bluuebunny/crossref_metadata_2025 huggingface dataset:
Effect of formaldehyde on the expression of adhesion molecules in nasal microvascular endothelial cells: the role of formaldehyde in the pathogenesis of sick building syndrome
Authors: Kim W. J. , Terada N. , Nomura T. , Takahashi R. , Lee S. D. , Park J. H. , Konno A.
Published: March 2003

Background Formaldehyde is associated with sick building syndrome (SBS), a set of diffuse and irritative symptoms predominantly involving the eyes and the respiratory tract. However, its pathophysiological mechanism in SBS has not yet been clarified.

Objective In this study we investigated the effect of formaldehyde on the expression of adhesion molecules on human mucosal microvascular endothelial cells (HMMECs). Furthermore, we investigated the effect of formaldehyde on adhesiveness of HMMECs to eosinophils.

Materials and methods HMMECs were incubated with various concentrations of formaldehyde (1 ng/mL−1 µg/mL) for 24 h, and the expressions of intercellular adhesion molecule‐1 (ICAM‐1) and vascular cell adhesion molecule (VCAM‐1) on HMMECs were evaluated by flow cytometry. The change in the expression of ICAM‐1 and VCAM‐1 mRNA was then evaluated by reverse transcriptase polymerase chain reaction. To understand the role of formaldehyde in eosinophilic inflammation of the nasal mucosa, we examined the effects of formaldehyde on the adhesiveness between HMMECs and eosinophils by eosinophil adhesion assay.

Results Formaldehyde increased the surface expressions of ICAM‐1 and VCAM‐1 on HMMECs. Formaldehyde also induced ICAM‐1 and VCAM‐1 mRNA. In addition, the adhesiveness between HMMECs and eosinophils was also increased by formaldehyde.

Conclusion These in vitro studies suggest that formaldehyde may play a role as the irritant of the nasal mucosa by increasing the expressions of adhesion molecules on HMMECs and by enhancing the adhesiveness between HMMECs and eosinophils.
Associations of FceεR1‐bβ polymorphisms with immunoglobin E antibody responses to common inhalant allergens in a rural population
Authors: Van Hage‐Hamsten M. , Johansson E. , Kronqvist M. , Loughry A. , Cookson W. O. C. M. , Moffatt M. F.
Published: March 2003

Background Polymorphisms within the β subunit of the high‐affinity receptor for IgE (FcεR1‐β) on chromosome 11q13 have been related to atopy and asthma and the lymphotoxin α (LTα) gene on chromosome 6 is implicated in asthma.

Objective To elucidate the association of polymorphisms in the FcεR1‐β and LTα genes to IgE responses and asthma in a family‐orientated rural population.

Methods A total of 461 adult farmers, who participated in an epidemiological follow‐up study on respiratory symptoms among farmers on the Swedish island of Gotland, were examined. The traits assessed included serum total IgE, IgE antibody responses to 21 common inhalant allergens and asthma.

Results The 237G mutation was only detected in seven persons. Atopy was found to be associated with the RsaI‐ex7 AB‐genotype (OR = 1.9; P = 0.04). The RsaI‐ex7 B allele had a significant influence on IgE responses to pollens and dust mites (OR = 5.5; P = 0.03 and OR = 5.2; P = 0.049, respectively). The influence of this allele was stronger when the association towards single dust mite species (Lepidoglyphus destructor) was estimated (OR = 7.1, P = 0.03) and the association increased even more when the major allergen of L. destructor (rLep d 2) was analysed (OR = 11.2, P = 0.02). These associations were independent of sex, age and smoking, and the estimates of RsaI‐in2 independent of RsaI‐ex7. RsaI‐in2, RsaI‐ex7 and LTα genotypes were unassociated with total serum IgE. No significant difference in the distribution of RsaI‐in2, RsaI‐ex7 and LTα genotypes was found among subjects with atopy or asthma compared to healthy controls.

Conclusion This study supports the notion that polymorphisms in the FcεR1‐β gene have significant effects on IgE responsiveness. Secondly, dust mites in rural populations influence the expression of genes on chromosome 11q13.
Genetic deficiency of human mast cell a‐tryptase
Authors: Soto D. , Malmsten C. , Blount J. L. , Muilenburg D. J. , Caughey G. H.
Published: March 2003

Background Human α‐ and β‐tryptases are proteases secreted by mast cells. Beta (but not α) tryptases are implicated in asthma. Genes encoding both types of tryptases cluster on chromosome 16p13.3.

Objective This study examines the hypothesis, generated from mapping data, that α‐alleles compete with some β‐alleles at one locus and that an adjacent locus contains β‐alleles exclusively. This hypothesis predicts that β‐alleles outnumber α and that some genomes lack α genes altogether.

Methods To test this hypothesis, we developed PCR‐based techniques to distinguish α from β genes. We then genotyped genomic DNA from individuals and tryptase‐expressing cell lines.

Results In support of our hypothesis, we find that α‐tryptase deficiency affects 80/274 (29%) of individuals surveyed. The genotype of the α‐deficient individuals is ββββ, due to inheritance of four β genes. The percentage of the population with the mixed genotypes ααββ and αβββ is 21% and 50%, respectively. Accounting for all α‐ and β‐alleles at the tandem loci on 16p13.3, overall α‐allele frequency is only 0.23, with β‐alleles considerably outnumbering α as hypothesized. In samples of defined ethnicity, α deficiency affects 45% of Caucasians, but a much lower percentage of other backgrounds, including African‐Americans and Asians. Examination of cell lines reveals that HMC‐1 and U‐937 lack α‐genes; thus, lack of α transcripts in these cells is due to absence of α‐genes rather than β‐selective transcription. By contrast, α‐transcribing Mono Mac 6 and KU812 cells contain α‐ and β‐genes.

Conclusions Genetic α‐tryptase deficiency is common and varies strikingly between ethnic groups. Because β‐tryptases are implicated in allergic disorders, inherited differences in α/β‐genotype may affect disease susceptibility, severity and response to tryptase inhibitor therapy.
Expression and activation of 15‐lipoxygenase pathway in severe asthma: relationship to eosinophilic phenotype and collagen deposition
Authors: Chu H. W. , Balzar S. , Westcott J. Y. , Trudeau J. B. , Sun Y. , Conrad D. J. , Wenzel S. E.
Published: March 2003

Background Although 15(S)‐hydroxyeicosatetraenoic acid (15(S)‐HETE), a product of 15‐lipoxygenase (15‐LO), may be involved in mild to moderate asthma, little is known about its potential roles in severe asthma.

Objectives This study was performed to evaluate 15(S)‐HETE levels in bronchoalveolar lavage fluid (BALF) from severe asthmatics with and without airway eosinophils and from the control groups. In addition, 15‐LO protein expression was examined in endobronchial biopsy, while its expression and activation were evaluated in BAL cells.

Results While 15(S)‐HETE levels in BALF were significantly higher in all severe asthmatics than normal subjects, severe asthmatics with airway eosinophils had the highest levels compared with mild, moderate asthmatics and normal subjects. 15(S)‐HETE levels were associated with tissue eosinophil numbers, sub‐basement membrane thickness and BALF tissue inhibitor of metalloproteinase‐1 levels, and were accompanied by increased 15‐LO expression in bronchial epithelium. In addition, activation of 15‐LO was suggested by the increased proportion of 15‐LO in the cytoplasmic membrane of alveolar macrophages from severe asthmatics.

Conclusion The data suggest that severe asthmatics with persistent airway eosinophils manifest high levels of 15(S)‐HETE in BALF, which may be associated with airway fibrosis. It is likely that 15‐LO expression and activation by airway cells explain the increased 15(S)‐HETE levels.
Non‐covalent presentation of sulfamethoxazole to human CD4+ T cells is independent of distinct human leucocyte antigen‐bound peptides
Authors: Burkhart C. , Britschgi M. , Strasser I. , Depta J. P. H. , Von Greyerz S. , Barnaba V. , Pichler W. J.
Published: March 2003

Background It has been shown that drugs comprise a group of non‐peptide antigens that can be recognized by human T cells in the context of HLA class II and that this recognition is involved in allergic reactions. Recent studies have demonstrated a MHC‐restricted but processing‐ and metabolism‐independent pathway for the presentation of allergenic drugs such as lidocaine and sulfamethoxazole (SMX) to drug‐specific T cells. However, there is little information so far on the precise molecular mechanisms of this non‐covalent drug presentation.

Objective The aim of this study was to evaluate the requirements for a specific peptide occupying the groove of the MHC class II molecule for the efficient presentation of non‐covalently bound drugs to CD4+ T cells.

Methods We analysed the effect of coincubation or prepulse of antigen presenting cells (APC) with different peptides on the proliferative responses of SMX‐specific CD4+ T cell clones. In a second series of experiments, we eluted HLA‐bound peptides from the surface of antigen presenting cells by mild acid treatment. Successful removal of peptides was tested directly using labelled peptides and functionally by monitoring activation and proliferation of peptide‐specific T cell clones. Finally, the presentation of SMX to SMX‐specific T cell clones before and after elution of MHC class II bound peptides was tested.

Results We found that neither peptide coincubation nor peptide prepulse of APC altered the proliferative response of SMX‐specific T cells. APC treated with the acid for a short time retained cell viability, MHC class II expression and antigen presenting cell function. However, defined peptides could be eluted from surface MHC class II molecules nearly quantitatively. Nevertheless, the chemically non‐reactive drug SMX could still be presented to specific T cells independent of the presence of distinct self‐peptides.

Conclusion Our data suggest that small molecules like drugs can bind to a multitude of HLA‐bound peptides or that, similar to superantigens, they might bind directly to HLA.
Intranasal application of chitin microparticles down‐regulates symptoms of allergic hypersensitivity to Dermatophagoides pteronyssinus and Aspergillus fumigatus in murine models of allergy
Authors: Strong P. , Clark H. , Reid K.
Published: March 2003

Background Previous studies have demonstrated that chitin in the form of microparticles that can be phagocytosed is a potent macrophage stimulator and promotes a Th1 cytokine response and it has been shown that oral administration of chitin microparticles is effective in down‐regulating serum IgE and lung eosinophilia in a mouse model of ragweed allergy. To date there have been no studies on the effectivness of directly applying chitin microparticles to the respiratory tract as a treatment for allergic symptoms.

Objective To test the effectivness of chitin microparticles when given intranasally as a treatment for the symptoms of respiratory allergy and allergic asthma and to compare its effectivness in two different mouse models of allergy, namely to Dermatophagoides pteronyssinus and Aspergillus fumigatus.

Results The intranasal application of microgram doses of chitin microparticles is an effective treatment for reducing serum IgE and peripheral blood eosinophilia, airway hyper‐responsiveness and lung inflammation in both allergy models results in elevation in Th1 cytokines IL‐12, IFN‐γ and TNF‐α and reduction in IL‐4 production during allergen challenge.

Conclusion Chitin microparticle suspensions have Th1 immunostimulatory properties and are effective when administered intranasally in mice. The stimulation of the nasal associated lymphoid tissue with chitin microparticles could offer a novel and natural approach to treating allergic disease in humans.
Increased airway inflammatory cells in endurance athletes: what do they mean?
Authors: Bonsignore M. R. , Morici G. , Vignola A. M. , Riccobono L. , Bonanno A. , Profita M. , Abate P. , Scichilone N. , Amato G. , Bellia V. , Bonsignore G.
Published: March 2003

Background Inflammatory cells are increased in the airways of endurance athletes, but their role in causing exercise‐induced respiratory symptoms and bronchoconstriction, or their possible long‐term consequences, are uncertain.

Aim To put the results of athlete studies in perspective, by analysing the pathogenesis of airway cell changes and their impact on respiratory function.

Results Athletes of different endurance sports at rest showed increased airway neutrophils. Elite swimmers and skiers also showed large increases in airway eosinophils and lymphocytes, possibly related to chronic, exercise‐related exposure to irritants or cold and dry air, respectively. Post‐exercise studies reported variable responses of airway cells to exercise, but found no evidence of inflammatory cell activation in the airways, at variance with exercise‐induced neutrophil activation in peripheral blood. The increase in airway inflammatory cells in athletes can result from hyperventilation‐induced increase in airway osmolarity stimulating bronchial epithelial cells to release chemotactic factors. Hyperosmolarity may also inhibit activation of inflammatory cells by causing shedding of adhesion molecules, possibly explaining why airway inflammation appears ‘frustrated’ in athletes. Data on exhaled nitric oxide are few and variable, not allowing conclusions about its usefulness as a marker of airway inflammation in athletes, or its role in modulating bronchial responsiveness.

Conclusions The acute and long‐term effects of exercise on airway cells need further study. Airway inflammatory cells are increased but not activated in athletes, both at rest and after exercise, and airway inflammation appears to regress in athletes quitting competitions. Altogether, these findings do not clearly indicate that habitual intense exercise may be detrimental for respiratory health. Rather, airway changes may represent chronic adaptive responses to exercise hyperventilation. An improved understanding of the effects of exercise on the airways will likely have a clinical impact on sports medicine, and on the current approach to exercise‐based rehabilitation in respiratory disease.
Endotoxin levels in Estonian and Swedish house dust and atopy in infancy
Authors: Böttcher M. F. , Björkstén B. , Gustafson S. , Voor T. , Jenmalm M. C.
Published: March 2003

Background Immune responses, including those to allergens, may be T helper (Th)2 skewed in newborns. In order to redress the fetal Th1/Th2 imbalance, Th1‐stimulating factors, such as bacterial endotoxin, may be required. The increasing prevalence and severity of atopic diseases in industrialized countries, which are in marked contrast with the low prevalence of allergy among children in the formerly socialist countries of Europe, have been suggested to be caused by a reduced microbial stimulation.

Aim To relate the endotoxin levels in house dust from two countries with a low (Estonia) and a high (Sweden) prevalence of allergy to the development of atopic disease and sensitization in the children during the first 2 years of life.

Methods The study included 108 children from Tartu, Estonia and 111 children from Linköping, Sweden. Skin prick tests were performed at 3, 6, 12 and 24 months of age, and questionnaires were distributed to the families. At 24 months, a paediatrician examined the children. Dust samples were collected from mattresses and carpets and the endotoxin concentration was determined by a chromogenic Limulus assay.

Results The endotoxin levels were higher in Estonian than in Swedish house dust (median levels 29 (range 0.25–280) and 14 (range 0.25–99) EU/mg dust, respectively, P < 0.001). Furthermore, the levels were inversely related to the development of atopic disease and sensitization in the Swedish, but not in the Estonian, children.

Conclusions The low prevalence of atopic disease in Estonia may, at least in part, be related to the high endotoxin levels in this country. The findings support that high levels of endotoxin, or other bacterial products with Th1‐stimulating properties, might protect children from developing atopic disease.
Association of alcohol consumption with total serum immunoglobulin E levels and allergic sensitization in an adult population‐based survey
Authors: González‐Quintela A. , Gude F. , Boquete O. , Rey J. , Meijide L. M. , Suarez F. , Fernández‐Merino M. C. , Pérez L. F. , Vidal C.
Published: March 2003

Background Chronic alcoholism is associated with increased total serum IgE levels.

Objective The study aimed to investigate the relationship between alcohol intake and both total serum IgE levels and allergic sensitization in a general adult population.

Materials and methods A total of 720 subjects was randomly selected (stratified by age) from the population older than 18 years of A‐Estrada (Spain) and invited to participate in the study. From 697 eligible subjects, 469 (67%, median age 54 years, range 18 to 92 years, 44% males, 75% of cases from a rural environment) agreed to participate. A battery of 13 skin prick tests to common aeroallergens was performed in all subjects. Cases with at least one positive test (n = 121, 26%) were considered to have allergic sensitization. The most frequent sensitisers were mites and pollens (24% and 10% of subjects, respectively). Total serum IgE was measured in 465 subjects (99%). Alcohol consumption was registered as the number of standard (approximately 10 g) drinking units habitually consumed per week. A total of 244 subjects (52%) were alcohol consumers (median intake, 14 units/week, range 1 to 147 units/week). Abstainers (n = 225, 48%) constituted the reference category.

Results Alcohol consumption of more than 14 units/week was associated with an increase in serum IgE levels after adjusting for age, gender, allergic sensitization and smoking (P = 0.02). Alcohol consumption was not significantly associated with either overall allergic sensitization or mite sensitization after adjusting for age, gender and smoking. However, alcohol consumption of more than 14 units/week was associated with an increased prevalence of pollen sensitization (adjusted OR 3.15, 95% CI 1.19 to 8.34, P = 0.02).

Conclusion Alcohol consumption above a certain threshold is associated with an increase in total serum IgE levels. Alcohol consumption may also be associated with an increased prevalence of pollen sensitization.
The health‐related quality of life effects of once‐daily cetirizine HCl in patients with seasonal allergic rhinitis: a randomized double‐blind, placebo‐controlled trial
Authors: Noonan M. J. , Raphael G. D. , Nayak A. , Greos L. , Olufade A. O. , Leidy N. K. , Champan D. , Kramer B.
Published: March 2003

Background Seasonal allergic rhinitis (SAR) is characterized by subjectively irritating symptoms that can have a substantial impact on the patient's health‐related quality of life (HRQL), adversely affecting physical and social or work activity, interpersonal relationships, and general psychological well‐being. The objective of this study was to test the effect of cetirizine HCl 10 mg once daily on the HRQL of adult patients 18–65 years of age with SAR, concurrently assessing safety and efficacy.

Methods Randomized double‐blind, placebo‐controlled, parallel group trial conducted during the 1999 spring SAR season at 19 centers in the US. Following a 1‐week placebo run‐in period, qualified patients were randomized to cetirizine 10 mg, or placebo once daily for a 2‐week treatment period. Change in Rhinoconjunctivitis Quality of Life Questionnaire (RQLQ) and Total Symptom Severity Complex (TSSC) scores from baseline were the primary outcomes of interest.

Results Of the 611 patients enrolled, 403 (66%) were randomized. Cetirizine‐treated patients reported significantly greater improvement in overall HRQL (P < 0.001) and in each of the seven domains of the RQLQ at all time‐points (P < 0.05 to < 0.001) than the placebo group. They also experienced significantly greater symptom relief (P < 0.001) and were more satisfied with treatment (65% vs. 44%) compared to the placebo group. Correlations between symptomatic relief and overall HRQL improvement were moderate to strong and statistically significant (r = 0.49–0.68, P < 0.01).

Conclusions Results of this study are consistent with previous investigations, suggesting cetirizine HCl 10 mg taken once daily in the morning offers symptomatic relief that improves the HRQL of adults suffering from SAR.
Breastfeeding and the prevalence of symptoms of allergic disorders in Japanese adolescents
Authors: Miyake Y. , Yura A. , Iki M.
Published: March 2003

Background Environmental factors acting early in life are key determinants of the incidence of allergic disease. Whether breastfeeding is protective against allergic disorders remains controversial.

Objective The present cross‐sectional study examined the relationship between feeding patterns in the first 3 months of life and the prevalence of symptoms of wheeze, atopic eczema, and rhinoconjunctivitis during the past 12 months in Japanese adolescents.

Methods The subjects were 5614 of 9008 students (62%) aged 12–15 years from all public junior high schools in Suita, Japan in 2001. This study used the diagnostic criteria of the International Study of Asthma and Allergies in Childhood. Adjustment was made for gender, grade, number of older siblings, and parental history of allergy.

Results Feeding pattern was unrelated to the prevalence of wheeze or rhinoconjunctivitis. The prevalence of atopic eczema was significantly higher in children who had been breastfed than in artificial milk feeders (adjusted odds ratios = 1.40 and 1.56, 95% confidence intervals: 1.01–1.98 and 1.13–2.22 for mixed milk intake and breastfeeding only vs. artificial milk consumption, respectively; P = 0.01 for linear trend). When children were divided according to a positive or negative allergic history in at least one parent, an increased prevalence of atopic eczema associated with breastfeeding was found in children with a negative parental allergic history compared with those with a positive parental allergic history.

Conclusion The findings suggest that breastfeeding may be associated with an increased prevalence of atopic eczema, especially among children without a parental history of allergy.
Widespread biphasic amyloidosis: response to acitretin
Authors: Hernández-Núñez A. , Daudén E. , Moreno de Vega M. J. , Fraga J. , Aragüés M. , García-Díez A.
Published: March 2003

We report a patient who presented with an intensely pruritic rash, with hyperpigmentation of the whole back and multiple hyperpigmented and hyperkeratotic papules over his forearms and lower extremities. Histologically, he showed deposits of amyloid that reacted positively with monoclonal antibodies against cytokeratins. The lesions were resistant to antihistamines and corticosteroids. Treatment with oral acitretin was started and great improvement of the lesion was noted; the patient became asymptomatic. We review the literature on the effect of retinoids in cutaneous amyloidosis.
Hand rash in trichinosis
Authors: Walsh D. S. , Jongsakul K. , Watt G.
Published: March 2003

Trichinosis patients may develop peri-orbital oedema, conjunctival haemorrhages, splinter haemorrhages of the fingernails, and nonspecific skin rashes. Here, we describe an unusual hand rash noted in several patients enrolled in a treatment study for trichinosis.
The management of porphyria cutanea tarda
Authors: Sarkany R. P. E.
Published: March 2003

Porphyria cutanea tarda (PCT), the commonest of all porphyrias, is usually characterized by blisters and fragility of skin in light-exposed areas. It can be clinically indistinguishable from other disorders including variegate porphyria and the diagnosis can only be made by rigorous biochemical analysis. PCT does not cause acute attacks of porphyria. It is usually an acquired condition caused by inhibition of the uroporphyrinogen decarboxylase enzyme in the liver. Hereditary haemochromatosis, hepatitis C virus infection, alcohol, oestrogens and a family history of PCT are the major risk factors for the condition and should be searched for specifically in all patients. Liver disease, including hepatocellular carcinoma, is common in patients with PCT, and should be investigated for at presentation by means of a liver biopsy where possible. Patients with severe hepatic pathology or longstanding untreated PCT need to be monitored for the development of hepatocellular carcinoma in the long term. Low dose twice weekly chloroquine is the mainstay of treatment, but venesection should be used in patients with severe iron overload or hepatitis C-related liver disease. Subsequently, long-term follow-up is needed in all patients to monitor for relapse.
Bacterial lipopolysaccharide (LPS)-specific antibodies in commercial human immunoglobulin preparations: superior antibody content of an IgM-enriched product
Authors: Trautmann M , Held T K , Susa M , Karajan M A , Wulf A , Cross A S , Marre R
Published: March 2003

The anti-LPS antibody content of commercial intravenous immunoglobulins was examined by quantitative ELISA using LPS preparations from Escherichia coli, Klebsiella and Pseudomonas aeruginosa O serotypes occurring most frequently in Gram-negative septicaemia. Three IgG products from different manufacturers and one IgM-enriched product were tested. Mean antibody levels were significantly higher in the IgM fraction of the IgM-enriched product compared with ‘pure’ IgG products, indicating that natural antibodies against bacterial LPS belong primarily to the IgM class. Immunoblotting studies showed that antibody specificities were directed mainly against O side chain epitopes. Antibodies against rough mutant LPS representing various chemotypes were detected in IgG but not in IgM products. The virtual absence of antibodies against Vibrio cholerae LPS indicated that human anti-LPS antibodies result from continuous environmental exposure to Gram-negative pathogens. These data support the further development of IgM-enriched preparations for prophylaxis and treatment of Gram-negative nosocomial infections.
Anti-C1q receptor/calreticulin autoantibodies in patients with systemic lupus erythematosus (SLE)
Authors: Van Den Berg R H , Siegert C E H , Faber-Krol M C , Huizinga T W J , Van ES L A , Daha M R
Published: March 2003

SLE is a disease characterized by the presence of multiple autoantibodies and high levels of circulating immune complexes. We studied the presence and functional relevance of autoantibodies directed against a receptor for the collagen-like stalks of the first subcomponent of complement, also known as calreticulin (cC1qR/CaR), in patients with SLE. In a cross-sectional study it was found that higher titres of antibodies against cC1qR/CaR are present in sera of SLE patients compared with normal donors. No association between anti-cC1qR/CaR titres and SLE disease activity was found. Following gel filtration of SLE serum it was found that anti-cC1qR/CaR reactivity is associated with the peak of monomeric IgG. Purified IgG from patients was able to specifically immunoprecipitate cC1qR/CaR. Since we have shown previously that cC1qR/CaR is able to inhibit the haemolytic activity of C1q, we determined a possible pathogenic role for anti-cC1qR/CaR on complement regulation. IgG derived from SLE serum reversed the inhibitory capacity of cC1qR/CaR in a dose-dependent fashion up to 63%, whereas IgG from normal donors had no significant effect. With respect to the capacity of anti-cC1qR/CaR antibodies to activate neutrophils, it was found that incubation of normal neutrophils with F(ab′)2 anti-cC1qR/CaR resulted in a very limited oxidative burst. However, cross-linking of F(ab′)2 anti-cC1qR/CaR on the neutrophils clearly induced neutrophil activation. Pre-incubation of the SLE-derived F(ab′)2 with cC1qR/CaR prevented activation of neutrophils up to 81 ± 5%. These results suggest that the presence of anti-cC1qR/CaR antibodies in patients with SLE may modulate complement and neutrophil activation.
Cytokine release and its modulation by dexamethasone in whole blood following exercise
Authors: Smits H H , Grünberg K , Derijk R H , Sterk P J , Hiemstra P S
Published: March 2003

Glucocorticoids (GC) play an important role in the treatment of inflammatory diseases like asthma. However, in selected patients a relative resistance to GC has been reported. Recently, it has been suggested that GC sensitivity of peripheral blood leucocytes may be regulated in a dynamic fashion during exercise, in association with activation of the hypothalamic–pituitary–adrenal (HPA) axis. The aim of the present study was to explore changes in the GC sensitivity of cytokine production by leucocytes following strenuous exercise by well trained oarsmen. These changes were studied using lipopolysaccharide (LPS)-induced and anti-CD2/anti-CD28 MoAb-stimulated cytokine release in whole blood and its modulation by dexamethasone. Following exercise, significant decreases in LPS-induced release of IL-6, tumour necrosis factor-alpha (TNF-α) and IL-10 and anti-CD2/anti-CD28 MoAb-stimulated secretion of interferon-gamma (IFN-γ) were observed. In addition, the inhibitory effect of dexamethasone on both IL-6 and TNF-α secretion was significantly reduced following exercise, whereas that on IL-10 and IFN-γ release was not affected. These exercise-induced changes were accompanied by activation of the HPA axis, as indicated by an increase in circulating adrenocorticotropic hormone (ACTH) levels immediately following exercise. The results from the present study suggest that GC sensitivity of whole blood cytokine release can be regulated in a dynamic fashion and that this can be assessed using an ex vivo stimulation assay. Moreover, since dexamethasone responsiveness of anti-CD2/anti-CD28 MoAb-induced IFN-γ secretion in whole blood is not affected by exercise, it may suggest that exercise differentially affects monocytes and lymphocytes. The dynamic regulation of steroid responsiveness of leucocytes, as observed in the present study, could have important consequences for the effectiveness of GC treatment in inflammatory diseases.
Differences in kappa to lambda (κ:λ) ratios of serum and urinary free light chains
Authors: Abe M , Goto T , Kosaka M , Wolfenbarger D , Weiss D T , Solomon A
Published: March 2003

Free light chains (FLC) are a natural product of B lymphocytes and, as such, represent a quantifiable biomarker of cellular proliferation. Accurate measurement of the concentrations of these components in serum and urine provides a unique means of ascertaining B cell immunoglobulin synthesis during physiologic and, especially, pathologic states, where such information has important diagnostic and therapeutic implications. Previously, use of such quantitative assays has been limited due to the lack of potent serologic reagents specific for these components. We have immunized mice with κ- and λ-type monoclonal human light chains (Bence Jones proteins (BJP)) and have obtained monoclonal antibodies (MoAbs) that differentiate between unbound and bound light chains. These highly specific MoAbs were used to measure by ELISA the concentrations of FLC in the serum of 22 normal individuals and in urine from 16 of these subjects. The mean serum κ and λ FLC concentrations were found to be 16.6 ± 6.1 μg/ml and 33.8 ± 14.8 μg/ml, respectively. In contrast, the values for urinary κ and λ FLC were 2.96 ± 1.84 μg/ml and 1.07 ± 0.69 μg/ml, respectively. In each case studied, the serum κ:λ ratio was consistently less than that of urine (mean values, serum ≈ 1:2; urine ≈ 3:1). That the rate of synthesis of λ-type FLC exceeded that of κ was evidenced in assays of culture fluid supernatants of unstimulated normal peripheral blood mononuclear cells (PBMC), where the mean κ:λ ratio was determined to be 1:1.4. Metabolic studies in which mice were injected with pools of κ- and λ-type BJP prepared in ratios of 1:1, 1:2 and 1:4 demonstrated that, regardless of the proportion, κ FLC were preferentially excreted. Our studies provide the first evidence that λ FLC are secreted by normal PBMC at a greater rate than are κ FLC, as evidenced in biosynthetic studies and by measurement of their serum concentrations. Further, we posit that quaternary structural differences between the two light-chain isotypes may account for the predominance of κversusλ components in urine.
Mutations of the p53 tumour suppressor gene in erosive rheumatoid synovial tissue
Authors: Rème T , Travaglio A , Gueydon E , Adla L , Jorgensen C , Sany J
Published: March 2003

Erosive rheumatoid arthritis (RA) is accompanied by synovial tissue hyperplasia associated with the proliferation of transformed-appearing synovial lining cells. In the present study we have analysed the expression of the p53 tumour suppressor gene in the synovial pannus tissue from patients at various stages of the disease. We used a combination of polymerase chain reaction (PCR) and single-strand conformation polymorphism (SSCP) on DNA and reverse transcription, PCR and sequencing on cDNAs from synovial tissues or purified synovial cell populations of 24 RA and three osteoarthritis (OA) patients. We also studied p53 expression by immunohistochemical analysis. Mutations suspected after SSCP were identified by systematic sequencing of the p53 exon 6, especially in the fibroblast-like, adherent synovial cell population, associated with an erosive disease. Some accumulation of the protein was detected in immunohistochemical analysis of the p53 tumour suppressor gene in the patients' synovial tissues. However, no sign of malignancy was seen in these patients after a 2-year survey. These results show some abnormalities in the p53 tumour suppressor gene in RA patients, but do not allow this to be related to characteristic proliferative features of the rheumatoid synovium.
Correlation of plasma monocyte chemoattractant protein-1 (MCP-1) and monocyte inflammatory protein-1α (MIP-1α) levels with disease activity and clinical course of sarcoidosis
Authors: Hashimoto S , Nakayama T , Gon Y , Hata N , Koura T , Maruoka S , Matsumoto K , Hayashi S , Abe Y , Horie T
Published: March 2003

MCP-1 and MIP-1α exhibit chemotactic activity toward macrophages/monocytes and induce the production of inflammatory cytokines affecting granuloma formation. Up-regulated expression of MCP-1 and MIP-1α in the affected organ of sarcoidosis has been shown; however, the relationship between their plasma levels and the clinical course of this disease has not been determined. In the present study we measured plasma MCP-1 and MIP-1α levels in 26 patients with active sarcoidosis by ELISA in order to assess the state of MCP-1 and MIP-1α in this disease. Most patients in this study (21/26) had clinical evidence of extrathoracic disease in addition to pulmonary involvement. In addition, a high proportion of patients (n = 15) showed spontaneous remission of disease, whereas five patients showed no spontaneous remission and six patients were treated with corticosteroids over the 2-year period of study. At the time of diagnosis, both plasma MCP-1 and MIP-1α levels in patients with active sarcoidosis were significantly higher than in the normal controls. The levels of these cytokines in patients with extrathoracic disease were compatible with those in patients without extrathoracic disease. A longitudinal evaluation of plasma MCP-1 and MIP-1α levels showed that the changes in both cytokines were closely related to the clinical course of sarcoidosis. These results suggest that plasma MCP-1 and MIP-1α may be useful parameters for monitoring the clinical course of sarcoidosis. In addition, plasma MCP-1 and MIP-1α may reflect subclinical evidence of extrathoracic sarcoidosis and may play a role in initiating monocyte migration into the tissue.
Isolation and phenotypic characterization of colonic macrophages
Authors: Rogler G , Hausmann M , Vogl D , Aschenbrenner E , Andus T , Falk W , Andreesen R , Schölmerich J , Gross V
Published: March 2003

Macrophages play an important role in the intestinal mucosal immune system. However, they are a poorly defined cell population. We therefore determined their phenotype in normal colonic mucosa. Macrophages were isolated from colonic biopsies and surgical specimens by collagenase digestion. Colonic macrophages were positively sorted by anti-CD33 magnetic beads. Flow cytometric triple fluorescence analysis was applied to study CD14, CD16, CD33, CD44, CD11b, CD11c, CD64, HLA-DR, CD80, CD86 and CD3/CD19 expression. CD33 was evaluated as a positive marker for intestinal macrophages. CD33+ cells isolated from normal colonic mucosa showed co-expression of the established intracellular macrophage marker CD68 in FACS analysis. CD33+ cells were capable of phagocytosis. Isolation of this cell population by magnetic anti-CD33 beads and culture resulted in a 4·2–40-fold increase in IL-1β and 4·5–44-fold increase in tumour necrosis factor-alpha (TNF-α) secretion compared with unsorted lamina propria mononuclear cells (LPMC). Of the CD33+ cells, 90·9 ± 6·9% (mean ± s.d.) were CD44+. However, macrophages from colonic mucosa showed only a low expression of CD14 (10·5 ± 3·8%), CD16 (10·1 ± 3·9%), HLA-DR (27·3 ± 9·2%), CD11b (17·4 ± 6·8%), CD11c (17·8 ± 10·4%). Furthermore, expression of CD80 (9·2 ± 4·2%) and CD86 (15·1 ± 7·3%) was low, suggesting a low ability of normal intestinal macrophages to activate T cells and T cell-mediated immune responses. We conclude that CD33 is useful for the isolation and flow cytometric characterization of colonic macrophages. These cells exhibit a single phenotype in normal mucosa (CD33++, CD44++, CD14−, CD16−, CD11b−, CD11c−, HLA-DRlow, CD80−, CD86−) lacking lipopolysaccharide (LPS) receptor and costimulatory molecules.
Polarized secretion of IL-6 and IL-8 by human retinal pigment epithelial cells
Authors: Holtkamp G M , Van Rossem M , DE Vos A F , Willekens B , Peek R , Kijlstra A
Published: March 2003

A number of cell types situated along interfaces of various tissues and organs such as the peritoneum and the intestine have been shown to secrete inflammatory cytokines in a polarized fashion. Retinal pigment epithelial (RPE) cells are positioned at the interface between the vascularized choroid and the avascular retina, forming part of the blood–retina barrier. These cells are potent producers of inflammatory cytokines and are therefore considered to play an important role in the pathogenesis of ocular inflammation. Whether cytokine secretion by these cells also follows a vectorial pattern is not yet known, and was therefore the subject of this study. Monolayers of human RPE cells (primary cultures and the ARPE-19 cell line) cultured on transwell filters were stimulated to produce IL-6 and IL-8 by adding IL-1β (100 U/ml) to either the upper or the lower compartment. After stimulation, the human RPE cell lines showed polarized secretion of IL-6 and IL-8 towards the basal side, irrespective of the side of stimulation. The ARPE-19 cell line also secreted IL-6 and IL-8 in a polarized fashion towards the basal side after basal stimulation; polarized secretion was, however, not apparent after apical stimulation. The observation that human RPE cells secrete IL-6 and IL-8 in a polarized fashion towards the choroid may represent a mechanism to prevent damage to the adjacent fragile retinal tissue.
IL-10 and IL-4 co-operate to normalizein vitroIgA production in IgA-deficient (IgAD) patients
Authors: Marconi M , Plebani A , Avanzini M A , Maccario R , Pistorio A , Duse M , Stringa M , Monafo V
Published: March 2003

In the present study we evaluated in vitro immunoglobulin production from IgAD individuals and healthy controls. Peripheral blood mononuclear cells (PBMC) from IgAD and controls were cultured with anti-CD40 MoAb presented on a CDw32-transfected fibroblast cell line (CD40 system) in the presence of IL-10, IL-2, IL-4, transforming growth factor-beta (TGF-β) alone as well as of IL-10 in combination with each of the other three cytokines. Only IL-10 added alone induced significant changes in baseline immunoglobulin production; marked increases in median supernatant levels of all three isotypes were observed in both groups. The most striking finding of this study was the synergizing effect of IL-4 on IgA production in the IgAD group when added with IL-10; median IgA supernatant level increased to a value superimposable on that found in the normal controls which remained about the same as when stimulated with IL-10 alone. The synergic effect of IL-4 and IL-10 was specific to the IgA isotype.
Colonic explant production of IL-1 and its receptor antagonist is imbalanced in inflammatory bowel disease (IBD)
Authors: Dionne S , D'agata I D , Hiscott J , Vanounou T , Seidman E G
Published: March 2003

IBD is associated with an increased activation of intestinal immune cells, which causes overproduction of proinflammatory cytokines such as IL-1β. IL-1β is implicated in mediating the sustained inflammatory response. IL-1 receptor antagonist (IL-1Ra), the naturally occurring inhibitor of IL-1, has been shown to have beneficial effects in experimental models of colitis. In this study we investigated the hypothesis that an imbalance between IL-1 and IL-1Ra exists in IBD by measuring their secretion by explant cultures of colonic biopsies. Freshly homogenized biopsies from involved tissue in IBD patients exhibited significantly lower IL-1Ra/IL-1β ratios than control and uninvolved IBD mucosal tissue. Using explant cultures, in vitro production of IL-1β and IL-1Ra increased progressively during the 4–18-h culture periods. IL-1β secretion was higher in supernatants from involved Crohn's disease (CD) and ulcerative colitis tissue compared with control tissue, and IL-1β levels increased with severity of inflammation. IL-1Ra secretion was not elevated in involved IBD samples, but significantly higher levels were released when moderate to severely involved tissue samples were compared with non-inflammatory controls. Similar to freshly homogenized tissue, explant studies showed that the IL-1Ra/IL-1β ratios were significantly decreased in involved IBD tissue, but not in uninvolved CD or inflammatory control specimens. These data support the hypothesis of an imbalance between IL-1β and IL-1Ra in IBD.
IL-6 receptor blockage inhibits the onset of autoimmune kidney disease in NZB/W F1 mice
Authors: Mihara M , Takagi N , Takeda Y , Ohsugi Y
Published: March 2003

In the present study, we examined the preventive effect of anti-mouse IL-6 receptor (IL-6R) antibody, MR16-1, on the development of autoimmune kidney disease in female NZB/W F1 (BWF1) mice. Immunological tolerance to MR16-1 or isotype-matched control antibody, KH-5, was induced by the simultaneous administration of anti-CD4 MoAb in mice. Thereafter, mice were intraperitoneally given 0.5 mg of MR16-1, 0.5 mg of KH-5 or saline once a week from 13 to 64 weeks of age. MR16-1 treatment dramatically suppressed proteinuria and prolonged the survival time of BWF1 mice. Only one out of 10 mice died with high levels of proteinuria throughout the experiment. MR16-1 almost completely suppressed the production of IgG forms of anti-DNA and anti-TNP antibodies, but not the IgM forms of these antibodies. In particular, all IgG subclasses (IgG1, IgG2a, IgG2b and IgG3) of anti-DNA antibody production were significantly suppressed. Moreover, serum IgG1, IgG2a and IgG3 levels in MR16-1-treated mice were lower than those in saline- and KH-5-treated mice, whereas serum IgM and IgA levels were not influenced. In conclusion, MR16-1 potently suppressed the development of autoimmune disease in BWF1 mice, and this was attributed to its effect of specific suppression of IgG class antibody production.
Contribution of the IL-2 and IL-10 genes to inflammatory bowel disease (IBD) susceptibility
Authors: Parkes M , Satsangi J , Jewell D
Published: March 2003

Although the importance of genetic susceptibility to IBD has been established by epidemiological studies, the genes involved remain poorly characterized. Important candidate genes include those encoding the immunoregulatory cytokines IL-2 and IL-10. The aim of this study was to assess the contribution of the IL-2 and IL-10 genes to IBD susceptibility. One hundred and ninety-eight pairs of siblings with IBD were genotyped at dinucleotide repeat polymorphisms within the IL-2 and IL-10 genes, and data analysed by the affected sib-pair method of linkage analysis and the transmission disequilibrium test (TDT). A subset of 89 affected sibling pairs was genotyped at markers flanking the IL-2 gene as part of a genome-wide search. The IL-2 polymorphism showed no linkage to IBD overall, but modest evidence for linkage to the ulcerative colitis (UC) data set (P = 0.028). A microsatellite 4 cM distal to the IL-2 gene showed a similar distortion in the ulcerative colitis subgroup (P = 0.006). The TDT showed some distortion of allelic transmission for the IL-2 polymorphism in the UC group, but this did not reach statistical significance (P = 0.09). Results for the IL-10 polymorphism were not significant. Thus the gene encoding IL-2 may contribute to UC susceptibility, but the effect is modest and must await replication in other data sets. The IL-10 gene does not appear to contribute to the risk of developing UC or Crohn's disease.
Mice incapable of making IL-4 or IL-10 display normal resistance to infection withMycobacterium tuberculosis
Authors: North R J
Published: March 2003

With a view to determining whether production of Th2 cytokines, IL-4 or IL-10, is responsible for the inability of mice to resolve infection with Mycobacterium tuberculosis, mice with targeted disruption of their IL-4 or IL-10 gene were compared with wild-type mice in terms of their ability to defend against an M. tuberculosis infection initiated via the respiratory route. The results show that mice that are unable to make either IL-4 or IL-10 are no more capable than wild-type mice at defending against tuberculosis (TB). Therefore, the results are inconsistent with the proposition that the inadequacy of Th1-mediated anti-tuberculosis immunity is due to its down-regulation by either of these Th2 cytokines.
Differential use of immunoglobulin light chain genes and B lymphocyte expansion at sites of disease in rheumatoid arthritis (RA) compared with circulating B lymphocytes
Authors: Moyes S P , Maini R N , Mageed R A
Published: March 2003

The presence of germinal centre-like structures and clonotypic expansion of lymphocytes in RA synovia may indicate a site-specific immune response to local antigens, rather than passively entrapped immune cells, that sustains synovial inflammation. In this study we compare the nature of immunoglobulin light chain variable region gene use in the synovium of RA patients with peripheral B cells to determine the nature of the synovial immune response. Using Vλ and Vκ gene fingerprinting, which relies on differences in CDR3 length, we demonstrate differences in the pattern of Vλ and Vκ use and clonotypic expansion of B cells between the synovium and peripheral blood of RA patients. Further, we show that some synovial rearrangements with long CDR3 are selectively expanded. These longer than usual CDR3 were generated by a number of mechanisms including N-additions. However, the observed differences were not uniform in different patients. These observations suggest that local synovial antigens drive significant numbers of T and B lymphocytes selected from an existing repertoire shaped by genetic and environmental factors. Further, the data argue against passive retention of most B cells in the synovium of RA patients.