bluuebunny/crossref_metadata_2025 huggingface dataset:
Degranulation influences heparin‐associated inhibition of RT‐PCR in human lung mast cells
Authors: Glaum M. C. , Wang Y. , Raible D. G. , Schulman E. S.
Published: March 2003

Background The study of gene expression from human lung mast cells (HLMC) has been limited by the ability to reliably detect mRNA transcripts from scant quantities of mast cell RNA contaminated with heparin.

Objective As heparin is granule‐associated within the mast cell, we examined the role of degranulation in altering the intrinsic ability of this proteoglycan to inhibit reverse transcription polymerase chain reaction (RT‐PCR) from HLMC RNA. We also explored alternative means of RNA isolation to eliminate primary heparin contamination.

Methods Purified HLMC (> 90% pure) were challenged for 2 h with buffer, anti‐IgE (3 µg/mL) and/or ionophore A23187 (100 ng/mL) or phorbol 12‐myristate 13‐acetate (50 ng/mL). Histamine release was measured using a spectroflourometric assay. Following challenge, RNA was isolated by either phenol‐chloroform extraction or by nitrocellulose spin column. Parallel samples were either treated with heparinase or placed on ice for 2 h prior to reverse transcription. PCR was performed using primers specific for the housekeeping gene, glyceraldehyde‐3‐phosphate dehydrogenase (GAPDH).

Results In each of five studies, GAPDH bands were detected at greater intensity in total cellular RNA (tcRNA) derived from degranulated than from non‐degranulated mast cells. However, when examining heparinase‐treated tcRNA from the same mast cell samples, the intensity of GAPDH bands normalized between degranulated and non‐degranulated cells. When comparing parallel samples of column‐purified tcRNA, subsequent treatment of samples with heparinase had no effect on the detection of GAPDH, indicating that endogenous heparin was effectively removed by the technique. Moreover, no variability was noted in GAPDH signal detected from resting vs. degranulated mast cells (n = 3).

Conclusions Degranulation influences the degree of heparin‐associated inhibition of RT‐PCR in HLMC, and that spin column purification of tcRNA is a time‐saving, effective alternative to heparinase pre‐treatment.
A polymorphism in the coding region of interleukin‐13 gene is associated with atopy but not asthma in Chinese children
Authors: Leung T. F. , Tang N. L. S. , Chan I. H. S. , Li A. M. , Ha G. , Lam C. W. K.
Published: March 2003

Background Interleukin (IL)‐13 is an important cytokine secreted from type 2 helper T lymphocytes. It is essential for modulating IgE synthesis by human B cells. Previous studies showed that polymorphisms in the IL‐13 gene were associated with serum total IgE or allergic asthma. The relationship of this marker with sensitization to individual aeroallergens has not been evaluated.

Objective We tested whether a polymorphism in the coding region of the IL‐13 gene is associated with asthma and atopy in asthmatic children in Hong Kong.

Methods We used restriction fragment length polymorphism to detect R130Q genotype in Chinese children with asthma and control subjects. Serum total IgE was measured by microparticle immunoassay and specific IgE to common aeroallergens was measured using fluorescent enzyme immunoassay. Pulmonary function studies were performed using spirometry.

Results One hundred and fifty‐seven patients and 54 control children were recruited. Their mean serum total IgE concentrations were 994 kIU/L and 473 kIU/L, respectively (P < 0.0001). Atopy as defined by ≥ 1 positive RAST was found in 141 patients and 32 control children. The GlnGln form of the R130Q polymorphism in the IL‐13 gene was associated with serum total IgE (P = 0.005) as well as specific IgE to Der p 1 (P = 0.021), mixed cockroaches (P = 0.03) and dog (P = 0.003) but not with physician‐diagnosed asthma (P = 0.621). In addition, the R130Q polymorphism did not correlate with subjective or objective indicators of asthma severity in our patients.

Conclusion Our results suggest that the R130Q polymorphism of the IL‐13 gene is associated with elevated serum total and allergen‐specific IgE but not asthma in Chinese children.
Selective protective effects of nitric oxide inhalation on allergen‐induced acute airway reactions in the pig
Authors: Middelveld R. J. M. , Alving K.
Published: March 2003

Background Nitric oxide (NO) is thought to be an important mediator of inflammatory processes during allergic reactions in the respiratory tract.

Objective This study was undertaken to investigate the effects of inhalation of NO on the allergen‐induced acute airway reactions in the pig.

Methods Specific pathogen‐free pigs were sensitized with Ascaris suum antigen and challenged with an allergen aerosol during mechanical ventilation and anaesthesia. One group (n = 8) was treated with inhaled NO (20 ppm) which was given from 30 min before allergen challenge until the experiments were completed at 120 min after challenge. A control group (n = 8) did not receive NO (< 0.001 ppm).

Results Inhalation of 20 ppm NO prevented the fall in arterial pO2/FiO2 levels that was observed in the control group (areas under the curve between 0 and 120 min were 3.7 ± 1.4 kPa/min in NO‐treated pigs vs. 15.9 ± 3.4 in controls, P < 0.01, Mann–Whitney U‐test) and it decreased baseline pulmonary arterial pressure (change from time‐point − 30–0 was 3.1 ± 5.3% in the control and − 19.9 ± 3.5% in the NO group, P < 0.01), which in turn resulted in a lower pulmonary arterial pressure during allergen challenge. NO also caused vasodilatation in the bronchial circulation, resulting in increased bronchial vascular conductance throughout the experiment. NO inhalation caused a small, but non‐significant, reduction in the allergen‐induced bronchoconstrictor response, whereas histamine release, as detected in urine, was not changed. Total protein levels in bronchoalveolar lavage (BAL) fluid were significantly decreased in the NO group at 120 min after challenge compared with 45 min (373 ± 101 µg/mL vs. 631 ± 184, respectively, P < 0.05, Wilcoxon matched pairs test), whereas levels in the control group did not change between these two time‐points (513 ± 282 vs. 599 ± 354, not significant).

Conclusion These findings indicate that NO inhalation improves ventilation/perfusion matching and causes some bronchodilatation during the allergen‐induced acute airway reaction, whereas histamine release is not affected. Moreover, NO inhalation enhanced the clearance of extravasated protein in the airways, possibly through increased bronchial blood flow. Even though some protective effects were seen, this study does not support a therapeutic role for exogenous NO in acute allergic reactions.
Grass pollen immunotherapy inhibits seasonal increases in basophils and eosinophils in the nasal epithelium
Authors: Wilson D. R. , Irani A.‐MA. , Walker S. M. , Jacobson M. R. , Mackay I. S. , Schwartz L. B. , Durham S. R.
Published: March 2003

Background Symptoms of allergic rhinitis are accompanied by infiltration of the nasal mucosa with inflammatory cells, predominantly eosinophils and metachromatic cells (basophils and mast cells). Specific immunotherapy (IT) reduces mucosal eosinophilia and numbers of metachromatic cells in the epithelium. A specific marker distinguishing basophils from mast cells was recently developed.

Objectives The basophil‐specific monoclonal antibody 2D7 was used to determine the influence of subcutaneous IT on numbers of nasal mucosal basophils compared with the effects of IT on neutrophils, eosinophils and mast cells.

Method During a randomized, placebo‐controlled trial of grass pollen IT in 44 adults with severe summer hay fever, nasal biopsies were taken at baseline, out of the pollen season, and at the peak of the pollen season following 2 years treatment. Biopsies were processed for immunohistochemistry for basophils (2D7+), mast cells (AA1+), eosinophils (MBP+) and neutrophils (neutrophil elastase+).

Results In placebo‐treated (PL) patients there were significant seasonal increases in basophils (P < 0.01), mast cells (P < 0.05) and eosinophils (P = 0.002) in the nasal submucosa. In IT‐treated patients significant increases in 2D7+ cells (P < 0.01) and eosinophils (P = 0.01) but not AA1+ cells (P = 0.9) were observed. These differences were significant between groups for eosinophils (P < 0.05). In the epithelium there were seasonal increases in AA1+ cells and eosinophils in both groups (PL: P < 0.01, IT: P < 0.05 for both). The between‐group difference was significant for eosinophils (P = 0.05). Basophils were observed in the epithelium of six out of 17 in the placebo group and one out of 20 in the IT group (P = 0.03). Neutrophil numbers remained constant in both epithelium and submucosa.

Conclusion Successful grass pollen immunotherapy was associated with inhibition of seasonal increases in basophils and eosinophils, but not mast cells or neutrophils within the nasal epithelium. Immunotherapy may act, at least in part, by reducing seasonal recruitment of basophils and eosinophils into the epithelium.
Crete: does farming explain urban and rural differences in atopy?
Authors: Barnes M. , Cullinan P. , Athanasaki P. , MacNeill S. , Hole A. M. , Harris J. , Kalogeraki S. , Chatzinikolaou M. , Drakonakis N. , Bibaki‐Liakou V. , Newman Taylor A. J. , Bibakis I.
Published: March 2003

Background Urban‐rural differences in the prevalence of atopy and associated allergic diseases have been reported in several non‐European countries. Within Europe, where such variations are less striking, a farm childhood has been postulated to be protective.

Objective We aimed to compare the prevalence of atopy in urban and rural children living in Crete and to examine the role of early exposure to a farming environment in explaining any measured differences.

Methods A cross‐sectional survey of children attending secondary schools in Iraklion, the capital city, and five villages 100 km to the south. Atopy was determined by the responses to skin prick tests with seven common aeroallergens.

Results Nine‐nundred and ninety‐seven children participated in the survey. 19.6% of those living in Iraklion, but only 9.6% of those from the villages, had a positive response to one or more skin prick tests. Among urban children there were significant gradients in the prevalence of atopy across several categories of animal contact and consumption of farm (unpasteurized) milk products – before the age of 5 years. These differences, however, were not apparent among the rural children.
The influence of inhalative corticosteroids on circulating Nerve Growth Factor, Brain‐Derived Neurotrophic Factor and Neurotrophin‐3 in allergic asthmatics
Authors: Noga O. , Hanf G. , Schäper C. , O'Connor A. , Kunkel G.
Published: March 2003

Background The neurotrophins Nerve Growth Factor (NGF), Brain‐Derived Neurotrophic Factor (BDNF) and Neurotrophin (NT)‐3 are produced, stored and released by various immunological cells. The influence of NTs upon the function of these cells is described. Elevated plasma levels were found in inflammatory, autoimmune and allergic diseases with the highest levels in allergic asthma. A connection between bronchial hyper‐responsiveness and serum levels has been reported.

Objective Little is known about the influence of treatment with inhaled corticosteroids (ICS) on serum NT levels and their influence on the asthmatic state.

Methods Eighty‐seven volunteers were studied. Thirty‐eight were stable allergic asthmatics with constant ICS doses, 29 were asthmatics not receiving anti‐asthmatic treatment and 20 were age‐ and sex‐matched healthy controls. Demographic and lung function data were evaluated. NT serum levels were determined by ELISA.

Results NGF and BDNF levels were significantly increased in untreated asthmatics compared to the control and the treated group, while NT‐3 demonstrated significantly higher levels in treated asthmatics compared to healthy controls. After stabilization of untreated subjects with ICS, the NT levels decreased significantly.

Conclusions These results suggest that NTs participate in allergic inflammation and asthma. Effective treatment leads to a decrease of circulating neurotrophic factors.
Hot, humid air increases cellular influx during the late‐phase response to nasal challenge with antigen
Authors: Assanasen P. , Baroody F. M. , Naureckas E. , Naclerio R. M.
Published: March 2003

Background Inhalation of hot, humid air (HHA: 37 °C, > 95% relative humidity (RH)) partially inhibits the early response to nasal challenge with antigen.

Objective To investigate whether HHA inhibited the late‐phase response to nasal challenge with antigen and increased hyper‐responsiveness of the nasal mucosa to histamine.

Methods Twenty subjects with seasonal allergic rhinitis, outside of their allergy season, participated in a randomized, 2‐way cross‐over study. The subjects continuously breathed room air (25 °C, 30% RH) or HHA delivered via a face mask during the entire experiment. Subjects were challenged intranasally with antigen 1 h after beginning conditioning. The response was monitored by symptoms and nasal lavage at 2‐h intervals after the last antigen challenge. Eight hours after antigen challenge, nasal challenge with histamine was performed.

Results Exposure to HHA significantly increased nasal mucosal temperature from baseline without affecting nasal secretion osmolality. HHA significantly inhibited antigen‐induced sneezes, congestion, pruritus, and human serum albumin levels during the early response to antigen challenge. HHA exposure, however, was associated with an 8‐fold increase in the eosinophil influx and a 15‐fold increase in the levels of eosinophil cationic protein during the late‐phase response compared to room air. There were no significant differences in nasal hyper‐responsiveness to histamine during either exposure.

Conclusion HHA partially decreases the early response to nasal challenge with antigen, but dramatically increases eosinophil influx. Increasing eosinophil number had no effects on the hyper‐responsiveness to histamine. We speculate that the physical conditions of air differentially impact the stages of allergic inflammation.
Breastfeeding duration is a risk factor for atopic eczema
Authors: Bergmann R. L. , Diepgen T. L. , Kuss O. , Bergmann K. E. , Kujat J. , Dudenhausen J. W. , Wahn U. ,   The MAS‐study group
Published: March 2003

Background The results of numerous studies on the influence of breastfeeding in the prevention of atopic disorders are often contradictory. One of the most important problems is confounding by other lifestyle factors.

Objective The aim of the present study was to analyse the effect of any breastfeeding duration on the prevalence of atopic eczema in the first seven years of life taking into account other risk factors.

Methods In an observational birth cohort study 1314 infants born in 1990 were followed‐up for seven years. At 3, 6, 12, 18, 24 months and every year thereafter, parents were interviewed and filled in questionnaires, children were examined and blood was taken for in vitro allergy tests. Generalized Estimation Equations (GEE)‐models were used to model risk factors for the prevalence of atopic eczema and for confounder adjustment

Results Breastfeeding was carried out for longer if at least one parent had eczema, the mother was older, did not smoke in pregnancy, and the family had a high social status. The prevalence of atopic eczema in the first seven years increased with each year of age (OR 1.05; 95% CI 1.01–1.09 for each year), with each additional month of breastfeeding (1.03; 1.00–1.06 for each additional month), with a history of parental atopic eczema (2.06; 1.38–3.08), and if other atopic signs and symptoms appeared, especially specific sensitization (1.53; 1.25–1.88), and asthma (1.41; 1.07–1.85). Although breastfeeding should be recommended for all infants, it does not prevent eczema in children with a genetic risk.

Conclusion Parental eczema is the major risk factor for eczema. But in this study, each month of breastfeeding also increased the risk
Human mast cells stimulate fibroblast proliferation, collagen synthesis and lattice contraction: a direct role for mast cells in skin fibrosis
Authors: Garbuzenko E. , Nagler A. , Pickholtz D. , Gillery P. , Reich R. , Maquart F.‐X. , Levi‐Schaffer F.
Published: March 2003

Background Mast cells, the key cells of immediate hypersensitivity type reactions, have also been postulated to have a central role in influencing tissue remodelling and fibrosis occurring in the skin.

Objective Our aim was to investigate the direct role of human mast cells (HMC) in skin fibrotic processes, by assessing the effects of the addition of the human mast cell line HMC‐1 to human skin fibroblasts, and to identify the responsible mediators.

Methods HMC‐1 sonicates were added to human skin fibroblasts and the following parameters were evaluated: proliferation ([3H]‐thymidine), collagen synthesis ([3H] proline), activity of matrix metalloproteinases (MMPs) (zymography) and tissue inhibitors of metalloproteinases (TIMPs) (reverse zymography), and collagen gel contraction.

Results HMC‐1 sonicate increased significantly both proliferation and collagen production in the human skin fibroblasts and these properties were not affected by heating of the sonicate (56 °C, 30 min, or 100 °C, 3 min). Two main mast cell mediators, histamine and tryptase, were found to be responsible for the increase in fibroblast proliferation and collagen production. HMC‐1 sonicate did not display any pre‐formed gelatinase activity, and its addition to the fibroblasts did not change their pro‐MMP‐2 and MMP‐2 activity. On the other hand, HMC‐1 were found to possess TIMP‐1 and TIMP‐2. Addition of HMC‐1 had no effect on fibroblasts TIMP‐1 but induced a dose‐dependent increase of TIMP‐2 activity. In addition, HMC‐1 sonicate seeded together with the fibroblasts in tri‐dimensional collagen gel significantly enhanced their contraction.

Conclusion We have shown that human mast cells, by granule‐stored and therefore quickly releasable mediators, increase human skin fibroblast proliferation, collagen synthesis, TIMP‐2 and collagen gel contraction. Therefore, mast cells have a direct and potentiating role in skin remodelling and fibrosis.
Association of body mass index with respiratory symptoms and atopy: results from the European Community Respiratory Health Survey
Authors: Jarvis D. , Chinn S. , Potts J. , Burney P. ,   the European Community Respiratory Health Survey
Published: March 2003

Background There are several reports showing that obese adults report more respiratory symptoms suggestive of asthma than those who are not obese.

Objective To determine the association of body mass index with respiratory symptoms and atopy in young adults

Method Information collected from 15 454 participants in the European Community Respiratory Health Survey, a multicentre cross‐sectional survey of young adults, was analysed to determine the association of body mass index with respiratory symptoms and atopy.

Results Men and women with a body mass index of greater than 30 were at an increased risk of wheeze with shortness of breath compared with those with a body mass of 20–24.99 (OR in men 1.85, 95% confidence interval 1.41–2.42; OR in women 2.03, 95% confidence interval 1.59–2.58). Similar associations were observed for other symptoms suggestive of asthma. Body mass index was not associated with ‘hayfever or nasal allergies’, specific IgE to house dust mite, grass or cat or with total IgE in men or women.

Conclusion Reported associations of body mass index with symptoms suggestive of asthma are unlikely to be explained by a higher risk of atopy in the obese. Alternative explanations must be sought.
Personal exposure to house dust mite allergen in bed: nasal air sampling and reservoir allergen levels
Authors: Gore R. B. , Hadi E. A. , Craven M. , Smillie F. I. , O'Meara T. J. , Tovey E. R. , Woodcock A. , Custovic A.
Published: March 2003

Background Assessment of personal exposure to dust mite allergen has relied on proxy measures. Only recently has a means to directly measure inhaled allergen particle number become available (the intra‐nasal air sampler).

Objective To quantify inspired dust mite group 1 and group 2 allergen‐bearing particles in bed in undisturbed conditions prior to sleep by nasal air sampling and to investigate the relationship between inhaled particles and reservoir allergen levels.

Methods Twelve volunteers wore nasal samplers in bed for 6 evenings, nose‐breathing in undisturbed conditions. Allergen‐bearing particles (‘halos’) were detected by immunostaining for Der p 1, Der p 2, or Der p 1 and Der p 2 together, and counted by light microscopy. Count data were square root transformed for analysis of variance. Mattress dust samples were assayed for Der p 1 and Der p 2 concentrations.

Results Square root detransformed mean particle counts per 30‐min sample were: Der p 1, 4.22; Der p 2, 5.9; Der p 1 + Der p 2, 4.87; and for all samples, 5.01, with no difference between the groups. With replicate samples, halo number correlated significantly with mattress allergen concentrations (Der p 1 r = 0.80, P < 0.01; Der p 2 r = 0.68, P < 0.02).

Conclusion Nasal air sampling can be used to quantify nocturnal Der p exposure in undisturbed conditions in an area with moderate exposure to mite allergen and can provide a direct measure of inhaled mite allergen. The choice of either Der p 1 or Der p 2 is appropriate for this purpose.
Eosinophil cationic protein is stored in, but not produced by, peripheral blood neutrophils
Authors: Byström J. , Garcia R. C. , Håkansson L. , Karawajczyk M. , Moberg L. , Soukka J. , Venge P.
Published: March 2003

Background Eosinophil cationic protein (ECP) is an eosinophil‐derived protein, which has been shown to be present in circulating neutrophils.

Objective To establish whether ECP is produced or internalized by peripheral blood neutrophils.

Methods This was done using microscopy, flow cytometry, fractionation of cells and RT‐PCR techniques.

Results No ECP mRNA was detected after extensive cell purification to eliminate all traces of contaminating eosinophils. Examination of immunostained neutrophils by light, confocal, electron microscopy together with cell fraction experiments, established that ECP is present intracellularly and is mostly associated to cell granules. Uptake studies by flow cytometry and by using both cold and radiolabelled ECP showed that it is internalized by neutrophils and stored in some proportion in their primary granules. Upon stimulation with serum‐treated Sephadex particles, the internalized ECP was partially released from cells.

Conclusion ECP is not produced but can be internalized by circulating neutrophils, which take it from the environment and partially store it in their primary granules.
Long‐range destruction of Der p 1 using experimental and commercially available ionizers
Authors: Goodman N. , Hughes J. F.
Published: March 2003

Background To reduce the risk of sensitization and the elicitation of allergy symptoms, it is important to reduce the level of allergens in the home. It has previously been demonstrated that corona discharge, the process by which ionizers produce ions, can destroy the major house dust mite allergen Der p 1.

Objective In this paper the denaturing efficacy of an experimental ionizer and two commercially available products are evaluated.

Methods The first test was conducted in an electrically grounded chamber with samples of Der p 1 placed in various positions for 1, 2 and 3 weeks. The second test was conducted in situ in an unoccupied, furnished office room for 1 week. Der p 1 concentration was quantified by two‐site monoclonal antibody enzyme‐linked immunosorbent assay (ELISA).

Results All ionizers in both tests caused significant reductions in allergen concentration (P < 0.05), reaching a maximum of 92% with the experimental ionizer in the chamber after 3 weeks. The percentage reductions observed in situ with the experimental and the larger commercial ionizer were similar, reaching a maximum of 32% at a distance of 4 m away from the experimental ionizer after 1 week of exposure.

Conclusion With a revised protocol for use, air ionizers may offer a simple, efficient and inexpensive way to reduce allergen levels in the domestic environment.
Exogenous interleukin‐16 inhibits antigen‐induced airway hyper‐reactivity, eosinophilia and Th2‐type cytokine production in mice
Authors: De Bie J. J. , Jonker E. H. , Henricks P. A. J. , Hoevenaars J. , Little F. F. , Cruikshank W. W. , Nijkamp F. P. , Van Oosterhout A. J. M.
Published: March 2003

Background IL‐16 has been described as a natural soluble CD4‐ligand with immunosuppressive effects in vitro. However, little is known about the effect of IL‐16 on immune responses in vivo.

Objective In the present study, we examined the effect of IL‐16 administration in a murine model of allergic asthma. Next, we determined whether these effects were mediated by modulation of CD4+ T lymphocytes.

Methods and results Intraperitoneal administration of IL‐16 completely inhibits antigen‐induced airway hyper‐responsiveness and largely decreases the number of eosinophils in bronchoalveolar lavage fluid (> 90%) and airway tissue of ovalbumin‐sensitized and challenged mice. Firstly, it appears that thoracic lymph node cells isolated from in vivo IL‐16‐treated ovalbumin‐challenged animals produce less IL‐4 (77%) and IL‐5 (85%) upon antigenic re‐stimulation, when compared to vehicle‐treated mice. Secondly, pre‐incubation of lymphocytes with IL‐16 in vitro reduces antigen‐induced proliferation (55%) and Th2‐type cytokine production (IL‐4; 56%, IL‐5; 77%). Thirdly, the presence of IL‐16 during priming cultures of TCR transgenic T cells (DO11.10), reduces IL‐4 (33%) and IL‐5 (35%), but not IL‐10 and IFNγ levels upon re‐stimulation.

Conclusion It can be concluded that IL‐16 has potent immunosuppressive effects on a Th2‐dominated allergic airway response.
Mast cells and T cells in Kimura's disease express increased levels of interleukin‐4, interleukin‐5, eotaxin and RANTES
Authors: Kimura Y. , Pawankar R. , Aoki M. , Niimi Y. , Kawana S.
Published: March 2003

Background Kimura's disease (KD) is a chronic inflammatory disorder characterized by tumours in the head and neck region, enlarged lymph nodes, increased eosinophil counts and high serum IgE. Mast cells are known to play a central role in IgE‐mediated allergic diseases through the release of inflammatory mediators like IL‐4, IL‐5 and chemokines. We hypothesized that mast cells may play a role in the pathogenesis of KD by regulating eosinophilic infiltration and IgE synthesis.

Objective In order to investigate the role of mast cells in the pathogenesis of KD, we examined the expression of cytokines/chemokines in the lesions of KD.

Methods We examined the number of tryptase+ cells, EG2+ cells, CD3+ cells, IL‐4+ cells, IL‐5+ cells, eotaxin+ cells, RANTES+ cells and CCR3+ cells in five specimens of KD versus normal tissues by immunohistochemistry. The sources of IL‐4, IL‐5, eotaxin and RANTES and the expression of CCR3 were examined by immunostaining of serial sections with antibodies to IL‐4, IL‐5, eotaxin, RANTES and CCR3, and antibodies to tryptase, ECP (EG2) and CD3.

Results Mast cells, activated eosinophils, T cells, IL‐4+ cells, IL‐5+ cells, eotaxin+ cells, RANTES+ cells and CCR3+ cells were all increased in the lesions of KD as compared with those in normal tissue. Mast cells and T cells were the major source of IL‐4, whereas mast cells, T cells and activated eosinophils were the main source of IL‐5. Mast cells, T cells and activated eosinophils were the main source of eotaxin and RANTES.

Conclusions The number of IL‐4, IL‐5, eotaxin and RANTES‐expressing mast cells and T cells were increased in the lesions of KD. As mast cells are lesional resident cells, these cells may play an important role in the pathogenesis of KD by regulating IgE synthesis and orchestrating eosinophilic infiltration.
The role of the epidermal growth factor receptor in sustaining neutrophil inflammation in severe asthma
Authors: Hamilton L. M. , Torres‐Lozano C. , Puddicombe S. M. , Richter A. , Kimber I. , Dearman R. J. , Vrugt B. , Aalbers R. , Holgate S. T. , Djukanović R. , Wilson S. J. , Davies D. E.
Published: March 2003

Background The extent of epithelial injury in asthma is reflected by expression of the epidermal growth factor receptor (EGFR), which is increased in proportion to disease severity and is corticosteroid refractory. Although the EGFR is involved in epithelial growth and differentiation, it is unknown whether it also contributes to the inflammatory response in asthma.

Objectives Because severe asthma is characterized by neutrophilic inflammation, we investigated the relationship between EGFR activation and production of IL‐8 and macrophage inhibitory protein‐1 alpha (MIP‐1α) using in vitro culture models and examined the association between epithelial expression of IL‐8 and EGFR in bronchial biopsies from asthmatic subjects.

Methods H292 or primary bronchial epithelial cells were exposed to EGF or H2O2 to achieve ligand‐dependent and ligand‐independent EGFR activation; IL‐8 mRNA was measured by real‐time PCR and IL‐8 and MIP‐1α protein measured by enzyme‐linked immunosorbent assay (ELISA). Epithelial IL‐8 and EGFR expression in bronchial biopsies from asthmatic subjects was examined by immunohistochemistry and quantified by image analysis.

Results Using H292 cells, EGF and H2O2 increased IL‐8 gene expression and release and this was completely suppressed by the EGFR‐selective tyrosine kinase inhibitor, AG1478, but only partially by dexamethasone. MIP‐1α release was not stimulated by EGF, whereas H2O2 caused a 1.8‐fold increase and this was insensitive to AG1478. EGF also significantly stimulated IL‐8 release from asthmatic or normal primary epithelial cell cultures established from bronchial brushings. In bronchial biopsies, epithelial IL‐8, MIP‐1α, EGFR and submucosal neutrophils were all significantly increased in severe compared to mild disease and there was a strong correlation between EGFR and IL‐8 expression (r = 0.70, P < 0.001).

Conclusions These results suggest that in severe asthma, epithelial damage has the potential to contribute to neutrophilic inflammation through enhanced production of IL‐8 via EGFR‐ dependent mechanisms.
Keratinocyte adhesion and the missing link: from Dowling–Meara to Hay–WellsSt John's Hospital Dermatological Society Annual Oration 2000
Authors: McGrath J. A.
Published: March 2003

Maintaining a protective barrier against the environment is an essential function of normal skin. Critical to this role are several structural proteins and glycoproteins that contribute to adhesive junctions linking adjacent keratinocytes and basal keratinocytes to the underlying dermis, as well as other regulatory proteins involved in aspects of epidermal development, differentiation and proliferation. Inherited abnormalities in the genes that encode these components may give rise to a range of genodermatoses, many of which are characterized structurally by a ‘missing’ or perturbed adhesive link and clinically by congenital skin blistering. This oration reviews some of the original clinical descriptions and observations made in this field, as well as providing an update on the corresponding recent molecular discoveries. The emphasis is on contributions made by past and present members of the St John's Hospital Dermatological Society.
Complexities of applying nasal tolerance induction as a therapy for ongoing relapsing experimental autoimmune encephalomyelitis (EAE) in DA rats
Authors: Bai X F , Li H L , Shi F D , Liu J Q , Xiao B G , Van Der Meide P H , Link H
Published: March 2003

EAE is an autoimmune disease of the central nervous system (CNS) that serves as an experimental model for the human inflammatory demyelinating disease multiple sclerosis (MS). Antigen-based immunotherapy including soluble antigen administration via feeding has been shown to be successful in treating EAE in rodents. In the present study, we explore nasal administration of small amounts of myelin basic protein (MBP) as a potential means of treatment of protracted, relapsing EAE (PR-EAE) in a novel DA rat system. We found that nasal administration of MBP prevented EAE induced with whole spinal cord homogenate + Freund's incomplete adjuvant (FIA), and strongly down-regulated levels of MBP-reactive interferon-gamma (IFN-γ)-secreting Th1-like cells. However, in rats with ongoing PR-EAE receiving the same regimen of MBP, a trend of aggravated disease was recorded, in conjunction with augmented levels of MBP-reactive IFN-γ-secreting Th1-like splenocytes during the acute phase of EAE. These data have implications for the clinical application of nasal tolerance to autoimmune diseases.
Blockade of IL-12 during the induction of collagen-induced arthritis (CIA) markedly attenuates the severity of the arthritis
Authors: Malfait A M , Butler D M , Presky D H , Maini R N , Brennan F M , Feldmann M
Published: March 2003

The effect of blocking IL-12, a potent inducer of interferon-gamma (IFN-γ) and promoter of Th1 cell responses, during the induction phase of CIA was investigated. Arthritis was elicited in male DBA/1 mice by immunizing with type II collagen (CII) in Freund's complete adjuvant. Neutralizing anti-IL-12 antibodies were administered twice weekly from CII immunization. It was found that administration of anti-IL-12 from immunization until the onset of clinical arthritis did not lower the incidence of arthritis, but dramatically attenuated the severity of the disease, both clinically and histopathologically. This regime was associated with reduced IFN-γ levels produced by ex vivo CII-stimulated draining lymph node cells, and with diminished spontaneous ex vivo production of tumour necrosis factor (TNF), IL-6 and IL-10 by freshly isolated synovial cells. Total anti-CII antibody serum levels in these mice were lower than in the controls, but there was no change in the IgG2a/IgG1 ratio. These findings confirm that IL-12 has a major role in the induction of murine CIA and suggests that this disease is propagated, in part, by cells of the Th1 phenotype.
Human CD4+ T lymphocytes recognize a highly conserved epitope of human T lymphotropic virus type 1 (HTLV-1)envgp21 restricted by HLA DRB1*0101
Authors: Kitze B , Usuku K , Yamano Y , Yashiki S , Nakamura M , Fujiyoshi T , Izumo S , Osame M , Sonoda S
Published: March 2003

HTLV-1 causes two distinct human diseases, HTLV-1-associated myelopathy/tropical spastic paraparesis (HAM/TSP) and adult T cell leukaemia/lymphoma (ATL). Persistently infected individuals carry a risk of < 1% of developing either disease. These basic epidemiological data imply that virus–host interactions, especially immunogenetic factors, influence the outcome of infection. Several studies showed that the HLA class II DR1 DQ5 haplotype is over-represented in HAM/TSP, but rare in ATL. Therefore, we selected four patients with HAM/TSP and one seronegative control who all carried the HLA DR1 DQ5 haplotype. We analysed the CD4+ T lymphocyte response against eight synthetic peptides of HTLV-1 envelope (env) glycoprotein gp21, a crucial target antigen in HAM/TSP. The first of two immunodominant epitopes corresponded to a domain of the HTLV-1 envelope protein which had previously been shown to be essential for HTLV-1 envelope function. The second immunodominant epitope overlapped a highly conserved sequence of the retroviral transmembrane envelope protein. DR1 (DRB1*0101)-restricted T lymphocytes were activated by the conserved peptide sequence in nanomolar concentrations. In contrast, this conserved sequence can also induce non-specific, cAMP-mediated immunosuppressive effects on T cells when added in micromolar concentrations to culture media, as shown by Haraguchi S, Good RA, James-Yarish M, Cianciolo GJ, Day NK, Proc Natl Acad Sci USA 1995; 92:5568–71. Hence, HTLV-1 env gp21 might exert either stimulating immunological or immunosuppressive effects in HTLV-1-infected individuals, depending on the level of its expression and the presence of HLA DRB1*0101.
Nasal tolerance in experimental autoimmune myasthenia gravis (EAMG): induction of protective tolerance in primed animals
Authors: Shi F-D , Bai X-F , LI H-L , Huang Y-M , Van Der Meide P H , Link H
Published: March 2003

Nasal administration of μg doses of acetylcholine receptor (AChR) is effective in preventing the development of B cell-mediated EAMG in the Lewis rat, a model for human MG. In order to investigate whether nasal administration of AChR modulates ongoing EAMG, Lewis rats were treated nasally with AChR 2 weeks after immunization with AChR and Freund's complete adjuvant. Ten-fold higher amounts of AChR given nasally (600 μg/rat) were required to ameliorate the manifestations of EAMG compared with the amounts necessary for prevention of EAMG. In lymph node cells from rats receiving 600 μg/rat of AChR, AChR-induced proliferation and interferon-gamma (IFN-γ) secretion were reduced compared with control EAMG rats receiving PBS only. The anti-AChR antibodies in rats treated nasally with 600 μg/rat of AChR had lower affinity, reduced proportion of IgG2b and reduced capacity to induce AChR degradation. Numbers of AChR-reactive IFN-γ and tumour necrosis factor-alpha (TNF-α) mRNA-expressing lymph node cells from rats treated nasally with 600 μg/rat of AChR were suppressed, while IL-4, IL-10 and transforming growth factor-beta (TGF-β) mRNA-expressing cells were not affected. Collectively, these data indicate that nasal administration of AChR in ongoing EAMG induced selective suppression of Th1 functions, i.e. IFN-γ and IgG2b production, but no influence on Th2 cell functions. The impaired Th1 functions may result in the production of less myasthenic anti-AChR antibodies and contribute to the amelioration of EAMG severity in rats treated with AChR 600 μg/rat by the nasal route.
B cell-deficient mice do not develop type II collagen-induced arthritis (CIA)
Authors: Svensson L , Jirholt J , Holmdahl R , Jansson L
Published: March 2003

To investigate the role of B cells in the development of CIA, a model for rheumatoid arthritis, we investigated susceptibility to CIA in mice lacking B cells due to the deletion of the IgM heavy chain gene (μMT). The μMT deletion was backcrossed into two different CIA-susceptible strains, B10.Q and B10.RIII. Two different variants of the CIA model are inducible in these strains: in B10.Q with rat type II collagen (CII) and in B10.RIII with bovine CII. Homozygous deletion of the IgM gene led to the absence of B cells and dramatically reduced immunoglobulin levels compared with wild-type mice. The deletion of IgM totally abrogated development of CIA in both strains, although the anti-CII T cell response did not differ between the μMT and wild-type controls. We conclude that B cells play a crucial role in the development of CIA.
Anti-ribosomal antibodies bind the Sm proteins D and B/B′
Authors: Caponi L , Bombardieri S , Migliorini P
Published: March 2003

In order to analyse the specificity of human anti-ribosomal P protein antibodies, anti-ribosomal P protein antibodies were affinity-purified from the sera of lupus patients. Their binding capacity towards recombinant SmD protein and recombinant SmB/B′ protein was evaluated by immunoblot and ELISA. Epitope mapping of SmD was performed by means of synthetic peptides. Anti-ribosomal P protein antibodies bound recombinant SmD (5/10) and SmB/B′ (4/10) on immunoblot; 6/10 showed binding capacity to SmD on ELISA. Inhibition experiments using ELISA confirmed the specificity of this binding. Our data indicate the cross-reactivity of spontaneously developed anti-ribosomal P protein antibodies with the B/B′ and D constituents of the Sm complex. The coexistence of anti-Sm and anti-ribosomal antibodies in lupus sera may therefore be due, at least in part, to the reactivity of a single autoantibody population.
Bystander suppression of murine collagen-induced arthritis by long-term nasal administration of a self type II collagen peptide
Authors: Bayrak Ş , Mitchison N A
Published: March 2003

Oral and more recently nasal tolerance have attracted attention as potential treatments of autoimmune disease. Arthritis induced by bovine type II collagen (CII) is a widely used animal model of rheumatoid arthritis, which is here used to investigate the efficacy of nasal treatment by a short peptide. The peptide spans residues 707–721 (designated p707), an epitope of mouse CII that is most strongly recognized after immunization of mice with this self-protein. The treatment was partially effective, but almost only when the peptide was administered in large doses over a prolonged period. Mice immunized with bovine CII respond mainly to other peptides, located in the CB11 fragment around amino acid residues 256–270. The tolerance effect therefore results from intramolecular suppression, between epitopes located in different parts of this large protein.
A vigorous virus-specific CD4+ T cell response may contribute to the association of HLA-DR13 with viral clearance in hepatitis B
Authors: Diepolder H M , Jung M-C , Keller E , Schraut W , Gerlach J-T , Grüner N , Zachoval R , Hoffmann R M , Schirren C-A , Scholz S , Pape G R
Published: March 2003

A strong virus-specific CD4+ and CD8+ T lymphocyte response to hepatitis B virus (HBV) has been associated with viral clearance, but little is known about factors determining the individual's ability to mount such a T cell response. Recently a strong association between the HLA class II allele DR13 and a self-limited course of HBV infection has been described. In the present study of 33 patients with acute hepatitis B we show that individuals carrying HLA-DR13 mount a more vigorous CD4+ T cell response to HBV core (5706 ct/min (25th/75th percentile 3239 ct/min; 10 552 ct/min)) than patients without HLA-DR13 (1365 ct/min (490 ct/min; 5334 ct/min); P = 0.006). However, peptide epitopes aa 50–69, aa 61–85, and aa 81–105 were recognized most frequently by both patient groups. Moreover, among 14 HBV core-specific CD4+ T cell clones from two patients with HLA-DR13, only one T cell clone was HLA-DR13-restricted. Our data suggest that the beneficial effect of the HLA-DR13 alleles on the outcome of HBV infection could be explained by a more vigorous HBV core-specific CD4+ T cell response, which may either be due to more proficient antigen presentation by the HLA-DR13 molecules themselves or a linked polymorphism in a neighbouring immunoregulatory gene.
Endogenous opioids modulate allograft rejection time in mice: possible relation with Th1/Th2 cytokines
Authors: Sacerdote P , Di San Secondo B E M R , Sirchia G , Manfredi B , Panerai A E
Published: March 2003

The continuous infusion of the opioid peptide β-endorphin prolongs skin allograft survival in mice, while the opiate receptor antagonist naloxone, administered together with the opioid at the time of transplantation, abolishes the effect of the opioid. Consistently, naloxone, when given alone at the time of transplantation, but not later, accelerates graft rejection and increases splenocyte IL-2 and interferon-gamma (IFN-γ) production. Splenocyte β-endorphin concentrations are lower in transplanted animals. The effects of exogenous β-endorphin and naloxone suggest that the endogenous opioid peptide β-endorphin exerts a tonic inhibitory effect over early events of T cell-mediated immune responses in vivo. The effects of β-endorphin and naloxone are consistent with the previously shown role of the opioid system in the modulation of the Th1/Th2 cytokine pattern.
mRNA cytokine profile in peripheral blood cells from chronic hepatitis C virus (HCV)-infected patients: effects of interferon-alpha (IFN-α) treatment
Authors: Shapiro S , Gershtein V , Elias N , Zuckerman E , Salman N , Lahat N
Published: March 2003

Natural immune responses, both cellular and humoral, are not capable of terminating HCV infection in most patients. A role has been suggested for peripheral blood leucocytes (PBL) in viral persistence and clinical implications, as these cells may serve as a viral reservoir and at the same time may be inadequate active participants in antiviral immune reactions. IFN-α administration, although only partially successful, is currently the main therapy available for chronic HCV patients. In addition to its antiviral effects, IFN-α regulates the function of cytokines, their receptors and other molecules of immune importance. The aim of this study was to determine cytokine mRNA expression in PBL derived from chronic HCV patients prior to and at termination of IFN-α treatment. HCV RNA was still observed in sera of most patients (10 out of 14 treated patients) at termination of treatment. In pretreated patients mRNA expression of Th2 (IL-4, IL-6 and IL-10) and Th3 (transforming growth factor-beta (TGF-β)) was observed in only a low percentage of PBL samples from patients, similar to controls. IFN-α treatment led to an elevation in the number of samples expressing these cytokines (significant for IL-4, IL-6, IL-10, tumour necrosis factor-alpha (TNF-α) and TGF-β), accompanied by reduction in liver enzymes but in serum viral load in only ≈ 30% of patients. Expression of TNF-α and TNF-β mRNA was observed in samples from patients but not controls, while no differences were observed for mRNA of classical Th1 cytokines (IL-2 and IFN-γ) between patients before or during treatment as well as controls. The cytokine mRNA profile following IFN-α treatment points to an anti-inflammatory response which does not appear to be involved in termination of the viral infection. The PBL cytokine profile observed in this study may explain the failure of the immune system to eradicate HCV chronic infection and suggests that early treatment in the acute phase of disease with agents that stimulate cytotoxic immune type 1 responses may lead to eradication of HCV infection.
Characterization of NK cells and extrathymic T cells generated in the liver of irradiated mice with a liver shield
Authors: Halder R C , Seki S , Weerasinghe A , Kawamura T , Watanabe H , Abo T
Published: March 2003

We previously reported that c-kit+ stem cells which give rise to extrathymic T cells are present in the liver of adult mice. Further characterization of extrathymic T cells in the liver of adult mice is conducted here. When mice with a liver shield were lethally (9.5 Gy) irradiated, all mice survived. All tested organs showed a distribution pattern of hepatic lymphocytes on day 7. The distribution pattern in the liver was characterized by an abundance of NK (CD3− IL-2Rβ+) and extrathymic T cells (CD3int IL-2Rβ+) before and after irradiation. To determine their function, post-irradiation allogeneic bone marrow transplantation (BMT) was performed in mice with or without a liver shield. Allogeneic BM cells were rejected in mice with a liver shield and specific activation of CD8+ CD3int IL-2Rβ+ cells was induced. At that time, potent cytotoxicity of liver mononuclear cells (MNC) against allogeneic thymocytes was induced. Both NK1.1+ and NK1.1− subsets of CD3int cells expanded in these mice. An in vivo elimination experiment of the subsets indicated that the NK1.1+ subset of CD3int cells (i.e. NK T cells) was much more associated with the rejection of allogeneic BM cells. However, even after the elimination of NK T cells, allogeneic BM cells were rejected. In this case, granulocytes expanded in parallel with NK1.1− subsets. Granulocytes may also be associated with the rejection of allogeneic BM cells. These results suggest that the liver is an important haematopoietic organ even in adult life.