Cross-Regulation of Innate and Adaptive Immunity: A New Perspective for the Pathogenesis of Inflammatory Bowel Disease

No abstract available.

Expression of TIM-3, Human beta-defensin-2, and FOXP3 and Correlation with Disease Activity in Pediatric Crohn's Disease with Infliximab Therapy

BACKGROUND/AIMS: This study investigated the expression of T cell immunoglobulin- and mucin-domain-containing molecule 3 (TIM-3), human beta-defensin (HBD)-2, forkhead box protein 3 (FOXP3), and the frequency of CD4+ CD25+ FOXP3+ regulatory T cells (Tregs) in children with Crohn's disease (CD) during infliximab therapy. METHODS: We enrolled 20 CD patients who received infliximab treatment for 1 year. Peripheral blood and colonic mucosal specimens were collected from all CD patients and from healthy control individuals. RESULTS: A significant difference in TIM-3 mRNA expression was evident in peripheral blood mononuclear cells and colonic mucosa between CD patients before infliximab therapy and the healthy controls (p<0.001 and p=0.005, respectively). A significant difference in HBD-2 mRNA expression was found in colonic mucosa between CD patients before infliximab therapy and the healthy controls (p=0.013). In the active phase of CD, at baseline, the median percentage of T cells that were CD25+ FOXP3+ was 1.5% (range, 0.32% to 3.49%), which increased after inflixmab treatment for 1 year to 2.2% (range, 0.54% to 5.02%) (p=0.008). CONCLUSIONS: Our study suggests that both the adaptive and innate immune systems are closely linked to each other in CD pathogenesis. And the results of our study indicate that it could be a useful therapeutic tool, where restoration of TIM-3, HBD-2 and the function of Tregs may repair the dysfunctional immunoregulation in CD.

Human beta defensin-4 and keratinocyte growth factor gene expression in cultured keratinocyte and fibroblasts of burned patients

PURPOSE: To evaluate KGF and human beta defensin-4 (HBD-4) levels produced by dermic fibroblasts and keratinocytes cultivated from burned patients' skin samples. METHODS: Keratinocytes and fibroblasts of 10 patients (four major burns, four minor burns and two controls) were primarily cultivated according to standard methods. HBD-4 and KGF genes were analyzed by quantitative PCR. RESULTS: In fibroblasts, KGF gene expression was 220±80 and 33.33±6.67 (M±SD; N=4), respectively for major and minor burn groups. In keratinocytes, KGF gene expression was 11.2±1.9 and 3.45±0.37 (M±SD; N=4), respectively for major and minor burn groups. In fibroblasts, HBD-4 gene expression was 15.0±4.0 and 11.5±0.5 (M±SD; N=4), respectively for major and minor burn. In keratinocyte, HBD-4 gene expression was 0.0±0.0 and 13.4±4.8 (M±SD; N=4), respectively for major and minor burn. CONCLUSIONS: KGF expression was increased in burn patient fibroblasts compared to control group. In keratinocytes culture, KGF suppression is inversely proportional to burn extension; it is active and increased in major burn but decreased in minor burn. HBD-4 expression was increased in fibroblasts and decreased in keratinocytes from all burned patients. .

Evaluation of hbeta;D-1 and hbeta; D-2 levels in saliva of patients with oral mucosal diseases / Evaluación de los niveles de hbeta; D-1 y hbeta;D-2 en la Saliva de pacientes con enfermedades de la mucosa oral

OBJECTIVE: This study aimed to determine a possible correlation between oral mucosal disease and salivary concentrations of the antimicrobial peptides human beta-defensin-1 (hβD-1) and human betadefensin- 2 (hβD-2). METHOD: The present work focussed on the establishment of a reversed phase-high performance liquid chromatography (RP-HPLC) procedure to quantify human beta-defensins (hβD-1 and hβD-2) in saliva samples of patients with oral diseases such as lichen planus (n = 10), Behçet (n = 10) and recurrent apthous stomatitis (n = 10). RESULTS: Linear calibration range for hβD-1 and hβD-2 defensins was 1.67−200 µg mL-1 and 3.13− 100 µg mL-1 with R2 values of 0.9998 and 0.996, correspondingly. The concentration of beta-defensins in saliva was determined by comparing the peak areas of eluted hβD-1 and hβD-2 with that of their standards. The variation of the amount of beta-defensins was evaluated by comparisons of the results obtained from the patients with oral mucosal diseases before and after treatments and the control subjects. The limit of detection (LOD) and limit of quantification (LOQ) were found to be 1.62 µg mL- 1 and 5.39 µg mL-1 for hβD-1 and 0.94 µg mL-1 and 3.13 µg mL-1 for hβD-2, respectively. CONCLUSION: The salivary beta-defensin concentration was significantly higher in patients with oral mucosal diseases than in healthy volunteers; furthermore, in patients with oral mucosal diseases, the concentration was significantly higher before treatment than after treatment.

OBJETIVO: Este estudio tuvo por objeto determinar una posible correlación entre la enfermedad de la mucosa oral y las concentraciones salivales de la beta-defensina humana 1 (hβD-1) y la beta-defensina humana 2 (hβD-2) de los péptidos antimicrobianos. MÉTODO: El presente trabajo estuvo encaminado al establecimiento de un procedimiento de cromatografía líquida de alta eficacia de fase reversa (RP-HPLC) para cuantificar las beta-defensinas humanas (hβD-1 y hβD-2) en muestras de saliva de pacientes con enfermedades orales como el liquen plano (n = 10), Behçet (n = 10), y la estomatitis aftosa recurrente (n = 10). RESULTADOS: El rango de calibración lineal de las defensinas hβD-1 y hβD-2 fue 1.67-200 µg mL-1 y 3.13-100 µg mL-1 con valores R2 de 0.9998 y 996, respectivamente. La concentración de beta-defensinas en la saliva se determinó utilizando el área de sus estándares. La variación de la cantidad de beta defensinas fue evaluada por comparaciones de los resultados obtenidos de los pacientes con enfermedades de la mucosa oral, antes y después de los tratamientos y los sujetos de control. Se halló que el límite de detección (LDD) y el límite de cuantificación (LDC) fueron 1.62 µg mL-1 y 5.39 µg mL- 1 para hβD-1 y 0.94 µg mL-1 y 3.13 µg mL-1 hβD-2, respectivamente. CONCLUSIÓN: La concentración de beta-defensina salival fue significativamente mayor en los pacientes con enfermedades de la mucosa oral que en los voluntarios sanos. Además, en pacientes con enfermedades de la mucosa oral, la concentración fue significativamente mayor antes del tratamiento que después del tratamiento.

Toll-like Receptors and Antimicrobial Peptides Expressions of Psoriasis: Correlation with Serum Vitamin D Level

To evaluate the association of Toll-like receptors (TLRs), antimicrobial peptides (AMPs) and vitamin D receptors (VDRs) in psoriasis, lesional (PP) and perilesional skin (PN) from psoriasis, atopic dermatitis (AD) patients and healthy controls (NN) were studied by immunohistochemistry. Compared with PN, AD and NN skin, dysregulated expression of TLRs, AMPs and VDR was detected in PP skin. Noteworthy, our results showed altered correlation between TLR2 and VDR expression in PP and PN skin. Human beta defensin 2 (HBD2) and cathelicidin (LL-37) expressions in the PP skin were higher in serum vitamin D sufficient (VDS) groups than serum vitamin D deficient (VDD) groups. Negative correlation was found between TLR2 and VDR expression in the PP skin of VDD groups. However, positive correlation was noted in the PP skin of VDS groups. Based on the present results, therapies targeting the activity of TLRs, AMPs and vitamin D, including modulation of the TLR-VDR pathways, might provide new therapeutic approaches to the psoriasis and other inflammatory skin diseases.

Study of rat beta-defensin-2 gene and protein expression in ventilator-associated pneumonia / 中华危重病急救医学

OBJECTIVE: To study the changes in the expression of rat beta-defensin-2 (RBD-2) gene in the lung tissue with P. aeruginosa (PA) pneumonia following tracheal mechanical ventilation (MV), and to evaluate the pathogenesis of ventilator-associated pneumonia (VAP). METHODS: A total of 58 normal healthy Sprague-Dawley rats, weighing between 280 and 320 g, were randomly divided into the control group and the conventional MV group (CMV). A tracheal catheter was inserted via mouth in every rat under urethane anesthesia. PA (1 MIC, 0.2 ml) was instilled into the tracheal in the control group. Rats of CMV group received MV (V(T)=12 ml/kg) through tracheal tube for 24 hours, and then were challenged intra-tracheally with PA (1 MIC, 0.2 ml). Fluid loss was replenished through intravenous infusion. The arterial catheter was used for hemodynamics, parameters were monitored, and arterial blood gases were determined. Samples of lung were harvested at 0 hours, 15 hours, 3 hours, 6 hours, 12 hours, 1 day, 3 days and 5 days, respectively, after bacterial challenge. The mRNA of RBD-2 was detected by reverse transcription-polymerase chain reaction (RT-PCR), and the protein levels were analyzed by Western blotting. RESULTS: Expression of RBD-2 mRNA and protein was lower in CMV group compared with the control 3 hours before instillation of bacteria. RBD-2 mRNA increased 3 hours after bacteria instillation, reaching the peak at 12-24 hours. No significant difference in RBD-2 expression between the control group and the CMV group within 3 hours, but it was significantly higher at 3 hours, 6 hours, 12 hours, 1 day, 3 days and 5 days in the control group than in the CMV group. The number of inflammatory cells infiltrating the bronchial submucous layer was significantly higher in the control group than in the CMV group (P<0.05). There was milder interstitial pulmonary edema and less red blood cells in the alveoli in the control group than in the CMV group. The mortality rate of the CMV group was 60%, which was significantly higher than that of the control group (20%, P<0.05). The positive rates of blood culture and bronchoalveolar lavage fluid (BALF) bacterial culture were also higher in the CMV group (P<0.05). The survival rate in CMV group (40%) was lower than that of the control group (P<0.05). CONCLUSION: The lowering of BD-2 gene and protein expression in the CMV group 3 hours after bacteria challenge might be one of the contributory factors in causing VAP.