interleukin-8 and Candidiasis--Oral

Candida albicans is an opportunistic pathogen that causes oral candidiasis and denture stomatitis. It has also been reported to infect oral mucositis lesions in patients who suffer from cancer affecting the head and neck and who receive chemotherapy and radiotherapy treatments. This study aimed to investigate the effects of two cinnamon bark fractions, i.e., an essential oil and an aqueous extract enriched in proanthocyanidins (Cinnulin PF®) on growth, biofilm formation, and adherence properties of C. albicans as well as on oral epithelial cells (barrier integrity, inflammatory response).. A microplate dilution assay was used to determine antifungal and anti-biofilm properties. A fluorescent assay was used to determine C. albicans adherence to oral epithelial cells. Cytotoxicity toward oral epithelial cells was assessed by determination of cell metabolic activity. Tight junction integrity of gingival keratinocytes was assessed by determination of transepithelial electrical resistance. IL-6 and IL-8 secretion by TNFα-stimulated oral epithelial cells was quantified by ELISA.. While Cinnulin PF® did not reduce C. albicans growth, the cinnamon bark oil exhibited high antifungal activity with minimum inhibitory concentrations and minimum fungicidal concentrations in the range of 0.039 to 0.078%. The cinnamon oil was also active against a pre-formed C. albicans biofilm. Interestingly, Cinnulin PF® prevented biofilm formation by C. albicans and attenuated its adherence to oral epithelial cells. At their effective concentrations, the cinnamon oil and the Cinnulin PF® displayed no significant cytotoxicity against oral epithelial cells. In an in vitro model, both cinnamon fractions reinforced the integrity of the oral epithelial barrier. Lastly, Cinnulin PF® inhibited the secretion of interleukin-6 and interleukin-8 by oral epithelial cells stimulated with TNF-α.. By their ability to attenuate growth, biofilm formation and adherence property of C. albicans, to reinforce the epithelial barrier function, and to exert anti-inflammatory properties the two cinnamon fractions (essential oil, Cinnulin PF®) investigated in the present study may be promising agents for treating oral infections involving C. albicans.

Topics: Antifungal Agents; Biofilms; Candida albicans; Candidiasis, Oral; Cell Line; Cinnamomum zeylanicum; Epithelial Cells; Humans; Interleukin-6; Interleukin-8; Mouth; Oils, Volatile; Plant Bark

Oral candidiasis is a common fungal disease mainly caused by Candida albicans. The aim of this study was to investigate the effects of A-type cranberry proanthocyanidins (AC-PACs) on pathogenic properties of C. albicans as well as on the inflammatory response of oral epithelial cells induced by this oral pathogen.. Microplate dilution assays were performed to determine the effect of AC-PACs on C. albicans growth as well as biofilm formation stained with crystal violet. Adhesion of FITC-labeled C. albicans to oral epithelial cells and to acrylic resin disks was monitored by fluorometry. The effects of AC-PACs on C. albicans-induced cytokine secretion, nuclear factor-kappa B (NF-κB) p65 activation and kinase phosphorylation in oral epithelial cells were determined by immunological assays.. Although AC-PACs did not affect growth of C. albicans, it prevented biofilm formation and reduced adherence of C. albicans to oral epithelial cells and saliva-coated acrylic resin discs. In addition, AC-PACs significantly decreased the secretion of IL-8 and IL-6 by oral epithelial cells stimulated with C. albicans. This anti-inflammatory effect was associated with reduced activation of NF-κB p65 and phosphorylation of specific signal intracellular kinases.. AC-PACs by affecting the adherence properties of C. albicans and attenuating the inflammatory response induced by this pathogen represent potential novel therapeutic agents for the prevention/treatment of oral candidiasis.

Topics: Anti-Inflammatory Agents; Biofilms; Candida albicans; Candidiasis, Oral; Cell Adhesion; Epithelial Cells; Interleukin-6; Interleukin-8; Interleukins; Mouth Mucosa; NF-kappa B; Phosphorylation; Phosphotransferases; Phytotherapy; Plant Extracts; Proanthocyanidins; Saliva; Vaccinium macrocarpon

Oral epithelial cells significantly influence host inflammatory responses against Candida albicans in oropharyngeal candidiasis. Pro-inflammatory cytokines function as an early innate immune system mediator during C. albicans infection in oral epithelial cells. We sought to elucidate the pattern of the molecular mechanisms governing the human gingival epithelial cells (HGECs) to C. albicans infection likely involve multiple converging signal transduction pathways.. Primary HGECs were cultured with C. albicans ATCC90029. Total RNA was extracted after 8 h of infection and monitored mRNA levels using Affymetrix GeneChip (Human Genome U133 plus 2.0 Array, 48 000 genes). GeneChip data was analyzed by GeneSpring software and Ingenuity Pathway Analysis system. Reverse transcription polymerase chain reaction (RT-PCR), real-time RT-PCR and immunohistochemistry were used to investigate gene expression changes.. The differentially expressed genes represented functions as diverse as immune response and inflammatory disease. IL-8, ICAM-1 and Cox-2 showed a greater than two fold change in expression relative to those in control cells. Altered mRNA levels in GeneChip analysis were confirmed by RT-PCR and real-time RT-PCR. Stronger immunoreactivity against ICAM-1 and Cox-2 was also observed in the infection with C. albicans in rat gingival epithelium. We have identified differential gene expression up-regulated or down-regulated with the up-regulation of IL-8 in C. albicans-treated cells.. These findings indicate that the molecular mechanisms underlying the IL-8 response of HGECs to C. albicans infection likely involve multiple converging signal transduction pathways.

Topics: Animals; Candidiasis, Oral; Cyclooxygenase 2; Epithelial Cells; Gene Expression Profiling; Gingiva; Humans; Intercellular Adhesion Molecule-1; Interleukin-8; Male; Oligonucleotide Array Sequence Analysis; Rats; Rats, Sprague-Dawley; RNA; RNA, Messenger; Signal Transduction

 Candida albicans is the predominant oral yeast associated with denture-induced stomatitis, and with an increasing population of denture wearers its incidence is increasing. Maintaining good oral and denture hygiene, through chemical and/or mechanical intervention, is essential to reducing this disease. The aim of this study, using a robust adherent C. albicans cell model system, was to evaluate and compare the efficacy of a novel denture cleanser to the efficacy of a commonly used dentifrice coupled with brushing.. Four C. albicans strains isolated from individuals diagnosed as having denture-induced stomatitis, were adhered to denture acrylic resin sections (1 cm(2) by 1 mm thickness) and after 4 hours of growth, challenged daily sequentially for 4 days with a denture cleanser (Polident) or intermittently with denture cleanser (day 1), then dentifrice (Colgate Cavity Protection Toothpaste) and brushing (days 2 and 3) and denture cleanser (day 4). Colony forming units were evaluated for each treatment, as were the levels of regrowth. Scanning electron microscopy (SEM) was also performed. Microbial susceptibility testing and time-kill studies were performed on biofilms. A coculture model was also used to assess interleukin-8 (IL-8) production from treated biofilms.. It was shown that sequential treatment with the denture cleanser killed and inhibited regrowth each day. Intermittent treatment showed that viable C. albicans biofilms were only retained rather than being dispersed, which could be visualized by SEM. Time-kill studies demonstrated that the novel denture cleanser was highly active and killed quickly, unlike the dentifrice. IL-8 was expressed in greater levels in 24-hour biofilms than in 4-hour biofilms, but treatment with denture cleanser reduced IL-8 output.. The data indicate that maintaining good oral health for denture wearers requires daily use of a denture cleanser rather than an alternating regimen. The inability of the denture cleanser to sterilize during intermittent treatments demonstrates the difficulty in controlling established biofilm. Moreover, the presence of mature biofilm may result in high levels of inflammation, but this can be controlled through denture cleansing.

Topics: Acrylic Resins; Biofilms; Candida albicans; Candidiasis, Oral; Coculture Techniques; Dentifrices; Denture Bases; Denture Cleansers; Humans; Interleukin-8; Oral Hygiene; Stomatitis, Denture; Toothbrushing

To investigate the expression of interleukin-1alpha (IL-1alpha), IL-1ra and IL-8 by the oral epithelium challenged by various Candida species.. In vitro candidiasis was induced by C. albicans wild type SC5314, its EFG1, CPH1 and secretory aspartyl proteinase (SAP) mutants and, ATCC isolates of C. albicans, C. tropicalis and C. dubliniensis using a reconstituted human oral epithelium (RHOE) model. IL-1alpha, IL-1ra and IL-8 levels in culture media were quantified by an enzyme-linked immunosorbent assay at 12, 24 and 48 h. Fungal invasion and IL-1ra expression in RHOE were detected by periodic acid-Schiff staining and immunohistochemistry.. Overall, the invasive Candida induced relatively higher levels of IL-1alpha, IL-1ra and IL-8 in the culture media than the noninvasive isolates. IL-1alpha and IL-1ra levels induced by Candida with hyphal invasion were significantly higher (P < 0.05) than those induced by the isolates without hyphal invasion at 12, 24 and 48 h. Candida albicans SC5314 induced IL-1ra expression in RHOE at 12 and 24 h but not at 48 h consistent with its hyphal invasion; while the noninvasive mutants and non-albicans Candida induced IL-1ra expression at 48 h.. The cytokine expression profiles in experimental oral candidiasis may be associated with the invasive potential of Candida.

Topics: Candida; Candida albicans; Candida tropicalis; Candidiasis, Oral; Cell Line; Culture Media; Epithelium; Humans; Hyphae; Immunohistochemistry; Interleukin-1alpha; Interleukin-8; Mouth Mucosa; Mutation; Receptors, Interleukin-1 Type I; Time Factors

Neutrophils are the main opponents of Candida albicans in chronic hyperplastic candidosis. They migrate from the circulation to the epithelium where they form microabscesses. We therefore hypothesized that the neutrophil chemokine interleukin-8 (IL-8) might play a role in the neutrophil-Candida interaction.. Biopsies from patients with chronic hyperplastic candidosis (n = 10) were stained using the avidin-biotin-peroxidase complex protocol for IL-8 and IL-8 receptor A and were compared to healthy control mucosa (n = 3). A set of C. albicans agar sections was similarly analysed.. In chronic hyperplastic candidosis lesions IL-8 was strongly expressed in both vascular endothelium and mucosal epithelium. Many resident and immigrant inflammatory cells, including intraepithelial neutrophils, were IL-8 receptor A positive. In addition, IL-8 (or an analogue) was found in the candidal mother cell in chronic hyperplastic candidosis and in agar, whereas the tips of the hyphae expressed IL-8 receptor A (or an analogue).. IL-8 may play a role in the recruitment of neutrophils from the vascular compartment to the epithelial microabscesses. C. albicans may have developed an ability to sense IL-8. The IL-8 ligand-receptor interaction may help to direct the growth of the IL-8-receptor-containing tips of the hyphae away from the IL-8-producing candidal cell body (a centrifugal growth pattern to facilitate host tissue penetration). Later, this ability might help to keep the vulnerable hyphal tips away from areas with high concentrations of host IL-8 and candidacidal neutrophils. We suggest that this phenomenon, in contrast to chemotropism, is named chemophobia.

Topics: Adult; Aged; Aged, 80 and over; Candida albicans; Candidiasis, Oral; Chemotaxis, Leukocyte; Chronic Disease; Endothelium, Vascular; Epithelium; Female; Humans; Hyperplasia; Immunoenzyme Techniques; Interleukin-8; Male; Middle Aged; Mouth Mucosa; Neutrophil Infiltration; Neutrophils; Palate; Receptors, Interleukin-8A; Tongue Diseases

The immune response and the anticandidal activity of keratinocytes and polymorphonuclear leukocytes (PMNs) play a key role in host defence against localized Candida albicans infection. An established model of oral candidosis based on reconstituted human oral epithelium (RHE) was supplemented with PMNs to study the effect of these immune cells during experimental oral candidosis. Infection of RHE with C. albicans induced a strong expression of the chemokine interleukin-8 (IL-8) and the cytokine granulocyte-macrophages colony-stimulating factor (GM-CSF), and a moderate stimulation of interleukin-1alpha (IL-1alpha), interleukin-1beta (IL-1beta), interleukin-6 (IL-6), interferon gamma (IFN-gamma) and tumour necrosis factor alpha (TNF-alpha) by keratinocytes. This immune response was associated with chemoattraction of PMNs to the site of infection, whereas uninfected RHE failed to induce cytokine expression or to attract PMNs. Growth of the pathogen and tissue damage of C. albicans-infected RHE were significantly reduced when PMNs were applied to the apical epithelial surface or when PMNs migrated through a perforated basal polycarbonate filter of the model. Notably, protection against epithelial tissue damage was also observed when PMNs were placed on the basal side of non-perforated filters, which prevented PMN migration into the RHE. Addition of PMNs enhanced a Th1-type immune response (IFN-gamma, TNF-alpha), down-regulated the expression of the Th2-type cytokine interleukin-10 (IL-10), and was associated with protection against Candida-induced tissue damage. This PMN-supplemented model of oral candidosis mimics the in vivo situation, and provides a promising tool for studying the immunological interactions between keratinocytes and C. albicans, as well as the influence of PMNs on C. albicans pathogenesis.

Topics: Candida albicans; Candidiasis, Oral; Cells, Cultured; Cytokines; Epithelium; Granulocyte-Macrophage Colony-Stimulating Factor; Humans; Interferon-gamma; Interleukin-1; Interleukin-6; Interleukin-8; Keratinocytes; Mouth Mucosa; Neutrophils; Tumor Necrosis Factor-alpha

Oropharyngeal candidiasis is a frequent opportunistic infection associated with immunocompromised hosts. Candida albicans is the principal species responsible for this infection. Production of interleukin-8 (IL-8), by oral epithelial cells can be expected to play a major role in the recruitment and activation of professional phagocytes at the infected site. The purpose of this study was to determine whether C. albicans triggers secretion of IL-8 by oral epithelial cells in vitro and investigate mechanisms of host cell-fungal interactions that trigger such responses. Oral epithelial cell lines (SCC4, SCC15, and OKF6/TERT-2) as well as primary gingival epithelial cells were used. Epithelial cells were cocultured with C. albicans, strains SC5314, ATCC28366 or ATCC32077, for 24-48 hr, and supernatants were analyzed for IL-8 content by ELISA. A germination-deficient mutant (efg1/efg1 cph1/cph1), otherwise isogenic to strain SC5314, was used to assess the requirement for germination in triggering IL-8 responses. In order to ascertain whether direct contact of yeast with host cells is required to trigger cytokine production, epithelial cells were separated from yeast using cell culture inserts. To test whether IL-8 secretion is dependent on IL-1alpha activity, epithelial cells were challenged with viable C. albicans in the presence or absence of neutralizing anti-IL-1alpha antibody or IL-1ra, and IL-8 secretion was measured in the supernatants. All cell lines and primary cultures responded to C. albicans with an increase in IL-8 secretion. IL-8 responses were contact-dependent, strain-specific, required yeast viability and germination into hyphae, and were in part autoregulated by IL-1alpha.

Topics: Candida albicans; Candidiasis, Oral; Cell Line; Cells, Cultured; Epithelial Cells; Humans; Interleukin 1 Receptor Antagonist Protein; Interleukin-1; Interleukin-8; Keratinocytes; Mouth Mucosa; Neutralization Tests; Sialoglycoproteins; Species Specificity

A defined and balanced immunomodulatory response is crucial for the protection of mucosal surfaces being in contact with pathogenic microorganisms. This study examined the local host response mechanisms of epithelial cells in experimental Candida albicans, C. tropicalis, and C. glabrata infections by measuring the expression of cytokines at the mRNA and protein level. During the course of infection with active but not with heat-killed C. albicans stimulation of the gene expression levels for interleukin-1alpha, interleukin-1beta, tumor necrosis factor, Exodus-2, P-selectin ligand, granulocyte-monocyte colony-stimulating factor, and interleukin-8 was observed by standard and quantitative reverse transcription-polymerase chain reaction. This cytokine pattern may favor a chemotactic and a T helper 1 response. Initial moderate or weak upregulation of these cytokine genes by reverse transcription-polymerase chain reaction was also observed in epithelial infection with the less virulent species C. tropicalis and C. glabrata. Heat-killed C. albicans failed to induce an epithelial immune response. At the protein level, expression of interleukin-8 protein was strongly enhanced during the course of C. albicans infection, whereas lower levels were seen with C. tropicalis and C. glabrata. The different expression patterns of cytokines were associated with differences in virulence of the Candida strains. This study's data, therefore, show a correlation between the virulence potential of pathogenic fungi, possibly mediated by specific virulence factors (such as proteinases), and the secretion of epithelial cytokines and chemokines, which may initiate in vivo a protective T helper 1 immunologic response and contribute to the recruitment of activated leukocytes and lymphocytes to the site of mucosal infection.

Topics: Candida albicans; Candidiasis, Oral; Chemokine CCL21; Chemokines, CC; Cytokines; Epithelium; Gene Expression; Granulocyte-Macrophage Colony-Stimulating Factor; Hot Temperature; Humans; In Vitro Techniques; Interleukin-1; Interleukin-8; Membrane Glycoproteins; RNA, Messenger; Tumor Necrosis Factor-alpha; Virulence

Candidiasis is the most commonly encountered opportunistic infection among HIV-positive subjects. The purpose of this study was to assess specific keratinocyte immune parameters in the pseudomembranous and erythematous forms of HIV-associated oral candidiasis.. This collaborative study from three centers analyzed 25 HIV-positive and 10 HIV-negative subjects with either pseudomembranous or erythematous candidiasis. Oral biopsy specimens from lesional tissues were procured, and histopathologic features were correlated with immunohistochemical and in situ hybridization investigations for the expression of interleukin 1 alpha, interleukin 8, antimicrobial calprotectin, lymphocyte populations, and Candida antigen.. Both pseudomembranous and erythematous candidiasis among HIV-infected subjects showed a mild interface lymphocytic mucositis with the presence of neutrophilic subcorneal abscesses in the latter. Erythematous candidiasis cases that failed to show surface mycelia, did yield positive results for Candida antigens in the parakeratinized layer. The expression of inflammatory chemokines were positive in all groups and calprotectin appeared to serve as a keratinocyte barrier to hyphal penetration.. The erythematous form of candidiasis is often devoid of hyphae yet the presence of Candida antigens in the surface epithelium implicates an immune or allergic process. The intactness of chemokines and antimicrobial calprotectin in keratinocytes may explain why disseminated candidiasis is rarely encountered in HIV-infected patients.

Topics: AIDS-Related Opportunistic Infections; Antigens, Fungal; Antigens, Surface; Calcium-Binding Proteins; Candida; Candidiasis, Oral; Chemokines; Erythema; Gene Expression Regulation; Gene Expression Regulation, Fungal; HIV Seronegativity; HIV Seropositivity; Humans; Hypersensitivity; Immunohistochemistry; In Situ Hybridization; Interleukin-1; Interleukin-8; Keratinocytes; Leukocyte L1 Antigen Complex; Lymphocytes; Mouth Mucosa; Neural Cell Adhesion Molecules; Neutrophils; Stomatitis