RESUMO
Ganoderma formosanum (GF) is a medicinal mushroom endemic to Taiwan. Previous research established the optimal culture conditions to produce exopolysaccharide rich in ß-glucan (GF-EPS) from submerged fermentation of GF. The present study investigated the antitumor effects of GF-EPS in a Lewis lung carcinoma cell (LLC1) tumor-bearing mice model. In the preventive model, GF-EPS was orally administered to mice before LLC1 injection. In the therapeutic model, GF-EPS oral administration was initiated five days after tumor cell injection. The tumor size and body weight of the mice were recorded. After sacrifice, the lymphocyte subpopulation was analyzed using flow cytometry. Spleen tissues were used to analyze cytokine mRNA expression. The results showed that GF-EPS (80 mg/kg) effectively suppressed LLC1 tumor growth in both the preventive and therapeutic models. GF-EPS administration increased the proportion of natural killer cells in the spleen and activated gene expression of several cytokines. Our results provide evidence that GF-EPS promotes tumor inhibition through immunomodulation in tumor-bearing mice.
Assuntos
Carcinoma Pulmonar de Lewis/tratamento farmacológico , Citocinas/genética , Polissacarídeos Fúngicos/administração & dosagem , Ganoderma/crescimento & desenvolvimento , Células Matadoras Naturais/metabolismo , Administração Oral , Animais , Peso Corporal/efeitos dos fármacos , Carcinoma Pulmonar de Lewis/genética , Carcinoma Pulmonar de Lewis/imunologia , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Fermentação , Polissacarídeos Fúngicos/imunologia , Polissacarídeos Fúngicos/farmacologia , Ganoderma/imunologia , Ganoderma/metabolismo , Regulação Neoplásica da Expressão Gênica , Imunomodulação , Células Matadoras Naturais/efeitos dos fármacos , Camundongos , Baço/imunologia , Carga Tumoral/efeitos dos fármacos , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
In the present study, we evaluated adjuvant potential of Poria cocos polysaccharide (PCP) on the Th1-type immune responses of C57/BL6 mice against ovalbumin (OVA). We first determined the effect of PCP on maturation of murine bone marrow derived dendritic cells (BMDCs), PCP significantly upregulated surface expression of MHCII, CD40, CD80, CD86 and enhanced production of IL-6 and IL-12p40. In addition, PCP affected receptor-mediated endocytosis, but not pinocytosis in BMDCs. Furthermore, OVA + PCP immunization induced specific cytotoxic CD8+ T cell killing of OVA (257-264) peptide pulsed cell. When mice were immunized subcutaneously in a week interval with OVA + PCP. Serum were collected for measuring OVA-specific antibody and splenocytes were harvested for analyzing CD69, IFN-γ ELISpot and cytokines production. The result indicated that OVA-specific IgG, IgG2a and IgG1 antibody levels in serum were significantly elevated by PCP compared with control. PCP increased OVA-specific IFN-γ-secreting CD8+, CD4+ T cells, promoted CD8+ T cell proliferation and up-regulated Th-1 type (IFN-γ, IL-2) cytokine production. In conclusion, data suggest that PCP enhanced cellular immune response and possess potential as a vaccine adjuvant for Th1 immune response.
Assuntos
Polissacarídeos Fúngicos/imunologia , Imunidade Celular/efeitos dos fármacos , Ovalbumina/imunologia , Células Th1/imunologia , Wolfiporia/química , Animais , Formação de Anticorpos/imunologia , Células da Medula Óssea/imunologia , Linfócitos T CD8-Positivos/imunologia , Citocinas/imunologia , Células Dendríticas/imunologia , Feminino , Polissacarídeos Fúngicos/química , Imunoglobulina G/imunologia , CamundongosRESUMO
Polysaccharides are the most abundant bioactive compounds in Ganoderma and have been widely used as dietary supplements in traditional Chinese medicine for thousands of years. Polysaccharides from Ganoderma exhibit unique biological properties, including anti-tumor, anti-inflammatory, and immunomodulatory activities. Herein, the sources and structures of polysaccharides from Ganoderma were presented. This work also reviews the immunomodulatory activities and possible mechanisms of polysaccharides from Ganoderma on different immune effector cells, including lymphocytes and myeloid cells. As an available adjunctive remedy, polysaccharides from Ganoderma can potentially be applied for the modulation of the host immune system, namely the innate immunity, the cellular immunity, and the humoral immunity.
Assuntos
Polissacarídeos Fúngicos/química , Polissacarídeos Fúngicos/farmacologia , Ganoderma/química , Sistema Imunitário/efeitos dos fármacos , Fatores Imunológicos/farmacologia , Animais , Antineoplásicos Imunológicos/farmacologia , Polissacarídeos Fúngicos/imunologia , Humanos , Imunidade Celular/efeitos dos fármacos , Imunidade Inata/efeitos dos fármacos , Linfócitos/efeitos dos fármacos , Linfócitos/imunologiaRESUMO
Medicinal mushrooms, Coriolus versicolor and Lentinus edodes are extremely attractive as nutraceuticals. Here we used fruiting bodies to prepare novel kombucha beverage. Microbiological, physicochemical and chemical properties were monitored for eleven days, while the immunological properties of kombucha polysaccharide extracts were determined in peripheral blood mononuclear cell (PBMC) cultures. FTIR analysis of polysaccharide extracts showed dominant presence of polysaccharides, in addition to phenols, lipids and proteins. C. versicolor kombucha extract displayed more complex polysaccharides, and a higher content of total polysaccharides, phenols and flavonoids compared to L. edodes kombucha extract. The extracts were not cytotoxic for PBMC in vitro up to 500 µg/ml, while immunomodulatory effects depended on their chemical compositions. The most prominent effect was on the reduction of Th2 cytokines and IL-10 in PBMC cultures. Based on these results, novel kombucha products could be recommended as functional beverages or nutraceuticals with potentially beneficial immunomodulatory effects in allergies.
Assuntos
Bebidas/microbiologia , Fermentação , Polissacarídeos Fúngicos/imunologia , Fatores Imunológicos/farmacologia , Polyporaceae/química , Polyporaceae/metabolismo , Polissacarídeos Fúngicos/química , Fatores Imunológicos/química , Leucócitos Mononucleares/efeitos dos fármacos , Leucócitos Mononucleares/imunologiaRESUMO
Even though both cellular and humoral immunities contribute to host defense, the role played by humoral immunity against the airborne opportunistic fungal pathogen Aspergillus fumigatus has been underexplored. In this study, we aimed at deciphering the role of the complement system, the major humoral immune component, against A. fumigatus Mass spectrometry analysis of the proteins extracted from A. fumigatus conidial (asexual spores and infective propagules) surfaces opsonized with human serum indicated that C3 is the major complement protein involved. Flow cytometry and immunolabeling assays further confirmed C3b (activated C3) deposition on the conidial surfaces. Assays using cell wall components of conidia indicated that the hydrophobin RodAp, ß-(1,3)-glucan (BG) and galactomannan (GM) could efficiently activate C3. Using complement component-depleted sera, we showed that while RodAp activates C3 by the alternative pathway, BG and GM partially follow the classical and lectin pathways, respectively. Opsonization facilitated conidial aggregation and phagocytosis, and complement receptor (CR3 and CR4) blockage on phagocytes significantly inhibited phagocytosis, indicating that the complement system exerts a protective role against conidia by opsonizing them and facilitating their phagocytosis mainly through complement receptors. Conidial opsonization with human bronchoalveolar lavage fluid (BALF) confirmed C3 to be the major complement protein interacting with conidia. Nevertheless, complement C2 and mannose-binding lectin (MBL), the classical and lectin pathway components, respectively, were not identified, indicating that BALF activates the alternative pathway on the conidial surface. Moreover, the cytokine profiles were different upon stimulation of phagocytes with serum- and BALF-opsonized conidia, highlighting the importance of studying interaction of conidia with complement proteins in their biological niche.
Assuntos
Aspergillus fumigatus/imunologia , Líquido da Lavagem Broncoalveolar/imunologia , Complemento C3/imunologia , Polissacarídeos Fúngicos/farmacologia , Macrófagos/efeitos dos fármacos , Soro/imunologia , Esporos Fúngicos/imunologia , Aspergilose/genética , Aspergilose/imunologia , Aspergilose/microbiologia , Aspergillus fumigatus/química , Líquido da Lavagem Broncoalveolar/química , Líquido da Lavagem Broncoalveolar/microbiologia , Parede Celular/química , Parede Celular/imunologia , Ativação do Complemento/efeitos dos fármacos , Complemento C3/genética , Citocinas/biossíntese , Citocinas/imunologia , Polissacarídeos Fúngicos/imunologia , Polissacarídeos Fúngicos/isolamento & purificação , Galactose/análogos & derivados , Interações entre Hospedeiro e Microrganismos/imunologia , Humanos , Imunidade Celular , Imunidade Humoral , Integrina alfaXbeta2/genética , Integrina alfaXbeta2/imunologia , Antígeno de Macrófago 1/genética , Antígeno de Macrófago 1/imunologia , Macrófagos/imunologia , Macrófagos/microbiologia , Mananas/imunologia , Mananas/isolamento & purificação , Mananas/farmacologia , Proteínas Opsonizantes/farmacologia , Fagocitose/efeitos dos fármacos , Cultura Primária de Células , Ligação Proteica , Espécies Reativas de Oxigênio , Soro/química , Soro/microbiologia , Esporos Fúngicos/química , beta-Glucanas/imunologia , beta-Glucanas/isolamento & purificação , beta-Glucanas/farmacologiaRESUMO
Fungal bioactive polysaccharides are well known and have been widely used in Asia as a part of the traditional diet and medicine. In fact, some biopolymers (mainly ß-glucans or glycoconjugate) have already made their way to the market as antitumor or immunostimulating drugs. In the last decades, the relationship between structure and activity of polysaccharides and their detailed mode of action have been the core of intense research to understand and utilize their medicinal properties. Most of the antitumor polysaccharides belong to conserved ß-glucans, with a linear ß-(1â3)-glucan backbone and attached ß-(1â6) branch. Structurally different ß-glucans appear to have different affinities toward their receptors and thus generate markedly different host responses. However, their antitumor activities are mainly influenced by molecular mass, degree of branching, conformation, and structure modification of the polysaccharides. ß-Glucans act on several immune receptors including Dectin-1, complement receptor (CR3) and TLR-2/6, then trigger both innate and adaptive response and enhance opsonic and nonopsonic phagocytosis. Various receptor interactions explain the possible mode of actions of polysaccharides.
Assuntos
Polissacarídeos Fúngicos/farmacologia , Animais , Antineoplásicos/farmacologia , Polissacarídeos Fúngicos/química , Polissacarídeos Fúngicos/imunologia , Humanos , Modelos Biológicos , Conformação Molecular , Relação Estrutura-Atividade , beta-Glucanas/químicaRESUMO
Current antifungal drugs present poor effectiveness and there is no available vaccine for fungal infections. Thus, novel strategies to treat or prevent invasive mycosis, such as cryptococcosis, are highly desirable. One strategy is the use of immunomodulators of polysaccharide nature isolated from mushrooms. The purpose of the present work was to evaluate the immunostimulatory activity of ß-(1,3)-glucan-containing exopolysaccharides (EPS) from the edible mushrooms Auricularia auricula in phagocytes and mice infected with Cryptococcus neoformans. EPS triggered macrophages and dendritic cell activation upon binding to Dectin-1, a pattern recognition receptor of the C-type lectin receptor family. Engagement of Dectin-1 culminated in pro-inflammatory cytokine production and cell maturation via its canonical Syk-dependent pathway signaling. Furthermore, upon EPS treatment, M2-like phenotype macrophages, known to support intracellular survival and replication of C. neoformans, repolarize to M1 macrophage pattern associated with enhanced production of the microbicidal molecule nitric oxide that results in efficient killing of C. neoformans. Treatment with EPS also upregulated transcript levels of genes encoding products associated with host protection against C. neoformans and Dectin-1 mediated signaling in macrophages. Finally, orally administrated ß-glucan-containing EPS from A. auricular enhanced the survival of mice infected with C. neoformans. In conclusion, the results demonstrate that EPS from A. auricula exert immunostimulatory activity in phagocytes and induce host protection against C. neoformans, suggesting that polysaccharides from this mushroom may be promising as an adjuvant for vaccines or antifungal therapy.
Assuntos
Agaricales/química , Criptococose/prevenção & controle , Polissacarídeos Fúngicos/imunologia , Fagócitos/efeitos dos fármacos , Fagócitos/imunologia , beta-Glucanas/imunologia , Animais , Criptococose/imunologia , Cryptococcus neoformans/imunologia , Citocinas/imunologia , Células Dendríticas/efeitos dos fármacos , Células Dendríticas/imunologia , Células Dendríticas/microbiologia , Fatores Imunológicos/farmacologia , Lectinas Tipo C/imunologia , Pneumopatias Fúngicas , Macrófagos/efeitos dos fármacos , Macrófagos/imunologia , Macrófagos/microbiologia , Camundongos , Camundongos Endogâmicos C57BL , Fagócitos/microbiologia , Transdução de Sinais , beta-Glucanas/farmacologiaRESUMO
The incidence and prevalence of serious fungal infections is rising, especially in immunosuppressed individuals. Moreover, co-administration of antibiotics and immunosuppressants has driven the emergence of new multidrug-resistant pathogens. The significant increase of multidrug-resistant pathogens, together with their ability to form biofilms, is associated with morbidity and mortality. Research on novel synthetically prepared immunomodulators as potential antifungal immunotherapeutics is of serious interest. Our study demonstrated the immunobiological activity of synthetically prepared biotinylated mannooligosaccharides mimicking Candida antigenic factors using RAW264.7 macrophages. Macrophage exposure to the set of eight structurally different mannooligosaccharides induced a release of Th1, Th2, Th17, and Treg cytokine signature patterns. The observed immune responses were tightly associated with structure, dose, exposure time, and selected signature cytokines. The viability/cytotoxicity of the mannooligosaccharide formulas was assessed based on cell proliferation. The structure-based immunomodulatory activity of the formulas was evaluated with respect to the length, branching and conformation of the various formulas. Glycoconjugate formulas with terminal ß-mannosyl-units tended to be more potent in terms of Candida relevant cytokines IL-12 p70, IL-17, GM-CSF, IL-6, and TNFα induction and cell proliferation, and this tendency was associated with structural differences between the studied glycoconjugate formulas. The eight tested mannooligosaccharide conjugates can be considered potential in vitro immunomodulative agents suitable for in vitro Candida diagnostics or prospectively for subcellular anti-Candida vaccine design.
Assuntos
Candida/imunologia , Candidíase/imunologia , Candidíase/microbiologia , Polissacarídeos Fúngicos/química , Polissacarídeos Fúngicos/imunologia , Imunomodulação , Macrófagos/imunologia , Oligossacarídeos/química , Oligossacarídeos/imunologia , Animais , Proliferação de Células , Citocinas/metabolismo , Polissacarídeos Fúngicos/síntese química , Ativação de Macrófagos , Macrófagos/microbiologia , Camundongos , Oligossacarídeos/síntese química , Células RAW 264.7 , Relação Estrutura-AtividadeRESUMO
Multiple exopolysaccharides (EPSs) have been isolated from various organisms in extreme environments and have yielded a variety of activities. The present study evaluated the immunomodulatory capabilities of an EPS (termed PHEPS) derived from the fungus Paecilomyces lilacinus PH0016, which was isolated from a tropical and hyperhaline environment in southern China. The macrophage RAW 264.7 cell line was used to investigate the mechanism of PHEPSinduced macrophage activation. The results indicated that RAW 264.7 macrophages were activated by PHEPS, in an effect slightly inferior to lipopolysaccharide (LPS), as evidenced by secretion of interleukin (IL)1ß, tumor necrosis factor (TNF)α and nitric oxide (NO), and by significantly increased phagocytosis in the cells treated with PHEPS. Nuclear factor (NF)κB p65 was significantly translocated into the nucleus in the PHEPStreated cells. In addition, expression of inducible NO synthase (iNOS) and IκBα degradation were enhanced in PHEPStreated cells. The phosphorylation levels of p38, JNK and ERK were also significantly increased in the PHEPStreated cells. Furthermore, IL1ß and TNFα production was markedly decreased in PHEPStreated cells when the mitogenactivated protein kinase (MAPK) pathways were blocked by the inhibitor Dectin1 and by antibodies against Tolllike receptor 4 (TLR4). The present results indicated that PHEPS from Paecilomyces lilacinus possessed the capability of activating RAW 264.7 cells via the TLR4/NFκB/MAPKs signaling pathway.
Assuntos
Polissacarídeos Fúngicos/imunologia , Sistema de Sinalização das MAP Quinases , Ativação de Macrófagos , NF-kappa B/imunologia , Paecilomyces/imunologia , Receptor 4 Toll-Like/imunologia , Animais , Macrófagos/imunologia , Macrófagos/microbiologia , Camundongos , Células RAW 264.7 , Transdução de SinaisRESUMO
Human thioredoxin (hTrx), which can be secreted from cells upon stress, functions in allergic skin inflammation as a T cell antigen due to homology and cross-reactivity with the fungal allergen Mala s13 of the skin-colonizing yeast Malassezia sympodialis. Recent studies have shown that cell wall polysaccharides of Malassezia are detected by the immune system via the C-type lectin receptors Dectin-1 and Dectin-2, which are expressed on myeloid cells. Therefore, this study aimed to investigate a putative interaction between Dectin-1, Dectin-2 and the allergens Mala s13 and hTrx. Stimulation of human monocyte-derived dendritic cells or macrophages with Mala s13 or hTrx resulted in remarkable secretion of IL-1ß and IL-23. Blocking experiments suggest that hTrx induces IL-23 by Dectin-1 binding and IL-1ß by binding to either Dectin-1 or Dectin-2. Regarding Mala s13, Dectin-1 appears to be involved in IL-1ß signaling. Interference of Syk kinase function was performed to investigate downstream signaling, which led to diminished hTrx responses. In our experiments, we observed rapid internalization of Mala s13 and hTrx upon cell contact and we were able to confirm direct interaction with Dectin-1 as well as Dectin-2 applying a fusion protein screening platform. We hypothesize that this cytokine response may result in a Th2/Th17-polarizing milieu, which may play a key role during the allergic sensitization in the skin, where allergen presentation to T cells is accompanied by microbial colonization and skin inflammation.
Assuntos
Alarminas/imunologia , Alérgenos/imunologia , Dermatite Atópica/imunologia , Polissacarídeos Fúngicos/imunologia , Lectinas Tipo C/metabolismo , Tiorredoxinas/imunologia , Apresentação de Antígeno/imunologia , Autoantígenos/imunologia , Buffy Coat/citologia , Reações Cruzadas/imunologia , Células Dendríticas/imunologia , Células Dendríticas/metabolismo , Humanos , Interleucina-1beta/metabolismo , Interleucina-23/metabolismo , Macrófagos/imunologia , Macrófagos/metabolismo , Malassezia/imunologia , Monócitos/imunologia , Monócitos/metabolismo , Moléculas com Motivos Associados a Patógenos/imunologia , Cultura Primária de Células , Proteínas Recombinantes/imunologia , Proteínas Recombinantes/isolamento & purificação , Transdução de Sinais/imunologia , Pele/imunologia , Pele/microbiologia , Linfócitos T Auxiliares-Indutores/imunologia , Linfócitos T Auxiliares-Indutores/metabolismoRESUMO
We previously found that Pleurotus ferulae polysaccharides (PFPS) improved the maturation and function of dendritic cells (DCs). In this study, we investigated the effects of PFPS on the antitumor efficacy of therapeutic human papillomavirus (HPV) DC-based vaccine. PFPS stimulated DCs pulsed with HPV E6/E7 peptides were used to treat tumor mice on day 5 & 12 (HPV + PFPS-DCs early) and day 12 & 19 (HPV + PFPS-DCs late) after TC-1 cell injection. Compared to control group, both HPV + PFPS-DCs early and HPV + PFPS-DCs late strategies significantly inhibited tumor growth, which was significantly correlated with the increased activation status of both CD4+ and CD8+ T cells, the decreased frequencies of myeloid-derived suppressor cells, and the induction of HPV-specific CD8+ T cell responses. The survival of tumor mice was also greatly improved by HPV + PFPS-DCs early. Moreover, HPV + PFPS-DCs early completely inhibited the growth of second challenged TC-1 cells in tumor free mice. The results showed that PFPS improved the antitumor efficacy of therapeutic HPV DC-based vaccine, suggesting that PFPS might be a potential adjuvant for DC-based vaccines. This study provides a potential strategy for developing the therapeutic DC-based vaccine against cervical cancer.
Assuntos
Vacinas Anticâncer/imunologia , Células Dendríticas/imunologia , Polissacarídeos Fúngicos/imunologia , Neoplasias/terapia , Vacinas contra Papillomavirus/imunologia , Pleurotus/química , Adjuvantes Imunológicos/administração & dosagem , Animais , Linfócitos T CD8-Positivos/imunologia , Vacinas Anticâncer/química , Linhagem Celular Tumoral , Células Dendríticas/efeitos dos fármacos , Feminino , Polissacarídeos Fúngicos/farmacologia , Humanos , Camundongos , Camundongos Endogâmicos C57BL , Neoplasias/imunologia , Papillomaviridae , Infecções por Papillomavirus/complicações , Infecções por Papillomavirus/prevenção & controle , Vacinas contra Papillomavirus/químicaRESUMO
ß-glucan, the most abundant fungal cell wall polysaccharide, has gained much attention from the scientific community in the last few decades for its fascinating but not yet fully understood immunobiology. Study of this molecule has been motivated by its importance as a pathogen-associated molecular pattern upon fungal infection as well as by its promising clinical utility as biological response modifier for the treatment of cancer and infectious diseases. Its immune effect is attributed to the ability to bind to different receptors expressed on the cell surface of phagocytic and cytotoxic innate immune cells, including monocytes, macrophages, neutrophils, and natural killer cells. The characteristics of the immune responses generated depend on the cell types and receptors involved. Size and biochemical composition of ß-glucans isolated from different sources affect their immunomodulatory properties. The variety of studies using crude extracts of fungal cell wall rather than purified ß-glucans renders data difficult to interpret. A better understanding of the mechanisms of purified fungal ß-glucan recognition, downstream signaling pathways, and subsequent immune regulation activated, is, therefore, essential not only to develop new antifungal therapy but also to evaluate ß-glucan as a putative anti-infective and antitumor mediator. Here, we briefly review the complexity of interactions between fungal ß-glucans and mononuclear phagocytes during fungal infections. Furthermore, we discuss and present available studies suggesting how different fungal ß-glucans exhibit antitumor and antimicrobial activities by modulating the biologic responses of mononuclear phagocytes, which make them potential candidates as therapeutic agents.
Assuntos
Polissacarídeos Fúngicos/imunologia , Sistema Fagocitário Mononuclear , beta-Glucanas/imunologia , Animais , Doença , HumanosRESUMO
Cryptococcus neoformans is a yeast-type opportunistic fungal pathogen with a capsule structure consisting of polysaccharides, such as glucuronoxylomannan and galactoxylomannan, and infects the lungs via an air-borne route. Most healthy individuals undergo asymptomatic infection with granulomatous lesions in the lungs caused by C. neoformans. However, immunocompromised hosts with severely impaired cellular immunity, such as those with acquired immune deficiency syndrome (AIDS), often suffer from disseminated infection into the central nervous system, leading to life-threatening meningoencephalitis. The recognition of pathogen-associated molecular patterns (PAMPs) by macrophages and dendritic cells plays an important role as the first line of host defense in the elimination of pathogens. Recently, numerous pattern recognition receptors (PRRs) that recognize these PAMPs have been identified. Also, the involvement of these PRRs, such as Toll-like receptors (TLRs), NOD-like receptors (NLRs), and C-type lectin receptors (CLRs), in cryptococcal infection has been analyzed. In particular, TLR9, NLR family pyrin domain-containing 3 (NLRP3), Dectin-2, mannose receptor (MR), and DC-SIGN have been found to recognize the DNA, cell wall components, intracellular polysaccharides, and mannoproteins, respectively. Future studies are expected to promote elucidation of the mechanisms of host immune response to C. neoformans, which will lead to the development of new vaccines and therapies for cryptococcal infection.
Assuntos
Criptococose/imunologia , Cryptococcus neoformans/imunologia , Moléculas com Motivos Associados a Patógenos/imunologia , Receptores de Reconhecimento de Padrão/imunologia , Parede Celular/imunologia , Cryptococcus neoformans/química , Cryptococcus neoformans/citologia , Cryptococcus neoformans/genética , DNA Fúngico/imunologia , Polissacarídeos Fúngicos/imunologia , Vacinas Fúngicas , Humanos , Hospedeiro Imunocomprometido , Lectinas Tipo C/imunologia , Glicoproteínas de Membrana/imunologia , Proteínas NLR/imunologia , Receptores Toll-Like/imunologiaRESUMO
Methamphetamine (METH) is a powerful and highly addictive stimulant that affects the central nervous system of users in the United States and worldwide, and its consumption is associated to the acquisition of HIV and AIDS-related infections. METH enhances cryptococcosis in mice, an opportunistic infection caused by the encapsulated fungus Cryptococcus neoformans. Due to its ability to survive within macrophages, C. neoformans is an ideal model to study pathogen-macrophage interactions. METH abrogates normal macrophage function, which might contribute to the higher rate and more rapid progression of infections in drug abusers. Hence, we investigated the role of complement and specific IgM to C. neoformans capsular polysaccharide on the function of J774.16 macrophage-like cells after exposure to METH. We found that complement and IgM significantly promotes complement-mediated phagocytosis of C. neoformans by J774.16 cells in comparison to co-incubation with complement alone. IgM enhances the expression of complement receptor 3 on the surface macrophages treated with the drug. Also, IgM-increased macrophage phagocytosis of C. neoformans may be associated with upregulation of GTPase-RhoA, a key regulator of the actin polymerization signaling cascade. Fungal cells incubated with complement and IgM in the presence of METH demonstrated higher number of cells per aggregate, a possible explanation for their enhanced ingestion by phagocytes. IgM increased killing of yeast cells by macrophages by inhibiting the alkalization of the phagosome and stimulating the intracellular production of nitric oxide. Together, our findings suggest that IgM stimulates the effector functions of macrophages against opportunistic pathogens in the setting of drug abuse.
Assuntos
Criptococose/imunologia , Cryptococcus neoformans/imunologia , Imunoglobulina M/metabolismo , Macrófagos/imunologia , Metanfetamina/farmacologia , Actinas/metabolismo , Animais , Apoptose , Linhagem Celular , Proteínas do Sistema Complemento/metabolismo , Polissacarídeos Fúngicos/imunologia , GTP Fosfo-Hidrolases/metabolismo , Humanos , Macrófagos/efeitos dos fármacos , Camundongos , Fagocitose , Fagossomos/metabolismo , Transdução de SinaisAssuntos
Aspergilose/imunologia , Aspergilose/microbiologia , Aspergillus fumigatus/fisiologia , Polissacarídeos Fúngicos/imunologia , Lectinas/imunologia , Pulmão/imunologia , Aspergilose/genética , Aspergillus fumigatus/genética , Aspergillus fumigatus/imunologia , Ativação do Complemento , Células Epiteliais/imunologia , Células Epiteliais/microbiologia , Humanos , Interleucina-8/genética , Interleucina-8/imunologia , Lectinas/genética , Pulmão/microbiologia , Ligação Proteica , beta-Glucanas/metabolismo , FicolinasRESUMO
Immunodetection is described in this chapter as a technique for producing specific antibodies for antigen detection of the major human fungal pathogens. In the case of Candida spp., heat-killed cells are used to immunize mice over a couple of weeks and then splenocytes are isolated and further fused with myelomas to easily propagate the antibodies produced in the mice. The resulting antibodies follow a purification process where antibody levels and concentrations are determined. Fungal cells are also lysed to obtain whole cell extracts as a prior step for identification of antigens using immunoprecipitation. Finally, this method permits the production of specific antibodies against fungi and the identification of the respective antigens in an in vivo model.
Assuntos
Antígenos de Fungos/imunologia , Candidíase Invasiva/diagnóstico , Polissacarídeos Fúngicos/imunologia , Animais , Anticorpos Monoclonais/isolamento & purificação , Candida/imunologia , Candidíase Invasiva/imunologia , Humanos , Hibridomas , Imunoensaio , Camundongos Endogâmicos BALB CRESUMO
The ability of mannose receptor (MR) to recognize the carbohydrate structures is well-established. Here, we reported that MR was crucial for the immune response to a Ganoderma atrum polysaccharide (PSG-1), as evidenced by elevation of MR in association with increase of phagocytosis and concentrations of interleukin-1ß (IL-1ß) and tumor necrosis factor-α (TNF-α) in normal macrophages. Elevation of MR triggered by PSG-1 also led to control lipopolysaccharide (LPS)-triggered inflammatory response via the increase of interleukin-10 (IL-10) and inhibition of phagocytosis and IL-1ß. Anti-MR antibody partly attenuated PSG-1-mediated anti-inflammatory responses, while it could not affect TNF-α secretion, suggesting that another receptor was involved in PSG-1-triggered immunomodulatory effects. MR and toll-like receptor (TLR)4 coordinated the influences on the TLR4-mediated signaling cascade by the nuclear factor-κB (NF-κB) pathway in LPS-stimulated macrophages subjected to PSG-1. Collectively, immune response to PSG-1 required recognition by MR in macrophages. The NF-κB pathway served as a central role for the coordination of MR and TLR4 to elicit immune response to PSG-1.
Assuntos
Polissacarídeos Fúngicos/farmacologia , Ganoderma/química , Lectinas Tipo C/imunologia , Macrófagos Peritoneais/efeitos dos fármacos , Macrófagos Peritoneais/imunologia , Lectinas de Ligação a Manose/imunologia , Receptores de Superfície Celular/imunologia , Animais , Células Cultivadas , Citocinas/metabolismo , Polissacarídeos Fúngicos/imunologia , Lectinas Tipo C/metabolismo , Macrófagos Peritoneais/metabolismo , Receptor de Manose , Lectinas de Ligação a Manose/metabolismo , Camundongos Endogâmicos BALB C , Fagocitose/efeitos dos fármacos , Fagocitose/imunologia , Receptores de Superfície Celular/metabolismo , Receptor 2 Toll-Like/metabolismo , Receptor 4 Toll-Like/metabolismo , Fator de Necrose Tumoral alfa/metabolismoRESUMO
Paracoccidioides brasiliensis and P. lutzii cause human paracoccidioidomycosis (PCM). They are dimorphic ascomycetes that grow as filaments at mild temperatures up to 28oC and as multibudding pathogenic yeast cells at 37oC. Components of the fungal cell wall have an important role in the interaction with the host because they compose the cell outermost layer. The Paracoccidioides cell wall is composed mainly of polysaccharides, but it also contains proportionally smaller rates of proteins, lipids, and melanin. The polysaccharide cell wall composition and structure of Paracoccidioides yeast cells, filamentous and transition phases were studied in detail in the past. Other cell wall components have been better analyzed in the last decades. The present work gives to the readers a detailed updated view of cell wall-associated proteins. Proteins that have been localized at the cell wall compartment using antibodies are individually addressed. We also make an overview about PCM, the Paracoccidioides cell wall structure, secretion mechanisms, and fungal extracellular vesicles.
Assuntos
Anticorpos Antifúngicos/biossíntese , Parede Celular/química , Proteínas Fúngicas/química , Paracoccidioides/química , Paracoccidioidomicose/imunologia , Parede Celular/metabolismo , Proteínas do Sistema Complemento/genética , Proteínas do Sistema Complemento/imunologia , Vesículas Extracelulares/química , Vesículas Extracelulares/imunologia , Polissacarídeos Fúngicos/química , Polissacarídeos Fúngicos/imunologia , Polissacarídeos Fúngicos/isolamento & purificação , Proteínas Fúngicas/imunologia , Proteínas Fúngicas/isolamento & purificação , Expressão Gênica , Interações Hospedeiro-Patógeno , Humanos , Imunidade Inata , Paracoccidioides/genética , Paracoccidioides/metabolismo , Paracoccidioides/patogenicidade , Paracoccidioidomicose/microbiologia , Paracoccidioidomicose/patologiaRESUMO
Protein-bound polysaccharides (PBP) isolated from Coriolus versicolor (CV) are classified as biological response modifiers capable of exhibiting various biological activities, such as anti-tumour and immunopotentiating activity. Since we have found in vivo studies that the tested PBP induced prolongation of endotoxin fever in rats, the aim of the present study was to investigate the in vitro effect of the PBP on the production of pro-inflammatory cytokines by the lipolysaccharide (LPS)-stimulated rat peripheral blood mononuclear cells (PBMCs). The results showed that the PBP affect the immunomodulating properties of the LPS-treated PBMCs by the enhancement of mitogenic activity and attenuation of the LPS-induced production of interleukin (IL)-1ß and IL-6. Moreover, the tested polysaccharides peptides themselves also exhibit immunomodulatory properties manifested in the increased cell proliferation and pro-inflammatory cytokine release from PBMCs. The effect of PBP on the both phenomena was time-dependent and occurred in the U-shaped dose response manner. These findings are significant when considering the use of commercially available PBP from CV extract by cancer patients suffering from immunodeficiency, who may experience microbial infections during therapy.
Assuntos
Basidiomycota/imunologia , Citocinas/metabolismo , Polissacarídeos Fúngicos/imunologia , Mediadores da Inflamação/metabolismo , Leucócitos Mononucleares/imunologia , Leucócitos Mononucleares/metabolismo , Lipopolissacarídeos/imunologia , Animais , Relação Dose-Resposta Imunológica , Feminino , Imunomodulação , RatosRESUMO
A coral-associated fungus Penicillium sp.gxwz446 that produced exopolysaccharde was isolated from the coral Echinogorgia flora in South China. Two neutral exopolysaccharides GX1-1 and GX2-1 were obtained from the fermented broth of the fungus and purified by anion-exchange and gel-permeation chromatography. Chemical and spectroscopic analyses showed that GX1-1 was a glucan, primarily composed of glucose, with a molecular weight of 5.0 kDa. GX1-1 mainly consists of (1â4)-linked α-d-glucopyranose units as the backbone, substituted at C-2 with a single α-d-glucopyranose on every sixth sugar residues. GX2-1 was a galactofuranose-containing mannogalactoglucan with a molecular weight of 9.5 kDa. The main linkages were composed of (1â4)-ß-d-Glcp, (1â5)-ß-d-Galf, (1â3,5)-ß-d-Galf, (1â6)-α-d-Manp and (1â2, 6)-α-d-Manp. GX1-1 showed RAW264.7 macrophage activation activity. After subjecting GX1-1 to sulfated modification, there was about one sulfate substitution on every sugar ring, primarily at O-6. The sulfated derivative of GX1-1 exhibited a more significant ability to promote the pinocytic activity of RAW264.7 cells and induce the production of NO.