Expression profiling and functional analysis of Toll-like receptors in primary healthy human nasal epithelial cells shows no correlation and a refractory LPS response.

BACKGROUND: Innate immune recognition via Toll-like receptors (TLRs) on barrier cells like epithelial cells has been shown to influence the regulation of local immune responses. Here we determine expression level variations and functionality of TLRs in nasal epithelial cells from healthy donors. METHODS: Expression levels of the different TLRs on primary nasal epithelial cells from healthy donors derived from inferior turbinates was determined by RT-PCR. Functionality of the TLRs was determined by stimulation with the respective ligand and evaluation of released mediators by Luminex ELISA. RESULTS: Primary nasal epithelial cells express different levels of TLR1-6 and TLR9. We were unable to detect mRNA of TLR7, TLR8 and TLR10. Stimulation with Poly(I:C) resulted in a significant increased secretion of IL-4, IL-6, RANTES, IP-10, MIP-1ß, VEGF, FGF, IL-1RA, IL-2R and G-CSF. Stimulation with PGN only resulted in significant increased production of IL-6, VEGF and IL-1RA. Although the expression of TLR4 and co-stimulatory molecules could be confirmed, primary nasal epithelial cells appeared to be unresponsive to stimulation with LPS. Furthermore, we observed huge individual differences in TLR agonist-induced mediator release, which did not correlate with the respective expression of TLRs. CONCLUSION: Our data suggest that nasal epithelium seems to have developed a delicate system of discrimination and recognition of microbial patterns. Hypo-responsiveness to LPS could provide a mechanism to dampen the inflammatory response in the nasal mucosa in order to avoid a chronic inflammatory response. Individual, differential expression of TLRs on epithelial cells and functionality in terms of released mediators might be a crucial factor in explaining why some people develop allergies to common inhaled antigens, and others do not.

Endoscopic treatment of inverted papilloma attached in the frontal sinus/recess.

BACKGROUND: Inverted papilloma (IP) is a benign sinonasal tumour for which endoscopic surgery, with complete removal of the underlying and surrounding mucoperiosteum at the attachment site followed by drilling and/or coagulation of this area, is the treatment of choice. This can be challenging in the frontal sinus. OBJECTIVES: To report on the outcome of treatment for IPs involving the frontal sinus. To propose the possible use of topical 5-fluorouracil 5% (5-FU) in the postoperative management of challenging IPs. METHODS: This is a retrospective cohort evaluation of patients with IPs attached in the frontal sinus or in the frontal recess and growing into the frontal sinus. Data on primary or revision surgery, uni- or bilaterality, attachment site, surgical procedure, 5-FU usage, recurrence and follow-up are provided. The end points are disease-free follow-up in months and recurrence. RESULTS: Twenty cases, including fifteen revision cases, were retrieved over a period of ten years. All cases were treated endoscopically. Two cases recurred (10%) and the intervention was repeated. In eight cases, 5-FU was applied at the end of surgery. None of these cases recurred. The mean follow-up after the last intervention was 42 months (standard deviation (SD) 22.1). CONCLUSION: IP involving the frontal sinus is a surgical challenge that can be successfully addressed endoscopically. The topical application of 5-FU could have a place in postoperative treatment when it is difficult to be absolutely sure that all diseased mucoperichondrium or mucoperiosteum at the attachment site(s) has been completely removed.

Mono-allergic and poly-allergic rhinitis patients have comparable numbers of mucosal Foxp3+CD4+ T lymphocytes.

BACKGROUND: We previously found that allergic rhinitis patients with an isolated pollen sensitization responded more strongly to a nasal provocation with grass pollen (GP) than patients who had an additional house dust mite (HDM) sensitization. To elucidate this phenomenon, we investigated the dynamics of Foxp3+CD4+ T lymphocytes in allergic rhinitis patients with distinct allergen sensitizations. METHODS: Three groups of allergic rhinitis patients with skin prick test confirmed allergic sensitizations were investigated and compared to 14 healthy controls: 14 subjects with an isolated grass pollen sensitization (Mono-GP); 9 subjects with isolated housedust mite sensitization (Mono-HDM); 29 subjects with grass pollen and house dust mite sensitization (poly-sensitized). Subjects in the Mono-GP group were challenged with grass pollen extract, subjects in the Mono-HDM group were challenged with house dust mite extract, subjects in the poly-sensitized group and the healthy controls were randomly challenged with either grass pollen or house dust mite. Nasal biopsies were taken before and after nasal provocation. We compared the distribution of FoxP3+CD4+ cells in nasal biopsies before and after nasal provocation using immunohistochemistry. RESULTS: There was no difference in the number of FoxP3+CD4+ cells between healthy and the three allergic groups at baseline.Nasal provocation did result in an increase in eosinophils in the three allergic groups, but did not result in a change in the number of FoxP3+CD4+ cells in any of the groups or induced differences between any of the groups. CONCLUSION: Clinical differences in the response between mono-GP and multiple-sensitized allergic individuals are not related to differences in the number of regulatory T cells in the nasal mucosa.

Acute sinusitis and blindness as the first presentation of chronic lymphocytic leukaemia.

Chronic lymphocytic leukaemia (CLL) is the most frequent form of leukaemia among adults in the Western world, presenting at a median age of 65 years. The diagnosis is usually made incidentally during routine blood examination while the disease is still in its early phase. We report a case of blindness of 24 hours due to acute sinusitis based on CLL localisation in a patient with undiagnosed CLL. Emergency endoscopic sinus surgery and intra- and extra-ocular orbital decompression were performed. The sinusitis resolved after surgery and intravenous antibiotics. Her vision improved within 24 hours and eventually recovered completely after six months. Her CLL remained in an indolent state, needing no active treatment. This case illustrates that blindness from a lymphoproliferative disorder may be treated with emergency endoscopic sinus surgery instead of conventional chemotherapy in order to salvage the vision first, even if the vision is lost for more than 24 hours.

Initial experiences with endoscopic rhino-neurosurgery in Amsterdam.

Endoscopic surgery of the skull base has been on the rise for several years. Endoscopic access for surgery can be achieved from the frontal sinus anteriorly along the skull base to the odontoid process posterior inferiorly. An endoscope is inserted through one nasal corridor and allows visualization of the working field and up to three surgical instruments can be used to address the lesion. This is called the "two nostrils-four hands technique". This is a retrospective study of 67 cases. Setting of the study is an Amsterdam University hospital. Cases were identified in the department of otorhinolaryngology and department of neurosurgery database. All patients operated between 1 January, 2008 and 1 February, 2012 with pituitary tumours that extend beyond the sella, sinonasal tumours and all non-pituitary skull-base tumours were included. Mean tumour diameter was 3.8 cm. We performed a near-to-gross total resection in 92% of cases where we intended to perform a total resection. The most frequent complication was CSF leakage. This study demonstrates that this technique is safe and reliable. What is needed is a dedicated team, which includes a dedicated anesthesiologist, endocrinologist, ophthalmologist, and radiation oncologist.

Long-term results of functional endoscopic sinus surgery in children with chronic rhinosinusitis with nasal polyps.

BACKGROUND: Chronic rhinosinusitis with nasal polyps (CRSwNP) is rare in children and has a major impact on Quality of Life (QoL). Functional endoscopic sinus surgery (FESS) has proven to be an effective treatment, but it is still unclear what long-term outcomes are in children with CRSwNP. The objective of this study was to assess long-term results of FESS in children with CRSwNP. METHODOLOGY: We performed a combined prospective and retrospective study. A QoL questionnaire was send to all children with CRSwNP who received FESS between the year 2000-2010. Almost half of these children also filled in this questionnaire preoperatively. RESULTS: Forty-four Children underwent FESS. From 18 patients, we also prospectively collected preoperative QoL questionnaires. The mean follow-up period was 4.0 years (+- 2.9). The mean age at surgery was 13 years (+-2.9). Of these children, 9 had CF and 10 children asthma. R-SOM scores showed a significant improvement both in general symptoms as well as several different domains when comparing pre- and postoperative questionnaires. Only 5 of 44 patients needed a subsequent intervention. In children with CF this was 3 of 9. CONCLUSION: This study demonstrates that long-term results of FESS in children with CRSwNP are good. QoL has improved significantly, especially in nasal symptoms, showing that FESS is a good treatment in children with CRSwNP. Furthermore, even children with CF show good results.

Nasal hyper-reactivity is a common feature in both allergic and nonallergic rhinitis.

BACKGROUND: Nasal hyper-reactivity is an increased sensitivity of the nasal mucosa to various nonspecific stimuli. Both allergic rhinitis (AR) and nonallergic rhinitis (NAR) patients can elicit nasal hyper-reactivity symptoms. Differences in the prevalence or type of nasal hyper-reactivity in AR and NAR patients are largely unknown. In this study, we quantitatively and qualitatively assessed nasal hyper-reactivity in AR and NAR. METHODS: In the first part, an analysis of a prospectively collected database was performed to reveal patient-reported symptoms of hyper-reactivity. In the second part, cold dry air provocation (CDA) was performed as a hyper-reactivity measure in AR and NAR patients and healthy controls, and symptoms scores, nasal secretions and peak nasal inspiratory flow were measured. Comparisons were made between AR and NAR patients in both studies. RESULTS: The database analysis revealed high hyper-reactivity prevalence in AR (63.4%) and NAR (66.9%). There were no differences between AR and NAR in terms of the number or type of hyper-reactivity stimuli. Hyper-reactivity to physical stimuli did not exclude a response to chemical stimuli, or vice versa. CDA provocation resulted in a significant increase in rhinitis symptoms and the amount of nasal secretions in AR and NAR patients, but not in controls. CONCLUSIONS: We found no quantitative or qualitative differences in nasal hyper-reactivity between AR and NAR patients. It is not possible to differentiate NAR subpopulations based on physical or chemical stimuli.

The microdebrider, a step forward or an expensive gadget?

BACKGROUND: Although the use of the microdebrider (shaver) is well known in endoscopic sinus surgery (FESS), there is lack of evidence from comparative studies focussing on the difference in operating time, intra-operative blood loss and user-friendliness between the microdebrider and traditional operating techniques. In this study we compared the use of the microdebrider to conventional instruments in FESS in these areas. METHODS: A prospective randomised double blind controlled trial in 60 patients with chronic rhinosinusitis with nasal polyposis (CRSwNP) undergoing bilateral FESS. Each subject received FESS using only traditional instruments (Blakseley forceps) on one side and the other side with the additional use of the microdebrider, this way serving as their own control. The primary outcome was operation time, intra-operative blood loss and user friendliness and secondly safety and postoperative healing with a follow-up period at different time points up to three months postoperative. RESULTS: We found a 37% longer operating time when operating without a microdebrider. This difference was highly significant. The microdebrider scored significantly higher on every different parameter of user friendliness, except on the preparation of the instrument needed before surgery. For estimated blood loss during surgery we found no differences. Also there was no significant difference in postoperative healing at any point of time. CONCLUSIONS: This study demonstrates that operating patient with CRSwNP with the microdebrider is efficient and that the microdebrider at the same time is safe and easy to use.

Long-term low-dose antibiotics in recalcitrant chronic rhinosinusitis: a retrospective analysis.

INTRODUCTION: In recalcitrant Chronic RhinoSinusitis (CRS) treatment with intranasal corticosteroids, short-term antibiotics and even sinus surgery is frequently insufficient. Long-term low-dose administration of antibiotics has been suggested as a treatment option in these patients. We analysed the outpatient clinic population treated with different long-term low-dose antibiotics at the AMC Amsterdam. PATIENTS AND METHODS: Eligible patients, who were treated with trimethoprim-sulfamethoxazole or macrolides, were retrospectively identified from our outpatient clinic in 2009. The two main outcome measures were sinonasal complaints and nasal endoscopic findings. A 5-point grading scale was used to score the results compared with the pre-treatment situation. This was measured at several time-points during, and after the antibiotic course, and at the end of the follow-up term. RESULTS: Seventy-six patients were included, 53 per cent had asthma and all of them had undergone sinus surgery. Seventy-eight per cent showed improvement of the symptoms, and 84 per cent demonstrated improvement of the sinonasal mucosa at the end of the course. No significant difference was found between the trimethoprim-sulfamethoxazole and macrolide group. DISCUSSION: Long-term low-dose treatment with antibiotics seems to improve CRS symptoms and the appearance of the sinonasal mucosa on nasal endoscopy. However, at this stage, strong conclusions are immature because no placebo-group has been included. Despite increasing use of long-term low-dose treatment of recalcitrant CRS in referral centres, hard clinical evidence is lacking. More research is urgently required.

Plasmacytoid dendritic cells in pulmonary lymphoid follicles of patients with COPD.

Plasmacytoid dendritic cells (pDCs) are professional antigen-presenting cells with antiviral and tolerogenic capabilities. Viral infections and autoimmunity are proposed to be important mechanisms in the pathogenesis of chronic obstructive pulmonary disease (COPD). The study aimed to quantify blood dendritic cell antigen 2-positive pDCs in lungs of subjects with or without COPD by immunohistochemistry and flow cytometry, combined with the investigation of the influence of cigarette smoke extract (CSE) on the function of pDCs in vitro. pDCs were mainly located in lymphoid follicles, a finding compatible with their expression of lymphoid homing chemokine receptors CXCR3 and CXCR4. pDC accumulated in the lymphoid follicles and in lung digests of patients with mild to moderate COPD, compared with smokers without airflow limitation and patients with COPD Global Initiative for Chronic Obstructive Lung disease (GOLD) stage III-IV. Exposing maturing pDC of healthy subjects to CSE in vitro revealed an attenuation of the expression of co-stimulatory molecules and impaired interferon-α production. Maturing pDC from patients with COPD produced higher levels of tumour necrosis factor (TNF)-α and interleukin (IL)-8 compared to pDC from healthy subjects. CSE significantly impairs the antiviral function of pDCs. In COPD, a GOLD stage dependent accumulation of pDC in lymphoid follicles is present, combined with an enhanced production of TNF-α and IL-8 by maturing pDCs.

Interactions between epithelial cells and dendritic cells in airway immune responses: lessons from allergic airway disease.

Micro-organisms constantly invade the human body and may form a threat to our health. Traditionally, concepts of defence mechanisms have included a protective outer layer of epithelia and a vigilant immune system searching for areas where the integrity of the outer layer may be compromised. Instead of considering these elements as two independent mechanisms, we should be treating them as a single integrated system. This review will present and discuss the role of local immune-competent cells and local epithelia in the recognition of potential pathogens and how the interaction between the two components may affect the initiation of the airway immune response. A concept emerges where airway mucosal dendritic cells act as integrators of both immunostimulatory and immunosuppressive signals that act within actively-involved mucosal tissue.

Presentation of two cases of nasal type NK/T-cell lymphoma.

Nasal type NK/T-cell lymphoma is a rare type of predominantly extranodal non-Hodgkin lymphoma. Early and correct diagnosis with prompt treatment of NK/T-cell lymphoma is important in view of its potentially aggressive behavior and poor response to treatment with additional sites of tumor developing sometimes weeks or months after initial diagnosis. Unfortunately diagnosis of NK/T-cell lymphomas often proves difficult. The diagnosis is essentially based on the clinical presentation of extranodal ulcerative lesions in the upper aero-digestive tract and histopathologic analysis of biopsies using immunohistochemistry. Here we present two cases with nasal-type NK/T-cell lymphoma that illustrate that definitive diagnosis is often delayed due to the atypical initial presentation. We will discuss the pitfalls in diagnosing this rare type of neoplasm and review the treatment options.

Desloratadine reduces systemic allergic inflammation following nasal provocation in allergic rhinitis and asthma patients.

BACKGROUND: Preclinical studies have demonstrated that some second-generation antihistamines have anti-inflammatory effects. It is not known whether these effects are also demonstrable in vivo. In this study we investigated the effect of treatment with desloratadine (DL) on systemic inflammation and on nasal and bronchial mucosal inflammation after nasal allergen provocation (NP) in subjects with grass-pollen-allergic rhinitis and asthma. METHODS: Twenty-six subjects with grass-pollen-allergic rhinitis and asthma were randomly allocated to 8 days of treatment with DL (n = 13) or placebo (n = 13) outside the grass pollen season. On day 7 they underwent nasal provocation with grass pollen allergen. Nasal and bronchial biopsies were taken for immunohistochemical evaluation, and blood samples were analysed. Rhinitis and asthma symptoms, peak nasal inspiratory flow and peak expiratory flow, were also measured at specified times. RESULTS: The number of circulating eosinophils decreased during DL treatment, and there was a reduced increase in circulating eosinophils after NP in these subjects. There was also a significant reduction in early bronchial clinical response. There was no significant lessening in the severity of the nasal symptoms. Nasal and bronchial mucosal inflammation parameters did not alter under DL treatment. CONCLUSION: These data suggest that treatment with DL reduces systemic eosinophilia and prevents the increase in circulating eosinophils after NP. DL also significantly reduces the early bronchial clinical response to NP. However, airway mucosal inflammation is not altered by 1 week of treatment.