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1.
PLoS Pathog ; 16(8): e1008733, 2020 08.
Artigo em Inglês | MEDLINE | ID: mdl-32817694

RESUMO

Staphylococcus aureus (S. aureus) is one of the most common bacterial infections worldwide, and antibiotic resistant strains such as Methicillin-Resistant S. aureus (MRSA) are a major threat and burden to public health. MRSA not only infects immunocompromised patients but also healthy individuals and has rapidly spread from the healthcare setting to the outside community. However, all vaccines tested in clinical trials to date have failed. Immunocompromised individuals such as patients with HIV or decreased levels of CD4+ T cells are highly susceptible to S. aureus infections, and they are also at increased risk of developing fungal infections. We therefore wondered whether stimulation of antifungal immunity might promote the type of immune responses needed for effective host defense against S. aureus. Here we show that vaccination of mice with a fungal ß-glucan particle (GP) loaded with S. aureus antigens provides protective immunity to S. aureus. We generated glucan particles loaded with the four S. aureus proteins ClfA, IsdA, MntC, and SdrE, creating the 4X-SA-GP vaccine. Vaccination of mice with three doses of 4X-SA-GP promoted protection in a systemic model of S. aureus infection with a significant reduction in the bacterial burden in the spleen and kidneys. 4X-SA-GP vaccination induced antigen-specific Th1 and Th17 CD4+ T cell and antibody responses and provided long-term protection. This work suggests that the GP vaccine system has potential as a novel approach to developing vaccines for S. aureus.


Assuntos
Saccharomyces cerevisiae/imunologia , Infecções Estafilocócicas/imunologia , Vacinas Antiestafilocócicas/imunologia , Staphylococcus aureus/imunologia , Animais , Anticorpos Antibacterianos/imunologia , Coagulase/administração & dosagem , Coagulase/genética , Coagulase/imunologia , Feminino , Humanos , Camundongos , Camundongos Endogâmicos C57BL , Saccharomyces cerevisiae/química , Infecções Estafilocócicas/microbiologia , Vacinas Antiestafilocócicas/administração & dosagem , Vacinas Antiestafilocócicas/genética , Staphylococcus aureus/genética , Células Th1/imunologia , Células Th17/imunologia , Vacinação , beta-Glucanas/administração & dosagem , beta-Glucanas/imunologia
2.
Sci Rep ; 10(1): 6740, 2020 04 21.
Artigo em Inglês | MEDLINE | ID: mdl-32317718

RESUMO

To protect the audiosensory organ from tissue damage from the immune system, the inner ear is separated from the circulating immune system by the blood-labyrinth barrier, which was previously considered an immune-privileged site. Recent studies have shown that macrophages are distributed in the cochlea, especially in the spiral ligament, spiral ganglion, and stria vascularis; however, the direct pathogen defence mechanism used by audiosensory receptor hair cells (HCs) has remained obscure. Here, we show that HCs are protected from pathogens by surrounding accessory supporting cells (SCs) and greater epithelial ridge (GER or Kölliker's organ) cells (GERCs). In isolated murine cochlear sensory epithelium, we established Theiler's murine encephalomyelitis virus, which infected the SCs and GERCs, but very few HCs. The virus-infected SCs produced interferon (IFN)-α/ß, and the viruses efficiently infected the HCs in the IFN-α/ß receptor-null sensory epithelium. Interestingly, the virus-infected SCs and GERCs expressed macrophage marker proteins and were eliminated from the cell layer by cell detachment. Moreover, lipopolysaccharide induced phagocytosis of the SCs without cell detachment, and the SCs phagocytosed the bacteria. These results reveal that SCs function as macrophage-like cells, protect adjacent HCs from pathogens, and provide a novel anti-infection inner ear immune system.


Assuntos
Células Ciliadas Auditivas Internas/fisiologia , Células Ciliadas Auditivas Externas/fisiologia , Células Labirínticas de Suporte/imunologia , Macrófagos/imunologia , Gânglio Espiral da Cóclea/fisiologia , Estria Vascular/fisiologia , Animais , Animais Recém-Nascidos , Escherichia coli/imunologia , Células Ciliadas Auditivas Internas/citologia , Células Ciliadas Auditivas Externas/citologia , Imunidade Inata , Interferon-alfa/biossíntese , Interferon-alfa/imunologia , Interferon beta/biossíntese , Interferon beta/imunologia , Células Labirínticas de Suporte/citologia , Células Labirínticas de Suporte/efeitos dos fármacos , Células Labirínticas de Suporte/virologia , Lipopolissacarídeos/farmacologia , Macrófagos/citologia , Macrófagos/efeitos dos fármacos , Macrófagos/virologia , Camundongos , Camundongos Endogâmicos ICR , Técnicas de Cultura de Órgãos , Fagocitose/efeitos dos fármacos , Saccharomyces cerevisiae/imunologia , Gânglio Espiral da Cóclea/citologia , Estria Vascular/citologia , Theilovirus/crescimento & desenvolvimento , Theilovirus/patogenicidade
3.
Surgery ; 167(6): 1012-1015, 2020 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-32098690

RESUMO

BACKGROUND: Hidradenitis suppurativa is a complex disorder, the pathogenesis of which is still unsolved. The known association between hidradenitis suppurativa and Crohn's disease, an autoimmune disease diagnosed with the presence of Anti-Saccharomyces cerevisiae antibodies of the IgG family, suggests that a much more complex mechanism than a simple infectious disorder is involved. The goal of this study is to report patients' characteristics and the outcome of 6 years of a yeast (Saccharomyces cerevisiae)-exclusion diet and surgery in the treatment of hidradenitis suppurativa. METHOD: We analyzed 185 patients with hidradenitis suppurativa with a self-evaluative questionnaire. Thirty-seven patients were treated in our center following our protocol. The other 148 were members of a support group for patients with hidradenitis suppurativa treated by other centers. RESULTS: In 80% of patients who had the onset of hidradenitis suppurativa before the age of 30, the female to male ratio was 3.34:1, 74% were active smokers, and 5% also had Crohn's disease. In the diet group, 70% had an improvement of hidradenitis suppurativa symptomatology, 81% of whom in less than 6 months. Also, 87% of patients demonstrated an immediate recurrence of skin lesions less than a week after consuming a food containing the yeast. Immunologic testing showed intolerance to yeast, wheat, and cow's milk in 20%, 29%, and 23% of patients, respectively. CONCLUSION: The analysis confirmed the stabilization and regression of hidradenitis suppurativa with our diet, presumably by decreasing the local and systemic inflammation, leading to a less invasive operative treatment. These new findings seem to link hidradenitis suppurativa to food intolerance and gut dysbiosis.


Assuntos
Hipersensibilidade Alimentar/complicações , Hidradenite Supurativa/dietoterapia , Saccharomyces cerevisiae/imunologia , Adulto , Animais , Doença de Crohn/epidemiologia , Feminino , Seguimentos , Hidradenite Supurativa/cirurgia , Humanos , Imunoglobulina G/sangue , Masculino , Leite/imunologia , Fumar/epidemiologia , Inquéritos e Questionários , Triticum/imunologia , Perda de Peso
4.
Int J Mol Sci ; 21(4)2020 Feb 18.
Artigo em Inglês | MEDLINE | ID: mdl-32085663

RESUMO

Individuals with intestinal barrier dysfunction are more prone to autoimmunity. Lipopolysaccharides (LPS) from gut bacteria have been shown to play a role in systemic inflammation, leading to the opening of the gut and blood-brain barrier (BBB). This study aims to measure antibodies against LPS and barrier proteins in samples positive for anti-Saccharomyces cerevisiae antibodies (ASCA) and anti-neutrophil cytoplasmic antibodies (ANCA) and compare them with these same antibodies in controls to determine whether a correlation between LPS and barrier proteins could be found. We obtained 94 ASCA- and 94 ANCA-positive blood samples, as well as 188 blood samples from healthy controls. Samples were assessed for antibodies to LPS, zonulin+occludin, S100B, and aquaporin-4 (AQP4). Results show significant elevation in antibodies in about 30% of ASCA- and ANCA-positive sera and demonstrate positive linear relationships between these antibodies. The findings suggest that individuals positive for ASCA and ANCA have increased odds of developing intestinal and BBB permeability compared to healthy subjects. The levels of LPS antibodies in both ASCA- and ANCA-positive and negative specimens showed from low and moderate to high correlation with antibodies to barrier proteins. This study shows that LPS, by damaging the gut and BBBs, contribute to the extra-intestinal manifestation of IBD. We conclude that IBD patients should be screened for LPS antibodies in an effort to detect or prevent possible barrier damage at the earliest stage possible to abrogate disease symptoms in IBS and associated disorders.


Assuntos
Anticorpos Anticitoplasma de Neutrófilos/sangue , Anticorpos Antifúngicos/sangue , Ácido Aspártico Endopeptidases/imunologia , Bactérias/imunologia , Lipopolissacarídeos/imunologia , Proteínas de Saccharomyces cerevisiae/imunologia , Saccharomyces cerevisiae/imunologia , Aquaporina 4/imunologia , Estudos de Casos e Controles , Humanos , Razão de Chances , Subunidade beta da Proteína Ligante de Cálcio S100/metabolismo
5.
mSphere ; 5(1)2020 02 12.
Artigo em Inglês | MEDLINE | ID: mdl-32051241

RESUMO

K1 represents a heterodimeric A/B toxin secreted by virus-infected Saccharomyces cerevisiae strains. In a two-staged receptor-mediated process, the ionophoric activity of K1 leads to an uncontrolled influx of protons, culminating in the breakdown of the cellular transmembrane potential of sensitive cells. K1 killer yeast necessitate not only an immunity mechanism saving the toxin-producing cell from its own toxin but, additionally, a molecular system inactivating the toxic α subunit within the secretory pathway. In this study, different derivatives of the K1 precursor were constructed to analyze the biological function of particular structural components and their influence on toxin activity as well as the formation of protective immunity. Our data implicate an inactivation of the α subunit during toxin maturation and provide the basis for an updated model of K1 maturation within the host cell's secretory pathway.IMPORTANCE The killer phenotype in the baker's yeast Saccharomyces cerevisiae relies on two double-stranded RNA viruses that are persistently present in the cytoplasm. As they carry the same receptor populations as sensitive cells, killer yeast cells need-in contrast to various bacterial toxin producers-a specialized immunity mechanism. The ionophoric killer toxin K1 leads to the formation of cation-specific pores in the plasma membrane of sensitive yeast cells. Based on the data generated in this study, we were able to update the current model of toxin processing, validating the temporary inactivation of the toxic α subunit during maturation in the secretory pathway of the killer yeast.


Assuntos
Fatores Matadores de Levedura/genética , Fatores Matadores de Levedura/imunologia , Mutagênese Sítio-Dirigida , Saccharomyces cerevisiae/genética , Saccharomyces cerevisiae/imunologia , Membrana Celular/patologia , Fenótipo , Vírus de RNA/patogenicidade , Saccharomyces cerevisiae/virologia , Proteínas de Saccharomyces cerevisiae/genética , Proteínas de Saccharomyces cerevisiae/imunologia
6.
Benef Microbes ; 11(2): 175-181, 2020 Mar 27.
Artigo em Inglês | MEDLINE | ID: mdl-31990221

RESUMO

Studies aiming at the development and evaluation of alternative methods to minimise losses caused by the gastrointestinal nematode Haemonchus contortus are extremely important. Such research is essential, given the high morbidity rates among sheep and the significant mortality rates of lambs, allied to the low efficacy of commercial products for the control of this parasite. The purpose of this study was to evaluate the effect of the Saccharomyces cerevisiae (YT001 - YEASTECH) on the control of H. contortus and its modulation of the immune response in experimentally infected sheep. Eighteen sheep were divided into two groups. Group 1, the control group, comprised animals infected with H. contortus and supplemented with distilled water, while Group 2, the treated group, consisted of animals infected and supplemented with S. cerevisiae (400 million cfu/day of suspension for 49 days). The following parasitological parameters were evaluated: number of eggs per gram of faeces, number of infective larvae (L3) recovered per faecal culture, and parasitic load of the abomasum. The following immunological parameters were quantified: immunoglobulin (Ig)A in the mucous secretions and serum IgG; cytokines interleukin (IL)-4, IL-5 and IL-10; number of eosinophils in the abomasal mucosa and groups of cells positive for the markers: MHCII, CD4+CD25+, CD5+CD8+, WC4, CD5+CD4+, CD8+CD11b+ and CD5+WC1 by whole blood flow cytometry. The results revealed a significant decrease (P<0.05) in the number of larvae and significantly higher serum IgG levels (P<0.05) in the group supplemented with S. cerevisiae. The supplemented animals showed significantly larger numbers of eosinophils (P<0.05), as well as more cells positive for MHCII, CD4+CD25+, CD5+CD8+ than the control animals. This study confirmed the beneficial action of S. cerevisiae on the host immune response to H. contortus, as evidenced mainly by the smaller number of L3 recovered from the faeces of sheep supplemented with S. cerevisiae.


Assuntos
Suplementos Nutricionais/microbiologia , Hemoncose/veterinária , Probióticos/administração & dosagem , Saccharomyces cerevisiae/imunologia , Doenças dos Ovinos/terapia , Ovinos/imunologia , Administração Oral , Animais , Anticorpos Anti-Helmínticos/sangue , Citocinas/imunologia , Eosinófilos/imunologia , Fezes/parasitologia , Hemoncose/imunologia , Hemoncose/terapia , Haemonchus , Interações entre Hospedeiro e Microrganismos/imunologia , Imunoglobulina A/análise , Imunoglobulina G/sangue , Contagem de Leucócitos , Masculino , Contagem de Ovos de Parasitas , Ovinos/parasitologia , Doenças dos Ovinos/imunologia , Doenças dos Ovinos/parasitologia
7.
Hepatology ; 71(2): 522-538, 2020 02.
Artigo em Inglês | MEDLINE | ID: mdl-31228214

RESUMO

Chronic alcohol consumption causes increased intestinal permeability and changes in the intestinal microbiota composition, which contribute to the development and progression of alcohol-related liver disease. In this setting, little is known about commensal fungi in the gut. We studied the intestinal mycobiota in a cohort of patients with alcoholic hepatitis, patients with alcohol use disorder, and nonalcoholic controls using fungal-specific internal transcribed spacer amplicon sequencing of fecal samples. We further measured serum anti-Saccharomyces cerevisiae antibodies (ASCA) as a systemic immune response to fungal products or fungi. Candida was the most abundant genus in the fecal mycobiota of the two alcohol groups, whereas genus Penicillium dominated the mycobiome of nonalcoholic controls. We observed a lower diversity in the alcohol groups compared with controls. Antibiotic or steroid treatment was not associated with a lower diversity. Patients with alcoholic hepatitis had significantly higher ASCA levels compared to patients with alcohol use disorder and to nonalcoholic controls. Within the alcoholic hepatitis cohort, patients with levels of at least 34 IU/mL had a significantly lower 90-day survival (59%) compared with those with ASCA levels less than 34 IU/mL (80%) with an adjusted hazard ratio of 3.13 (95% CI, 1.11-8.82; P = 0.031). Conclusion: Patients with alcohol-associated liver disease have a lower fungal diversity with an overgrowth of Candida compared with controls. Higher serum ASCA was associated with increased mortality in patients with alcoholic hepatitis. Intestinal fungi may serve as a therapeutic target to improve survival, and ASCA may be useful to predict the outcome in patients with alcoholic hepatitis.


Assuntos
Disbiose/etiologia , Disbiose/imunologia , Hepatite Alcoólica/complicações , Hepatite Alcoólica/imunologia , Intestinos/microbiologia , Micobioma , Adulto , Idoso , Anticorpos Antifúngicos/sangue , Candida/imunologia , Estudos de Coortes , Disbiose/sangue , Feminino , Hepatite Alcoólica/sangue , Humanos , Fenômenos do Sistema Imunológico , Masculino , Pessoa de Meia-Idade , Saccharomyces cerevisiae/imunologia
8.
Reumatol. clín. (Barc.) ; 15(6): 360-362, nov.-dic. 2019. tab
Artigo em Espanhol | IBECS | ID: ibc-189654

RESUMO

OBJETIVO: Valorar la relación entre la calprotectina fecal, ASCA y marcadores de enfermedad en espondiloartritis (Spa). MÉTODOS: Se evaluaron pacientes ≥18años que cumplían criterios ASAS o criterios modificados de Nueva York. Se analizan criterios de actividad, función física, analíticos y datos demográficos. RESULTADOS: Se incluyeron 33 pacientes. Todos excepto uno tenían valores normales de ASCA. Encontramos correlación significativa entre calprotectina y PCR pero no con otros parámetros, ni entre los niveles de calprotectina y la toma de AINE (p = 0,001). Tampoco entre los niveles de PCR y el uso de AINE. Después de retirar los AINE no encontramos diferencias en los niveles de calprotectina (p = 0,9). CONCLUSIÓN: La calprotectina fecal está elevada en pacientes con Spa y se correlaciona positivamente con la PCR. La elevación de la calprotectina fecal no se ve alterada por el uso de AINE. La cantidad de ASCA no cambia y no se correlaciona con ningún parámetro clínico en el estudio poblacional


OBJECTIVE: To assess the relationship between the increase of fecal calprotectin, anti-Saccharomyces cerevisiae antibodies (ASCA) and disease markers in a group of patients with spondyloarthritis. METHODS: We evaluated patients who were at least 18-years-old and met the Assessment in Spondyloarthritis International Society (ASAS) criteria for spondyloarthritis or the New York modified criteria. We analyzed activity criteria, physical function, analytical criteria (human leukocyte antigen [HLA] B27, fecal calprotectin, presence of ASCA, among others) and demographic data. RESULTS: We included 33 patients. All but one patient had normal ASCA values. We found statistical significance in the correlation of calprotectin with C-reactive protein (CRP) but not with other parameters. We also found a relationship between calprotectin levels and nonsteroidal anti-inflammatory drug (NSAID) intake (P=.001). We found no relationship between CRP levels and NSAID use. After discontinuation of NSAIDs for one month, we found no significant differences in calprotectin levels (P=.9). CONCLUSION: Fecal calprotectin is elevated in patients with spondyloarthritis and correlates positively with CRP. Level of fecal calprotectin is not altered by NSAID use. The amount of ASCA present does not change and does not correlate with any clinical parameters in the study population


Assuntos
Humanos , Adulto , Pessoa de Meia-Idade , Anticorpos Antifúngicos/sangue , Fezes/química , Complexo Antígeno L1 Leucocitário/análise , Saccharomyces cerevisiae/imunologia , Espondilartrite/diagnóstico , Espondilartrite/metabolismo , Biomarcadores/análise , Proteína C-Reativa/análise , Espondilartrite/sangue
9.
Protein Eng Des Sel ; 32(5): 219-230, 2019 12 31.
Artigo em Inglês | MEDLINE | ID: mdl-31769480

RESUMO

Yeast display immunoprecipitation is a combinatorial library screening platform for the discovery and engineering of antibodies against membrane proteins using detergent-solubilized membrane fractions or cell lysates as antigen sources. Here, we present the extension of this method for the screening of antibodies that bind to membrane protein complexes, enabling discovery of antibodies that target antigens involved in a functional protein-protein interaction of interest. For this proof-of-concept study, we focused on the receptor-mediated endocytosis machinery at the blood-brain barrier, and adaptin 2 (AP-2) was chosen as the functional interaction hub. The goal of this study was to identify antibodies that bound to blood-brain barrier (BBB) membrane protein complexes containing AP-2. Screening of a nonimmune yeast display antibody library was carried out using detergent-solubilized BBB plasma membranes as an antigen pool, and antibodies that could interact with protein complexes containing AP-2 were identified. Downstream characterization of isolated antibodies confirmed targeting of proteins known to play important roles in membrane trafficking. This functional yeast display immunoprecipitation screen may be applied to other systems where antibodies against other functional classes of protein complexes are sought.


Assuntos
Antígenos/imunologia , Barreira Hematoencefálica/imunologia , Proteínas de Membrana/imunologia , Complexos Multiproteicos/imunologia , Saccharomyces cerevisiae , Anticorpos de Cadeia Única , Animais , Células HEK293 , Humanos , Masculino , Ratos , Ratos Sprague-Dawley , Saccharomyces cerevisiae/genética , Saccharomyces cerevisiae/imunologia , Anticorpos de Cadeia Única/genética , Anticorpos de Cadeia Única/imunologia
10.
Genes (Basel) ; 10(9)2019 08 31.
Artigo em Inglês | MEDLINE | ID: mdl-31480411

RESUMO

Many pathogens (virus, bacteria, fungi, or parasites) have developed a wide variety of mechanisms to evade their host immune system. The budding yeast Saccharomyces cerevisiae has successfully been used to decipher some of these immune evasion strategies. This includes the cis-acting mechanism that limits the expression of the oncogenic Epstein-Barr virus (EBV)-encoded EBNA1 and thus of antigenic peptides derived from this essential but highly antigenic viral protein. Studies based on budding yeast have also revealed the molecular bases of epigenetic switching or recombination underlying the silencing of all except one members of extended families of genes that encode closely related and highly antigenic surface proteins. This mechanism is exploited by several parasites (that include pathogens such as Plasmodium, Trypanosoma, Candida, or Pneumocystis) to alternate their surface antigens, thereby evading the immune system. Yeast can itself be a pathogen, and pathogenic fungi such as Candida albicans, which is phylogenetically very close to S. cerevisiae, have developed stealthiness strategies that include changes in their cell wall composition, or epitope-masking, to control production or exposure of highly antigenic but essential polysaccharides in their cell wall. Finally, due to the high antigenicity of its cell wall, yeast has been opportunistically exploited to create adjuvants and vectors for vaccination.


Assuntos
Evasão da Resposta Imune/genética , Imunidade/genética , Saccharomyces cerevisiae/genética , Transgenes , Animais , Doenças Transmissíveis/genética , Doenças Transmissíveis/imunologia , Herpesvirus Humano 4/genética , Herpesvirus Humano 4/imunologia , Humanos , Saccharomyces cerevisiae/imunologia
11.
Front Immunol ; 10: 1977, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31475017

RESUMO

The immune defensive mechanisms active in the solitary ascidian Ciona robusta include phagocytic and encapsulating activity, largely brought about by phagocytic cells within the haemocyte population, the presence of complement components, which have been molecularly and functionally identified, and expression of a number of immune-related genes and pathways, identified by genome-based homology with vertebrate counterparts. Since C. robusta only displays highly conserved innate immune mechanisms, being devoid of an adaptive immune system, this organism is an excellent model for studying the features of innate memory, i.e., the capacity of the innate immune system to re-programming its responsiveness to potentially dangerous agents upon repeated exposure. In this study, we have developed an in vivo model for assessing the establishment and molecular/functional features of innate memory, by sequentially exposing C. robusta to a priming stimulus (microbial molecules), followed by a period of resting to return to basal conditions, and a challenge with microbial agents in homologous or cross-stimulation. The endpoints of immune activation were a functional activity (phagocytosis) and the molecular profiles of immune-related gene expression. The results show that exposure of C. robusta to microbial agents induces a reaction that primes animals for developing a different (expectedly more protective) response to subsequent challenges, showing the effective establishment of an immune memory. This immune memory relies on the modulation of a number of different mechanisms, some of which are priming-specific, others that are challenge-specific, and others that are non-specific, i.e., are common to all priming/challenge combinations (e.g., up-regulation of the Tnf and Lbp genes). Memory-dependent expression of the humoral immunity-related gene C3ar inversely correlates with memory-dependent variations of phagocytic rate, suggesting that complement activation and phagocytosis are alternative defensive mechanisms in C. robusta. Conversely, memory-dependent expression of the cellular immunity-related gene Cd36 directly correlates with variations of phagocytic rate, suggesting a direct involvement of this gene in the functional regulation of phagocytosis.


Assuntos
Ciona intestinalis/imunologia , Imunidade Inata/imunologia , Memória Imunológica/imunologia , Animais , Bacillus cereus/imunologia , Escherichia coli K12/imunologia , Hemócitos/imunologia , Lipopolissacarídeos/imunologia , Fagocitose/imunologia , Saccharomyces cerevisiae/imunologia
12.
Mycopathologia ; 184(4): 461-478, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31230200

RESUMO

The details of how gut-associated lymphoid tissues such as Peyer's patches (PPs) in the small intestine play a role in immune surveillance, microbial differentiation and the mucosal barrier protection in response to fungal organisms such as Candida albicans are still unclear. We particularly focus on PPs as they are the immune sensors and inductive sites of the gut that influence inflammation and tolerance. We have previously demonstrated that CD11c+ phagocytes that include dendritic cells and macrophages are located in the sub-epithelial dome within PPs sample C. albicans. To gain insight on how specific cells within PPs sense and respond to the sampling of fungi, we gavaged naïve mice with C. albicans strains ATCC 18804 and SC5314 as well as Saccharomyces cerevisiae. We measured the differential gene expression of sorted CD45+ B220+ B-cells, CD3+ T-cells and CD11c+ DCs within the first 24 h post-gavage using nanostring nCounter® technology. The results reveal that at 24 h, PP phagocytes were the cell type that displayed differential gene expression. These phagocytes were able to sample C. albicans and discriminate between strains. In particular, strain ATCC 18804 upregulated fungal-specific pro-inflammatory genes pertaining to innate and adaptive immune responses. Interestingly, PP CD11c+ phagocytes also differentially expressed genes in response to C. albicans that were important in the protection of the mucosal barrier. These results highlight that the mucosal barrier not only responds to C. albicans, but also aids in the protection of the host.


Assuntos
Candida albicans/imunologia , Perfilação da Expressão Gênica , Interações Hospedeiro-Patógeno , Inflamação/patologia , Nódulos Linfáticos Agregados/imunologia , Nódulos Linfáticos Agregados/patologia , Administração Oral , Animais , Antígenos CD/análise , Linfócitos B/química , Linfócitos B/imunologia , Células Dendríticas/química , Células Dendríticas/imunologia , Feminino , Camundongos , Saccharomyces cerevisiae/imunologia , Linfócitos T/química , Linfócitos T/imunologia
13.
Fish Shellfish Immunol ; 90: 118-125, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31054358

RESUMO

The present study reveals purification and characterization of the lectin from the haemolymph of Metapenaeus dobsoni. The Md-Lec was purified by affinity chromatography with mannose coupled sepharose CL-4B column and it exhibits single band with a molecular weight of 68 kDa in SDS-PAGE. Furthermore, the molecular mass was confirmed by MALDI-TOF and functional groups present were analysed by FTIR. The surface morphology of purified Md-Lec displays the homogeneous nature of protein. The X-ray diffraction (XRD) analysis expresses three peaks at 10.7716̊, 21.6258̊ and 31.7523̊which indicate the crystalline nature of the protein and the retention time of 3.068 min evident from HPLC reveals the purity of the sample. Functional analysis of purified Md-Lec exhibits yeast agglutination activity against Saccharomyces cerevisiae and has the ability to agglutinate the human erythrocytes, which was observed by light microscopy. It also exhibited phenoloxidase activation, encapsulation and phagocytic activities. In addition, purified Md-Lec showed the broad spectrum of bacterial agglutination activity against Gram negative Vibrio parahaemolyticus and Aeromonas hydrophila, important fish pathogens. Antiviral potential and anticancer activity of purified Md-Lec against CyHV-2 virus and MDA-MB-231 breast cancer cell lines were also evaluated in this study.


Assuntos
Anti-Infecciosos/farmacologia , Proteínas de Artrópodes/imunologia , Lectinas/imunologia , Monofenol Mono-Oxigenase/metabolismo , Penaeidae/imunologia , Aeromonas hydrophila/imunologia , Aglutinação/fisiologia , Animais , Lectinas/metabolismo , Penaeidae/enzimologia , Penaeidae/metabolismo , Saccharomyces cerevisiae/imunologia , Vibrio parahaemolyticus/imunologia
14.
Br J Dermatol ; 181(4): 866-869, 2019 10.
Artigo em Inglês | MEDLINE | ID: mdl-30980721
15.
Appl Microbiol Biotechnol ; 103(13): 5183-5192, 2019 07.
Artigo em Inglês | MEDLINE | ID: mdl-31020381

RESUMO

Recombinant Saccharomyces cerevisiae strains expressing HIV antigens have shown promising pre-clinical results. Probiotic S. cerevisiae strains naturally induce gut immunity; thus, genetically engineered probiotic strains could be used to stimulate immune responses against HIV in the mucosa. Probiotic strains have a higher rate of heterologous protein production, meaning higher antigen's epitope expression levels per yeast cell. We expressed HIV-1 Gag protein in the probiotic yeasts' surface, which was eagerly phagocytosed by and induced type 1 polarization of human monocyte-derived dendritic cells (DCs) from healthy donors in vitro. We further matured DCs derived from HIV-1+ donors with transformed yeasts and incubated them with autologous T cells. Only DCs matured with Gag-expressing probiotic strains were able to efficiently present antigen to CD8+ T cells and induced their clonal expansion. Our results show that genetically engineered probiotic S. cerevisiae strains are a promising vaccination strategy against HIV.


Assuntos
Linfócitos T CD8-Positivos/imunologia , Células Dendríticas/imunologia , Memória Imunológica , Probióticos , Produtos do Gene gag do Vírus da Imunodeficiência Humana/imunologia , Vacinas contra a AIDS/imunologia , Apresentação do Antígeno , Diferenciação Celular , Engenharia Genética , HIV-1/genética , HIV-1/imunologia , Humanos , Masculino , Fagocitose , Saccharomyces cerevisiae/genética , Saccharomyces cerevisiae/imunologia , Células Th1/imunologia , Produtos do Gene gag do Vírus da Imunodeficiência Humana/genética
16.
Autoimmunity ; 52(1): 37-47, 2019 02.
Artigo em Inglês | MEDLINE | ID: mdl-30884988

RESUMO

A clear correlation exists between microbiota and the dysregulation of the immune response in Inflammatory Bowel Diseases (IBD), which comprise Crohn's disease (CD) and ulcerative colitis (UC). These unbalanced reactions also involve humoral responses, with antibodies against Saccharomyces cerevisiae. Thus, here we aimed to quantify IgA and IgG specific to S. cerevisiae (ASCA) in quiescent CD and UC, to correlate the production of these antibodies with patient's inflammatory response and disease clinical presentation. Twenty-nine subjects (16 CD and 13 UC) and 45 healthy controls were enrolled in this study and had plasma samples tested for ASCA and cytokines (IL-2, IL-4, IL-6, IL-10, IFN-γ, TNF-α), besides clinical evaluation. IBD patients had increase IgA and IgG ASCA, especially those with colonic (L2) and fistulizing (B3) CD. Similarly, patients who dropped out the treatment had augmented ASCA, while IgG was reduced in those receiving sulfasalazine treatment. Furthermore, the quiescent CD patients had elevated IL-6 on plasma, especially in the absence of treatment, together with increased counter regulatory response of IL-10. There was a positive correlation between IgA and IgG on CD but not UC, as well as between IgA and TNF in total IBD patients. In addition, the levels of IgG x TNF, IgA x IL-10 and IgG x IL-10 were also correlated in CD, indicating that ASCA production may be influenced by the inflammatory response. Finally, we concluded that ASCA could be pointed as relevant biomarker of CD presentation and residual inflammation, even in clinical remission patients.


Assuntos
Anticorpos Antifúngicos , Doença de Crohn , Imunoglobulina A , Imunoglobulina G , Saccharomyces cerevisiae/imunologia , Adulto , Idoso , Anticorpos Antifúngicos/sangue , Anticorpos Antifúngicos/imunologia , Doença de Crohn/sangue , Doença de Crohn/imunologia , Citocinas/sangue , Citocinas/imunologia , Feminino , Humanos , Imunoglobulina A/sangue , Imunoglobulina A/imunologia , Imunoglobulina G/sangue , Imunoglobulina G/imunologia , Masculino , Pessoa de Meia-Idade
17.
J Pediatr Gastroenterol Nutr ; 69(1): 82-87, 2019 07.
Artigo em Inglês | MEDLINE | ID: mdl-30789863

RESUMO

OBJECTIVE: Although anti-Saccharomyces cerevisiae antibodies (ASCAs) could be a useful biomarker in differentiating Crohn disease (CD) from ulcerative colitis (UC), their role as prognostic markers in children with CD has been underinvestigated. This longitudinal prospective observational study aimed to assess the prognostic value of ASCA status among children with CD managed using biologics. METHODS: The study population comprised children with inflammatory bowel disease diagnosed with CD from 2012 to 2018. Cox regression model with adjustment for a priori covariates was used to examine the response to anti-tumor necrosis factor (TNF) biological therapy among ASCA-positive patients in comparison to ASCA-negative patients. RESULTS: There were 273 measurements available from the study cohort comprising children with CD, who were followed up for a median duration of 14 months (interquartile range 5-42). ASCA-positive patients had a higher risk for moderate to severe clinical disease (odds ratio 2.88; 95% confidence interval [CI] 1.2-7.55) and extensive endoscopic distribution (odds ratio 3.30; CI 1.12-9.74) at baseline in comparison to ASCA-negative patients, respectively. In comparison to ASCA immunoglobulin G (IgG)-negative patients, ASCA IgG-positive patients who were treated with biologics had a significantly lower relapse rate (adjusted hazard ratio 0.12; CI 0.02-0.93). Ten (14%) patients had an unstable ASCA value with either ASCA immunoglobulin A or ASCA IgG status changing from positive to negative or vice versa. CONCLUSIONS: ASCA-positive children with CD present with more extensive (endoscopic) and clinically severe disease. ASCA IgG is a useful prognostic marker among children with CD who receive biologics.


Assuntos
Anticorpos Antifúngicos/sangue , Doença de Crohn/sangue , Doença de Crohn/tratamento farmacológico , Imunoglobulina G/sangue , Adalimumab/uso terapêutico , Adolescente , Anticorpos Anticitoplasma de Neutrófilos/sangue , Biomarcadores/sangue , Criança , Feminino , Fármacos Gastrointestinais/uso terapêutico , Humanos , Imunoglobulina A/sangue , Infliximab/uso terapêutico , Estudos Longitudinais , Masculino , Prognóstico , Estudos Prospectivos , Recidiva , Saccharomyces cerevisiae/imunologia , Índice de Gravidade de Doença , Fator de Necrose Tumoral alfa/antagonistas & inibidores
18.
AIDS Res Hum Retroviruses ; 35(5): 461-472, 2019 05.
Artigo em Inglês | MEDLINE | ID: mdl-30784316

RESUMO

The mechanisms causing HIV-associated immune activation remain incompletely understood. Alteration of intestinal integrity with subsequent translocation of bacterial products appears to play an important role; however, little is known about the impact of fungal translocation. We assessed the effect of fungal translocation and its association with immune activation in people living with HIV (PLWH) compared with uninfected controls. We measured serum levels of ß-D-glucan (BDG) and anti-Saccharomyces cerevisiae antibodies (ASCA) immunoglobulin G (IgG) and immunoglobulin A (IgA) and markers of systemic inflammation and immune activation in virally suppressed PLWH on antiretroviral therapy (ART) and uninfected controls. T-test and Mann-Whitney tests were used to compare markers by HIV status and correlation and regression analyses were used to assess associations of fungal translocation markers with markers of inflammation. One hundred seventy-six participants were included (128 HIV+ and 48 HIV-); 72% male, 65% African American, median age was 50 years, and CD4 was 710 cells/cm3. Levels of BDG tended to be lower in HIV+ when compared with controls (p = .05). No significant difference in levels of ASCA IgG and IgA was seen between groups (p > .75). There was a significant correlation between BDG and several markers of inflammation and immune activation in PLWH, not seen in uninfected controls. In contrast, no correlations were seen between levels of ASCA IgG and IgA with inflammatory markers. PLWH on ART do not have higher levels of BDG or ASCA when compared with uninfected controls, however, the association found between BDG and several inflammation markers suggests a potential role of fungal translocation in the heightened immune activation seen in treated HIV.


Assuntos
Anticorpos Antifúngicos/sangue , Glucanos/sangue , Infecções por HIV/imunologia , Infecções por HIV/microbiologia , Inflamação , Ativação Linfocitária , Adulto , Antivirais/uso terapêutico , Biomarcadores/sangue , Estudos de Coortes , Estudos Transversais , Feminino , Infecções por HIV/sangue , Infecções por HIV/tratamento farmacológico , Humanos , Imunoglobulina A/sangue , Imunoglobulina G/sangue , Masculino , Pessoa de Meia-Idade , Saccharomyces cerevisiae/imunologia
19.
J Immunol Methods ; 466: 24-31, 2019 03.
Artigo em Inglês | MEDLINE | ID: mdl-30611766

RESUMO

The variable lymphocyte receptor B (VLRB) of jawless vertebrates has a similar function to the antibodies produced by jawed vertebrates, and has been considered as an alternative source to mammalian antibodies for use in biological research. We developed a modified yeast display vector system (pYD8) to display recombinant hagfish VLRB proteins on the extracellular surface of yeast for the isolation of antigen-specific VLRBs. After observing an up-regulation in the VLRB response in hagfish immunized with hemagglutinin 1 of avian influenza virus H9N2 subtype (H9N2-HA1), the antigen-specific VLRBs decorated on the yeast's surface were selected by quantitative library screening through magnetic-activated cell sorting (MACS) and fluorescent-activated cell sorting (FACS). We also demonstrated a strong specificity of the antigen-specific VLRBs, when expressed as a secreted protein using a mammalian expression system. Together, our findings suggest that the pYD8 vector system could be useful for screening antigen-specific hagfish VLRBs, and the specificity of secreted VLRB may have potential for a variety of biological applications.


Assuntos
Antígenos/imunologia , Receptores de Antígenos de Linfócitos B/imunologia , Saccharomyces cerevisiae/imunologia , Animais , Linfócitos B/imunologia , Feiticeiras (Peixe) , Receptores de Antígenos de Linfócitos B/genética
20.
Rheumatology (Oxford) ; 58(7): 1176-1187, 2019 07 01.
Artigo em Inglês | MEDLINE | ID: mdl-30649509

RESUMO

OBJECTIVES: To examine faecal calprotectin (F-calprotectin) levels and presence of anti-Saccharomyces cerevisiae antibodies (ASCA) and their associations with disease subtype and current status in axial SpA (axSpA). METHODS: F-calprotectin and ASCA in serum were compared between consecutive patients with a clinical axSpA diagnosis, classified as non-radiographic axSpA (nr-axSpA; n = 40) or AS (n = 90), and with healthy controls (n = 35). Furthermore, standard axSpA outcome measures were compared between axSpA patients (nr-axSpA and AS combined) with elevated vs normal F-calprotectin, ASCA IgA and IgG, respectively. RESULTS: Elevated F-calprotectin (⩾50 mg/kg) was observed in 27% of nr-axSpA patients, 38% of AS patients and 6% of controls. F-calprotectin was significantly higher in AS vs nr-axSpA [AS: geometric mean 41 (95% CI 32, 54) mg/kg; nr-axSpA: 24 (95% CI 16, 38) mg/kg; P = 0.037], and in each axSpA subtype vs controls. Overall, worse disease activity and physical function scores were observed among axSpA patients with elevated vs normal F-calprotectin levels, with significant differences regarding patient's visual analogue scale for global health, ASDAS using CRP, and BASFI (adjusted for age, sex, NSAID use, anti-rheumatic treatments, and CRP). ASCA titres and seropositivity (⩾10 U/ml) were similar in nr-axSpA (IgA/IgG-seropositivity: 8%/26%) and AS (7%/28%), and clinical outcome measures did not differ between patients with elevated vs normal ASCA IgA or IgG, respectively. Compared with controls (IgA/IgG-seropositivity: 0%/17%), ASCA IgA was significantly higher in both axSpA subtypes, and IgG was significantly higher in the AS group. CONCLUSION: In patients with axSpA, gut inflammation measured by elevated F-calprotectin is associated with worse disease activity and physical function, and may be a marker of more severe disease.


Assuntos
Fezes/química , Complexo Antígeno L1 Leucocitário/análise , Espondilartrite/diagnóstico , Adulto , Idoso , Anticorpos Antifúngicos/sangue , Antirreumáticos/uso terapêutico , Biomarcadores/análise , Estudos de Casos e Controles , Estudos de Coortes , Feminino , Humanos , Imunoglobulina A/sangue , Imunoglobulina G/sangue , Masculino , Pessoa de Meia-Idade , Saccharomyces cerevisiae/imunologia , Índice de Gravidade de Doença , Espondilartrite/tratamento farmacológico , Espondilartrite/metabolismo , Espondilite Anquilosante/diagnóstico , Espondilite Anquilosante/tratamento farmacológico , Espondilite Anquilosante/metabolismo , Escala Visual Analógica
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