Keratinocyte Response to Infection with Sporothrix schenckii.

Sporotrichosis is a subacute, or chronic mycosis caused by traumatic inoculation of material contaminated with the fungus Sporothrix schenckii which is part of the Sporothrix spp. complex. The infection is limited to the skin, although its progression to more severe systemic or disseminated forms remains possible. Skin is the tissue that comes into contact with Sporothrix first, and the role of various cell lines has been described with regard to infection control. However, there is little information on the response of keratinocytes. In this study, we used the human keratinocyte cell line (HaCaT) and evaluated different aspects of infection from modifications in the cytoskeleton to the expression of molecules of the innate response during infection with conidia and yeast cells of Sporothrix schenckii. We found that during infection with both phases of the fungus, alterations of the actin cytoskeleton, formation of membrane protuberances, and loss of stress fibers were induced. We also observed an overexpression of the surface receptors MR, TLR6, CR3 and TLR2. Cytokine analysis showed that both phases of the fungus induced the production of elevated levels of the chemokines MCP-1 and IL-8, and proinflammatory cytokines IFN-α, IFN-γ and IL-6. In contrast, TNF-α production was significant only with conidial infection. In late post-infection, cytokine production was observed with immunoregulatory activity, IL-10, and growth factors, G-CSF and GM-CSF. In conclusion, infection of keratinocytes with conidia and yeast cells of Sporothrix schenckii induces an inflammatory response and rearrangements of the cytoskeleton.

Bacillus licheniformis and Bacillus subtilis, Probiotics That Induce the Formation of Macrophage Extracellular Traps.

Probiotics are considered living microorganisms that help preserve the health of the host who uses them. Bacillus are a genus of Gram-positive bacteria used as probiotics for animal and human consumption. They are currently distributed in various commercial forms. Two of the species used as probiotics are B. licheniformis and B. subtilis. Macrophages are central cells in the immune response, being fundamental in the elimination of microbial pathogens, for which they use various mechanisms, including the formation of extracellular traps (METs). There have been very few studies carried out on the participation of macrophages in response to the interaction of probiotics of the genus Bacillus with the host. In this work, we used macrophages from the J774A mouse cell line.1, and we found that they are susceptible to infection by the two Bacillus species. However, both species were eliminated as the infection progressed. Using confocal microscopy, we identified the formation of METs from the first hours of infection, which were characterized by the presence of myeloperoxidase (MPO) and citrullinated histone (Hit3Cit). Quantitative data on extracellular DNA release were also obtained; release was observed starting in the first hour of infection. The induction of METs by B. licheniformis caused a significant decrease in the colony-forming units (CFU) of Staphylococcus aureus. The induction of METS by bacteria of the Bacillus genus is a mechanism that participates in controlling the probiotic and potentially pathogenic bacteria such as S. aureus. The induction of METs to control pathogens may be a novel mechanism that could explain the beneficial effects of probiotics of the genus Bacillus.

Analysis of inflammatory cytokine expression in the urinary tract of BALB/c mice infected with Proteus (P.) mirabilis and enteroaggregative Escherichia (E.) coli (EAEC) strains.

This study aimed to analyze the proinflammatory cytokine mRNA expression in the urinary tract of BALB/c mice infected with bacterial strains with uropathogenic potential. Groups of four 6-week-old female BALB/c mice were intraurethrally inoculated with 5 × 107 colony-forming units (CFU) of P. mirabilis ATCC29906, EAEC O42, P. mirabilis RTX339, or sterile saline (control group) and then sacrificed at 0, 2, 4, 7, or 10 days post-infection (p.i.). Samples were cultured to determine the CFU/mL in urine or CFU/g in the bladders and kidneys. Cytokine expression (tumor necrosis factor (TNF)-α and interleukin (IL)-1ß, -6, and -8) was evaluated in the target organs using real-time PCR and immunohistochemistry; histology was examined with hematoxylin and eosin staining. The results are presented as the means and standard deviations and were compared using one-way ANOVA, with p < 0.05 indicating significant differences. Bacteriuria was not detected in the infected groups; bacterial colonization occurred in the target organs at all time points, but was higher in mice infected with EAEC O42 or P. mirabilis RTX339 at 7 days p.i. The expression of all cytokine mRNAs was seen, but only the levels of the IL-8 protein increased in situ at 7 days p.i. in the P. mirabilis RTX339 and EAEC O42 groups in both organs. Morphological alterations, observed in all of the infected groups, were more prominent in the EAEC O42 and P. mirabilis RTX339 groups. The findings provide insights into the uropathogenicity and inflammatory cytokine expression in the urinary tract of mice infected with three previously untested bacterial strains.

Keratinocyte infection by Actinomadura madurae triggers an inflammatory response.

BACKGROUND: Actinomycetoma is a syndrome of the skin characterized by chronic inflammation and lesions with nodular grain-like structures. The most common aetiological agents are Nocardia brasiliensis and Actinomadura madurae. In response to infection with these organisms the body produces an inflammatory immune response in the skin. The aim of the present study was to determine the production of chemokines, pro-inflammatory cytokines, antimicrobial peptides and the expression of Toll-like receptors (TLRs) in keratinocytes infected by A. madurae. METHODS: A cell line of HaCaT keratinocytes was infected with A. madurae at a multiplicity of infection of 20:1 for 2 h and the samples were collected from 2 to 72 h post-infection. Intracellular replication of the bacterium was evaluated by counting of colony-forming units, the TLR expression and antimicrobial peptide production were assayed by confocal microscopy and chemokine and pro-inflammatory cytokine levels were determined by enzyme-linked immunosorbent assay. RESULTS: Early in the infection, A. madurae was able to achieve intracellular replication in keratinocytes, however, the cells eventually controlled the infection. In response to the infection, keratinocytes overexpressed TLR2 and TLR6, produced high concentrations of cytokines monocyte chemoattractant protein-1, interleukin 8, human ß-defensin-1, human ß-defensin-2 and LL37 and low levels of tumour necrosis factor α. CONCLUSIONS: The human keratinocytes contribute to the inflammatory process in response to A. madurae infection by overexpressing TLRs and producing chemokines, pro-inflammatory cytokines and antimicrobial peptides.

[Presence of lactoferrin (LF) in lymphocutaneous sporotrichosis. Yeast-bound antimicrobial peptide]. / Presencia de lactoferrina (LF) en la esporotricosis linfocutánea. Péptido antimicrobiano unido a levaduras.

Sporotrichosis is a common subcutaneous mycosis in Latin America, produced by dimorphic fungi belong to Sporothrix schenckii complex of cryptic species. Infection is acquired by traumatic inoculation with contaminated organic material. Host immune response includes polymorphonuclear neutrophils chemotaxis and release of granular components. Lactoferrin is a protein member of the transferrin family of iron-binding proteins, present inside polymorphonuclear granular structure, and has been reported to affect growth and development of infectious agents, including fungal organisms. Nevertheless, lactoferrin expression in sporotrichosis infections has not been reported yet. OBJECTIVE: To determine the expression of lactoferrin using immunohistochemical staining in sporotrichosis human infection. MATERIAL AND METHODS: The dermatology department's files during a period of five years were reviewed; cases with a diagnosis of sporotrichosis were selected and lactoferrin immunostaining was performed when enough biological material was available. RESULTS: Three cases with a diagnosis of sporotrichosis and adequate biological material on paraffin block were identified. In all cases, lactoferrin immunostaining was positive around yeast cell.

Activation and IL-1ß secretion of human peripheral phagocytes infected with Actinomadura madurae, Nocardia asteroides and Candida albicans.

OBJECTIVE: To evaluate the ability of Actinomadura madurae (A. madurae) and Nocardia asteroides (N. asteroides), using Candida albicans (C. albicans) as prototypic control, to elicit the activation and IL-1ß secretion of blood phagocytic cells from healthy donors. METHODS: Microscopic evaluation of phagocytosis/activation, cell viability and spectrophotometric quantitation of endocytosis/activation, were assessed by using formazan blue test in human blood phagocytes infected with C. albicans, A. madurae or N. asteroides treated with either normal human serum (NHS) or with decomplemented NHS. Interlukin-1ß from culture supernatants of infected polymorphonuclear was tested by ELISA kit assay. RESULTS: Microscopic assay showed that phagocytosis and activation of adherent mononuclear phagocytes were greater with C. albicans followed by A. madurae and then by N. asteroides. Spectrophotometric assay in polymorphonuclear phagocytes infected with NHS-treated pathogens indicated that activation was similarly higher by C. albicans and A. madurae and lower by N. asteroides. Kinetic assays in infected polymorphonuclear cells showed that viability was decreased by C. albicans and N. asteroides or unaffected with A. madurae. Levels of IL-1ß at 8 h of incubation were higher with C. albicans followed by A. madurae whereas lower levels were found with N. asteroides. CONCLUSIONS: The extent of cell-viability and activation as well IL-1ß secretion may be related with the virulence of C. albicans and N. asteroides and other parameters remain to be explored for assessing the virulence of A. madurae.

Tratamiento periodontal no quirúrgico en pacientes con gingivitis y periodontitis moderada. Respuesta bioquímica y microbiológica / Non surgical periodontal treatment in patients with gingivitis and moderate periodontitis. Biochemical and microbiological response

Objetivo: Conocer la respuesta inflamatoria a través de la presencia de interleucina 1β e identificar microorganismos patógenos como posibles marcadores inmunológicos y microbiológicos en el diagnóstico y tratamiento periodontal no quirúrgico en sujetos con gingivitis y periodontitis crónica moderada en población mexicana. Material y métodos: En este estudio prospectivo de cohortes, se seleccionaron 18 pacientes con signos clínicos de gingivitis y 17 pacientes con periodontitis crónica moderada, se recolectaron las muestras de biopelícula subgingival y de fluido gingival crevicular. Se cuantificó la interleucina 1β durante las fases pretratamiento, postratamiento y de mantenimiento del tratamiento periodontal no quirúrgico. Resultados: Las variables de respuesta microbiológica mostraron que Porphyromonas gingivalis, Prevotella intermedia, Fusobacterium nucleatum, Aggregatibacter actinomycetemcomitans disminuyeron significativamente en individuos con gingivitis. Así como Porphyromonas gingivalis, Tannerella forsythia, Fusobacterium nucleatum, Aggregatibacter actinomycetemcomitans y Actinomyces sp. en periodontitis crónica moderada. Las variables de respuesta bioquímica mostraron una disminución significativa en la concentración y cuenta total de interleucina 1β en los individuos con periodontitis crónica moderada en la fase de mantenimiento del tratamiento así como de las variables de respuesta clínica. Conclusión: Hay reducción de los niveles de interleucina 1β con la disminución de la microflora. Los niveles de interleucina 1β son marcadores sensibles para el diagnóstico y severidad de la enfermedad periodontal.

Objective: To ascertain inflammatory response through interleucina 1β presence and identify pathogenic microorganisms as possible immunological and microbiological markers in diagnosis and treatment non-surgical periodontal in patients with gingivitis and moderate chronic periodontitis in a sample of Mexican population. Material and methods: In the present prospective cohort study, 18 patients with signs of gingivitis and 17 patients with moderate chronic periodontitis were selected. Samples of subgingival biofilm and of crevicular gingival fluid were collected. Interleukine 1β was quantified during the pre-treatment, post-treatment and maintenance phases of the nonsurgical periodontal treatment. Continuous variables were analyzed with the Student test, as well as categorical variables which were analyzed with the TurkeyKramer test. For independent groups the Pearson test was used. Results: Microbiological response variables showed that Porphyromonas gingivalis, Prevotella intermedia , Fusobacterium nucleatum , Aggregatibacter actinomycetemcomitans significantly decreased in subjects with gingivitis. Porphyromonas gingivalis , Tannerella forsythia , Fusobacterium nucleatum , Aggregatibacter actinomycetemcomitans and Actinomyces ssp . decreased in cases. Biochemical response variables showed significant decrease in IL-1β concentration and total count in individuals with moderate chronic periodontitis in treatment maintenance phase. The same result applied to clinical response variables. Conclusions: There is a decrease in Interleukin 1β levels with decrease in microflora. Interleukin 1β are sensitive markers for diagnosis of periodontal disease and assessment of its severity.

Actinomicetoma / Actinomycetoma

El actinomiocetoma es una enfermedad crónica de la piel y tejidos subcutáneos de evolución muy lenta y por tanto poco invalidante durante los primeros meses o años de su aparición. Esto ocasiona que su diagnóstico y tratamiento resulten una desventaja para el individuo que la padece y favorece al progreso de la enfermedad. Un problema adicional es que la incidencia de esta patología en nuestro país es baja y se presenta preferentemente en poblaciones que trabajan en el campo; por esta razón, el presente trabajo de revisión bibliográfica tiene como principal objetivo actualizar la información acerca de las principales características de este padecimiento, las estrategias de diagnóstico y tratamiento, así como las evidencias experimentales que permiten su estudio en un modelo animal