bluuebunny/crossref_metadata_2025 huggingface dataset:
Airway function and nasal inflammation in seasonal allergic rhinitis and asthma
Authors: Ciprandi G. , Cirillo I. , Vizzaccaro A. , Milanese M. , Tosca M. A.
Published: June 2004

Background
Allergic rhinitis (AR) and asthma are frequently associated and characterized by a Th2‐dependent inflammation. Nasal and bronchial obstruction may be objectively measured.

Objective
The aim of this study was to evaluate the relationships among upper and lower airway function and nasal inflammation in subjects with seasonal allergic rhinitis (SAR) and asthma.

Methods
Twenty out‐patients (12 males and eight females, mean age: 23.4+3.6 years) with SAR and asthma were evaluated during the pollen season. All of them showed a moderate–severe grade of nasal obstruction. Total symptom score, rhinomanometry, spirometry, nasal lavage, and nasal scraping were obtained in all subjects. Eosinophils were counted by conventional staining; IL‐4 and IFN‐γ were measured by immunoassay on fluids recovered from nasal lavage.

Results
Functional parameters, i.e. nasal airflow and forced expiratory volume in 1 s (FEV1), were correlated with nasal eosinophils (R2>0.83, P<0.001). Inflammatory parameters, i.e. eosinophils were correlated with immunological parameters, i.e. IL‐4 and IFN‐γ levels (R2=0.93, P<0.001). Nasal symptoms were correlated with nasal airflow (ρ=−0.71, P0.01) and eosinophils (ρ=0.72, P<0.01). Nasal airflow was correlated with FEV1 (r=0.89, P<0.0001).

Conclusions
This study demonstrates the close connection between Th2 cytokines and eosinophil infiltration in the nose. There is also clear evidence concerning the relationships between eosinophils infiltration and cytokines levels. Nasal eosinophils can be regarded as the most important predictors of upper and lower airway functions. These findings constitute first evidence of a relationship among nasal Th2‐related inflammation and nasal and bronchial airflow in patients with SAR and asthma.
Functional expression of high‐affinity receptor for immunoglobulin E on mast cells precedes that of tryptase during differentiation from human bone marrow‐derived CD34 progenitors cultured in the presence of stem cell factor and interleukin‐6
Authors: Shimizu Y. , Suga T. , Maeno T. , Aoki F. , Tsukagoshi H. , Kawata T. , Sakai K. , Narita T. , Takahashi T. , Ishikawa S. , Morishita Y. , Nakajima T. , Hara F. , Miura T. , Kurabayashi M.
Published: June 2004

Background
CD34+ progenitor cells develop into tryptase+, CD117+ mast cells when cultured in the presence of recombinant human stem cell factor (rhSCF). However, spontaneous IgE receptor (FcɛRI) expression during human mast cell development is not well examined.

Objective
Here, the expression and function of FcɛRI in and on human bone marrow‐derived mast cells (HBMMCs) during development were investigated.

Methods and Results
At 4 weeks of culture, predominant cells expressed high‐affinity IgE receptor α chain (FcɛRIα) on the cell surface determined by flow cytometry, but CD117 was less expressed. Immunocytochemistry with antitryptase mAb and anti‐FcɛRIα mAb revealed intracellular and surface expression of FcɛRIα at 2 weeks of culture, but tryptase was less expressed. FcɛRIα mRNA transcript preceded that of tryptase mRNA at 2 weeks of culture determined by real‐time RT‐PCR, and FcɛRIα, FcɛRIβ, FcɛRIγ, and tryptase mRNA increased along with differentiation. FcɛRIα cross‐link on HBMMC and 4‐week‐old mast cells/mast cell precursors induced the release of IL‐5 and granulocyte macrophage‐colony stimulating factor, which was enhanced by rhSCF.

Conclusion
These data indicated that HBMMC constitutively and spontaneously expressed functional FcɛRI subunits at the early stage of differentiation, probably because of the differences in the ability and functional property of progenitors.
Permeability of synthetic and feather pillows to live house dust mites and house dust
Authors: Siebers R. , Nam H.‐S. , Crane J.
Published: June 2004

Background
Previous studies have demonstrated significantly higher house dust mite (HDM) allergen levels from synthetic pillows, compared to feather pillows. Reasons for these differences could be lower permeability of feather pillow coverings to allergen in dust, greater HDM penetration of synthetic pillow covering, or both.

Objectives
To determine the permeability of synthetic and feather pillow coverings to live HDMs and house dust.

Methods
Twenty live adult HDMs were seeded on top of two types of synthetic pillow covering (one standard polyester and one newer polyester/cotton type) and one type of feather pillow coverings with adequate food supply below in sealed culture dishes, kept at 23°C and 70% relative humidity. After 24 and 48 h live HDM numbers remaining on top of the coverings were enumerated microscopically. Three aliquots of fine house dust (each in triplicate) were placed on top of the synthetic and feather pillow coverings, shaken gently for 30 min and penetrated dust was collected and weighed.

Results
After 24 h, all 20 HDMs had penetrated the standard synthetic pillow coverings, and no HDMs had penetrated either the feather pillow or the new synthetic pillow coverings after 24 or 48 h. Dust permeability (% of applied dust) for the standard synthetic, new type synthetic and feather pillow coverings were 0.88%, 0.07%, and 0.07%, respectively. This compared to 0.02% for a commercial occlusive pillow cover.

Conclusions
These findings of total permeability of standard synthetic pillow coverings to live HDMs, and their greater permeability to house dust could explain their reported higher HDM allergen levels, compared to feather pillow coverings. Newer types of synthetic pillow coverings are similar to feather pillow coverings in their permeability to live HDMs and house dust.
Interleukin‐18 levels in induced sputum are reduced in asthmatic and normal smokers
Authors: McKay A. , Komai‐Koma M. , MacLeod K. J. , Campbell C. C. , Kitson S. M. , Chaudhuri R. , Thomson L. , McSharry C. , Liew F. Y. , Thomson N. C.
Published: June 2004

Background
IL‐18 is a cytokine which is known to have an important role in the development of a Th1 lymphocyte response. As such, it may have a regulatory role in asthma by modifying Th2 lymphocyte responses. Cigarette smoking may amplify the airway inflammation associated with asthma.

Objective
This study investigated if IL‐18 could be detected in induced sputum from asthmatics and normal subjects and if smoking altered IL‐18 levels.

Methods
Induced sputum was obtained from asthmatic (31 smokers, 35 non‐smokers) and normal (20 smokers, 20 non‐smokers) subjects. All smokers had a smoking history of 15 pack years. IL‐18 levels in sputum supernatant were measured by ELISA. IL‐18 mRNA expression and cellular localization were assessed by quantitative PCR and immunocytochemistry, respectively.

Results
Smoking was associated with a significant reduction in IL‐18 levels (median (interquartile range) – smokers 20 (0–102) pg/mL vs. non‐smokers 358 (50–876) pg/mL, P<0.001). This was more pronounced in asthmatics (smokers, 47 (40–64) pg/mL vs. non‐smokers, 530 (30–1484) pg/mL; P<0.001) than in normal subjects (smokers, 25 (0–78) pg/mL vs. non‐smokers, 247 (50–656) pg/mL; P<0.01). Within each of the smoking and non‐smoking groups there was no significant difference in IL‐18 levels between asthmatic and normal subjects. There was no correlation between sputum IL‐18 levels and any specific cell type in the sputum samples nor serum IgE levels. IL‐18 mRNA expression was reduced in asthmatic smokers compared with non‐smokers. IL‐18 production was localized to sputum macrophages by immunocytochemistry.

Conclusions
IL‐18 is detectable in induced sputum samples from both asthmatic and normal subjects. Cigarette smoking significantly reduces sputum IL‐18 levels. This effect is more pronounced in asthmatics than in normal subjects.
The expression of stem cell factor and c‐kit receptor in human asthmatic airways
Authors: Al‐Muhsen S. Z. , Shablovsky G. , Olivenstein R. , Mazer B. , Hamid Q.
Published: June 2004

Background
Asthmatic airways are characterized by infiltration with a variety of inflammatory cells such as mast cells and eosinophils. Stem cell factor (SCF) is an important activating and chemotactic factor for both mast cells and eosinophils. In addition, it is a critical growth and differentiation factor for mast cells.

Objectives
To investigate the contribution of SCF to the pathogenesis of asthma, we examined the expression of SCF and its receptor c‐kit in bronchial biopsies and bronchoalveolar lavage (BAL) specimens obtained from asthmatic subjects (n=13) and non‐asthmatic control subjects (n=10).

Methods
SCF and c‐kit were detected by in situ hybridization (ISH) and immunocytochemistry (ICC). In order to phenotype the cells expressing SCF and c‐kit in asthmatic tissue and BAL cells, combined ISH and ICC were also performed.

Results
There was a significant difference (P<0.001) in the SCF mRNA expression in asthmatic airway epithelium (70.38±12.33% positive cells) compared with controls (12.7±17.21% positive cells). There was also a significant difference in subepithelial SCF‐mRNA expression, being higher in asthmatics (P<0.001). A significant difference was also found in c‐kit receptor mRNA expression in asthmatic biopsies both in epithelium (P<0.001) and subepithelium (P<0.05) compared with controls. ICC results were consistent with the ISH for both SCF and c‐kit receptor from asthmatics and controls. The SCF and c‐kit receptor mRNA and immunoreactivity in cells recovered from bronchial washing were also significantly higher in asthmatics compared with controls (P<0.05). While SCF expression was localized predominantly in the epithelial layer in bronchial biopsy tissues, alveolar macrophages were found to be the major source of SCF in bronchial washing from asthmatic subjects.

Conclusion
The results of this study demonstrate the increased expression of SCF and its receptor, c‐kit within human asthmatic airways, which suggests an important role of this cytokine in the pathophysiology of asthma.
Cow's milk‐specific immunoglobulin E levels as predictors of clinical reactivity in the follow‐up of the cow's milk allergy infants
Authors: García‐Ara M. C. , Boyano‐Martínez M. T. , Díaz‐Pena J. M. , Martín‐Muñoz M. F. , Martín‐Esteban M.
Published: June 2004

Background
IgE‐mediated cow's milk proteins (CMPs) allergy shows a tendency to disappear with age. The sooner tolerance is detected, the earlier the substitute diets can be suspended and the quicker family emotional hardship is alleviated.

Objective
To analyse the specific IgE levels to cow's milk and its proteins, which help to separate tolerant from no tolerant children in the follow‐up of infants with allergy to cow's milk.

Patients and methods
Sixty‐six infants diagnosed with IgE‐mediated allergy to CMPs were included in this prospective follow‐up study. Periodic reassessments were carried out every 6 months until they were 2‐years old and then, annually, until tolerance arose or until the last reassessment in which tolerance had not been achieved. Non‐tolerant infants were followed, at least, for a period of 3 years. In each visit, the same skin tests and determination of specific IgE (CAP System FEIA) for milk and its proteins were carried out. The open challenge test was repeated unless a clear transgression to milk, which came to be positive, had taken place within the previous 3 months in each of the follow‐up visits. Specific IgE levels to milk and its proteins, in different moments of the follow‐up were analysed by means of the receiver‐operating characteristic curve to predict clinical reactivity.

Results
Throughout the follow‐up 45 (68%) infants became tolerant. The follow‐up mean for tolerant infants was 21.2 months whereas for non‐tolerant infants it was 58 months. The specific IgE levels which were predictors of the clinical reactivity (positive predictive value (PPV)90%), grew as the age of the infants increased: 1.5, 6 and 14 kUA/L for milk in the age range 13–18 and 19–24 months and in the third year, respectively. Specific IgE levels to casein: 0.6, 3 and 5 kUA/L, respectively, predicted clinical reactivity (PPV90%) in the different analysed moments of the follow‐up. The cut‐off points: 2.7, 9 and 24 kUA/L for milk and 2, 4.2 and 9 kUA/L for casein, respectively, predicted clinical reactivity with an accuracy 95% corresponding to a specificity of 90%.

Conclusions
Monitorization of specific IgE concentration for milk and casein by means of the CAP system in allergic children to CMPs allows us to predict, to a high degree of probability, clinical reactivity. Age factor must be taken into account to evaluate the specific IgE levels which are predictors of tolerance or clinical reactivity.
ADAM33 polymorphism: association with bronchial hyper‐responsiveness in Korean asthmatics
Authors: Lee J. H. , Park H.‐S. , Park S. W. , Jang A. S. , Uh S. T. , Rhim T. , Park C.‐S. , Hong S.‐J. , Holgate S. T. , Holloway J. W. , Shin H. D.
Published: June 2004

Background
A disintegrin and metalloprotease 33 (ADAM33) is expressed in the lung by fibroblasts and bronchial smooth muscle cells. Given its structure and cellular provenance, ADAM33 may be associated with airway remodelling and bronchial hyper‐responsiveness. Single nucleotide polymorphisms (SNPs) and haplotypes of the ADAM33 gene have previously been associated with asthma susceptibility in the Caucasian population.

Objective and Methods
To assess whether genetic variants of ADAM33 are related to asthma in a Korean population, we conducted an association study of the ADAM33 gene with asthma susceptibility, bronchial hyper‐reactivity and serum IgE in Korean asthmatics (n=326) and normal controls (n=151). Five of the 14 polymorphisms originally reported to be associated with asthma development (S1 G>A, T1 T>C, V‐1 C>A, V1 T>A, V4 C>G) were genotyped using single base extension and electrophoresis. Haplotypes and their frequencies were inferred using the algorithm implemented by the software Arlequin. Allele frequencies of each SNP and haplotypes were compared between the patients and the normal controls using logistic regression analysis.

Results
There was no significant difference in the distribution of SNPs and the six haplotypes between asthmatics and normal controls. All single SNPs and six haplotypes in ADAM33 were also analysed for the association with level of PC20 using general linear models. The distribution of the T1 T>C SNP and one haplotype (ht4: GCGG) showed significant association with log‐transformed PC20 methacholine level in the asthma patients (P=0.03 and 0.0007, respectively, using a co‐dominant model).

Conclusion
Polymorphism of ADAM33 may contribute to development of BHR in asthma.
Effect of cysteinyl leukotriene receptor antagonist on CD11b and CD23 expression in asthmatic children
Authors: Gagro A. , Aberle N. , Rabatić S. , Ajduk J. , Jelačić J. , Dekaris D.
Published: June 2004

Background
Cysteinyl leukotrienes are potent pro‐inflammatory mediators that contribute to the pathophysiologic features observed in allergic asthma. Inhibitors of leukotriene receptors represent novel therapy in asthma treatment. In addition to the protection from early asthmatic responses, these drugs have recently been shown to protect from late airway responses too.

Methods
We studied the effect of treatment with an oral antagonist of cysteinyl leukotriene receptors on the increased expression of the low‐affinity IgE receptor, CD23, on B cells, and of its ligands, CD11b and CD11c, on CD4+ T cells and monocytes in peripheral blood of patients with allergic asthma. In this uncontrolled open‐label study, 14 children with allergic asthma received montelukast, a cysteinyl leukotrine receptor antagonist, for a period of 6 weeks after demonstrating forced expiratory volume in 1 s (FEV1) of less than 80% of the predicted value. Samples of peripheral heparinized blood and sera were obtained before and after therapy completion. Three‐colour immunofluorescence analysis was performed, and expression of CD11b and CD11c on CD4+ T lymphocytes and monocytes as well as the expression of CD21 and CD23 on B cells were determined (n=12). Peripheral blood eosinophil count, changes in FEV1 and peak expiratory flow rate (PEFR), asthma exacerbations, and as‐needed use of β‐agonist were also monitored.

Results
Montelukast improved FEV1 and PEFR, and decreased peripheral eosinophil counts in all study patients. There was no significant change in the expression of CD21 and CD23 on B cells. The expression of CD11c on CD4+ T cells and of both CD11b and CD11c on monocytes remained similar to the pretreatment expression. However, the percentage of CD11b+CD4+ T lymphocytes significantly decreased after treatment with montelukast. This was accompanied by a significant decrease in the levels of total IgE.

Conclusion
The capacity of 6‐week montelukast therapy to reduce the percentage of CD11b CD4+ T cells might be a mechanism leading to the immune response modulation on this T cell subset interaction with CD23‐expressing B cells and subsequent down‐regulation of IgE synthesis.
Rosmarinic acid in perilla extract inhibits allergic inflammation induced by mite allergen, in a mouse model
Authors: Sanbongi C. , Takano H. , Osakabe N. , Sasa N. , Natsume M. , Yanagisawa R. , Inoue K.‐i. , Sadakane K. , Ichinose T. , Yoshikawa T.
Published: June 2004

Background
Perilla and its constituent rosmarinic acid have been suggested to have anti‐allergic activity. However, few studies have examined the effects on allergic asthma.

Objective
The purpose of this study was to evaluate the effect of oral administration of perilla leaf extract, which contains high amount of rosmarinic acid, on a murine model of allergic asthma induced by house dust mite allergen.

Methods
C3H/He mice were sensitized by intratracheal administration of Dermatophagoides farinae (Der f). Mice were orally treated with rosmarinic acid in perilla extract (PE) (1.5 mg/mouse/day).

Results
Der f challenge of sensitized mice elicited pulmonary eosinophilic inflammation, accompanied by an increase in lung expression of IL‐4 and IL‐5, and eotaxin. Daily treatment with rosmarinic acid in PE significantly prevented the increases in the numbers of eosinophils in bronchoalveolar lavage fluids and also in those around murine airways. Rosmarinic acid in PE treatment also inhibited the enhanced protein expression of IL‐4 and IL‐5, and eotaxin in the lungs of sensitized mice. Der f challenge also enhanced allergen‐specific IgG1, which were also inhibited by rosmarinic acid in PE.

Conclusion
These results suggest that oral administration of perilla‐derived rosmarinic acid is an effective intervention for allergic asthma, possibly through the amelioration of increases in cytokines, chemokines, and allergen‐specific antibody.
Mechanical strain enhances proteoglycan message in fibroblasts from asthmatic subjects
Authors: Ludwig M. S. , Ftouhi‐Paquin N. , Huang W. , Pagé N. , Chakir J. , Hamid Q.
Published: June 2004

Background
Remodelling of the asthmatic airway includes increased deposition of proteoglycan (PG) molecules. One of the stimuli driving airway remodelling may be excessive mechanical stimulation.

Objective
We hypothesized that fibroblasts from asthmatic patients would respond to excessive mechanical strain with up‐regulation of message for PGs.

Methods
We obtained fibroblasts from asthmatic patients (AF) and normal volunteers (NF) using endobronchial biopsy. Cells were maintained in culture until the fifth passage and then grown on a flexible collagen‐coated membrane. Using the Flexercell device, cells were then subjected to cyclic stretch at 30% amplitude at 1 Hz for 24 h. Control cells were unstrained. Total RNA was extracted from the cell layer and quantitative RT‐PCR performed for decorin, lumican and versican mRNA.

Results
In unstrained cells, the expression of decorin mRNA was greater in AF than NF. With strain, NF showed increased expression of versican mRNA and AF showed increased expression of versican and decorin mRNA. The relative increase in versican mRNA expression with strain was greater in AF than NF.

Conclusions
These data support the hypothesis that proteoglycan message is increased in asthmatic fibroblasts subject to mechanical strain. This finding has implications for the mechanisms governing airway wall remodelling in asthma.
Th1‐dominant shift of T cell cytokine production, and subsequent reduction of serum immunoglobulin E response by administration in vivo of plasmid expressing Txk/Rlk, a member of Tec family tyrosine kinases, in a mouse model
Authors: Takeno M. , Yoshikawa H. , Kurokawa M. , Takeba Y. , Kashiwakura J.‐i. , Sakaguchi M. , Yasueda H. , Suzuki N.
Published: June 2004

Background
Th1 and Th2 cells, resulting from antigenic stimulation in the presence of IL‐12 and IL‐4, respectively, are implicated in the pathology of various diseases including allergic and autoimmune diseases. Txk/Rlk is a member of Tec family tyrosine kinases. We reported that Txk acts as a Th1‐specific transcription factor in the T lymphocytes.

Objective
In this study we have asked whether administration of txk expression plasmid brings about a Th1/Th2 shift in vivo of the mice, and subsequent reduction of circulating IgE.

Methods
Mice were administered a txk expression plasmid with hemagglutinating virus of Japan (HVJ) envelope vector. Txk expressions in spleen cells were assessed by immunoblotting and immunocytochemical staining. Cytokine productions by the spleen cells and serum Ig concentrations were studied by ELISA.

Results
Administration of a txk expression plasmid with HVJ vector induced expression of Txk in the spleen cells. The spleen cells showed enhanced Th1‐specific cytokine production; spleen cells from the txk administered mice produced more IFN‐γ as compared with those from control plasmid‐administered mice in an antigen‐specific manner. IL‐2 and IL‐4 secretions of the spleen cells were comparable between the two mouse groups. Txk administration did not reduce serum IgG concentration. It markedly reduced total IgE level and an IgG1/IgG2a ratio, reflection of Th1/Th2 balance, in sera. Furthermore, txk administration reduced ovalbumin (OVA)‐specific IgE levels in sera of the OVA sensitized mice.

Conclusion
Thus, Txk enhances IFN‐γ secretion and thus modulates Th1/Th2 cytokine balance, leading to reduction of serum IgE.
Non‐linear short‐term effects of airborne pollen levels with allergenic capacity on asthma emergency room admissions in Madrid, Spain
Authors: Tobías A. , Galán I. , Banegas J. R.
Published: June 2004

Background
Several time‐series studies have shown positive associations between pollen levels and asthma morbidity. However, few studies have included air pollution as a potential confounder when examining this relationship, and they have only done so on a linear basis.

Objective
To investigate the potential non‐linear short‐term effects of the types of pollen with allergenic capacity across the whole range of exposure on the daily number of asthma‐related hospital emergencies in Madrid for the period 1995–1998.

Methods
Data were collected on the daily number of asthma emergency room admissions and daily average levels of major types of pollen with allergenic capacity: Olea, Plantago, Poaceae and Urticaceae. We used Poisson regression with generalized additive models, controlling for trend and seasonality, meteorological variables, acute respiratory infections and air pollutants. To study the non‐linear effect of pollen levels on asthma emergency room admissions, pollens were categorized into five groups defined on the basis of their respective distributions.

Results
The strongest associations were registered in respect of a lag of 1 day for Urticaceae, a lag of 2 days for Plantago, and a day lag of 3 days for Poaceae. Adjustment for the different types of pollen failed to result in major changes. Non‐linearities were evident for pollen levels. The greatest increase in risk vs. the reference category (minimum value – 50th percentile) was observed for the categories between the 95th and 99th percentiles, with increases of 32.0% for Plantago, 32.2% for Poaceae and 24.6% for Urticaceae, and between the 99th percentile and the maximum value, with increases of 31.3%, 78.7% and 49.8%, respectively. Olea was not related with asthma emergency room admissions.

Conclusions
Pollens with allergenic capacity in Madrid are positively associated with asthma‐related hospital emergencies. These associations remain stable when simultaneous adjustment is made for the four types of pollen and for air pollutants and meteorological variables. In terms of non‐linear effects, Poaceae register the strongest association with asthma emergency room admissions.
Presence of staphylococcal exfoliative toxin A in sera of patients with atopic dermatitis
Authors: Yagi S. , Wakaki N. , Ikeda N. , Takagi Y. , Uchida H. , Kato Y. , Minamino M.
Published: June 2004

Background
It has been reported that the toxins that Staphylococcus aureus produces are associated with the exacerbation of atopic dermatitis (AD). It has been shown in many studies that staphylococcal enterotoxin (SE) A and SEB contribute to AD by humoral immunity through IgE production as a superantigen. On the other hand, little attention has been paid to the relationship between AD and exfoliative toxin x (ETx).

Objective
We investigated the toxins that are frequently detected from the skin of patients and how these toxins affect AD.

Methods
S. aureus, isolated from the skin of 100 patients with mild to severe AD, were examined for the producibility of toxins by polymerase chain reaction. Serum samples were obtained from 21 patients with mild and moderate AD. The levels of SEB, ETA, total IgE, specific IgE, and specific IgG in sera were measured by ELISA.

Results
SEB was most frequently detected from S. aureus on the skin of these patients as previously reported. And ETx, to which little attention has been paid so far, was frequently detected next to SEB. Furthermore, ETA was detected from the sera of almost all the AD patients. SEB was not detected at all. Although the level of ETA in the AD group was significantly higher than that of controls, ETA‐specific IgE was not detected from their sera. High levels of ETA tended to be detected from infantile patients. Although there were no significant differences in the levels of ETA‐IgG between AD and the controls, its prevalence was more than twice as high as the controls in AD.

Conclusion
These results suggest that many AD patients were exposed to ETx. We conclude that ETx may contribute to exacerbation of AD, particularly in infants, by a mechanism that is not through specific IgE production, unlike SEB.
Acute infection with influenza virus enhances susceptibility to fatal pneumonia followingStreptococcus pneumoniaeinfection in mice with chronic pulmonary colonization withPseudomonas aeruginosa
Authors: SEKI M , HIGASHIYAMA Y , TOMONO K , YANAGIHARA K , OHNO H , KANEKO Y , IZUMIKAWA K , MIYAZAKI Y , HIRAKATA Y , MIZUTA Y , TASHIRO T , KOHNO S
Published: June 2004

We established a mouse model in which fatal pneumonia was induced by pneumococcal superinfection following influenza virus infection in chronic Pseudomonas aeruginosa infected mice. In this mouse model, influenza virus infection caused a significant increase in inflammatory cells, cytokines and severe tissue damage in the lungs of these P. aeruginosa infected mice, before pneumococcal infection. Intrapulmonary virus titres were significantly increased in mice with chronic P. aeruginosa infection, compared with control mice. Neutrophil function analysis showed significant reduction of myeloperoxidase (MPO) activity and lysozyme secretion by influenza virus infection in these mice. Our results suggest that influenza virus infection may play an important role in inducing pneumococcal pneumonia in chronic P. aeruginosa infected mice. Our results suggested that our mouse model is useful for investigating the pathogenesis of influenza virus infection in patients with chronic lung infection.
Dietary fructooligosaccharides up-regulate immunoglobulin A response and polymeric immunoglobulin receptor expression in intestines of infant mice
Authors: NAKAMURA Y , NOSAKA S , SUZUKI M , NAGAFUCHI S , TAKAHASHI T , YAJIMA T , TAKENOUCHI-OHKUBO N , IWASE T , MORO I
Published: June 2004

We examined whether or not dietary fructooligosaccharides (FOS) in infancy can have a beneficial effect on the mucosal immune system. Newborn BALB/c mice, accompanied by their dams until 21 days of age, were fed either a control diet based on casein [FOS(–) diet group] or a FOS(–) diet supplemented with 5% (w/w) FOS [FOS(+) diet group]. Total IgA levels in tissue extracts from the intestines of mice in the FOS(+) diet group at 38 days of age were about twofold higher (P < 0·05) than those in the FOS(–) diet group in the jejunum, ileum and colon. Ileal and colonic polymeric immunoglobulin receptor (pIgR) expression in the FOS(+) diet group at 36 days of age was 1·5-fold higher than in the FOS(–) diet group (P < 0·05). Consistent with these results, the ileal IgA secretion rate of the FOS(+) diet group at 37 days of age was twofold higher than that of the FOS(–) diet group (P < 0·05). Moreover, the percentage of B220+IgA+ cells in Peyer's patches (PP) was significantly higher in the FOS(+) diet group than in the FOS(–) diet group (6·2%versus 4·3%, P < 0·05), suggesting that isotype switching from IgM to IgA in PP B cells might be enhanced in vivo. Taken together, our findings suggest that dietary FOS increases the intestinal IgA response and pIgR expression in the small intestine as well as the colon in infant mice.
Anti-desmoglein 1 antibodies in Tunisian healthy subjects: arguments for the role of environmental factors in the occurrence of Tunisian pemphigus foliaceus
Authors: KALLEL SELLAMI M , BEN AYED M , MOUQUET H , DROUOT L , ZITOUNI M , MOKNI M , CERRUTI M , TURKI H , FEZZA B , MOKHTAR I , BEN OSMAN A , ZAHAF A , KAMOUN M R , JOLY P , MASMOUDI H , MAKNI S , TRON F , GILBERT D
Published: June 2004

Pemphigus foliaceus is an autoimmune blistering skin disease mediated by autoantibodies directed against desmoglein 1 and occurs as a sporadic form throughout the world, or as an endemic form called fogo selvagem in Brazil. Healthy subjects living in Brazilian endemic areas produce antidesmoglein 1 antibodies, suggesting the role of environmental factors in the initiation of the autoimmune response. Tunisia was described recently as an endemic area where the disease is characterized by its high rate among young people, especially women. An enzyme-linked immunosorbent assay using recombinant desmoglein 1 as antigen was used to detect antibodies against desmoglein 1 and calibrated with sera from 67 French healthy blood donors, 20 French pemphigus foliaceus patients and patients with other bullous skin diseases. When sera from 179 healthy Tunisian blood donors were tested, 31 (17%) were found positive. The desmoglein 1 binding activity of these 31 sera was confirmed in 10 cases by indirect immunofluorescence analysis and/or immunoblotting using human epidermal extract. Subclass analysis of antidesmoglein 1 antibodies showed that they were almost exclusively of the IgG2 subclass in positive normal sera and of IgG4 subclass in patients with PF. Thus, antibodies against desmoglein 1 are prevalent in normal subjects living in Tunisia which, along with their IgG2 isotype, suggests the role of the environment in the pathogenesis of this endemic type of pemphigus foliaceus and the need for additional factors to switch from a subclinical to a clinical form of the disease.
Down-regulation of CXCR-4 and CCR-5 expression by interferon-γis associated with inhibition of chemotaxis and human immunodeficiency virus (HIV) replication but not HIV entry into human monocytes
Authors: CREERY D , WEISS W , LIM W T , AZIZ Z , ANGEL J B , KUMAR A
Published: June 2004

Alterations in the expression of CXCR4 and CCR5, the co-receptors for HIV entry, may be associated with susceptibility of monocytic cells to HIV infection. Interferon (IFN)-γ has been shown to inhibit HIV replication in monocytic cells, but the molecular mechanism involved is not well understood. To determine if IFN-γ regulates HIV replication by altering CXCR-4/CCR-5 expression and hence virus entry into monocytic cells, we investigated the effects of IFN-γ on CXCR-4 and CCR-5 expression and its biological implications with respect to HIV entry, replication and chemotaxis towards the CXCR-4 and CCR-5 ligands SDF-1 and MIP-1α, respectively. IFN-γ decreased CXCR-4 and CCR-5 expression on monocytes derived from HIV-negative adults, HIV-positive adults and HIV-negative cord blood. This down-regulation of chemokine receptor expression did not result in a corresponding change in mRNA expression but was associated with elevated levels of the endogenously produced chemokines SDF-1 and RANTES. Furthermore, IFN-γ inhibited chemotaxis in response to SDF-1 and MIP-1α, inhibited HIV replication, but failed to inhibit virus entry in monocytic cells. These results suggest that although IFN-γ-induced down-regulation of CXCR-4 and CCR-5 expression is associated with an inhibition of SDF-1-/MIP-1α-mediated chemotaxis, IFN-γ-induced inhibition of HIV replication may be mediated at levels subsequent to the virus entry.
Oligoclonal expansion of circulating and tissue-infiltrating CD8+ T Cells with killer/effector phenotypes in juvenile dermatomyositis syndrome
Authors: MIZUNO K , YACHIE A , NAGAOKI S , WADA H , OKADA K , KAWACHI M , TOMA T , KONNO A , OHTA K , KASAHARA Y , KOIZUMI S
Published: June 2004

Although triggering by infectious agents and abnormal immune responses may play some role in the pathogenesis of juvenile dermatomyositis syndrome (JDMS), the precise mechanism of muscle destruction and vascular damage is largely unknown. In this study, we tried to elucidate the role of cytotoxic T cells in two patients with JDMS, who were diagnosed based on the characteristic symptoms, laboratory data, MRI findings and electromyographic patterns. Peripheral blood T cell phenotypes were determined by flow cytometry, using mAbs against specific T cell receptor (TCR) Vβs. Complementarity-determining region3 (CDR3) size analysis was performed by gene scanning of CDR3 polymerase chain reaction (PCR) amplification products specific for each Vβ. Subsequently, CDR3 nucleotide sequences were obtained after cloning of the predominant products. The distribution of lymphocytes infiltrating the muscle tissue was analysed by immunohistochemistry. In both patients examined, a unique combination of TCR Vβ repertoires was increased within the CD8+ T cells. These subpopulations expressed a characteristic phenotype, indicating that they are memory/effector T cells with killer functions. At the same time, immunohistological and molecular biological examinations of the biopsied muscle samples revealed that identical CD8+ T cell clones with identical phenotypes/TCR Vβ infiltrated within the inflammatory tissue, in particular around vessels. These findings indicate that oligoclonal expansion of CD8+ T cells plays a central role in the pathogenesis of muscle injury in the juvenile form of dermatomyositis syndrome and may provide a useful clinical parameter of disease activity and responsiveness to anti-inflammatory therapy.
Murine viral hepatitis involves NK cell depletion associated with virus-induced apoptosis
Authors: LEHOUX M , JACQUES A , LUSIGNAN S , LAMONTAGNE L
Published: June 2004

Mouse hepatitis virus type 3 (MHV3), a coronavirus, is an excellent animal model for the study of immunological disorders related to acute and chronic hepatitis. In this study, we have verified if the fulminant hepatitis induced by MHV3 could be related to an impairment of innate immunity. Groups of three C57BL/6 mice were infected with the pathogenic L2-MHV3 or attenuated YAC-MHV3 viruses, and the natural killer (NK) cell populations from liver, spleen and bone marrow were analysed. The percentage of intrahepatic NK1·1+T cell receptor (TCR)− cells did not increase while NK1·1+TCRinter cells decreased in both L2-MHV3- and YAC-MHV3-infected mice. Concurrently, splenic and myeloid NK1·1+ cells decreased in L2-MHV3-infected mice. However, the cytotoxic activity of NK cells increased in liver and decreased in bone marrow from pathogenic L2-MHV3-infected mice while no modification was detected in YAC-MHV3-infected mice. Flow cytometric analysis revealed that both normal and larger splenic or myeloid NK cells decreased more in pathogenic L2-MHV3-infected mice than in attenuated YAC-MHV3-infected mice. In vitro viral infections of interleukin (IL)-15-stimulated lymphoid cells from liver and bone marrow revealed that L2-MHV3 induced higher decreases in cell viability of NK1·1+ cells than the YAC-MHV3 variant. The NK cell decreases were due to the viral permissivity leading to cytopathic effects characterized by cell rounding, syncytia formation and apoptosis. Larger NK+ syncytia were observed in L2-MHV3-infected cells than in YAC-MHV3-infected cells. These results suggest that NK cell production is impaired by viral infection favouring fulminant hepatitis.
Expression of CTLA-4 (CD152) in peripheral blood T cells of children with influenza virus infection including encephalopathy in comparison with respiratory syncytial virus infection
Authors: AYUKAWA H , MATSUBARA T , KANEKO M , HASEGAWA M , ICHIYAMA T , FURUKAWA S
Published: June 2004

Influenza virus and respiratory syncytial virus (RSV) are the most common causes of acute severe respiratory infection in children during the winter. There have been few reports about peripheral blood T cell activation in vivo in influenza virus infection and conflicting results concerning peripheral blood T cells activation in RSV infection. Cytotoxic T lymphocyte-associated antigen 4 (CTLA-4, CD152) is a receptor present on T cells that plays a critical role in the down-regulation of antigen-activated immune responses. To clarify the status of peripheral blood T cells, we investigated intracellular CTLA-4 expression in T cells in patients with influenza virus and RSV infection. We collected blood samples from 15 patients with influenza virus infection, including three with complications of influenza virus-associated encephalopathy and 18 patients with RSV infection, as well as 44 healthy children. We determined the intracellular expression of CTLA-4 in CD4+ and CD8+ T cells by flow cytometry. There were no significant differences in the percentages of intracellular CTLA-4-positive CD4+ T cells and CD8+ T cells by age. The percentages of intracellular CTLA-4-positive CD4+ T cells in the patients with influenza virus infection were significantly higher than those in healthy children (P < 0·01). In particular, the patients with influenza virus-associated encephalopathy had sevenfold higher percentages of CTLA-4-positive CD4+ T cells than influenza patients without encephalopathy (P < 0·05). The patients with influenza virus-associated encephalopathy had increased percentages of CTLA-4-positive CD8+ cells at the acute stage in comparison with the convalescent stage and in control subjects (P < 0·01, respectively). RSV patients showed no increase in CTLA-4-positive CD4+ T cells or CD8+ T cells. The immunological status of peripheral T cell activation is substantially different in influenza virus infection and RSV infection. The patients with RSV infection did not show any increase in CTLA-4-positive peripheral blood T cells. There was a remarkable increase in intracellular CTLA-4 in CD4+ and CD8+ T cells in influenza virus-associated encephalopathy. Down-regulation of antigen-activated peripheral blood T cell activation might play an important role in the pathogenesis of influenza virus-associated encephalopathy and host defence against influenza virus infection.
Mast cells at the host–pathogen interface: host-protectionversusimmune evasion in leishmaniasis
Authors: SAHA B , TONKAL A M D J , CROFT S , ROY S
Published: June 2004

Infection of a susceptible host with Leishmania, a protozoan parasite, causes the disease leishmaniasis, which is characterized by neutrophil, eosinophil, macrophage, lymphocyte and mast cell infiltration into the infected tissue followed by parasite growth. Although the roles played by other cells in leishmaniasis are known, the role of mast cells remains to be ascertained. Here, we demonstrate that Leishmania regulates mast cell infiltration to the site of infection, mast cell production and mast cell function resulting in differential growth of the parasite in resistant (C57BL/6 or CBA/T6T6) and susceptible (BALB/c) macrophages. An interleukin-3-dependent augmentation in mast cell committed progenitors is observed in BALB/c but not in C57BL/6 mice during Leishmania infection. The mast cell supernatants inhibit IFN-γ-dependent restriction of Leishmania growth in macrophages in BALB/c mice whereas the reverse phenomenon occurs in C57BL/6 mice. Our data reveals a different facet of host–pathogen interaction.
B and T cell responses elicited by monoclonal anti-idiotypic antibody (Ab2β) mimicking gp43 fromParacoccidioides brasiliensis
Authors: SOUZA E B , LOPES J D , ALMEIDA S R
Published: June 2004

Paracoccidioidomycosis is a systemic mycosis endemic in Latin America, with a high prevalence in Brazil, Argentina, Colombia and Venezuela. The aetiological agent of disease is the thermal dimorphic fungus, Paracoccidioides brasiliensis. A glycoprotein of 43 kD (gp43) is the major antigen of P. brasiliensis. Antibodies directed to this antigen are detected in the sera of all patients with paracoccidioidomycosis (PCM). Recently, it has been shown that mice immunized with anti-gp43 monoclonal antibodies (MAbs) (Ab1), induce the idiotypic cascade in the gp43 system, which produced both, anti-Id antibodies (Ab2) and anti-anti-Id antibodies (Ab3). To further characterize the idiotypic cascade modulation in mice immunized with anti-gp43 MAb 17c, hybridomas were produced. Ab2 MAbs named 7.B12 inhibited (>95%) the binding of gp43 to MAb 17c (Ab1), suggesting that this anti-Id MAb bind to the idiotope, thus fulfilling the internal image criteria. To elucidate whether Ab2 MAb could act as antigen in serological assays, instead of gp43, sera from PCM patients were tested. Using an ELISA test, it was observed that antibodies from patients and not normal serum bound to Ab2. However, the ELISA test using Ab2 bound to the solid phase made possible to serologically monitor the patients after antifungal therapy, showing an equivalent curve when compared with ELISA test employing purified gp43. Our results also showed that, when mice were immunized with Ab2β and their cells were exposed to gp43 in vitro, a T cell proliferation response was observed.
Lymphocyte apoptosis in acute respiratory syncytial virus bronchiolitis
Authors: ROE M F E , BLOXHAM D M , WHITE D K , ROSS-RUSSELL R I , TASKER R T C , O'DONNELL D R
Published: June 2004

Respiratory syncytial virus (RSV) infection may have an effect on the development of T cell memory responses. RSV bronchiolitis in infants is associated with a transient decline in circulating lymphocytes. We hypothesized that the mechanism underlying this lymphopenia is apoptosis. Blood was taken from 32 infants during primary RSV bronchiolitis and three months later. Using flow cytometry, we found that absolute numbers of both CD3+/CD4+ T-helper lymphocytes (P = 0·029) and CD3+/CD8+ cytotoxic lymphocytes (CTL) (P = 0·043) were significantly reduced during acute infection. Up-regulated expression both of Fas (P < 0·001) and tumour necrosis factor-related apoptosis-inducing ligand (TRAIL) receptor (P < 0·001) was found during acute illness on both CD3+/CD4+ and CD3+/CD8+ lymphocytes, when compared with convalescent samples. Expression of Fas on CD4+ lymphocytes was inversely related to CD4+ number (P = 0·03). Plasma levels of soluble Fas ligand (P = 0·028) and caspase-1 (P = 0·037), determined by enzyme-linked immunosorbent assay, were increased during bronchiolitis. Plasma interleukin-18, a product of caspase-1 activity, was not raised. Taken together, these data suggest that in acute RSV infection, CD4+ helper lymphocytes and CD8+ cytotoxic lymphocytes are primed to undergo apoptosis. This is a mechanism through which lymphopenia may occur and T cell memory may be altered.
Dendritic cells overexpressing Fas-ligand induce pulmonary vasculitis in mice
Authors: BUONOCORE S , FLAMAND V , CLAESSEN N , HEERINGA P , GOLDMAN M , FLORQUIN S
Published: June 2004

Dendritic cells (DC) genetically engineered to express Fas (CD95) ligand (FasL-DC) have been proposed as immunotherapeutic tools to induce tolerance to allografts. However, we and others recently showed that FasL-DC elicit a vigorous inflammatory response involving granulocytes and can promote Th1-type CD4+ and cytotoxic CD8+ T lymphocytes. This prompted us to evaluate the pathology induced by intravenous injection of FasL-DC in mice. We observed that FasL-DC obtained after retroviral gene transfer of bone marrow precursors derived from Fas-deficient C57Bl/6 mice induce massive pulmonary inflammation and pleuritis one day after a single intravenous injection in C57Bl/6 mice. Two months later, all mice presented granulomatous vasculitis of small to medium sized vessels, alveolar haemorrhage and pleuritis. In these lesions, apoptotic bodies were found in large number. Anti-neutrophilic cytoplasmic and anti-myeloperoxidase autoantibodies were not detected. This study documents that intravenous injection of FasL-DC causes severe lung granulomatous vasculitis. This new animal model for vasculitis is inducible, highly reproducible and shares many features with human Wegener granulomatosis. This model may be an appropriate tool to further investigate the pathogenesis of vasculitis and test new therapeutic strategies. Moreover, our findings highlight the potential severe complications of FasL-DC-based immunotherapy.
RecombinantMycobacterium bovisbacillus Calmette-Guérin (BCG) expressing mouse IL-18 augments Th1 immunity and macrophage cytotoxicity
Authors: LUO Y , YAMADA H , CHEN X , RYAN A A , EVANOFF D P , TRICCAS J A , O'DONNELL M A
Published: June 2004

Interleukin-18 (IL-18) has been demonstrated to synergize with BCG for induction of a T-helper-type 1 (Th1) immune response. Since successful treatment of superficial bladder cancer with BCG requires proper induction of Th1 immunity, we have developed a recombinant (r) BCG strain that functionally secretes mouse (m) IL-18. This rBCG-mIL-18 strain significantly increased production of the major Th1 cytokine IFN-γ in splenocyte cultures, at levels comparable to that elicited by control BCG plus exogenous rIL-18. IFN-γ production by splenocytes was eliminated by addition of neutralizing anti-IL-18 antibody. Endogenous IL-12 played a favourable role whereas IL-10 played an adverse role in rBCG-mIL-18-induced IFN-γ production. Enhanced host antimycobacterial immunity was observed in mice infected with rBCG-mIL-18 which showed less splenic enlargement and reduced bacterial load compared to control mice infected with BCG. Further, splenocytes from rBCG-mIL-18-infected mice, in response to BCG antigen, displayed increased production of IFN-γ and GMCSF, decreased production of IL-10, elevated cellular proliferation and higher differentiation of IFN-γ-secreting cells. rBCG-mIL-18 also enhanced BCG-induced macrophage cytotoxicity against bladder cancer MBT-2 cells in a dose-dependent manner. Neutralizing all endogenous macrophage-derived cytokines tested (IL-12, IL-18 and TNF-α) as well as IFN-γ severely diminished the rBCG-mIL-18-induced macrophage cytolytic activity, indicating a critical role for these cytokines in this process. Cytokine analysis for supernatants of macrophage-BCG mixture cultures manifested higher levels of IFN-γ and TNF-α in rBCG-mIL-18 cultures than in control BCG cultures. Taken together, this rBCG-mIL-18 strain augments BCG's immunostimulatory property and may serve as a better agent for bladder cancer immunotherapy and antimycobacterial immunization.