RESUMEN
BACKGROUND: Staphylococcus aureus (S. aureus) is a pathogen responsible for a wide range of clinical manifestations and potentially fatal conditions. There is a paucity of information on the influence of androgens in the immune response to S. aureus infection. In this study, we evaluated the influence of the hormone 5α-dihydrotestosterone (DHT) on mouse peritoneal macrophages (MPMs) and human peripheral blood monocytes (HPBMs) induced by S. aureus. METHODS: An in vitro model of MPMs from BALB/c sham males, orchiectomised (OQX) males, and females was used. Cells were inoculated with 10 µL of S. aureus, phage-type 80 or sterile saline (control) for 6 h. The MPMs of OQX males and females were pre-treated with 100 µL of 10-2 M DHT for 24 h before inoculation with S. aureus. The concentration of the cytokines TNF-α, IL-1α, IL-6, IL-8, and IL-10; total nitrites (NO-2); and hydrogen peroxide (H2O2) were measured in the supernatant of MPM cultures. In addition, the toll-like receptor 2 (TLR2) and nuclear factor kappa B (NF-kB) genes that are involved in immune responses were analysed. For the in vitro model of HPBMs, nine men and nine women of childbearing age were selected and HPBMs were isolated from samples of the volunteers' peripheral blood. In women, blood was collected during the periovulatory period. The HPBMs were inoculated with S. aureus for 6 h and the supernatant was collected for the analysis of cytokines TNF-α, IL-6, IL-12; and GM-CSF, NO-2, and H2O2. The HPBMs were then removed for the analysis of 84 genes involved in the host's response to bacterial infections by RT-PCR array. GraphPad was used for statistical analysis with a p value < 0.05. RESULTS: Our data demonstrated that MPMs from sham males inoculated with S. aureus displayed higher concentrations of inflammatory cytokines and lower concentrations of IL-10, NO-2, and H2O2 when compared with MPMs from OQX males and females. A similar result was observed in the HPBMs of men when compared with those of women. Previous treatment with DHT in women HPBMs increased the production of pro-inflammatory cytokines and decreased the levels of IL-10, NO-2, and H2O2. The analysis of gene expression showed that DHT increased the activity of the TLR2 and NF-kB pathways in both MPMs and HPBMs. CONCLUSIONS: We found that DHT acts as an inflammatory modulator in the monocyte/macrophage response induced by S. aureus and females exhibit a better immune defence response against this pathogen.
Asunto(s)
Infecciones Estafilocócicas , Staphylococcus aureus , Masculino , Humanos , Femenino , Animales , Ratones , Staphylococcus aureus/metabolismo , Dihidrotestosterona/farmacología , FN-kappa B/genética , FN-kappa B/metabolismo , Interleucina-10 , Monocitos/metabolismo , Receptor Toll-Like 2/metabolismo , Factor de Necrosis Tumoral alfa , Peróxido de Hidrógeno , Interleucina-6 , Citocinas/metabolismo , Infecciones Estafilocócicas/microbiología , Macrófagos/metabolismoRESUMEN
Staphylococcus aureus is a Gram-positive bacterium that is considered an important human pathogen. Due to its virulence and ability to acquire mechanisms of resistance to antibiotics, the clinical severity of S. aureus infection is driven by inflammatory responses to the bacteria. Thus, the present study aimed to investigate the modulating role of citral in inflammation caused by S. aureus infection. For this, we used an isolate obtained from a nasal swab sample of a healthy child attending a day-care centre in Vitória da Conquista, Bahia, Brazil. The role of citral in modulating immunological factors against S. aureus infection was evaluated by isolating and cultivating human peripheral blood mononuclear cells. The monocytes were treated with 4%, 2%, and 1% citral before and after inoculation with S. aureus. The cells were analysed by immunophenotyping of monocyte cell surface molecules (CD54, CD282, CD80, HLA-DR, and CD86) and cytokine dosage (IL-1ß, IL-6, IL-10, IL-12p70, IL-23, IFN-γ, TGF-ß, and TNF-α), and evaluated for the expression of 84 genes related to innate and adaptive immune system responses. GraphPad Prism software and variables with P values < 0.05, were used for statistical analysis. Our data demonstrated citral's action on the expression of surface markers involved in recognition, presentation, and migration, such as CD14, CD54, and CD80, in global negative regulation of inflammation with inhibitory effects on NF-κB, JNK/p38, and IFN pathways. Consequently, IL-1ß, IL-6, IL-12p70, IL-23, IFN-γ, and TNF-α cytokine expression was reduced in groups treated with citral and groups treated with citral at 4%, 2%, and 1% and infected, and levels of anti-inflammatory cytokines such as IL-10 were increased. Furthermore, citral could be used as a supporting anti-inflammatory agent against infections caused by S. aureus. There are no data correlating citral, S. aureus, and the markers analysed here; thus, our study addresses this gap in the literature.
Asunto(s)
Monoterpenos Acíclicos/farmacología , Factores Inmunológicos/farmacología , Monocitos/efectos de los fármacos , Infecciones Estafilocócicas/inmunología , Staphylococcus aureus/inmunología , Adulto , Brasil , Células Cultivadas , Niño , Humanos , Inflamación/complicaciones , Inflamación/tratamiento farmacológico , Inflamación/inmunología , Masculino , Monocitos/inmunología , Infecciones Estafilocócicas/complicaciones , Infecciones Estafilocócicas/tratamiento farmacológico , Staphylococcus aureus/efectos de los fármacos , Adulto JovenRESUMEN
OBJECTIVE: To study the inflammatory profile and genes involved in the response to bacterial infections in women who developed spontaneous abortion in the presence of Ureaplasma parvum. DESIGN: Cross-sectional study. SETTING: A maternal and child referral center. PATIENT(S): Eighty-nine women with spontaneous abortion and 20 women with normal vaginal delivery (control group) were studied. INTERVENTION(S): Samples of biopsied placental tissue were collected for Mollicutes detection. MAIN OUTCOME MEASURE(S): The samples were subjected to histologic analysis, immunohistochemical evaluation for macrophages and lymphocytes, cytokine quantification, and quantitative polymerase chain reaction array to evaluate the expression of 84 genes related to the innate and adaptive immune responses. RESULT(S): The presence of U. parvum in the abortion group was positively associated with the influx of polymorphonuclear cells in the placental tissue and increased concentrations of interleukin-6 and interleukin-12p70. U. parvum caused downregulation of genes involved in the immune response, such as attraction of immune cells, activation of an inflammatory response, T-helper cell 17 response activation, and activation of the complement system at the beginning and end of pregnancy. CONCLUSION: The direct action of U. parvum on placental tissue altered the gestational tolerogenic state, reducing the immune response against pathogens and activating the extrinsic apoptotic pathway, causing spontaneous abortion.
Asunto(s)
Aborto Espontáneo/microbiología , Histocompatibilidad Materno-Fetal , Tolerancia Inmunológica , Placenta/microbiología , Complicaciones Infecciosas del Embarazo/microbiología , Infecciones por Ureaplasma/microbiología , Ureaplasma/patogenicidad , Aborto Espontáneo/diagnóstico , Aborto Espontáneo/inmunología , Inmunidad Adaptativa , Apoptosis , Proteínas Reguladoras de la Apoptosis/genética , Estudios de Casos y Controles , Estudios Transversales , Citocinas/genética , Femenino , Regulación de la Expresión Génica , Histocompatibilidad Materno-Fetal/genética , Interacciones Huésped-Patógeno , Humanos , Tolerancia Inmunológica/genética , Inmunidad Innata , Placenta/inmunología , Embarazo , Complicaciones Infecciosas del Embarazo/diagnóstico , Complicaciones Infecciosas del Embarazo/genética , Complicaciones Infecciosas del Embarazo/inmunología , Factores de Riesgo , Ureaplasma/inmunología , Infecciones por Ureaplasma/diagnóstico , Infecciones por Ureaplasma/genética , Infecciones por Ureaplasma/inmunologíaRESUMEN
BACKGROUND: Probiotics are important tools in therapies against vaginal infections and can assist traditional antibiotic therapies in restoring healthy microbiota. Recent research has shown that microorganisms belonging to the genus Lactobacillus have probiotic potential. Thus, this study evaluated the potential in vitro probiotic properties of three strains of Lactiplantibacillus plantarum, isolated during the fermentation of high-quality cocoa, against Gardnerella vaginalis and Neisseria gonorrhoeae. Strains were evaluated for their physiological, safety, and antimicrobial characteristics. RESULTS: The hydrophobicity of L. plantarum strains varied from 26.67 to 91.67%, and their autoaggregation varied from 18.10 to 30.64%. The co-aggregation of L. plantarum strains with G. vaginalis ranged from 14.73 to 16.31%, and from 29.14 to 45.76% with N. gonorrhoeae. All L. plantarum strains could moderately or strongly produce biofilms. L. plantarum strains did not show haemolytic activity and were generally sensitive to the tested antimicrobials. All lactobacillus strains were tolerant to heat and pH resistance tests. All three strains of L. plantarum showed antimicrobial activity against the tested pathogens. The coincubation of L. plantarum strains with pathogens showed that the culture pH remained below 4.5 after 24 h. All cell-free culture supernatants (CFCS) demonstrated activity against the two pathogens tested, and all L. plantarum strains produced hydrogen peroxide. CFCS characterisation in conjunction with gas chromatography revealed that organic acids, especially lactic acid, were responsible for the antimicrobial activity against the pathogens evaluated. CONCLUSION: The three strains of L. plantarum presented significant probiotic characteristics against the two pathogens of clinical importance. In vitro screening identified strong probiotic candidates for in vivo studies for the treatment of vaginal infections.
Asunto(s)
Antibiosis/fisiología , Cacao/microbiología , Alimentos Fermentados/microbiología , Gardnerella vaginalis/fisiología , Lactobacillus plantarum/fisiología , Neisseria gonorrhoeae/fisiología , Probióticos , Fermentación , Humanos , Lactobacillus plantarum/aislamiento & purificaciónRESUMEN
In recent years, certain Lactobacillus sp. have emerged in health care as an alternative therapy for various diseases. Based on this, this study is aimed at evaluating in vitro the potential probiotics of five lactobacilli strains isolated from pulp of cupuaçu fruit fermentation against Gardnerella vaginalis and Neisseria gonorrhoeae. Our lactobacilli strains were classified as safe for use in humans, and they were tolerant to heat and pH. Our strains were biofilm producers, while hydrophobicity and autoaggregation varied from 13% to 86% and 13% to 25%, respectively. The coaggregation of lactobacilli used in this study with G. vaginalis and N. gonorrhoeae ranged from 15% to 36% and 32% to 52%, respectively. Antimicrobial activity was present in all tested Lactobacillus strains against both pathogens, and the growth of pathogens in coculture was reduced by the presence of our lactobacilli. Also, all tested lactobacilli reduced the pH of the culture, even in incubation with pathogens after 24 hours. The cell-free culture supernatants (CFCS) of all five lactobacilli demonstrated activity against the two pathogens with a halo presence and CFCS characterization assay together with gas chromatography revealed that lactic acid was the most abundant organic acid in the samples (50% to 62%). Our results demonstrated that the organic acid production profile is strain-specific. This study revealed that cupuaçu is a promising source of microorganisms with probiotic properties against genital pathogens. We demonstrated by in vitro tests that our Lactobacillus strains have probiotic properties. However, the absence of in vivo tests is a limitation of our work due to the need to evaluate the interaction of our lactobacilli with pathogens in the vaginal mucosa. We believe that these findings may be useful in developing a product containing our lactobacilli and their supernatants in order to support with vaginal health.