Fungus Paracoccidioides Brasiliensis Research Articles

HeNe laser increases the pro-inflammatory cytokines production and the in vitro phagocytosis of P. brasiliensis by macrophages.

Event Abstract Back to Event HeNe laser increases the pro-inflammatory cytokines production and the in vitro phagocytosis of P. brasiliensis by macrophages. Thiago Alves Da Costa1, Rodolfo Thomé1, Rosaria Di Gangi1, Carolina Francelin1 and Liana Verinaud1* 1 State University of Campinas, Department of Structural and Functional Biology; Institute of Biology., Brazil Cutaneous lesions caused by the fungus Paracoccidioides brasiliensis (Pb) lead to painful ulcerative wounds, which are difficult to be cured by conventional treatment with anti-fungal drugs due to their adverse effects and the multidrug resistance that some fungal isolates present (Andriole, 1998, Trent & Kirsner, 2003). We have previously showed that Helium-Neon (HeNe) laser irradiation of paracoccidioidomycotic lesions in mice accelerates the wound healing as well as the fungal clearance, which is very probably linked to the death of the fungus by macrophages (Nagib et al., 2010, Alves da Costa et al., 2013). In this study we evaluated, in vitro, whether HeNe laser irradiation was capable of modulating the phenotypic and functional profile of human and murine macrophages. Peritoneal macrophages from BALB/c mice were irradiated with different dosages of HeNe laser (0.836; 1.5 and 3J/cm2) and cultivated with Pb yeast cells (1:4, macrophage:fungi) for 6 hours. Flow cytometry analysis of macrophages showed that laser irradiation promoted an increase in the expression of TLR-4, which are molecules involved in yeast recognition and internalization, but did not alter other markers, like CD11b, F4/80, MHC-II, for example. Also, co-cultures of irradiated macrophages and Pb showed a higher phagocytic index and a higher pro-inflammatory (IFN-γ, IL-6, TNF-α) cytokines production. Similar results were obtained with a human macrophage cell line (THP1) even under the lowest incidence energy of the laser. Collectively, our results indicate that the accelerated wound healing process, observed after the treatment of paracoccidioidomycotic lesions with the HeNe laser, must be associated with a direct action of irradiation on macrophages. Acknowledgements Supported by FAPESP (#2010/08485-5, #2012/22131-7) and CAPES. References Alves da Costa, T., Di Gangi, R., Costa, M. R. S. N., Thomé, R., Paulino, L. C., Verinaud, L. (2013) . Effect of Helium-Neon (HeNe) laser irradiation on lesions in experimental paracoccidioidomycosis. Trends In Photochemistry And Photobiology, Accepted manuscript. Andriole, V. T. (1999). The 1998 Garrod lecture. Current and future antifungal therapy: new targets for antifungal agents. J. Antimicrobial Chemotherapy 44, 151-162. Nagib, P. R. N., Gameiro, J., Alves da Costa, T., Di Gangi, R., Ribeiro, J. S., Paulino, L. C., Verinaud, L. (2010). Effect of HeNe laser irradiation on extracellular matrix deposition and expression of cytokines and chemokines in paracoccidioidomycotic lesions. Photochem. Photobiol. 86, 920-924. Trent, J. T., Kirsner, R. S. (2003). Identifying and treating mycotic skin infections. Adv Skin Wound Care 16, 122-9. Keywords: Paracoccidioides brasiliensis, paracoccidioidomycotic lesions, Phagocytosis, Pro-inflammatory cytokines, HeNe Laser Conference: 15th International Congress of Immunology (ICI), Milan, Italy, 22 Aug - 27 Aug, 2013. Presentation Type: Abstract Topic: Host-pathogen interactions Citation: Alves Da Costa T, Thomé R, Di Gangi R, Francelin C and Verinaud L (2013). HeNe laser increases the pro-inflammatory cytokines production and the in vitro phagocytosis of P. brasiliensis by macrophages.. Front. Immunol. Conference Abstract: 15th International Congress of Immunology (ICI). doi: 10.3389/conf.fimmu.2013.02.01003 Copyright: The abstracts in this collection have not been subject to any Frontiers peer review or checks, and are not endorsed by Frontiers. They are made available through the Frontiers publishing platform as a service to conference organizers and presenters. The copyright in the individual abstracts is owned by the author of each abstract or his/her employer unless otherwise stated. Each abstract, as well as the collection of abstracts, are published under a Creative Commons CC-BY 4.0 (attribution) licence (https://creativecommons.org/licenses/by/4.0/) and may thus be reproduced, translated, adapted and be the subject of derivative works provided the authors and Frontiers are attributed. For Frontiers’ terms and conditions please see https://www.frontiersin.org/legal/terms-and-conditions. Received: 29 Jun 2013; Published Online: 22 Aug 2013. * Correspondence: Prof. Liana Verinaud, State University of Campinas, Department of Structural and Functional Biology; Institute of Biology., Campinas, SP, Brazil, verinaud@unicamp.br Login Required This action requires you to be registered with Frontiers and logged in. To register or login click here. Abstract Info Abstract The Authors in Frontiers Thiago Alves Da Costa Rodolfo Thomé Rosaria Di Gangi Carolina Francelin Liana Verinaud Google Thiago Alves Da Costa Rodolfo Thomé Rosaria Di Gangi Carolina Francelin Liana Verinaud Google Scholar Thiago Alves Da Costa Rodolfo Thomé Rosaria Di Gangi Carolina Francelin Liana Verinaud PubMed Thiago Alves Da Costa Rodolfo Thomé Rosaria Di Gangi Carolina Francelin Liana Verinaud Related Article in Frontiers Google Scholar PubMed Abstract Close Back to top Javascript is disabled. Please enable Javascript in your browser settings in order to see all the content on this page.

Alterations in stromal cell compartment during thymus atrophy induced by P. brasiliensis infection.

Event Abstract Back to Event Alterations in stromal cell compartment during thymus atrophy induced by P. brasiliensis infection. Thiago Alves Da Costa1, Rosaria Di Gangi1, Carolina Francelin1, Rodolfo Thomé1, Ronei L. Mamoni2 and Liana Verinaud1* 1 State University of Campinas, Department of Structural and Functional Biology; Institute of Biology, Brazil 2 State University of Campinas, Department of Clinical Patology; Medical Sciences Faculty, Brazil Thymus is the development site of T lymphocytes, which are important cells of the specific immune response. To perform its function, the thymic microenvironment must be preserved (Ciofani & Zuniga-Pflucker, 2007). Despite its importance, the thymus is a target organ for many infectious diseases, including paracoccidioidomycosis, the most prevalent systemic mycosis in Latin America, caused by the fungus Paracoccidioides brasiliensis (Pb) (Brito et al., 2003). We have already demonstrated in experimental models that Pb invades the thymic microenvironment, inducing severe atrophy characterized by organ weight decrease and loss of corticomedullary delimitation (Brito et al., 2003, Souto et al. 2003). We have also reported that such atrophy is related to the increased death of immature lymphocytes and to the premature migration of these cells to the periphery. However, the possibility that thymic atrophy during the paracoccidioidomycosis infection is also associated to alterations in non-lymphoid cells, which include dendritic cells (DCs), thymic cortical (cTECs) and medullary epithelial cells (mTECs) cannot be discarded. In this study, we examined the stromal cell compartment of thymuses from BALB/c mice infected with the Pb virulent isolate (Pb18) during the acute phase of the disease. By immunofluorescence technique, our results show an altered pattern of distribution of DCs as well as mTECs (Keratin-5+ cells) and cTECs (Keratin-8+) in thymuses from Pb-infected animals. Also, by flow cytometry technique, we observed phenotypic alterations in DCs and a reduction in the number of mTECs and cTECs. Altogether, these findings show that the stromal compartment is also deeply affected during thymic atrophy caused by Pb infection, complementing previous studies of our laboratory. Acknowledgements Supported by FAPESP(#2012/22131-7 & #2013/08194-9) and CAPES. References Brito, V. N., Souto, P. C. S., Cruz-Höfling, M. A., Ricci, L. C., Verinaud, L. (2003). Thymus invasion and atrophy induced by Paracoccidioides brasiliensis in BALB/c mice. Medical Mycology 41, 83-87. Ciofani, M., Zuniga-Pflucker, J. C. (2007). The thymus as an inductive site for T lymphopoiesis. Annu Rev Cell Dev Biol 23, 463-93. Souto, P. C. S., Brito, V. N., Gameiro, J., Cruz Höfling, M. A., Verinaud, L. (2003). Programmed cell death in thymus during experimental paracoccidioidomycosis. Med Microbiology and Immunology 192, 225-229. Keywords: Thymus Gland, thymus atrophy, stromal cell compartment, Paracoccidioides brasiliensis, Paracoccidioidomycosis Conference: 15th International Congress of Immunology (ICI), Milan, Italy, 22 Aug - 27 Aug, 2013. Presentation Type: Abstract Topic: Host-pathogen interactions Citation: Alves Da Costa T, Di Gangi R, Francelin C, Thomé R, Mamoni RL and Verinaud L (2013). Alterations in stromal cell compartment during thymus atrophy induced by P. brasiliensis infection.. Front. Immunol. Conference Abstract: 15th International Congress of Immunology (ICI). doi: 10.3389/conf.fimmu.2013.02.01002 Copyright: The abstracts in this collection have not been subject to any Frontiers peer review or checks, and are not endorsed by Frontiers. They are made available through the Frontiers publishing platform as a service to conference organizers and presenters. The copyright in the individual abstracts is owned by the author of each abstract or his/her employer unless otherwise stated. Each abstract, as well as the collection of abstracts, are published under a Creative Commons CC-BY 4.0 (attribution) licence (https://creativecommons.org/licenses/by/4.0/) and may thus be reproduced, translated, adapted and be the subject of derivative works provided the authors and Frontiers are attributed. For Frontiers’ terms and conditions please see https://www.frontiersin.org/legal/terms-and-conditions. Received: 29 Jun 2013; Published Online: 22 Aug 2013. * Correspondence: Prof. Liana Verinaud, State University of Campinas, Department of Structural and Functional Biology; Institute of Biology, Campinas, SP, Brazil, verinaud@unicamp.br Login Required This action requires you to be registered with Frontiers and logged in. To register or login click here. Abstract Info Abstract The Authors in Frontiers Thiago Alves Da Costa Rosaria Di Gangi Carolina Francelin Rodolfo Thomé Ronei L Mamoni Liana Verinaud Google Thiago Alves Da Costa Rosaria Di Gangi Carolina Francelin Rodolfo Thomé Ronei L Mamoni Liana Verinaud Google Scholar Thiago Alves Da Costa Rosaria Di Gangi Carolina Francelin Rodolfo Thomé Ronei L Mamoni Liana Verinaud PubMed Thiago Alves Da Costa Rosaria Di Gangi Carolina Francelin Rodolfo Thomé Ronei L Mamoni Liana Verinaud Related Article in Frontiers Google Scholar PubMed Abstract Close Back to top Javascript is disabled. Please enable Javascript in your browser settings in order to see all the content on this page.

Inhibition of PGE2 and LTB4 production by dendritic cells induced by Paracoccidioides brasiliensis is associated with the non maturation of these cells

Event Abstract Back to Event Inhibition of PGE2 and LTB4 production by dendritic cells induced by Paracoccidioides brasiliensis is associated with the non maturation of these cells Reginaldo K. Fernandes1*, Tatiana F. Bachiega1, Daniela R. Rodrigues1, Márjorie A. Golim2, Luciane A. Dias-Melicio3, Helanderson A. Balderramas1, Ramon Kaneno4 and Ângela M. Soares1* 1 Universidade Estadual Paulista "Júlio de Mesquita Filho" (UNESP) - Câmpus de Botucatu, Microbiologia e Imunologia - Instituto de Biociências, Brazil 2 Universidade Estadual Paulista "Júlio de Mesquita Filho" (UNESP), Hemocentro, Brazil 3 Universidade Estadual Paulista "Júlio de Mesquita Filho" (UNESP), Depto. Patologia - FMB - Botucatu, Brazil 4 Universidade Estadual Paulista "Júlio de Mesquita Filho" (UNESP), Microbiologia e Imunologia, Brazil The fungus Paracoccidioides brasiliensis is the etiological agent of paracoccidioidomycosis, a systemic mycosis endemic in Latin America. Dendritic cells (DCs) play a crucial role in the detection of pathogens, triggering an initial response from the host as well as the instruction of the adaptive immune response. Mediators released by DCs, by an autocrine way, modulate their functions. Among these, we highlight the prostaglandins and leukotrienes. We therefore assessed whether human DCs produce PGE2 and LTB4 in response to fungus, the participation of recognition receptors of molecular patterns (PRRs) in this production, as well as the modulating role of these eicosanoids on phenotypic maturation of these cells. Human immature DCs (CD14-/ CD1ahigh/CD83low) derived from the differentiation of monocytes cultured with GM-CSF and IL-4 (7 days) released substantial concentrations of PGE2 and LTB4 which increased significantly in the case of PGE2, with incubation with LPS for 1h, 2h, 4h, 8h, 12h, 18h, 24h or 48h. However, the levels of PGE2 and LTB4 were significantly inhibited after challenge with two different strains of the fungus. Assays using monoclonal antibodies to block PRRS showed the involvement of mannose receptor in PGE2 inhibition by the fungus. Additionally, phenotyping assays showed that after challenge with the fungus, DCs did not change their phenotype of immature cells to mature ones. Accordingly, we detected a clear association between inhibition of the production of PGE2 and LTB4 fungus and inability to induce maturation of DCs. Assays using exogenous PGE2 and LTB4 confirmed this association. Acknowledgements Financial Support: FAPESP 2011/12477-0 / 2011/04556-8 Keywords: Eicosanoids, Dendritic Cells, prostaglandin, leukotriene, Paracoccidioides brasiliensis Conference: 15th International Congress of Immunology (ICI), Milan, Italy, 22 Aug - 27 Aug, 2013. Presentation Type: Abstract Topic: Host-pathogen interactions Citation: Fernandes RK, Bachiega TF, Rodrigues DR, Golim MA, Dias-Melicio LA, Balderramas HA, Kaneno R and Soares ÂM (2013). Inhibition of PGE2 and LTB4 production by dendritic cells induced by Paracoccidioides brasiliensis is associated with the non maturation of these cells. Front. Immunol. Conference Abstract: 15th International Congress of Immunology (ICI). doi: 10.3389/conf.fimmu.2013.02.00536 Copyright: The abstracts in this collection have not been subject to any Frontiers peer review or checks, and are not endorsed by Frontiers. They are made available through the Frontiers publishing platform as a service to conference organizers and presenters. The copyright in the individual abstracts is owned by the author of each abstract or his/her employer unless otherwise stated. Each abstract, as well as the collection of abstracts, are published under a Creative Commons CC-BY 4.0 (attribution) licence (https://creativecommons.org/licenses/by/4.0/) and may thus be reproduced, translated, adapted and be the subject of derivative works provided the authors and Frontiers are attributed. For Frontiers’ terms and conditions please see https://www.frontiersin.org/legal/terms-and-conditions. Received: 25 Apr 2013; Published Online: 22 Aug 2013. * Correspondence: Mr. Reginaldo K Fernandes, Universidade Estadual Paulista "Júlio de Mesquita Filho" (UNESP) - Câmpus de Botucatu, Microbiologia e Imunologia - Instituto de Biociências, Botucatu-SP, Brazil, regiskeller@msn.com Prof. Ângela M Soares, Universidade Estadual Paulista "Júlio de Mesquita Filho" (UNESP) - Câmpus de Botucatu, Microbiologia e Imunologia - Instituto de Biociências, Botucatu-SP, Brazil, acsoares@ibb.unesp.br Login Required This action requires you to be registered with Frontiers and logged in. To register or login click here. Abstract Info Abstract The Authors in Frontiers Reginaldo K Fernandes Tatiana F Bachiega Daniela R Rodrigues Márjorie A Golim Luciane A Dias-Melicio Helanderson A Balderramas Ramon Kaneno Ângela M Soares Google Reginaldo K Fernandes Tatiana F Bachiega Daniela R Rodrigues Márjorie A Golim Luciane A Dias-Melicio Helanderson A Balderramas Ramon Kaneno Ângela M Soares Google Scholar Reginaldo K Fernandes Tatiana F Bachiega Daniela R Rodrigues Márjorie A Golim Luciane A Dias-Melicio Helanderson A Balderramas Ramon Kaneno Ângela M Soares PubMed Reginaldo K Fernandes Tatiana F Bachiega Daniela R Rodrigues Márjorie A Golim Luciane A Dias-Melicio Helanderson A Balderramas Ramon Kaneno Ângela M Soares Related Article in Frontiers Google Scholar PubMed Abstract Close Back to top Javascript is disabled. Please enable Javascript in your browser settings in order to see all the content on this page.

Important aspects of oral paracoccidioidomycosis–a literature review

Paracoccidioidomycosis is a deep mycosis endemic to Latin America, with considerable morbidity and mortality. It is caused by the dimorphic fungus Paracoccidioides brasiliensis, which affects, among other organs in the human body, the oral cavity. Fungus virulence and immunocompetence of the host determine the establishment of infection or active disease, whose severity and clinical behaviour depend mostly on the cellular immune response of the host. Often, oral lesions constitute the first sign and site of confirmation of diagnosis, which in most cases is delayed. The success of the treatment depends on early and correct diagnosis, as well as on the patient's adherence to the drug therapy.

Osteomyelitis and pyoarthritis resulting from local paracoccidioidomycosis in an immunocompetent patient: a case report

IntroductionParacoccidioidomycosis is a type of mycosis that is endemic to Brazil and is triggered by the fungus Paracoccidioides brasiliensis. Isolated bone involvement in this disease is very rare, especially in children. To the best of our knowledge this report documents the first case of an immunocompetent pediatric patient in which paracoccidioidomycosis of the hip articulation was the sole manifestation of the disease (that is, there were no pulmonary or skin lesions).Case presentationAn 11-year-old Brazilian Caucasian boy from a rural area was examined in the orthopedic ward of our emergency department. Our patient reported a three-month history of pain in the right hip with intermittent claudication and also complained of recurring episodes of intense pain and an inability to walk, which he had been experiencing for the previous five days. He additionally presented with a fever that had persisted for two days. Our patient’s medical history did not include any clinical respiratory manifestations, skin lesions, history of trauma or immunosuppression risk factors.ConclusionsThis is one of the very few reported cases of isolated articular involvement in osteomyelitis in a pediatric immunocompetent patient. Paracoccidioidomycosis should be considered among the differential diagnoses in such cases, especially in cases of patients who reside in rural areas where the condition is considered to be endemic, in order to administer the proper course of treatment in a timely fashion and improve the chances of a favorable prognosis.

A Paracoccidioides brasiliensis glycan shares serologic and functional properties with cryptococcal glucuronoxylomannan

The cell wall of the yeast form of the dimorphic fungus Paracoccidioides brasiliensis is enriched with α1,3-glucans. In Cryptococcus neoformans, α1,3-glucans interact with glucuronoxylomannan (GXM), a heteropolysaccharide that is essential for fungal virulence. In this study, we investigated the occurrence of P. brasiliensis glycans sharing properties with cryptococcal GXM. Protein database searches in P. brasiliensis revealed the presence of sequences homologous to those coding for enzymes involved in the synthesis of GXM and capsular architecture in C. neoformans. In addition, monoclonal antibodies (mAbs) raised to cryptococcal GXM bound to P. brasiliensis cells. Using protocols that were previously established for extraction and analysis of C. neoformans GXM, we recovered a P. brasiliensis glycan fraction composed of mannose and galactose, in addition to small amounts of glucose, xylose and rhamnose. In comparison with the C. neoformans GXM, the P. brasiliensis glycan fraction components had smaller molecular dimensions. The P. brasiliensis components, nevertheless, reacted with different GXM-binding mAbs. Extracellular vesicle fractions of P. brasiliensis also reacted with a GXM-binding mAb, suggesting that the polysaccharide-like molecule is exported to the extracellular space in secretory vesicles. An acapsular mutant of C. neoformans incorporated molecules from the P. brasiliensis extract onto the cell wall, resulting in the formation of surface networks that resembled the cryptococcal capsule. Coating the C. neoformans acapsular mutant with the P. brasiliensis glycan fraction resulted in protection against phagocytosis by murine macrophages. These results suggest that P. brasiliensis and C. neoformans share metabolic pathways required for the synthesis of similar polysaccharides and that P. brasiliensis yeast cell walls have molecules that mimic certain aspects of C. neoformans GXM. These findings are important because they provide additional evidence for the sharing of antigenically similar components across phylogenetically distant fungal species. Since GXM has been shown to be important for the pathogenesis of C. neoformans and to elicit protective antibodies, the finding of similar molecules in P. brasiliensis raises the possibility that these glycans play similar functions in paracoccidiomycosis.

PARACOCIDIOIDOMICOSIS DISEMINADA CRONICA COMO PRIMERA MANIFESTACION DE SIDA

Paracoccidioidomycosis (PCM) is a systemic fungal infection endemic in subtropical areas of Central and South America and caused by the dimorphic fungus Paracoccidioides brasiliensis. PCM is a prevalent disease in Latin America, from Mexico (20 degrees northern latitude) to Argentina (35 degrees southern latitude). This distribution includes tropical and subtropical regions and endemic countries with mild temperatures and high and constant humidity throughout the year. Approximately 10 million people are infected by this fungus, however only 2% may develop a disseminated form of the disease. The constant migration from rural to urban areas increases the risk of reactivation of quiescent forms of PCM outside the endemic areas in immunocompetent as well as in immunosupressed patients. The chronic disseminated form of this disease is the most frequent clinical manifestation, characterized by the pulmonary and mucocutaneous involvement. We present the case of an HIV seropositive male with chronic disseminated paracoccidioidomycosis as the first manifestation of retroviral infection. Diagnosis was made based on the epidemiological, clinical, histopathological and microbiological findings.

New β-carboline alkaloids from Galianthe ramosa (Rubiaceae)

Galianthe Griseb. (Rubiaceae, tribe Spermacoceae) is a genus comprising 50 endemic species in South America, its main center of diversity is located in central and southern Brazil. In our continuing chemical and biological investigations concerning Rubiaceae species from Cerrado, we report the isolation of new compounds 1 and 2, together with the known epi-catechin (3), ursolic and oleanolic acid. The alkaloid 2 inhibited the enzyme malato synthase from pathogenic fungus Paracoccidioides brasiliensis wich is considered an important target since it is not found in human.

Paracoccidioides brasiliensis lipids modulate macrophage activity via Toll-dependent or independent mechanisms

The macrophages are the first host cells that interact with the fungus Paracoccidioides brasiliensis, but the main mechanisms that regulate this interaction are not well understood. Because the role played by P. brasiliensis lipids in macrophage activation was not previously investigated, we aimed to assess the influence of diverse lipid fractions from P. brasiliensis yeasts in this process. The possible participation of TLR2 and TLR4 signaling was also evaluated using TLR2- and TLR4-defective macrophages. Four lipid-rich fractions were studied as follows: F1, composed by membrane phospholipids and neutral lipids, F2 by glycolipids of short chain, F3a by membrane glycoproteins anchored by glycosylphosphatidylinositol (GPI) groups, and F3b by glycolipids of long chain. All assayed lipid fractions were able to activate peritoneal macrophages and induce nitric oxide (NO) production. Importantly, the F1 and F3a fractions exerted opposite effects in the control of P. brasiliensis uptake and killing, but both fractions inhibited cytokines production. Furthermore, the increased NO production and expression of costimulatory molecules induced by F3a was shown to be TLR2 dependent although F1 used Toll-independent mechanisms. In conclusion, our work suggests that lipid components may play a role in the innate immunity against P. brasiliensis infection using Toll-dependent and independent mechanisms to control macrophage activation.

High IL-4 expression and IL-4 gene polymorphisms are associated with susceptibility to human paracoccidioidomycosis (164.21)

Abstract Paracoccidioidomycosis is caused by the dimorphic fungus Paracoccidioides brasiliensis. After invasion, the fungi are immediately destroyed or overcome the local defenses of the host and begin multiplication, causing initial injury. Previous studies have demonstrated that the severity of disease is associated with a Th2 immune response characterized by high IL-4 production. In the present study, we analyzed two polymorphisms in the IL-4 gene, -590 C/T and the intron-3 microsatellite, in 76 patients with paracoccidioidomycosis and in 73 control individuals from an endemic area. A significant correlation was observed between the RP2/RP2 intron-3 genotype and infection with P. brasiliensis (p=0.011), while the RP1/RP1 genotype was correlated with resistance. The IL-4 promoter polymorphism showed a significant correlation of the presence of T allele with the disease (p=0.030). Furthermore, high IL-4 expression observed on the patient group compared with control and was associated with TT genotype. This result suggests that IL-4 polymorphisms are associated with the ability of the host to control P. brasiliensis infection. The potential biological role of this polymorphism is supported by our observation that patients with disease produce high levels of IL-4 after mitogen or antigen stimulation and the association of high IL-4 levels with TT genotype. IL-4 gene is located in the cytokine cluster region of chromosome 5, where other polymorphisms have been described.

Production of TNF-α, nitric oxide and hydrogen peroxide by macrophages from mice with paracoccidioidomycosis that were fed a linseed oil-enriched diet

Omega-3 polyunsaturated fatty acids (n-3 PUFA) can modulate the immune system and their primary effect is on macrophage function. Paracoccidioidomycosis (PCM) is an endemic systemic mycosis in Latin America that is caused by the dimorphic fungus Paracoccidioides brasiliensis (Pb). Macrophages are the main defence against this pathogen and have microbicidal activity that is dependent on interferon-Γ and tumour necrosis factor (TNF)-α. These cytokines stimulate the synthesis of nitric oxide (NO) and hydrogen peroxide (H₂O₂), leading to the death of the fungus. To study the effect of n-3 PUFA on the host immune response during experimental PCM, macrophages that were obtained from animals infected with Pb18 and fed a diet enriched by linseed (LIN) oil were cultured and challenged with the fungus in vitro. The macrophage function was analysed based on the concentrations of TNF-α, NO and H₂O₂. LIN oil seems to influence the production of TNF-α during the development of disease. A diet enriched with LIN oil influences the microbicidal activity of the macrophages by inducing the production of cytokines and metabolites such as NO and H₂O₂, predominantly in the chronic phase of infection.

Surface-expressed enolase contributes to the adhesion of Paracoccidioides brasiliensis to host cells

Paracoccidioidomycosis is a systemic mycosis caused by the dimorphic fungus Paracoccidioides brasiliensis. Understanding the interactions between P. brasiliensis and the host tissue depends on the study of the different steps of the process of colonization, especially adhesion, in which the pathogen recognizes ligands on the surface of host cells. This study aimed to verify the role of enolase in the host cell-fungus interaction and the ability of enolase to bind to extracellular matrix components, to determine its subcellular localization, and to study the P. brasiliensis enolase amino acid sequence. The data revealed that fibronectin is the major ligand of enolase. Evaluation of the location of enolase at an ultrastructural level revealed that it is distributed in various cellular compartments, but at a high level in the cell wall. The analysis of the amino acid sequence revealed an internal plasminogen-binding motif ((254)FYKADEKKY(262)), which is conserved in most organisms and described as an important interaction site of the enolase with the host cell surface. This suggests that enolase performs additional functions related to the glycolytic pathway and also plays a role of adhesion in P. brasiliensis. Therefore, this study increases the knowledge about the characteristics of enolase and its influence on the binding process of P. brasiliensis.

Experimental Model of Arthritis Induced by Paracoccidioides brasiliensis in Rats

Paracoccidioidomycosis (PCM), a disease caused by the fungus Paracoccidioides brasiliensis (Pb), is highly prevalent in Brazil, where it is the principal cause of death by systemic mycoses. The disease primarily affects men aged 30-50 year old and usually starts as a pulmonary focus and then may spread to other organs and systems, including the joints. The present study aimed to develop an experimental model of paracoccidioidomycotic arthritis. Two-month-old male Wistar rats (n = 48) were used, divided in 6 groups: test groups EG/15 and EG/45 (received one dose of 100 μl of saline containing 10(5) Pb viable yeasts in the knee); heat killed Pb-group HK/15 and HK/45 (received a suspension of 10(5) Pb nonviable yeasts in the knee) and control groups CG/15 and CG/45 (received only sterile saline in the knee). The rats were killed 15 and 45 days postinoculation. In contrast with the control rats, the histopathology of the joints of rats of the test groups (EG/15 and EG/45) revealed a picture of well-established PCM arthritis characterized by extensive sclerosing granulomatous inflammation with numerous multiple budding fungal cells. The X-ray examination revealed joint alterations in these groups. Only metabolic active fungi evoked inflammation. The experimental model was able to induce fungal arthritis in the knees of the rats infected with metabolic active P. brasiliensis. The disease tended to be regressive and restrained by the immune system. No evidence of fungal dissemination to the lungs was observed.

Antifungal activity of altenusin isolated from the endophytic fungus Alternaria sp. against the pathogenic fungus Paracoccidioides brasiliensis

BackgroundAltenusin is a biphenyl derivative isolated from different species of fungi, which presents several biological activities. AimsWe report the antifungal activity of the altenusin isolated from the endophytic fungus Alternaria sp., against clinical isolates of Paracoccidioides brasiliensis, and its action on cell walls of P. brasiliensis and the nonpathogenic yeast Schizosaccharomyces pombe. MethodsIn vitro antifungal activity of altenusin was evaluated using the broth microdilution method against 11 strains of P. brasiliensis and one strain of S. pombe. The effects of the altenusin on the cell wall were estimated using the sorbitol protection assay. ResultsThe altenusin presented strong activity against P. brasiliensis with MIC values ranging between 1.9 and 31.2μg/ml, and 62.5μg/ml for S. pombe. Our results demonstrated that the MIC values for altenusin were increased for P. brasiliensis Pb18 and for S. pombe when the medium was supplemented with sorbitol. Additionally, S. pombe cells treated with altenusin were more rounded in shape than untreated cells. ConclusionsAltenusin showed activity against clinical strains of P. brasiliensis at the concentration tested, and this compound probably affects fungal cell walls. These findings suggest that altenusin could act through the inhibition of cell wall synthesis or assembly in P. brasiliensis and S. pombe, and could be considered as a lead compound for the design of new antifungals.

Comparative transcriptome analysis of Paracoccidioides brasiliensis during in vitro adhesion to type I collagen and fibronectin: identification of potential adhesins

Paracoccidioidomycosis is caused by the dimorphic fungus Paracoccidioides brasiliensis. The extracellular matrix (ECM) plays an important role in regulation of cell adhesion, differentiation, migration and proliferation of cells. An in vitro binding assay of P. brasiliensis yeast cells adhering to type I collagen and fibronectin was performed in order to identify novel adhesins. Representational difference analysis (RDA) was employed to identify genes upregulated under adhesion-inducing conditions. Expressed sequence tags (ESTs) from cDNA libraries generated by the RDA technique were analyzed. Genes related to functional categories, such as metabolism, transcription, energy, protein synthesis and fate, cellular transport and biogenesis of cellular components were upregulated. Transcripts encoding the P. brasiliensis protein enolase (PbEno) and the high-affinity cooper transporter (PbCtr3) were identified and further characterized. The recombinant enolase (rPbEno) and a synthetic peptide designed for PbCtr3 were obtained and demonstrated to be able to bind ECM components. Immunofluorescence assays demonstrated that rPbEno specifically binds to the macrophage surface, reinforcing the role of this molecule in the P. brasiliensis interaction with host cells. In addition, upregulation of selected genes was demonstrated by qRT-PCR. In synthesis, the strategy can be useful in characterization of potential P. brasiliensis adhesins.

Synthetic Peptides Mimic gp75 from Paracoccidioides brasiliensis in the Diagnosis of Paracoccidioidomycosis

Paracoccidioidomycosis (PCM) is a systemic granulomatous disease, endemic in Latin America, caused by the thermal dimorphic fungus Paracoccidioides brasiliensis. Although some fungal antigens have already been characterized and used for serological diagnosis, cross-reactions have been frequently observed. Thus, the examination of fungal forms in clinical specimens or isolation of P. brasiliensis by culture is still the most frequent method for the diagnosis of this mycosis. In this study, a random peptide phage display library was used to select mimotopes of P. brasiliensis, which were employed as antigens in an indirect enzyme-linked immunosorbent assay. The protective monoclonal antibody against experimental PCM (anti-gp75) was used as molecular target to screen a phage display library. That approach led to a synthetic peptide named P2, which was synthesized and tested against PCM patients’ sera to check whether it was recognized. There was significant recognition of P2 by sera of untreated PCM patients when compared with normal human sera. Sera from treated PCM group, patients with other mycosis or co-infected with HIV had much lower recognition of P2 than untreated patient group. The test showed a sensitivity of 100 and 94.59% of specificity in relation to human sera control. These data indicate a potential use of P2 as diagnostic tool in PCM. Its application for serological diagnosis of PCM may contribute to the development and standardization of simpler, faster and highly reproducible immunodiagnostic tests at low cost.

Paracoccidioides brasiliensis GP43-derived peptides are potent modulators of local and systemic inflammatory response

Paracoccidioidomycosis is a systemic granulomatous disease caused by the dimorphic fungus Paracoccidioides brasiliensis. Its major antigen is a 43 kDa glycoprotein whose peptides embody different functions: P10 peptide, a T-cell epitope, induces protective response while P4 and P23 peptides inhibit both, macrophage functions and inflammatory reaction, thus facilitating infection. Here we investigated the modulating mechanisms of the immune response exerted by P4 and P23 involved in the latter inhibitory effect on macrophages. Moreover we analyzed the peptides effects in different models in vivo. While evaluating whether P4 and P23 present systemic anti-inflammatory effects in vivo, we showed that their intraperitonial administration decreased footpad swelling in mice infected with either P. brasiliensis or Mycobacterium bovis. Both, qPCR and ELISA assays suggested that this anti-inflammatory effect depended on alterations in the kinetics of production of innate immunity modulators such as TNF-α, IL6, IL10 and TLR2. IL10 seems to be early produced than TNF-α and IL6, produced later in presence of peptides. Higher doses or intravenously given P4 and P23 resulted in earlier and more prolonged anti-inflammatory effects. Moreover, continuous treatment with P4 and P23 sustained the anti-inflammatory activity throughout.

A retrospective study of oral manifestations in patients with paracoccidioidomycosis

South American blastomycosis, paracoccidioidomycosis (Pb mycosis) or Lutz disease is an endemically fungal infection in Latin America. It is caused by the dimorphic fungus Paracoccidioides brasiliensis and may cause oral mucosal lesions. The incidence of Pb mycosis oral lesions was evaluated in patients assisted at a Brazilian Dental School's Specialized Oral Diagnosis Service with special focus on the different clinical forms of these lesions, its location, patients' occupation, deleterious habits, and diagnosis methodology. Students' and professionals' initial diagnoses were compared with the definitive diagnosis. Lesions were detected 31 cases (18 patients). The results show that 88.8% of the patients were male with a mean age of 50 years and 39% work(ed) with activities related to agriculture. As much as 88.9% were smokers and 72.2% were alcohol users. Exfoliative cytology was performed in 66.6% of the patients. Oral mucosa (30%), gingiva (16.6%) and lips 16.6% were the most common sites of Pb mycosis oral lesions. Comparing the initial with the definitive diagnosis made by the professionals their accuracy was 33% (6 out of 18 patients). Students' diagnosis was more accurate demonstrating 72.5% of initial correct diagnosis (13 out of 18). Statistical analysis by ANOVA (α=0.05, SPSS WIN) demonstrated a significant difference between the diagnosis of Pb mycosis made by students and professionals when considering initial diagnosis and final diagnosis (after histopathological analysis) (p=0.25). Incisional biopsy and exfoliate cytology are efficient for an early diagnosis of this disease in mouth. Students' training in diagnosis of oral pathologies to recognize lesions is urgent to improve public health.

The role of adjuvants in therapeutic protection against paracoccidioidomycosis after immunization with the P10 peptide

Paracoccidioidomycosis (PCM), a common chronic mycosis in Latin America, is a granulomatous systemic disease caused by the thermo-dimorphic fungus Paracoccidioides brasiliensis. The glycoprotein gp43 is the main antigen target of P. brasiliensis and a 15-mer internal peptide (QTLIAIHTLAIRYAN), known as P10, defines a major CD4+-specific T cell epitope. Previous results have indicated that, besides having a preventive role in conventional immunizations prior to challenge with the fungus, protective anti-fungal effects can be induced in P. brasiliensis-infected mice treated with P10 administered with complete Freund’s adjuvant (CFA). The peptide elicits an IFN-γ-dependent Th1 immune response and is the main candidate for effective immunotherapy of patients with PCM, as an adjunctive approach to conventional chemotherapy. In the present study we tested the therapeutic effects of P10 combined with different adjuvants [aluminum hydroxide, CFA, flagellin, and the cationic lipid dioctadecyl-dimethylammonium bromide (DODAB)] in BALB/c mice previously infected with the P. brasiliensis Pb18 strain. Significant reductions in the number of colony forming units of the fungus were detected in lungs of mice immunized with P10 associated with the different adjuvants 52 days after infection. Mice treated with DODAB and P10, followed by mice treated with P10 and flagellin, showed the most prominent effects as demonstrated by the lowest numbers of viable yeast cells as well as reductions in granuloma formation and fibrosis. Concomitantly, secretion of IFN-γ and TNF-α, in contrast to interleukin (IL)-4 and IL-10, was enhanced in the lungs of mice immunized with P10 in combination with the tested adjuvants, with the best results observed in mice treated with P10 and DODAB. In conclusion, the present results demonstrate that the co-administration of the synthetic P10 peptide with several adjuvants, particularly DODAB, have significant therapeutic effects in experimental PCM.

ArtinM offers new perspectives in the development of antifungal therapy

The thermally dimorphic fungus Paracoccidioides brasiliensis is the causative agent of paracoccidioidomycosis (PCM), the most frequent systemic mycosis that affects the rural populations in Latin America. Despite significant developments in antifungal chemotherapy, its efficacy remains limited since drug therapy is prolonged and associated with toxic side effects and relapses. In response to these challenges, it is now recognized that several aspects of antifungal immunity can be modulated to better deal with fungal infections. A common idea for halting fungal infections has been the need to activate a cell-based, pro-inflammatory Th1 immune response to improve the fungal elimination. ArtinM, a D-mannose binding lectin from Artocarpus heterophyllus, has the property of modulating immunity against several intracellular pathogens. Here, we review the immunomodulatory activity of ArtinM during experimental PCM in mice. Both prophylactic and therapeutic protocols of ArtinM administration promotes a Th1 immune response balanced by IL-10, which outstandingly reduces the fungal load in organs of the treated mice while maintaining a controlled inflammation at the site of infection. A carbohydrate recognition-based interaction of ArtinM with Toll-like receptor 2 (TLR2) accounts for initiating the immunomodulatory effect of the lectin. The precise identification of the TLR2 N-glycan(s) targeted by ArtinM may support novel basis for the development of antifungal therapy.