitory concentration) values of 3,4±0,1 and 7,5±0,8 µM, respectively. LQB-118 induced promastigote phosphatidylserine externalization accompanied by increased reactive oxygen species production and ATP depletion. Intracellular amastigote DNA fragmentation was also observed, without affecting the viability of macrophages. The treatment of L. braziliensis-infected hamsters with LQB-118, either orally or intralesionally, was effective in the control of lesion size, parasite load and increase intradermal reaction to parasite antigen. Taken together, these results show that the antileishmanial effect of LQB-118 extends to L. braziliensis in the hamster model, involves the induction of parasite apoptosis and shows promising therapeutic option by oral or local routes in leishmaniasis.


Subject(s)
Antiprotozoal Agents/pharmacology , Apoptosis/drug effects , Leishmania braziliensis/drug effects , Leishmaniasis, Cutaneous/parasitology , Naphthoquinones/pharmacology , Pterocarpans/pharmacology , Animals , Cricetinae , Female , Leishmaniasis, Cutaneous/pathology , Macrophages/drug effects , Macrophages/parasitology , Membrane Potential, Mitochondrial/drug effects , Mesocricetus , Phosphatidylserines/metabolism
10.
Trials ; 14: 379, 2013 Nov 11.
Article in English | MEDLINE | ID: mdl-24216069

ABSTRACT

BACKGROUND: Chronic chagasic cardiomyopathy is an inflammatory disease that occurs in approximately 30% of patients infected by the protozoan Trypanosoma cruzi, and it has a profile of high morbidity and mortality. The worst prognosis and the progression of this cardiomyopathy are associated with an exacerbated immune response and the production of proinflammatory cytokines, which also occur in other cardiomyopathies. Some nutrients, including omega-3 polyunsaturated fatty acids (PUFAs), promote the inhibition and/or stimulation of cytokine production. The objective of this trial is to study the effects of omega-3 PUFA supplementation on the inflammatory response and lipid profile in patients with chronic chagasic cardiomyopathy. METHODS/DESIGN: This is a parallel, randomized, placebo-controlled, double-blind clinical trial with 40 patients that will be conducted at a reference unit for Chagas disease patients, where the patients will be selected. The study will include patients with chronic chagasic cardiomyopathy who are 18 years of age or older. The exclusion criteria are (a) ongoing diarrheal disease, (b) inflammatory bowel disease, (c) diabetes or other endocrine disease, (d) use of fibrates, niacin, or statins, (e) use of anti-inflammatory drugs, (f) pregnant and lactating women, (g) use of vitamin, mineral, or omega-3 supplementation during the previous 30 days, (h) hospital admission during the study, and (i) other associated cardiomyopathies. The intervention will be treatment with omega-3 PUFAs at a dose of 3 g/day for 8 weeks, compared to placebo (corn oil). The primary endpoints will be the concentrations of inflammatory markers (interleukin (IL)-1, IL-2, IL-4, IL-6, IL-10, tumor necrosis factor (TNF)α, interferon (IFN)γ, and transforming growth factor (TGF)ß). Secondary endpoints will be the fasting glucose, lipid, and anthropometric profiles. For statistical analysis, we plan to run either a t test or Wilcoxon test (numerical variables) and Pearson's χ2 or Fisher's exact test (categorical data), as appropriate. DISCUSSION: Evidence suggests that the anti-inflammatory action of omega-3 PUFAs may have beneficial effects on chronic chagasic cardiomyopathy, as shown for other cardiomyopathies, due to improved control of the inflammatory response. At the end of the study, we predict that patients will have lower inflammatory markers and an improved metabolic and anthropometric profile. TRIAL REGISTRATION: Current Controlled Trials NCT01863576.


Subject(s)
Anti-Inflammatory Agents/therapeutic use , Chagas Cardiomyopathy/drug therapy , Dietary Supplements , Fatty Acids, Omega-3/therapeutic use , Research Design , Biomarkers/blood , Brazil , Chagas Cardiomyopathy/blood , Chagas Cardiomyopathy/diagnosis , Chi-Square Distribution , Chronic Disease , Clinical Protocols , Double-Blind Method , Female , Humans , Inflammation Mediators/blood , Lipids/blood , Male , Nutrition Assessment , Nutritional Status , Time Factors , Treatment Outcome
11.
Microbes Infect ; 11(2): 181-90, 2009 Feb.
Article in English | MEDLINE | ID: mdl-19070676

ABSTRACT

We investigated the role of autophagy in infection of macrophages by Leishmania amazonensis. Induction of autophagy by IFN-gamma or starvation increased intracellular parasite load and the percentages of infected macrophages from BALB/c but not from C57BL/6 mice. In contrast, starvation did not affect the replication of either Leishmania major or Trypanosoma cruzi in BALB/c macrophages. In BALB/c macrophages, starvation resulted in increased monodansylcadaverine staining and in the appearance of double-membrane and myelin-like vesicles characteristic of autophagosomes. Increased parasite load was associated with a reduction in NO levels and was attenuated by wortmannin, an inhibitor of autophagy. In infected macrophages from BALB/c, but not from C57BL/6 mice, starvation increased the number of lipid bodies and the amounts of PGE(2) produced. Exogenous PGE(2) increased parasite load in macrophages from BALB/c, but not C57BL/6 mice. The cyclooxygenase inhibitor indomethacin prevented the increase of parasite load in starved BALB/c macrophages, and actually induced parasite killing. These results suggest that autophagy regulates the outcome of L. amazonensis infection in macrophages in a host strain specific manner.


Subject(s)
Autophagy , Leishmania mexicana/immunology , Macrophages/immunology , Macrophages/parasitology , Animals , Female , Male , Mice , Mice, Inbred BALB C , Mice, Inbred C57BL , Nitric Oxide/biosynthesis , Prostaglandins/metabolism , Trypanosoma cruzi/immunology
12.
An Acad Bras Cienc ; 76(3): 519-27, 2004 Sep.
Article in English | MEDLINE | ID: mdl-15334250

ABSTRACT

Leishmania amazonensis is the main agent of diffuse cutaneous leishmaniasis, a disease associated with anergic immune responses. In this study we show that the crude antigen of Leishmania amazonensis (LaAg) but not L. braziliensis promastigotes (LbAg) contains substances that suppress mitogenic and spontaneous proliferative responses of T cells. The suppressive substances in LaAg are thermoresistant (100 degrees C/1h) and partially dependent on protease activity. T cell anergy was not due to a decreased production of growth factors as it was not reverted by addition of exogenous IL-2, IL-4, IFN-gamma or IL-12. LaAg did not inhibit anti-CD3-induced T cell activation, suggesting that anergy was due to a defect in antigen presentation. It was also not due to cell necrosis, but was accompanied by expressive DNA fragmentation in lymph node cells, indicative of apoptosis. Although pre-incubation of macrophages with LaAg prevented their capacity to present antigens, this effect was not due to apoptosis of the former. These results suggest that the T cell anergy found in diffuse leishmaniasis may be the result of parasite antigen-driven apoptosis of those cells following defective antigen presentation.


Subject(s)
Antigens, Protozoan/immunology , Apoptosis/immunology , Clonal Anergy/immunology , Leishmania/immunology , Leishmaniasis, Diffuse Cutaneous/immunology , T-Lymphocytes/immunology , Animals , Leishmaniasis, Diffuse Cutaneous/parasitology , Lymph Nodes/cytology , Lymph Nodes/immunology , Mice , Mice, Inbred BALB C
13.
An. acad. bras. ciênc ; 76(3): 519-527, Sept. 2004. ilus, graf
Article in English | LILACS | ID: lil-364481

ABSTRACT

A Leishmania amazonensis é o principal agente etiológico da leishmaniose cutânea difusa, uma doença associada a respostas imunes anérgicas. Neste estudo nós mostramos que o extrato bruto de promastigotas de Leishmania amazonensis (LaAg), mas não de L. braziliensis (LbAg), contém substâncias que suprimem respostas proliferativas, espontâneas e mitogênicas, de células T. As substâncias supressoras no LaAg são termo-resistentes (100§C/1h) e parcialmente dependentes da atividade de proteases. A anergia de células T não foi devida à diminuição na produção de fatores de crescimento, uma vez que não foi revertida pela adição de: IL-2, IL-4, IFN-g ou IL-12. O LaAg não inibiu a ativação de células T induzida por anti-CD3, sugerindo que a anergia é devida a um defeito na apresentação de antígenos. A anergia não foi devida à necrose celular, mas foi acompanhada de uma expressiva fragmentação de DNA nas células de linfonodos, indicativo de apoptose. Apesar da pré-incubação de macrófagos com LaAg ter inibido sua capacidade de apresentação de antigenos, este efeito não foi devido à apoptose dos primeiros. Estes resultados sugerem que a anergia de células T encontrada na leishmaniose difusa deve ser devida à apoptose dessas células que se segue à apresentação defeituosa de antígenos pelo antígeno do parasito.


Subject(s)
Animals , Mice , Antigens, Protozoan , Apoptosis , Leishmania , Leishmaniasis, Diffuse Cutaneous , T-Lymphocytes , Lymph Nodes , Mice, Inbred BALB C
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