New Insights on the Inflammatory Role of Lutzomyia longipalpis Saliva in Leishmaniasis

When an haematophagous sand fly vector insect bites a vertebrate host, it introduces its mouthparts into the skin and lacerates blood vessels, forming a hemorrhagic pool which constitutes an intricate environment of cell interactions. In this scenario, the initial performance of host, parasite, and vector “authors” will heavily influence the course of Leishmania infection. Recent advances in vector-parasite-host interaction have elucidated “co-authors” and “new roles” not yet described. We review here the stimulatory role of Lutzomyia longipalpis saliva leading to inflammation and try to connect them in an early context of Leishmania infection

Lutzomyia longipalpis Saliva Triggers Lipid Body Formation and Prostaglandin E2 Production in Murine Macrophages

After the injection of saliva into the host's skin by sand flies, a transient erythematous reaction is observed, which is related to an influx of inflammatory cells and the release of various molecules that actively facilitate the blood meal. It is important to understand the specific mechanisms by which sand fly saliva manipulates the host's inflammatory responses. Herein, we report that saliva from Lutzomyia (L.) longipalpis, a widespread Leishmania vector, induces early production of eicosanoids. Intense formation of intracellular organelles called lipid bodies (LBs) was noted within those cells that migrated to the site of saliva injection. In vitro and ex vivo, sand fly saliva was able to induce LB formation and PGE2 release by macrophages. Interestingly, PGE2 production induced by L. longipalpis saliva was dependent on intracellular mechanisms involving phosphorylation of signaling proteins such as PKC-α and ERK-1/2 and subsequent activation of cyclooxygenase-2. Thus, this study provides new insights into the pharmacological properties of sand fly saliva and opens new opportunities for intervening with the induction of the host's inflammatory pathways by L. longipalpis bites

Cooperation between Apoptotic and Viable Metacyclics Enhances the Pathogenesis of Leishmaniasis

Mimicking mammalian apoptotic cells by exposing phosphatidylserine (PS) is a strategy used by virus and parasitic protozoa to escape host protective inflammatory responses. With Leishmania amazonensis (La), apoptotic mimicry is a prerogative of the intramacrophagic amastigote form of the parasite and is modulated by the host. Now we show that differently from what happens with amastigotes, promastigotes exposing PS are non-viable, non-infective cells, undergoing apoptotic death. As part of the normal metacyclogenic process occurring in axenic cultures and in the gut of sand fly vectors, a sub-population of metacyclic promastigotes exposes PS. Apoptotic death of the purified PS-positive (PSPOS) sub-population was confirmed by TUNEL staining and DNA laddering. Transmission electron microscopy revealed morphological alterations in PSPOS metacyclics such as DNA condensation, cytoplasm degradation and mitochondrion and kinetoplast destruction, both in in vitro cultures and in sand fly guts. TUNELPOS promastigotes were detected only in the anterior midgut to foregut boundary of infected sand flies. Interestingly, caspase inhibitors modulated parasite death and PS exposure, when added to parasite cultures in a specific time window. Efficient in vitro macrophage infections and in vivo lesions only occur when PSPOS and PS-negative (PSNEG) parasites were simultaneously added to the cell culture or inoculated in the mammalian host. The viable PSNEG promastigote was the infective form, as shown by following the fate of fluorescently labeled parasites, while the PSPOS apoptotic sub-population inhibited host macrophage inflammatory response. PS exposure and macrophage inhibition by a subpopulation of promastigotes is a different mechanism than the one previously described with amastigotes, where the entire population exposes PS. Both mechanisms co-exist and play a role in the transmission and development of the disease in case of infection by La. Since both processes confer selective advantages to the infective microorganism they justify the occurrence of apoptotic features in a unicellular pathogen

Estudo protéico da saliva de Lutzomyia longipalpis (Lutz & Neiva, 1912)(Diptera:Phlebotominae) e o efeito de seus extratos utilizando o modelo do bolsão inflamatório

Submitted by Repositório Arca ([email protected]) on 2019-07-17T17:07:26Z No. of bitstreams: 1 license.txt: 1748 bytes, checksum: 8a4605be74aa9ea9d79846c1fba20a33 (MD5)Approved for entry into archive by Ana Maria Fiscina Sampaio ([email protected]) on 2019-07-19T12:45:48Z (GMT) No. of bitstreams: 2 Deboraci Brito Prates Estudo...2005.pdf: 29648134 bytes, checksum: 3d7ec65618760ab28757b7ec5a99f542 (MD5) license.txt: 1748 bytes, checksum: 8a4605be74aa9ea9d79846c1fba20a33 (MD5)Made available in DSpace on 2019-07-19T12:45:48Z (GMT). No. of bitstreams: 2 Deboraci Brito Prates Estudo...2005.pdf: 29648134 bytes, checksum: 3d7ec65618760ab28757b7ec5a99f542 (MD5) license.txt: 1748 bytes, checksum: 8a4605be74aa9ea9d79846c1fba20a33 (MD5) Previous issue date: 2005CAPES, CNPq e INSTITUTO DO MILÊNIO.Universidade Federal da Bahia. Faculdade de Medicina. Salvador, BA, Brasil / Fundação Oswaldo Cruz. Centro de Pesquisas Gonçalo Moniz. Salvador, BA, Brasil.O conteúdo protéico da saliva de Lutzomyia longipalpis (Lutz & Neiva, 1912) (Diptera: Psychodidae: Phlebotominae) foi estudado. Este flebotomíneo é o principal vetor da Leishmaniose Visceral Americana em várias regiões do Brasil. A saliva de vetores desempenha um importante papel na obtenção do alimento sanguíneo, na lubrificação das peças bucais e na transmissão dos parasitos. O conteúdo e o perfil protéico da glândula salivar de fêmeas de Lu. longipalpis em jejum sanguíneo e alimentadas com sangue foram analisados em diferentes dias. O conteúdo protéico da glândula salivar de fêmeas em jejum aumentou de 1,955 µg/mL no primeiro dia após emergência do inseto adulto para 7,457 µg/mL no terceiro dia de vida, não variando muito até o sétimo dia. Um dia após a alimentação sanguínea, o conteúdo protéico diminuiu para 5,046 µg/mL, porém essa concentração aumentou nos dias subseqüentes, atingindo 8,172 µg/mL no sétimo dia. O conteúdo de proteínas em machos foi menor que em fêmeas. O perfil eletroforético das proteínas da saliva de fêmeas foi analisado por SDS-PAGE. Bandas correspondentes a 45 KDa foram mais intensas. A maioria dos polipeptídeos presente na saliva das fêmeas em jejum estava também presente em fêmeas alimentadas. Contudo, em fêmeas com um dia de vida não foi possível observar as bandas de 6, 24, 27 e 130 KDa e em fêmeas com um dia após a alimentação sanguínea, não foi possível observar as bandas de 6, 16, 27, 32, 37, 49, 61 e 130 KDa. O perfil eletroforético bidimensional (20) de proteínas da saliva de fêmeas com 3, 5 e 7 dias de vida revelou um predomínio de peptídeos entre 14 e 45 KDa. O gel 20 de proteínas da saliva de fêmeas com 3 dias revelou 82 spots, o gel de 5 dias revelou 30 spots e o gel de 7 dias, 48 spots; todos os spots foram correspondentes. Além disso, o efeito da saliva e de 6 frações suas, obtidas por HPLC, no recrutamento de leucócitos foi estudado utilizando o modelo do bolsão de ar inflamatório em camundongos BALB/c. Os neutrófilos foram as células predominantes no exsudato. Porém, o recrutamento de macrófagos foi diferencial entre os grupos. O SGS induziu maior recrutamento de macrófagos e os extratos 5, 9 e 10 tiveram uma capacidade de recrutamento de macrófagos acima de 50% com relação ao SGS. Eosinófilos também estiveram presentes no exsudato inflamatório, principalmente quando utilizado o SGS e os extratos 5 e 9 como estímulos.The amount of salivary gland protein in Lutzomyia longipalpis (Lutz & Neiva, 1912) (Diptera: Psychodidae: Phlebotominae) was studied. This phletotomine is the main vector of the American Visceral Leishmaniasis in many Brazilian areas. The saliva of vector plays an important role in the taking of blood meal, in the lubrication of the mouthparts and in the parasite transmission. The salivary gland protein content and profile from unfed and blood fed female of Lu. longipalpis were analyzed at different times. The amount of salivary gland protein from sugar-fed female increased from 1.955 |ag/mL in the first day after adult emergence to 7.457 |ag/mL in the third day, without change until the seventh day. One day after blood feeding, the protein content decreased to 5.046 ^g/mL, however this concentration increased at following days to reach 8.172 ).ig/mL. The male protein content was lower than female protein content. The electrophoretic profile of the saliva proteins from female insects was analyzed by SDS-PAGE. Bands correspondents to 45 KDa were more intense. The most of polypeptides present in the saliva of sugar fed female also were present in the saliva of blood fed females. However, in female with one day after emergence was not possible to observe the bands 6, 24, 27 and 130 KDa and in female with one day after blood feeding was not possible to observe the bands 6, 16, 27, 32, 37, 49, 61 and 130 KDa. The bidimensional electrophoretic (2D) profile of the saliva proteins from female with 3, 5 and 7 days after emergence revealed a predominance of peptides between 14 and 45 KDa. The saliva protein 2D gel from female with 3 days revealed 82 spots, the 2D gel from 5 days revealed 30 spots and the 2D gel from 7 days revealed 48 spots; all spots were correspondent. Moreover, the effect of the saliva and six of its fractions, obtained by HPLC, in the leukocyte recruitment was studied using the inflammatory air pouch model in BALB/c mice. The neutrophils were the predominant cells in the exsudate. But, the macrophage recnjitment was different between groups. The SGS induced major macrophage recruitment and the extracts 5, 9 e 10 had a capacity of macrophage recruitment above 50% regarding the SGS. Eosinophils were also present in the inflammatory exsudate, mainly when the SGS and the extracts 5 and 9 were used as stimuli

Teaching sequences in sciences, health and intersectionalities: experience reports

Programa de Pós-graduação Patologia Humana e ExperimentalPrograma de Pós-graduação Biotecnologia em Saúde e Medicina InvestigativaCoordenação de Aperfeiçoamento de Pessoal de Nível Superior (Capes)Resum

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

Vaccination with the Leishmania major ribosomal proteins plus CpG oligodeoxynucleotides induces protection against experimental cutaneous leishmaniasis in mice

Submitted by Ana Maria Fiscina Sampaio ([email protected]) on 2014-03-13T19:19:59Z No. of bitstreams: 1 Borra S Vaccination with....pdf: 578715 bytes, checksum: e301f276716710b41f51f2b9d13736c5 (MD5)Made available in DSpace on 2014-03-13T19:19:59Z (GMT). No. of bitstreams: 1 Borra S Vaccination with....pdf: 578715 bytes, checksum: e301f276716710b41f51f2b9d13736c5 (MD5) Previous issue date: 2008Instituto de Salud Carlos III. Centro Nacional de Microbiología. Unidad de Inmunología Viral. Majadahonda, Madrid, SpainUniversidad Autónoma de Madrid. Facultad de Ciencias. Centro de Biología Molecular Severo Ochoa. Departamento de Biología Molecular. Madrid, SpainUniversidad Autónoma de Madrid. Facultad de Ciencias. Centro de Biología Molecular Severo Ochoa. Departamento de Biología Molecular. Madrid, SpainUniversidad Autónoma de Madrid. Facultad de Ciencias. Centro de Biología Molecular Severo Ochoa. Departamento de Biología Molecular. Madrid, SpainFundação Oswaldo Cruz. Centro de Pesquisa Gonçalo Moniz. Salvador, BA, BrasilFundação Oswaldo Cruz. Centro de Pesquisa Gonçalo Moniz. Salvador, BA, BrasilFundação Oswaldo Cruz. Centro de Pesquisa Gonçalo Moniz. Salvador, BA, BrasilUniversidad Autónoma de Madrid. Facultad de Ciencias. Centro de Biología Molecular Severo Ochoa. Departamento de Biología Molecular. Madrid, SpainUniversidad Autónoma de Madrid. Facultad de Ciencias. Centro de Biología Molecular Severo Ochoa. Departamento de Biología Molecular. Madrid, SpainIn the present work we analyze the antigenicity of Leishmania major ribosomal proteins (LRP) in infected BALB/c mice.We show that BALB/c mice vaccinated with LRP in the presence of CpG oligodeoxynucleotides (CpG-ODN) were protected against the development of dermal pathology and showed a reduction in the parasite load after challenge with L. major. This protection was associated with the induction of an IL-12 dependent specific-IFN-g response mediated mainly by CD4þ T cell, albeit a minor contribution of CD8þ T cells cannot be ruled out. Induction of Th1 responses against LRP also resulted in a reversion of the Th2 responses associated with susceptibility. A marked reduction of IgG1 antibody titer against parasite antigens besides an impaired IL-4 and IL-10 cytokine production by parasite specific T cells was observed. In addition, we show that the administration of the LRP plus CpG-ODN preparation also conferred protection in the naturally resistant C57BL/6 mice. In this strain protection was associated with a LRP specific IFN-g production in lymph nodes draining the challenge site. We believe that these evolutionary conserved proteins, combined with adjuvants that favor Th1 responses, may be relevant components of a pan-Leishmania vaccine. 2008 Elsevier Masson SAS. All rights reserve

Inflammatory cell infiltration and high antibody production in balb/c mice caused by natural exposure to Lutzomyia longipalpis bites

Submitted by Ana Maria Fiscina Sampaio ([email protected]) on 2011-09-14T18:12:44Z No. of bitstreams: 1 Silva F Inflammatory cell infiltration....pdf: 468424 bytes, checksum: 27dce6a49eb7b55d0e2786e519a3cff9 (MD5)Made available in DSpace on 2011-09-14T18:12:44Z (GMT). No. of bitstreams: 1 Silva F Inflammatory cell infiltration....pdf: 468424 bytes, checksum: 27dce6a49eb7b55d0e2786e519a3cff9 (MD5) Previous issue date: 2005Fundação Oswaldo Cruz. Centro de Pesquisas Gonçalo Moniz. Salvador, Bahia, BrazilFundação Oswaldo Cruz. Centro de Pesquisas Gonçalo Moniz. Salvador, Bahia, BrazilFundação Oswaldo Cruz. Centro de Pesquisas Gonçalo Moniz. Salvador, Bahia, BrazilFundação Oswaldo Cruz. Centro de Pesquisas Gonçalo Moniz. Salvador, Bahia, BrazilFundação Oswaldo Cruz. Centro de Pesquisas Gonçalo Moniz. Salvador, Bahia, BrazilFundação Oswaldo Cruz. Centro de Pesquisas Gonçalo Moniz. Salvador, Bahia, BrazilFundação Oswaldo Cruz. Centro de Pesquisas Gonçalo Moniz. Salvador, Bahia, BrazilSand flies inject saliva into the mammalian host when probing for a blood meal. Understanding the initial vertebrate reactions against sand fly saliva is important for possible interventions because these insects transmit diseases to humans and other animals. Little is known of these reactions to New World sand flies. Repeated exposure of BALB/c mice to Lutzomyia longipalpis bites leads to local inflammatory cell infiltration comprised of neutrophils, macrophages, and eosinophils. Total IgG and IgG1 antibodies react predominantly with three major protein bands (45, 44, and 16 kD) of the insect saliva by Western blot. The injection of immune serum previously incubated with salivary gland homogenate induced an early infiltration with neutrophils and macrophages, suggesting the participation of immune complexes in triggering inflammation

Changes in amounts of total salivary gland proteins of Lutzomyia longipallpis (Diptera: Psychodidae) according to age and diet

Submitted by Ana Maria Fiscina Sampaio ([email protected]) on 2014-03-13T16:57:01Z No. of bitstreams: 1 Prates DB Changes in amounts....pdf: 207901 bytes, checksum: d38c782c49e631a15a95ecc58be75e12 (MD5)Made available in DSpace on 2014-03-13T16:57:01Z (GMT). No. of bitstreams: 1 Prates DB Changes in amounts....pdf: 207901 bytes, checksum: d38c782c49e631a15a95ecc58be75e12 (MD5) Previous issue date: 2008Fundação Oswaldo Cruz. Centro de Pesquisa Gonçalo Moniz. Salvador, BA, Brasil / Universidade Federal da Bahia. Faculdade de Medicina. Salvador, BA, BrasilUniversidade Estadual Paulista de Rio Claro. Instituto de Biociências de Rio Claro. Departamento de Biologia. Laboratório de Biologia Estrutural e Zooquímica. Rio Claro, SP, 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. Salvador, BA, BrasilInstituto de Investigação em Imunologia iii. Salvador, BA, BrasilFundação Oswaldo Cruz. Centro de Pesquisa Gonçalo Moniz. Salvador, BA, Brasil / Universidade Federal da Bahia. Faculdade de Medicina. Salvador, BA, Brasil / Instituto de Investigação em Imunologia iii. Salvador, BA, BrasilFundação Oswaldo Cruz. Centro de Pesquisa Gonçalo Moniz. Salvador, BA, Brasil / Universidade Federal da Bahia. Faculdade de Medicina. Salvador, BA, Brasil / Instituto de Investigação em Imunologia iii. Salvador, BA, BrasilSaliva plays important roles in facilitation of a bloodmeal, lubrication of mouthparts, and parasite transmission for some vector insects. Salivary composition changes during the lifetime of an insect, and differences in the salivary proÞle may inßuence its functions. In this report, the amount and proÞle of salivary gland protein of the American visceral leishmaniasis vector Lutzomyia longipalpis (Lutz & Neiva, 1912) were analyzed at different times of insect development and diet. Protein content from unfed female sand ßies increased signiÞcantly with age, and a signiÞcant difference was observed in sugar-fed females during the Þrst 10 d of adult life. Salivary protein content sharply decreased 1 d after blood feeding, with gradual increase in concentration the following days. SDSpolyacrylamide gel electrophoresis analysis revealed that most polypeptides present in the saliva of sugar-fed also were present in the saliva of blood-fed females. Understanding changes in sand ßyÕs saliva contents at distinct days after emergence and the inßuence of a bloodmeal in this aspect may reveal the role played by saliva during leishmaniasis transmission

Scientific Reports

p. 1-6Leishmania infantum chagasi causes visceral leishmaniasis (VL); it is transmitted by the sand fly Lutzomyia longipalpis that injects saliva and parasites into the host's skin during a blood meal. Chickens represent an important blood source for sand flies and their presence in the endemic area is often cited as a risk factor for VL transmission. However, the role of chickens in VL epidemiology has not been well defined. Here, we tested if chicken antibodies against Lu. longipalpis salivary gland sonicate (SGS) could be used as markers of exposure to sand fly bites. All naturally exposed chickens in a VL endemic area in Brazil developed anti-SGS IgY antibodies. Interestingly, Lu. longipalpis recombinant salivary proteins rLJM17 and rLJM11 were also able to detect anti-SGS IgY antibodies. Taken together, these results show that chickens can be used to monitor the presence of Lu. longipalpis in the peri-domiciliary area in VL endemic regions, when used as sentinel animals