Does cytokine profiling of aspirate from jaw cysts and tumors have a role in diagnosis?

PURPOSE: The objective of the present study was twofold: first, to assess aspirates for use in cytokine profiling and second, to initiate pilot analyses to determine whether the cytokine profiling can serve as an aid in the diagnosis of jaw lesions. MATERIALS AND METHODS: The aspirates from 12 benign odontogenic cysts and tumors of the jaw were collected and randomized, and a formal incisional biopsy was performed to establish the tissue diagnosis. The biopsies revealed keratocystic odontogenic tumor, ameloblastoma, and dentigerous cyst. The cystic aspirate was analyzed using the Q-Plex Human Cytokine Screen to detect cytokine expression and determine the level of expression for each pathologic entity. An array of 16 cytokines was investigated, including interleukin (IL)-1α, IL-1ß, IL-2, IL-4, IL-5, IL-6, IL-8, IL-10, IL-12, IL-13, IL-15, IL-17, IL-23, interferon-γ, tumor necrosis factor (TNF)-α, and TNF-ß. Tables were developed to determine the ratio of expression for the candidate cytokine pairs that were differentially expressed among the 3 pathologic entities encountered. One-way analysis of variance was used to search for significant differences in the ratio of expression of the candidate pairs among the 3 entities. RESULTS: Cytokines expressed by the 3 distinct jaw lesions were detected in the aspirate without the need for tissue biopsy. Cytokine profiling of these entities is possible owing to differential expression of the various cytokines studied. The ratio of expression was significant (P < .05) for 15 pairs of cytokines: IL-5/IL-1α, IL-4/IL-2, IL-8/IL-4, TNF-ß/IL-6, IL-23/IL-6, TNF-α/IL-23, TNF-α/TNF-ß, TNF-α/IL-8, TNF-ß/IL-5, TNF-ß/TNF-α, TNF-ß/IL-13, IL-12/IL-23, IL-13/IL-15, IL-15/IL-2, and IL-6/IL-2. A comparison of the mean values indicated a "high/low" expression value for each lesion type for the 15 cytokine pairs. CONCLUSIONS: Cytokines, expressed by the 3 groups of jaw lesions, can be detected in the cystic aspirate, and a comparison of the ratio of the expression of the aspirates demonstrated a differential expression pattern of cytokines among the 3 groups. These ratios could assist in establishing a prompt and accurate diagnosis of lesions that might be difficult to discern clinically and radiographically. The use of a simple, minimally invasive aspiration procedure can help to establish an accurate diagnosis.

Influence of smoking on gingival crevicular fluid cytokines in severe chronic periodontitis.

AIM: The aim of this study was to compare the expression of 22 chemokines and cytokines in gingival crevicular fluid (GCF) from smokers and non-smokers with periodontitis and periodontally healthy control subjects. MATERIALS AND METHODS: Forty subjects with generalized severe chronic periodontitis (20 smokers and 20 non-smokers) and 12 periodontally healthy control subjects participated in this study. Four diseased and two healthy sites were selected from each of the periodontitis subjects. GCF samples were collected and cytokines analysed utilizing a multiplexed immunoassay (Luminex(®) ). Statistical analyses employed non-parametric tests including the Mann-Whitney and Wilcoxon matched-pairs signed-rank tests. RESULTS: Compared with healthy control subjects, GCF in subjects with chronic periodontitis contained significantly higher amounts of interleukin (IL)-1α, IL-1ß, IL-6, IL-12(p40) (pro-inflammatory cytokines); IL-8, macrophage chemotactic protein (MCP)-1, macrophage inflammatory protein (MIP)-1α, regulated on activation normal T-cell expressed and secreted (RANTES) (chemokines); IL-2, IFN-γ, IL-3, IL-4 (Th1/Th2 cytokines); IL-15 [regulator of T-cells and natural killer (NK) cells]. Smokers displayed decreased amounts of pro-inflammatory cytokines [IL-1α, IL-6, IL-12(p40)], chemokines (IL-8, MCP-1, MIP-1, RANTES), and regulators of T-cells and NK cells (IL-7, IL-15). CONCLUSIONS: Periodontitis subjects had significantly elevated cytokine and chemokine profiles. Smokers exhibited a decrease in several pro-inflammatory cytokines and chemokines and certain regulators of T-cells and NK-cells. This reflects the immunosuppressant effects of smoking which may contribute to an enhanced susceptibility to periodontitis.

Poor Oral Hygiene and High Levels of Inflammatory Cytokines in Saliva Predict the Risk of Overweight and Obesity.

The study aimed to determine if oral hygiene influences not only oral health but also potentially metabolic disorders such as overweight or obesity. Participants were 94 patients: 40 with increased body mass and 54 with normal body mass. The methods included dental examination, a questionnaire concerning hygienic habits and an assessment of selected salivary inflammatory markers. The new parameter named "cleaning index" (describing the interaction between average time of tooth brushing in minutes and its frequency per day) significantly correlated with Body Mass Index (RSpearman = 0.300). The multivariate regression model incorporating cleaning index, approximal plaque index, receptor 1 for tumor necrosis factor-alpha (TNFα-R1) and interleukin-15 (IL-15) had a high power to predict overweight or obesity (AUC = 0.894). Patients with poor oral hygiene (approximal plaque index >40%) were more than eight times more likely to suffer from obesity than patients with good oral hygiene. Cleaning index higher than 4 decreased the odds by about 85%. Oral hygiene habits, adjusted by salivary concentrations of selected inflammatory markers may allow predicting effectively overweight or obesity risk. Early proper dental prophylaxis and treatment could lead to the better prevention of metabolic disorders.

The contribution of interleukin-13 and -15 to the cytokine network within normal and diseased gingiva.

BACKGROUND: There are no comparisons of concentrations of interleukin (IL)-13 and -15 and gingival sulcular depth within healthy or inflamed gingiva. Study of non-oral inflammations indicated that IL-15 has similar functions to IL-2, and IL-13 has similar functions to IL-4 and -10. Thus, IL-13 and -15 likely have a function in gingival inflammation. METHODS: We compared the concentrations of IL-13 and -15 within healthy or inflamed human gingiva to gingival sulcular depth and the concentration of various T-helper 1 and T-helper 2 cytokines in a group of Hispanic subjects. Gingival papillae were grouped by the depth of the adjacent gingival sulcus: 6 mm (diseased). The gingival concentrations of IL-2, -4, -6, -10, -13, and -15 and interferon (IFN)-gamma were assessed by enzyme-linked immunosorbent assay. RESULTS: IL-2, -4, -6, -10, and -13 and IFN-gamma concentrations were higher within gingiva adjacent to 3 to 6 mm diseased compared to normal; IL-15 concentrations were significantly greater within normal gingiva than within diseased gingiva (P <0.001). Within gingiva adjacent to >6 mm sulci, IL-6 concentrations were significantly higher than in normal gingiva, and the concentrations of the other cytokines were similar to those in normal gingiva. Gingival IL-6 concentration correlated significantly with the adjacent gingival sulcular depth, whereas the IL-15 concentration correlated inversely with the adjacent gingival sulcular depth (P <0.001). CONCLUSIONS: IL-13, -4, and -10 had a similar concentration pattern within normal and diseased tissue; however, the concentration pattern of IL-15 was not similar to IL-2. Relatively low IL-15 concentrations within diseased gingiva suggest that IL-15 might have anti-inflammatory properties.

Evaluation of p53, bcl-2, and interleukin-15 levels in gingival crevicular fluid of cyclosporin A-treated patients.

BACKGROUND: Apoptosis plays an important role in the maintenance of tissue homeostasis. Considering that apoptosis mediators may play a role in the pathogenesis of drug-induced gingival overgrowth, this study was conducted to evaluate p53, bcl-2, and interleukin-15 (IL-15) levels in gingival crevicular fluid (GCF) of cyclosporin A (CsA)-treated patients. METHODS: Twenty renal transplant patients exhibiting CsA-induced gingival overgrowth and 15 systemically healthy gingivitis patients were included in the study; 15 systemically and periodontally healthy volunteer subjects served as the healthy control group. GCF samples were obtained from one interdental site with gingival overgrowth (GO+) and one site without (GO-) from each CsA-treated patient; hyperplasia index, probing depth, papilla bleeding index, and plaque presence were recorded. One site from each gingivitis patient and healthy control was selected, GCF samples were obtained, and the same clinical parameters were recorded. GCF p53, bcl-2, and IL-15 levels were analyzed by enzyme-linked immunosorbent assay. The results were tested statistically. RESULTS: p53 and bcl-2 levels were below the minimum detectable level in all GCF samples analyzed. CsA GO+ and CsA GO- sites, as well as gingivitis sites, exhibited significantly higher GCF levels of IL-15 compared to healthy controls (P<0.05). The difference between CsA GO+ sites and gingivitis sites was not statistically significant, although the total amount of IL-15 in CsA GO+ sites was lower than gingivitis sites (P>0.05). The total amount of IL-15 in CsA GO- sites was significantly lower than gingivitis sites (P<0.05). No significant correlation was found between the clinical parameters and GCF IL-15 levels (P>0.05). CONCLUSIONS: The pathogenesis of CsA-induced gingival overgrowth is multifactorial. The findings of the present study indicate that IL-15 may play a role in the pathogenesis of CsA-induced gingival overgrowth due to its interactions with CsA and its role in apoptosis and inflammation.

Immune responses induced by Pelargonium sidoides extract in serum and nasal mucosa of athletes after exhaustive exercise: modulation of secretory IgA, IL-6 and IL-15.

The evidence that exhaustive exercise may compromise the immune response is mainly confirmed by upper respiratory tract infections which are probably related to the decrease in secretory immunoglobulin A in the upper airway mucosa and/or profile changes of systemic cytokines as well as local cytokines of the upper respiratory tract. An extract from Pelargonium sidoides roots is currently used to treat infections in the upper airways. The aim of the present study was to evaluate the action of this herbal medicine on the immune response of athletes submitted to an intense running session by analyzing the production of immunoglobulin A in their saliva and of cytokines both locally and systemically, using a placebo as control. The results show that Pelargonium sidoides extract modulates the production of secretory immunoglobulin A in saliva, both interleukin-15 and interleukin-6 in serum, and interleukin-15 in the nasal mucosa. Secretory immunoglobulin A levels were increased, while levels of IL-15 and IL-6 were decreased. Based on this evidence, we suggest that this herbal medicine can exert a strong modulating influence on the immune response associated with the upper airway mucosa in athletes submitted to intense physical activity.

Role of smoking and type 2 diabetes in the immunobalance of advanced chronic periodontitis.

BACKGROUND: This study evaluates the tissue levels of interleukin (IL)-17(+), IL-15(+), Foxp3(+) cells, fibrosis, and plasma B-cell infiltration in sites with chronic periodontitis in smokers and subjects with type 2 diabetes. METHODS: Gingival biopsies were harvested from the following groups: systemically and periodontally healthy subjects (healthy group, n = 10); non-smokers and subjects with advanced periodontitis and without diabetes (non-risk factor/periodontitis group, n = 10); heavy smokers with advanced periodontitis and without diabetes (smoking/periodontitis group, ≥20 cigarettes per day for at least the past 5 years, n = 10); and non-smoking poorly controlled subjects with diabetes (glycated hemoglobin levels ≥9%) with advanced periodontitis (diabetes mellitus/periodontitis group [DMP], n = 10). The number of IL-17(+), IL-15(+), and Foxp3(+) cells was analyzed by immunohistochemistry, whereas the amount of fibrosis and plasma B-cell infiltration in gingival tissue was analyzed by histomorphometry. RESULTS: The number of Foxp3(+) cells was significantly higher in the periodontitis groups compared to the healthy group (P <0.05). The DMP group presented higher levels of Foxp3(+) cells than other periodontitis groups (P <0.05). The levels of IL-15(+) and IL-17(+) cells and the amount of fibrosis were higher in the DMP group than in the other groups (P <0.05). There was a trend for a decreased B-cell infiltration in the DMP group (P >0.05). There was a slightly significant negative correlation between B-cell infiltration and the amount of fibrosis (P <0.05). CONCLUSION: Upregulation of IL-17(+), IL-15(+), and Foxp3(+) cells and increased amounts of fibrosis were observed in chronic periodontitis sites in subjects with type 2 diabetes, suggesting that periodontitis development in these subjects may be influenced by the T helper 17/T regulatory axis.