Tom Godthelp

Dynamics of mast cells in the nasal mucosa of patients with allergic rhinitis and non‐allergic controls: a biopsy study

Mast cell degranulation is thought to be an important component of the pathogenesis of allergic rhinitis. Quantitative studies on mast cells in nasal mucosa after allergen exposure have given widely divergent results, ranging from an overall decrease via redistribution to an overall increase. We investigated this problem by employing a combination of anti‐IgE and toluidine blue staining of biopsy specimens. In allergic patients anti‐IgE was found to identify all mast cells and toluidine blue to detect mast cells that were not (totally) degranulated.

Antigen presenting cells in the nasal mucosa of patients with allergic rhinitis during allergen provocation

Background The role of antigen presenting cells (APC) in allergic rhinitis is underexposed. Allergen presentation to T lymphocytes is probably an important aspect of the pathophysiological mechanism of allergic rhinitis.

Dynamics of nasal eosinophils in response to a nonnatural allergen challenge in patients with allergic rhinitis and control subjects: a biopsy and brush study

BACKGROUND Eosinophils are thought to play an important role in the symptomatology and pathophysiology of allergic rhinitis. Most quantitative studies on eosinophils in nasal mucosa have focused on the dynamics of eosinophils in the acute and late phases of the allergic reaction by using different cell sampling techniques. Little is known about the dynamics of eosinophils during a more prolonged period of allergen exposure and the activation of eosinophils induced by allergen challenge. OBJECTIVE The aim of this study was to investigate the dynamics and activation of the eosinophils in the nasal mucosa of patients with an isolated grass pollen allergy during an out-of-season 2-week allergen exposure, mimicking the natural grass pollen season. METHODS Seventeen patients with isolated grass pollen allergy and four control subjects were challenged daily with the allergen during a 2-week period in the winter. Nasal brush specimens were obtained before provocation and each day during the provocation period. Biopsy specimens were obtained once before, six times during, and once after the provocation period. Preparations made of nasal brush and nasal biopsy specimens were stained with the monoclonal antibody BMK 13 and Giemsa stain as paneosinophil markers and with the monoclonal antibody EG2 to identify activated eosinophils. RESULTS We found significant increases in the total number of eosinophils and the number of activated eosinophils in the epithelium and lamina propria. These increases were most explicit in the second week. BMK 13 was found to be a paneosinophil marker superior to Giemsa staining. CONCLUSION Eosinophils are not only involved in the acute and late phases of the allergic reaction but are probably even more involved in the chronic phase.

Effect of 3 months' nasal steroid therapy on nasal T cells and Langerhans cells in patients suffering from allergic rhinitis

The effect of nasal corticosteroid therapy on allergic rhinitis is uncertain. In a double‐blind, placebo‐controlled study over 3 months, we investigated the influence of a new corticosteroid spray, fluticasone propionate aqueous nasal spray (FPANS), on Langerhans cells (CD1a+ cells), HLA‐DR+ cells, and T cells in nasal mucosa. Efficacy was evaluated by nasal symptom score. This treatment significantly decreased the number of CDla+ cells and HLA‐DR+ cells in the nasal mucosa. Furthermore, a clear trend of decreasing numbers of T cells in nasal epithelium was found. No change in nasal symptom score was found after the treatment period. These findings suggest that fluticasone propionate aqueous nasal spray decreases the antigen presentation in nasal allergy.

The effect of nasal steroid aqueous spray on nasal complaint scores and cellular infiltrates in the nasal mucosa of patients with nonallergic, noninfectious perennial rhinitis ☆ ☆☆ ★

Topical corticosteroids are the therapy of choice for nonallergic, noninfectious perennial rhinitis (NANIPER). However, the efficacy of steroid therapy in NANIPER is controversial, as is its mode of action. To our surprise, of 300 patients initially diagnosed as having NANIPER, only 65 reached threshold nasal symptom scores. Patients were randomized into four different treatment regimens: placebo administered twice daily (BD) for 8 weeks, fluticasone propionate aqueous nasal spray (FPANS) (200 microg) once daily (OD) and placebo OD for 8 weeks, FPANS (200 microg) OD and placebo OD for 4 weeks followed by FPANS (200 microg) BD for 4 weeks, and FPANS (200 microg) BD for 8 weeks. A small decrease in nasal symptoms was found, which only reached significance for sneezing in the FPANS 200 microg BD group. A significant dose-dependent decrease in immunocompetent cells was found in nasal biopsy specimens obtained before, after 4 weeks, and after 8 weeks of treatment. We conclude that FPANS did not significantly reduce nasal symptoms in this group of selected NANIPER patients, even though a significant effect on cells in the nasal mucosa was seen.

Allergen binding to specific IgE in the nasal mucosa of allergic patients

BACKGROUND Until now, it has not been possible to identify specific IgE locally in the airway mucosa. In this study we investigated the possibility of determining specific allergen binding locally in the nasal mucosa. METHODS Nasal mucosal biopsy specimens were taken from 11 patients with symptoms of an isolated grass pollen allergy, 10 patients with symptoms of perennial allergic rhinitis in response to house dust mite allergen, and 10 nonallergic control subjects. Sections of these biopsy specimens were stained by using commercially available biotinylated allergens (AlaSTAT, Diagnostic Products Corp.). RESULTS Staining with biotinylated grass pollen (GP1) demonstrated positive cells only in patients with grass pollen allergy. Biotinylated Dermatophagoides pteronyssinus (D1) only stained cells in patients with perennial allergy. Specific binding of allergen to cells of patients with allergy and the blocking experiments proved the method to be highly specific. Allergen-positive cells stained double with IgE, the high-affinity receptor for IgE (Fc epsilon RI), CD1, HLA-DR, tryptase, and chymase. Most allergen-positive cells proved to be mast cells. CONCLUSION This immunohistochemical study shows the presence of specific IgE against grass pollen and house dust mite allergens locally on cells in the airway mucosa.

Mast cells, eosinophils and IgE-positive cells in the nasal mucosa of patients with vasomotor rhinitis: an immunohistochemical study

Vasomotor rhinitis (VMR) is a disorder of unknown pathogenesis. Forty patients with VMR were carefully selected on the basis of inclusion and exclusion criteria proposed by Mygind and Weeke. Nasal biopsy specimens were taken in the patient group as well as in a group of ten controls. Brush cytology was also taken in the VMR group. Inflammatory cells were identified and counted in the nasal mucosa, with the use of immunohistochemical techniques and a panel of monoclonal antibodies. Eosinophils were studied with the use of BMK13, EG2, and Giemsa. Mast cells were studied with anti-chymase (B7), anti-tryptase (G3) and toluidine blue. Sections were stained with IgE as well. There was no significant difference in the number of eosinophils, mast cells and IgE-positive cells between the two groups. Additionally, in contrast with other reports, in sections that were double-stained with anti-chymase and anti-tryptase, single chymase-positive cells were found.

Long-term effects of corticosteroid nasal spray on nasal inflammatory cells in patients with perennial allergic rhinitis

The effect of long‐term topical nasal corticosteroid therapy on nasal inflammatory cells is unclear.

Local corticosteroid treatment: the effect on cells and cytokines in nasal allergic inflammation

Regular and prophylactic use of topical corticosteroids is a well tolerated and effective treatment for allergic rhinitis. The symptomatology of allergic rhinitis is considered to be the result of the accumulation and activation of infiltrating inflammatory cells, releasing mediators, and cytokines. Corticosteroids can suppress many stages of the allergic inflammatory process. This may explain their potent effect on allergic symptomatology. The reduction in cell numbers and probably also cytokines by local corticosteroid therapy differs from cell to cell. Some cells, such as antigen presenting (Langerhans) cells and eosinophils, are highly sensitive to corticosteroid treatment. Others, like T cells, are only significantly reduced in exaggerated situations, for instance after provocation with a high allergen dose or after treatment with a high dose of corticosteroids. Some cells, like macrophages, are not influenced at all.

Fixation with Carnoy's fluid reduces the number of chymase-positive mast cells : not all chymase-positive mast cells are also positive for tryptase

Mast cells in the nasal mucosa can be studied by means of monoclonal antibodies (mAb) against tryptase (T+MC) and chymase (C+MC). Fixation with acetone gives more positive cells than does fixation with Carnoys fluid. In frozen biopsy specimens of allergic nasal mucosa fixed with acetone, the number of T+MC equals that of C+MC. When fixed with Carnoys fluid, however, the number of T+MC is larger than the number of C+MC. The decrease in both T+MC and C+MC resulting from fixation with Carnoys fluid is time‐related and depends on the type of mAb used. Carnoy fixation time gives a decrease in the number of C+MC within 1 min, whereas the number of T+MC decreases only after 10 min. Within 1 min, the number of C+MC decreases to a level where continued fixation no longer gives further decreases in the number of cells. Two populations of mast cells can be distinguished here: one sensitive and the other insensitive to Carnoys fluid. When double‐staining is used, fixation with acetone gives three populations of mast cells: one positive for tryptase (T+C‐MC), another positive for tryptase and chymase (T+C+MC), and a third one positive for chymase (T‐C+MC). These three populations were found in lymph node, spleen, thymus, dermis, lung parenchyma, small intestinal submucosa, and nasal mucosa.