Papel da proteína heme oxigenase 1 na infecção de macrófagos por leishmania chagasi

Submitted by Ana Maria Fiscina Sampaio ([email protected]) on 2012-08-01T21:22:36Z No. of bitstreams: 1 Nivea Farias Luz Papel da proteína hemeoxigenase....pdf: 1624634 bytes, checksum: 6f830105c144bc3c799743a490e5a511 (MD5)Made available in DSpace on 2012-08-01T21:22:36Z (GMT). No. of bitstreams: 1 Nivea Farias Luz Papel da proteína hemeoxigenase....pdf: 1624634 bytes, checksum: 6f830105c144bc3c799743a490e5a511 (MD5) Previous issue date: 2011Fundação Oswaldo Cruz. Centro de Pesquisas Gonçalo Moniz. Salvador, BA, BrasilA leishmaniose visceral (LV) apresenta ampla distribuição geográfica e é fatal caso não seja tratada. As manifestações hematológicas são constantes na LV e em casos não tratados os pacientes evoluem à óbito por sangramento maciço ou anemia grave. Neste cenário, mecanismos ligados à hemólise, metabolismo do heme e atividade da enzima heme oxigenase podem estar envolvidos na imunopatogênese da LV, no entanto essa perspectiva ainda não foi explorada. A heme oxigenase (HO) tem importantes propriedades regulatórias e está envolvida em processos fisiológicos e patofisiológicos como citoproteção e inflamação. Apesar de sua sugestiva participação no contexto da infecção por Leishmania, uma rápida pesquisa no PubMed com as palavras heme oxigenase e Leishmania remete a somente três trabalhos até a presente data. Nesse projeto testaremos a hipótese de que a ativação da enzima heme oxigenase-1 (HO-1) favorece a infecção por Leishmania (L) chagasi, principal agente etiológico da LV humana no Brasil. Nossas observações nesse trabalho indicam que a enzima HO-1 é induzida em macrófagos durante a infecção por L. chagasi e que a indução farmacológica da HO-1, pela CoPP aumenta a carga parasitária de macrófagos infectados por L. chagasi e reduz a produção de mediadores pró-inflamatórios frente à estimulação por LPS, tais como TNF, NO, PGE2, MCP-1, IL-1β e IL-6. Além disso, a HO-1 favorece um ambiente anti-inflamatório onde prevalece a presença de IL-10 sobre a de TNF. Macrófagos derivados de medula óssea de camundongos deficientes no gene HO-1 tem menor carga parasitária, quando infectados por L. chagasi em comparação aos macrófagos de camundongos selvagens. Esses achados indicam um potencial deletério para a HO-1 na infecção por L. chagasi, bem como sugerem possíveis mecanismos envolvidos na imunopatogênese da LV.Visceral leishmaniasis (VL) is a widespread disease and is fatal if left untreated. Hematological manifestations are common in VL and untreated patients evolve to death from massive bleeding and severe anemia. In this scenario, mechanisms related to hemolysis, heme metabolism and enzyme activity of heme oxygenase may be involved in the immunopathogenesis of VL. But that panorama has not been explored. Heme oxygenase (HO) has important regulatory properties and is involved in patho-physiological processes such as cytoprotection and inflammation. Despite HO participation in the context of Leishmania infection is suggestive, a quick search on PubMed with the words heme oxygenase and Leishmania refers to only three papers to date. This project will test the hypothesis that heme oxygenase- 1 (HO-1) activation favors Leishmania (L) chagasi, the main etiology agent of human VL in Brazil. Our observations indicate that HO-1 is induced in macrophages during L. chagasi infection and pharmacological induction of HO-1 by CoPP increases parasite load of infected macrophages and results in inhibition of TNF- α, IL-1β, IL-6, MCP-1, PGE2 and Nitrite levels upon LPS stimulation and simultaneously induced a higher IL-10/TNF-α ratio in peritoneal macrophages contributing to the anti inflammatory pathway that favors L. chagasi replication. Beyond this, we observed that bone marrow derived macrophages knockout to HO-1 gene have a significant low parasite load when infected by L. chagasi than their wild type counterparts. In summary, our findings suggest that this enzyme can play a deleterious role in VL and clarify one of the immunoregulatory mechanisms involved in VL

Ativação da heme oxigenase-1 e via da necroptose como mecanismos imunopatogênicos na infecção de macrófagos por Leishmania infantum

Submitted by Ana Maria Fiscina Sampaio ([email protected]) on 2016-02-04T14:23:59Z No. of bitstreams: 1 Nivea Farias Luz Ativação...2015.pdf: 10567971 bytes, checksum: 1479b4bbd75d3db2ffcf895208002d81 (MD5)Approved for entry into archive by Ana Maria Fiscina Sampaio ([email protected]) on 2016-02-04T14:24:19Z (GMT) No. of bitstreams: 1 Nivea Farias Luz Ativação...2015.pdf: 10567971 bytes, checksum: 1479b4bbd75d3db2ffcf895208002d81 (MD5)Made available in DSpace on 2016-02-04T14:24:19Z (GMT). No. of bitstreams: 1 Nivea Farias Luz Ativação...2015.pdf: 10567971 bytes, checksum: 1479b4bbd75d3db2ffcf895208002d81 (MD5) Previous issue date: 2015Fundação Oswaldo Cruz, Centro de Pesquisas Gonçalo Moniz. Salvador, BA, BrasilA Leishmaniose visceral (LV) apresenta ampla distribuição geográfica e é fatal caso não seja tratada. As manifestações hematológicas são constantes na LV e em casos não tratados os pacientes evoluem à óbito por sangramento maciço ou anemia grave. Neste cenário, mecanismos ligados à morte celular, hemólise, metabolismo do heme e atividade da enzima heme oxigenase podem estar envolvidos na imunopatogênese da LV. A heme oxigenase (HO) tem importantes propriedades regulatórias e está envolvida em processos fisiológicos e patofisiológicos como citoproteção e inflamação. Nesse projeto testamos a hipótese de que a ativação da enzima heme oxigenase-1 (HO-1) favorece a infecção por Leishmania infantum chagasi, principal agente etiológico da LV humana no Brasil e de que mecanismos de morte celular inflamatória induzida por heme estão associados com a resistência ao parasita. Nossas observações nesse trabalho indicam que a enzima HO-1 é induzida em macrófagos durante a infecção por L. chagasi e que a indução farmacológica da HO-1, pela CoPP aumenta a carga parasitária de macrófagos infectados por L. chagasi e reduz a produção de mediadores próinflamatórios. Além disso, a HO-1 favorece um ambiente anti-inflamatório onde prevalece a presença de IL-10 sobre a de TNF. Macrófagos derivados de medula óssea de camundongos deficientes no gene HO-1 têm menor carga parasitária, quando infectados por L. chagasi em comparação aos macrófagos de camundongos selvagens. Além disso, pacientes com LV apresentam maiores níveis de heme-oxigenase 1 e de heme no soro. Nossas observações indicam que heme é capaz de induzir necroptose em macrófagos humanos, e de que moléculas da via da necroptose estão associadas com a resistência na infecção por Leishmania. A molécula RIPK1 controla a replicação de Leishmania por um mecanismo independente da produção de IL-1β, enquanto que a molécula PGAM5 depende de IL-1βpara controlar o crescimento do parasita. Por fim, encontramos que essas proteínas participam do controle da replicação por Leishmania em um modelo experimental de Leishmaniose cutânea. Esses achados indicam um potencial deletério para a HO-1 na infecção por L. chagasi, e um papel protetor da necroptose na infecção porLeishmania.Visceral leishmaniasis (VL) is a widespread disease and is fatal if left untreated. Hematological manifestations are common in VL and untreated patients evolve to death from massive bleeding and severe anemia. In this scenario, mechanisms related to cell death pathways, hemolysis, heme metabolism and enzymatic activity of heme oxygenase may be involved in the immunopathogenesis of the disease. Heme oxygenase (HO) has important regulatory properties and is involved in patho-physiological processes such as cytoprotection and inflammation. This project tested the hypothesis that heme oxygenase- 1 (HO-1) activation favors Leishmania infantum chagasi infection, the main etiologic agent of human VL in Brazil, we also tested whether heme induced inflammatory cell death pathways are involved in resistance to Leishmania infection. Our observations indicate that HO-1 is induced in macrophages infected with L. infantum chagasi and pharmacological induction for HO-1 by CoPP increases parasite load of infected macrophages and reduces production on inflammatory mediators. In addition, HO-1 contributes to the anti inflammatory pathway that favors L. chagasi replication through a higher IL-10/TNF-α ratio in macrophages. We also observed that bone marrow derived macrophages knockout to HO-1 gene have a significant lower parasite load when infected by L. infantum chagasi than their wild type counterparts. Beyond this, we found that patients with VL presented higher systemic concentrations of HO- 1 and heme than healthy individuals. We found that heme is able to induce programmed necrosis “necroptosis” in human cells and that molecular players from necroptosis pathway contribute to resistance to Leishmania infection. RIPK1 controls Leishmania replication through a mechanism independent of IL-1β production, while PGAM5 requires IL-1β to control Leishmania replication. Finally, we found that RIPK1 and PGAM5 play an important role in controlling Leishmania replication in a cultaneous leishmaniasis experimental model. Our findings argue that HO-1 has a critical role in L. chagasi replication and necroptosis pathway is involved in resistance against Leishmania infection

Programmed necrosis in the cross talk of cell death and inflammation

Submitted by Ana Maria Fiscina Sampaio ([email protected]) on 2015-05-27T14:16:06Z No. of bitstreams: 1 Chan FKM Programmed....pdf: 1513332 bytes, checksum: 357651763eff241bde315ead53b8b817 (MD5)Approved for entry into archive by Ana Maria Fiscina Sampaio ([email protected]) on 2015-05-27T16:41:58Z (GMT) No. of bitstreams: 1 Chan FKM Programmed....pdf: 1513332 bytes, checksum: 357651763eff241bde315ead53b8b817 (MD5)Made available in DSpace on 2015-05-27T16:41:58Z (GMT). No. of bitstreams: 1 Chan FKM Programmed....pdf: 1513332 bytes, checksum: 357651763eff241bde315ead53b8b817 (MD5) Previous issue date: 2014University of Massachusetts Medical School. Department of Pathology, Immunology and Microbiology Program. Worcester, MassachusettsUniversity of Massachusetts Medical School. Department of Pathology, Immunology and Microbiology Program. Worcester, Massachusetts / Fundação Oswaldo Cruz. Centro de Pesquisas Gonçalo Moniz. Salvador, BA, BrasilUniversity of Massachusetts Medical School. Department of Pathology, Immunology and Microbiology Program. Worcester, MassachusettsCell proliferation and cell death are integral elements in maintaining homeostatic balance in metazoans. Disease pathologies ensue when these processes are disturbed. A plethora of evidence indicates that malfunction of cell death can lead to inflammation, autoimmunity, or immunodeficiency. Programmed necrosis or necroptosis is a form of nonapoptotic cell death driven by the receptor interacting protein kinase 3 (RIPK3) and its substrate, mixed lineage kinase domain-like (MLKL). RIPK3 partners with its upstream adaptors RIPK1, TRIF, or DAI to signal for necroptosis in response to death receptor or Toll-like receptor stimulation, pathogen infection, or sterile cell injury. Necroptosis promotes inflammation through leakage of cellular contents from damaged plasma membranes. Intriguingly, many of the signal adaptors of necroptosis have dual functions in innate immune signaling. This unique signature illustrates the cooperative nature of necroptosis and innate inflammatory signaling pathways in managing cell and organismal stresses from pathogen infection and sterile tissue injury

Nanomedicine: Nanotechnology, Biology and Medicine

Texto completo: acesso restrito. p. 985–995We recently demonstrated that immunization with polyester poly(lactide-co-glycolide acid) (PLGA) nanoparticles loaded with the 11-kDa Leishmania vaccine candidate kinetoplastid membrane protein 11 (KMP-11) significantly reduced parasite load in vivo. Presently, we explored the ability of the recombinant PLGA nanoparticles to stimulate innate responses in macrophages and the outcome of infection with Leishmania braziliensis in vitro. Incubation of macrophages with KMP-11-loaded PLGA nanoparticles significantly decreased parasite load. In parallel, we observed the augmented production of nitric oxide, superoxide, TNF-α and IL-6. An increased release of CCL2/MCP-1 and CXCL1/KC was also observed, resulting in macrophage and neutrophil recruitment in vitro. Lastly, the incubation of macrophages with KMP-11-loaded PLGA nanoparticles triggered the activation of caspase-1 and the secretion of IL-1β and IL-18, suggesting inflammasome participation. Inhibition of caspase-1 significantly increased the parasite load. We conclude that KMP-11-loaded PLGA nanoparticles promote the killing of intracellular Leishmania parasites through the induction of potent innate responses

Journal of Immunology

Texto completo: acesso restrito. p. 1196-1204In many hemolytic disorders, such as malaria, the release of free heme has been involved in the triggering of oxidative stress and tissue damage. Patients presenting with severe forms of malaria commonly have impaired regulatory responses. Although intriguing, there is scarce data about the involvement of heme on the regulation of immune responses. In this study, we investigated the relation of free heme and the suppression of anti-inflammatory mediators such as PGE2 and TGF-β in human vivax malaria. Patients with severe disease presented higher hemolysis and higher plasma concentrations of Cu/Zn superoxide dismutase (SOD-1) and lower concentrations of PGE2 and TGF-β than those with mild disease. In addition, there was a positive correlation between SOD-1 concentrations and plasma levels of TNF-α. During antimalaria treatment, the concentrations of plasma SOD-1 reduced whereas PGE2 and TGF-β increased in the individuals severely ill. Using an in vitro model with human mononuclear cells, we demonstrated that the heme effect on the impairment of the production of PGE2 and TGF-β partially involves heme binding to CD14 and depends on the production of SOD-1. Aside from furthering the current knowledge about the pathogenesis of vivax malaria, the present results may represent a general mechanism for hemolytic diseases and could be useful for future studies of therapeutic approaches

Association between the haptoglobin and heme oxygenase 1 genetic profiles and soluble CD163 in susceptibility to and severity of human malaria

Submitted by Ana Maria Fiscina Sampaio ([email protected]) on 2014-07-14T12:34:54Z No. of bitstreams: 1 Mendonça VRR and Association between....pdf: 1483842 bytes, checksum: 789494313a7e9e5c126a3356d3e09691 (MD5)Made available in DSpace on 2014-07-14T12:34:54Z (GMT). No. of bitstreams: 1 Mendonça VRR and Association between....pdf: 1483842 bytes, checksum: 789494313a7e9e5c126a3356d3e09691 (MD5) Previous issue date: 2012Fundação Oswaldo Cruz. Centro de Pesquisa Gonçalo Moniz. Salvador, BA, Brasil / Universidade Federal da Bahia. Faculdade de Medicina da Bahia. Salvador, BA, BrasilFundação Oswaldo Cruz. Centro de Pesquisa Gonçalo Moniz. Salvador, BA, Brasil / Universidade Federal da Bahia. Faculdade de Medicina da Bahia. Salvador, BA, BrasilFundação Oswaldo Cruz. Centro de Pesquisa Gonçalo Moniz. Salvador, BA, BrasilFundação Oswaldo Cruz. Centro de Pesquisa Gonçalo Moniz. Salvador, BA, Brasil / Universidade Federal da Bahia. Faculdade de Medicina da Bahia. Salvador, BA, Brasil / Instituto Nacional de Ciência e Tecnologia de Investigação em Imunologia (iii-INCT). São Paulo, SP, BrasilFundação Oswaldo Cruz. Centro de Pesquisa Gonçalo Moniz. Salvador, BA, Brasil / Universidade Federal da Bahia. Faculdade de Farmácia. Salvador, BA, BrasilNational Institute of Allergy and Infectious Diseases. National Institutes of Health. Immunobiology Section. Laboratory of Parasitic Diseases. Bethesda, Maryland, USAeFundação Oswaldo Cruz. Centro de Pesquisa Gonçalo Moniz. Salvador, BA, Brasil / Universidade Federal da Bahia. Faculdade de Medicina da Bahia. Salvador, BA, Brasil / Instituto Nacional de Ciência e Tecnologia de Investigação em Imunologia (iii-INCT). São Paulo, SP, BrasilIntravascular hemolysis is a hallmark event in the immunopathology of malaria that results in increased systemic concentrations of free hemoglobin (Hb). The oxidation of Hb by free radicals causes the release of heme, which amplifies inflammation. To circumvent the detrimental effects of free heme, hosts have developed several homeostatic mechanisms, including the enzyme haptoglobin (Hp), which scavenges cell-free Hb, the monocyte receptor CD163, which binds to Hb-Hp complexes, and heme oxygenase-1 (HO-1), which degrades intracellular free heme. We tested the association between these three main components of the host response to hemolysis and susceptibility to malaria in a Brazilian population. The genetic profiles of the HMOX1 and Hp genes and the plasma levels of a serum inflammatory marker, the soluble form of the CD163 receptor (sCD163), were studied in 264 subjects, including 78 individuals with symptomatic malaria, 106 individuals with asymptomatic malaria, and 80 uninfected individuals. We found that long (GT)n repeats in the microsatellite polymorphism region of the HMOX1 gene, the Hp2 allele, and the Hp2.2 genotype were associated with symptomatic malaria. Moreover, increased plasma concentrations of heme, Hp, HO-1, and sCD163 were associated with susceptibility to malaria. The validation of these results could support the development of targeted therapies and aid in reducing the severity of malari

Cell Host and Microbe

Texto completo: acesso restrito. p. 693–704Disease tolerance is a defense strategy that limits the fitness costs of infection irrespectively of pathogen burden. While restricting iron (Fe) availability to pathogens is perceived as a host defense strategy, the resulting tissue Fe overload can be cytotoxic and promote tissue damage to exacerbate disease severity. Examining this interplay during malaria, the disease caused by Plasmodium infection, we find that expression of the Fe sequestering protein ferritin H chain (FtH) in mice, and ferritin in humans, is associated with reduced tissue damage irrespectively of pathogen burden. FtH protection relies on its ferroxidase activity, which prevents labile Fe from sustaining proapoptotic c-Jun N-terminal kinase (JNK) activation. FtH expression is inhibited by JNK activation, promoting tissue Fe overload, tissue damage, and malaria severity. Mimicking FtH’s antioxidant effect or inhibiting JNK activation pharmacologically confers therapeutic tolerance to malaria in mice. Thus, FtH provides metabolic adaptation to tissue Fe overload, conferring tolerance to malaria

Prostaglandin E2/Leukotriene B4 balance induced by Lutzomyia longipalpis saliva favors Leishmania infantum infection

Made available in DSpace on 2015-05-27T13:39:48Z (GMT). No. of bitstreams: 2 license.txt: 1914 bytes, checksum: 7d48279ffeed55da8dfe2f8e81f3b81f (MD5) patrícia_bozza2etal_IOC_2014.pdf: 3705278 bytes, checksum: 7aaa8c4b38527574249dc03af0a5e20e (MD5) Previous issue date: 2014Fundação Oswaldo Cruz. Centro de Pesquisas Gonçalo Muniz. Salvador, BA, Brasil / Universidade Federal da Bahia Ocidental. Centro de Ciências Biológicas e da Saúde. Barreiras, BA, Brasil.Fundação Oswaldo Cruz. Centro de Pesquisas Gonçalo Muniz. Salvador, BA, Brasil / Universidade Federal da Bahia. Instituto de Ciências da Saúde. Departamento de Biomorfologia. Salvador, BA, Brasil.Fundação Oswaldo Cruz. Centro de Pesquisas Gonçalo Muniz. Salvador, BA, Brasil / Universidade Federal da Bahia. Salvador, BA, Brasil.Fundação Oswaldo Cruz. Centro de Pesquisas Gonçalo Muniz. Salvador, BA, Brasil / Universidade Federal da Bahia. Salvador, BA, Brasil.National Institutes of Health. National Institute of Allergy and Infectious Diseases. Laboratory of Parasitic Diseases. Section of Immunobiology. Bethesda, MD, USA.Fundação Oswaldo Cruz. Centro de Pesquisas Gonçalo Muniz. Salvador, BA, Brasil.Fundação Oswaldo Cruz. Centro de Pesquisas Gonçalo Muniz. Salvador, BA, Brasil / Universidade Federal da Bahia. Salvador, BA, Brasil / Instituto Nacional de Ciência e Tecnologia (INCT). Instituto de Investigação em Imunobiologia. São Paulo, SP, Brasil.Fundação Oswaldo Cruz. Centro de Pesquisas Gonçalo Muniz. Salvador, BA, Brasil / Instituto Nacional de Ciência e Tecnologia (INCT). Instituto de Investigação em Imunobiologia. São Paulo, SP, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Centro de Pesquisas Gonçalo Muniz. Salvador, BA, Brasil / Instituto Nacional de Ciência e Tecnologia (INCT). Instituto de Investigação em Imunobiologia. São Paulo, SP, Brasil.Background Eicosanoids and sand fly saliva have a critical role in the Leishmania infection. Here, we evaluated the effect of Lutzomyia longipalpis salivary gland sonicate (SGS) on neutrophil and monocyte recruitment and activation of eicosanoid production in a murine model of inflammation. Methods C57BL/6 mice were inoculated intraperitonealy with Lutzomyia longipalpis SGS or Leishmania infantum or both, followed by analyses of cell recruitment, parasite load and eicosanoid production. Results Intraperitoneal injection of Lutzomyia longipalpis SGS together with Leishmania infantum induced an early increased parasite viability in monocytes and neutrophils. L. longipalpis SGS increased prostaglandin E2 (PGE2), but reduced leukotriene B4 (LTB4) production ex vivo in peritoneal leukocytes. In addition, the pharmacological inhibition of cyclooxygenase 2 (COX-2) with NS-398 decreased parasite viability inside macrophages during Leishmania infection in the presence of L. longipalpis SGS arguing that PGE2 production is associated with diminished parasite killing. Conclusions These findings indicate that L. longipalpis SGS is a critical factor driving immune evasion of Leishmania through modulation of PGE2/LTB4 axis, which may represent an important mechanism on establishment of the infection

Plos one

Acesso restrito: Texto completo. p. 1-9.Background: Areas that are endemic for malaria are also highly endemic for hepatitis B virus (HBV) infection. Nevertheless, it is unknown whether HBV infection modifies the clinical presentation of malaria. This study aimed to address this question. Methodology and Findings: An observational study of 636 individuals was performed in Rondoˆ nia, western Amazon, Brazil between 2006 and 2007. Active and passive case detections identified Plasmodium infection by field microscopy and nested Polymerase Chain Reaction (PCR). HBV infections were identified by serology and confirmed by real-time PCR. Epidemiological information and plasma cytokine profiles were studied. The data were analyzed using adjusted multinomial logistic regression. Plasmodium-infected individuals with active HBV infection were more likely to be asymptomatic (OR: 120.13, P,0.0001), present with lower levels of parasitemia and demonstrate a decreased inflammatory cytokine profile. Nevertheless, co-infected individuals presented higher HBV viremia. Plasmodium parasitemia inversely correlated with plasma HBV DNA levels (r =20.6; P = 0.0003). Conclusion: HBV infection diminishes the intensity of malaria infection in individuals from this endemic area. This effect seems related to cytokine balance and control of inflammatory responses. These findings add important insights to the understanding of the factors affecting the clinical outcomes of malaria in endemic regions