bluuebunny/crossref_metadata_2025 huggingface dataset:
Validity of specific IgE antibodies in children with egg allergy
Authors: Boyano Martínez T. , García‐Ara C. , DÍaz‐Pena J. M. , Muñoz F. M. , García Sánchez G. , Esteban M. M.
Published: March 2003

Background The demonstration of specific IgE antibodies to egg supports the existence of allergy to this food, but a correct diagnosis can only be obtained after a challenge test. Several studies have assessed different cut‐off points in the level of these antibodies as predictors of clinical reactivity.

Objective Validation of the specific IgE antibodies measured by the CAP System Fluorescence enzyme immunoassay (FEIA) technique in the diagnosis of egg allergy in children under 2 years of age.

Methods A prospective study of 81 children with suspected egg allergy was performed. Specific IgE antibodies was quantified for egg white, egg yolk, ovoalbumin and ovomucoid. The diagnostic challenge test was carried out following the previously established criteria. The validity of the specific IgE antibodies was analysed using children with a negative diagnostic challenge test as control group.

Results The prevalence of egg allergy in the group studied was 79% and egg white was the allergen that showed the greatest diagnostic efficacy. The sensitivity and positive predictive value of the prick test and of the CAP to egg white were excellent and the specificity and the negative predictive value had lower values. A level of ≥ 0.35 KUA/L for specific IgE antibodies to egg white predicted the existence of reaction in 94% of the cases.

Conclusions Quantification of the specific IgE antibodies to egg white is useful in the diagnosis of egg allergy. In children under 2 years of age with a background of immediate hypersensitivity after egg ingestion and presence of specific IgE antibodies to egg white of ≥ 0.35 KUA/L, diagnostic challenge test is not necessary to establish the diagnosis of allergy to this food.
Increased occurrence of asthma and allergy: critical appraisal of studies using allergic sensitization, bronchial hyper‐responsiveness and lung function measurements
Authors: Wieringa M. H. , Vermeire P. A. , Brunekreef B. , Weyler J. J.
Published: March 2003

Background Many studies have reported an increase in the occurrence of asthma and respiratory allergies in recent decades, but this increase is mostly based on studies using rather subjective measurements of asthma and allergies, such as questionnaires and doctor's diagnosis. None of the reviews specifically focused on studies using more ‘objective’ measurements, such as sensitization (specific IgE or skin prick testing (SPT)), bronchial hyper‐responsiveness (BHR) or lung function (LF).

Objective To review articles studying a time trend of occurrence of these ‘more objective’ measurements.

Methods A MEDLINE‐search (1966–February 2000) was performed. The following criteria were used: population‐based, using IgE, SPT, BHR or LF measurements in the same age‐group at least twice, with at least 2 years between and using similar methods.

Results The MEDLINE‐search resulted in only 16 articles, performed in 13 populations in seven different countries. Nine articles used the same objective measurements twice in the whole population. Three of these reported a non‐significant increase or decrease. The other six articles found a significant increase in at least one objective measurement and of these only three reported a consistent significant increase.

Conclusions The increase in the occurrence of reported asthma and allergy is supported by only a few articles confirming these results with ‘more objective measurements’.
Allergen‐induced Th1 and Th2 cytokine secretion in relation to specific allergen sensitization and atopic symptoms in children
Authors: Jenmalm M. C. , Van Snick J. , Cormont F. , Salman B.
Published: March 2003

Background Allergic diseases are believed to be due to T helper (Th)2‐like immunity to allergens in affected tissues, and immune responses to allergens are characterized by a cross‐regulation between Th1 and Th2 cells. Atopic individuals may develop IgE antibodies to only one or more allergens. However, the mechanisms behind sensitization to a specific allergen, e.g. why an individual develops IgE to cat but not birch, are not known. Our aim was to study birch‐ and cat‐induced Th1 and Th2 cytokine secretion in children who were sensitized to birch but not to cat, and vice versa.

Materials and methods The subjects in the study were 60 12‐year‐old children. Seventeen of the children were sensitized (skin prick test and circulating IgE positive) to birch but not cat, 13 were sensitized to cat but not birch, 11 were sensitized both to birch and cat, and 19 children were skin prick test and circulating IgE negative. Forty‐six children had a history of atopic symptoms, and 42 of them had current symptoms. Peripheral blood mononuclear cells were separated from venous blood and stimulated with cat or birch allergen. The levels of IL‐4, IL‐5, IL‐9, IL‐10, IL‐13 and IFN‐γ in the cell supernatants were analysed by ELISA.

Results Sensitized children produced more of the Th2 cytokines IL‐4, IL‐5, IL‐9 and IL‐13 than non‐sensitized atopic and non‐atopic children in response to stimulation with the allergen they were sensitized to. High levels of the Th2 cytokines IL‐4 and IL‐5 and low levels of the anti‐inflammatory cytokine IL‐10 were associated with atopic symptoms, and high cat‐induced IL‐9 levels with asthma.

Conclusions The Th2 cytokines IL‐4, IL‐5, IL‐9 and IL‐13 were all commonly detected in sensitized children after stimulation with the specific, in contrast to an unrelated, allergen. Atopic symptoms were associated with increased levels of IL‐4 and IL‐5 and tended to be associated with low levels of IL‐10, and asthma with high cat‐induced IL‐9 levels.
Allergen‐specific T cell reactivity in cord blood: the influence of maternal cytokine production
Authors: Kopp M. V. , Zehle C. , Pichler J. , Szépfalusi Z. , Moseler M. , Deichmann K. , Forster J. , Kuehr J.
Published: March 2003

Background Successful pregnancy is dependent upon T helper (Th)2‐type‐dominated immunological responsiveness in gestation‐associated compartments.

Objective In our study we observed the influence of the maternal Th2‐associated cytokine pattern on the naive fetal T cell phenotype and asked if circulating Th2 cytokines of atopic mothers affects the Th1/Th2 differentiation of the fetus.

Methods Cord blood mononuclear cells (CBMC) and peripheral blood mononuclear cells (PBMC) of the corresponding mothers were isolated. The proliferative response of CBMC and PBMC to Betalactoglobulin (BLG) was assessed by liquid scintillation counting. The cytokines interferon (IFN)‐γ, and interleukin (IL)‐5, IL‐10 and IL‐13 in the cell culture supernatants were measured using the ELISA technique. We then defined two subgroups based on maternal levels of specific IgE against aeroallergens: sensitized mothers (MA+) and their neonates (NMA+) (n = 18) and non‐sensitized mothers (MA−) and their neonates (NMA−) (n = 29).

Results Nearly all mothers (98%) and neonates (92%) had a positive proliferation response after stimulation with BLG (mean stimulation index (10–90 percentile): neonates: 7 (2–15); mothers 14 (5–29)). In supernatants of BLG‐stimulated cell cultures, sensitized mothers showed a significantly lower IFN‐γ concentration in comparison to non‐sensitized mothers (MA+ = 25; MA− = 123 IU/L; P < 0,05), whereas the neonates did not differ significantly (NMA+ = 306; NMA− = 224 IU/L; n. s.). Nor was any difference found in the IL‐13 concentration between the two groups of sensitized and non‐sensitized mothers (MA+ = 48; MA− = 125 pg/mL; n. s.). CBMC of neonates with a sensitized mother showed significantly higher IL‐13 concentrations in response to BLG than neonates of non‐sensitized mothers (NMA+ = 1442, NMA− 738 pg/mL; P < 0.05). The IL‐5 and IL‐10 concentrations did not differ significantly within the neonatal and the maternal subgroups.

Conclusions Our data suggests that maternal sensitization to allergens is associated with the reduced maternal production of the Th2 antagonist IFN‐γ and elevated production of the Th2 cytokine IL‐13 in the offspring.
Pets and vermin are associated with high endotoxin levels in house dust
Authors: Heinrich J. , Gehring U. , Douwes J. , Koch A. , Fahlbusch B. , Bischof W. , Wichmann H. E.
Published: March 2003

Background Previous studies have shown that the risk for allergic sensitization is lower in children who grew up on farms and in young adults who were exposed to dogs in early childhood. A higher microbial exposure in general and in particular to endotoxin in early childhood might contribute to this lower risk of atopy.

Objective We examined whether the presence of pets or vermin in the home is associated with higher endotoxin concentrations in settled house dust.

Methods House dust was sampled in a standardized manner on the living room floors of 454 homes of German children aged 5–10 years (participation rate 61%). Endotoxin was assessed with a quantitative kinetic chromogenic Limulus Amebocyte Lysate (LAL) method. Associations between endotoxin levels, pets and vermin are presented as ratios of the crude and confounder adjusted geometric means (means ratios) in the category of study vs. a reference category using multiple linear regression models.

Results Endotoxin concentrations in living room floor dust sampled in homes without pets and vermin were lower (1246 ng per square meter, 1519 ng endotoxin/g dust, n = 157) than those sampled in homes with pets or vermin (2267 ng per square meter, 2200 ng endotoxin/g dust, n = 296). After adjustment for city of residence, season of dust sampling, age of the building and story of the dwelling, means ratios for endotoxin expressed per gram of dust were statistically significantly increased for dog (1.64, 95% CI 1.09–2.46), for cat (1.50, 95% CI 1.03–2.18) and for cockroach (3.01, 95% CI 1.37–6.60), whereas no major statistically significant associations were found for other pets, ants and mice.

Conclusion Keeping a dog or a cat in the home is consistent with higher exposure to endotoxin and might therefore contribute to the lower risk of atopy in later life.
Local lung responses following local lung challenge with recombinant lungworm antigen in systemically sensitized sheep
Authors: Collie D. D. S. , MacAldowie C. N. , Pemberton A. D. , Woodall C. J. , McLean N. , Hodgson C. , Kennedy M. W. , Miller H. R. P.
Published: March 2003

Background Chronic mast cell‐mediated inflammation may contribute significantly towards the extensive tissue remodelling that is a feature of lungworm infection in ruminants. Understanding the factors that control tissue remodelling is a necessary step toward effective management and treatment of conditions that feature such pathology.

Objective We sought to define in a novel ovine model system, the cellular, immune and mast cell phenotypic events that occur following local lung challenge with a recombinant protein antigen, DvA‐1, derived from the ruminant lungworm nematode, Dictyocaulus viviparus.

Methods Two spatially disparate lung segments in systemically sensitized sheep were challenged on three occasions with DvA‐1 (3xDVA) and two further segments were challenged with saline (3xSAL). Two months after the third challenge, one of the two segments previously repeatedly challenged with DvA‐1 was challenged again with DvA‐1 (3xDVA:DVA) whilst the other was challenged with saline (3xDVA:SAL). A similar protocol was followed with the saline challenged segments (3xSAL:SAL and 3xSAL:DVA). Bronchoalveolar lavage fluid (BALF) (n = 16) and tissue (n = 3) were collected after the last challenge.

Results Cellular changes 24 h after the fourth challenge were characterized by an increase in the absolute numbers of neutrophils and eosinophils in BALF from 3xDVA:DVA and 3xSAL:DVA segments. Local antibody production was implied through increased levels of antibody in both 3xDVA:DVA and 3xDVA:SAL segments, with the latter being unaffected by inflammation. Levels of active transforming growth factor beta‐1 (TGF‐β1) were significantly increased in 3xDVA:SAL segments and a trend towards an increase was apparent in 3xDVA:DVA segments. Total TGF‐β1 levels were significantly correlated with eosinophil counts in all except the 3xDVA:SAL segments. Such changes in the bronchoalveolar space were complemented by increased ratios of sheep mast cell proteinase‐1 expressing cells and tryptase expressing cells, to toluidine blue positive cells in airways from 3xDVA:DVA segments.

Conclusion Mast cell phenotypic events occurring as a consequence of antigen challenge were limited to segments in which changes in BALF were characterized by neutrophil influx and increased local antibody production.
Effects of bβ‐endorphin on nasal allergic inflammation
Authors: Baumgarten C. R. , Schmitz P. , O'Connor A. , Kunkel G.
Published: March 2003

Background πβ‐endorphin is a derivative of pro‐opiomelanocortin. Cells of the immune system can also synthesize and secrete β‐endorphin. Its concentration is increased during the allergic reaction and during stress. Increased reactivity during psychological stress of allergic subjects is also well known.

Objective Is β‐endorphin one physiological link between stress and an exacerbation of the allergic reaction?

Methods First, intranasal β‐endorphin challenges with subsequent lavages to determine histamine and albumin levels and measurements of nasal flow and resistance in dose‐response and time course experiments were performed. Secondly, we examined whether β‐endorphin pre‐treatment increased the antigen‐induced release of histamine and albumin in nasal lavages and the clinical symptoms.

Results Exogenous β‐endorphin (100 pM−10 µM/mL) induced a dose‐dependent increase in nasal symptoms in asymptomatic allergic subjects with rhinitis (n = 14) as well as in non‐allergic controls (n = 10), but did not release any mediators into nasal secretion. However, comparing the antigen‐evoked release of mediators into nasal secretions with that of a β‐endorphin pre‐treated antigen challenge we could note a significant enhancement of human serum albumin influx (P < 0.05) and histamine liberation (P < 0.05) 10 min after antigen challenge compared with the allergen challenge alone, with also a correlation with the more pronounced decrease in nasal flow (P < 0.05).

Conclusion These results suggest that β‐endorphin‐induced increase in nasal congestion is mediated through direct neuroendocrine receptor activation independent of mast cell activation and that during the allergic reaction there is a β‐endorphin/mast cell interaction that enhances the mediator response to nasal allergen challenge.
A lower prevalence of atopy symptoms in children with type 1 diabetes mellitus
Authors: Meerwaldt R. , Odink R. J. , Landaeta R. , Aarts F. , Brunekreef B. , Gerritsen J. , Van Aalderen W. M. C. , Hoekstra M. O.
Published: March 2003

Background The Th1/Th 2 concept is a model to understand the pathophysiology of certain diseases. Atopic diseases (asthma, eczema and hayfever) are characterized by a chronic inflammatory reaction that is dominated by Th 2 cells, and type 1 diabetes mellitus (DM) is Th1 cell dominated. Because it is known that Th1 and Th 2 cells reciprocally counteract each other, it can be speculated that the prevalence of Th 2‐mediated disease is lower in patients with Th1‐mediated disease.

Objective To compare the prevalence of atopic diseases between children with DM and age‐matched controls.

Methods Parents of children with DM were requested by Dutch paediatricians to complete the International Study of Asthma and Allergies in Childhood (ISAAC) questionnaire on the prevalence of atopic diseases. A control group was derived from a Dutch cross‐sectional survey (the ISAAC2 study).

Results We received 555 completed questionnaires, which is estimated to be 25% of the total number of Dutch children with DM. The control group consisted of 777 children. After age‐matching, the questionnaires of 188 DM patients were used. Symptoms of asthma, hayfever and eczema were reported less in the group of children with DM compared with the control group (wheeze last year, OR 0.796, 95% CI 0.408–1.554; hayfever symptoms last year, OR 0.642, 95% CI 0.369–1.118; eczema symptoms last year, OR 0.693, 95% CI 0.430–1.115).

Conclusion The lower prevalence of astma, hayfever and eczema symptoms in DM patients compared with age‐matched controls, although not statistically significant, is consistent with the Th1/Th 2 concept.
Migration through basement membrane modulates eosinophil expression of CD44
Authors: Dallaire M.‐J. , Ferland C. , Lavigne S. , Chakir J. , Laviolette M.
Published: March 2003

Background Tissue eosinophils express more membrane receptors and release more mediators than blood eosinophils, suggesting that migration from blood to tissue modulates eosinophil phenotype and functions.

Objective We postulated that eosinophil passage through endothelial basement membrane, an important step of eosinophil migration into tissue, may be responsible for some of these changes.

Method We previously showed that 5‐oxo‐6, 8, 11, 14‐eicosatetraenoic acid (5‐oxo‐ETE) in combination with IL‐5 promotes eosinophil migration through Matrigel®, a mouse tumour cell‐derived basement membrane. Using this model, we evaluated the effect of trans‐Matrigel migration on purified human blood eosinophil expressions of CD44, CD69 and HLA‐DR that either increase or appear on activated eosinophils, and releases of peroxidase (EPO), leukotriene (LT) C4 and granulocyte‐monocyte colony stimulating factor (GM‐CSF).

Results IL‐5, but not 5‐oxo‐ETE, increased eosinophil expression of CD44 and CD69. Migration of eosinophils through Matrigel significantly increased CD44 expression level over the one induced by IL‐5 (P = 0.0001). Migration through Matrigel did not modify CD69 expression compared with the one obtained in the presence of IL‐5 alone; however, incubation of eosinophils on Matrigel decreased IL‐5‐induced CD69 (P = 0.0001). Trans‐Matrigel migration did not modify HLA‐DR expression, nor EPO, LTC4 and GM‐CSF releases.

Conclusion These data show that in vitro trans‐Matrigel migration and Matrigel contact modulate eosinophil membrane receptor expression. Consequently, they suggest that migration through basement membrane mediates changes in cell‐surface phenotype observed on activated eosinophils and probably prepares them for interactions with tissue components and cells.
Characterization and comparison of commercially available German and American cockroach allergen extracts1
Authors: Patterson M. L. , Slater J. E.
Published: March 2003

Background In this study we examine the variability among unstandardized cockroach allergen extracts.

Methods We obtained 24 aqueous and glycerinated cockroach allergen extracts from nine manufacturers. We used previously characterized cockroach extracts, E2‐Cg and E2‐Ca, as references. The modified ninhydrin assay was used to determine protein concentration of each extract. Relative potencies of extracts were determined by competition ELISA, using a human allergic serum pool. Bla g 1 and Bla g 2 levels of glycerinated German cockroach extracts were determined by ELISA using monoclonal antibodies. Extracts were also analysed by SDS‐PAGE.

Results Commercial cockroach allergen extracts had highly variable protein contents that were lower than the protein contents of the references. Electrophoretic data confirmed the presence of a variable number and intensity of protein bands in extracts among manufacturers. The relative potencies of the commercial extracts were between 10 and 782 BAU/mL for German cockroach and 10–250 BAU/mL for American cockroach. The mean Bla g 1 content of the commercial extracts was significantly lower than that of the reference (P = 0.001). The mean Bla g 2 content of the commercial extracts was higher than that of the E2‐Cg reference but the Bla g 2 levels were more variable compared to Bla g 1. In glycerinated German cockroach extracts, protein concentrations, relative potencies and specific allergen levels were significantly correlated (P < 0.001).

Conclusion Our tests indicate that commercially available cockroach allergen extracts are variable in protein content, electrophoretic banding patterns, relative potency and Bla g 2 levels. In glycerinated German cockroach extracts, protein concentrations, relative potencies and specific allergen levels were significantly correlated.
Molecular cloning and characterization of a new Japanese cedar pollen allergen homologous to plant isoflavone reductase family
Authors: Kawamoto S. , Fujimura T. , Nishida M. , Tanaka T. , Aki T. , Masubuchi M. , Hayashi T. , Suzuki O. , Shigeta S. , Ono K.
Published: March 2003

Background Japanese cedar (Cryptomeria japonica) pollen is a major cause of seasonal pollinosis, and more than 10% of Japanese people suffer from this allergic disorder. However, only two major pollen allergens, Cry j 1 and Cry j 2, have been identified and exclusively characterized.

Objective The aim of this study was to explore and identify important Japanese cedar pollen allergens other than Cry j 1 or Cry j 2.

Methods C. japonica
cDNA library was immunoscreened by rabbit antiserum raised against a partially purified cedar pollen allergen fraction. An isolated cDNA clone was inserted into a glutathione S‐transferase (GST)‐tagged
Escherichia coli expression vector to obtain recombinant GST fusion protein. Non‐fusion recombinant protein was purified by glutathione Sepharose affinity chromatography in conjunction with factor Xa cleavage of the GST moiety. IgE‐binding ability of the recombinant protein was then evaluated by western blot analysis and enzyme‐linked immunosorbent assay (ELISA).

Results The cDNA encodes 306 amino acids with significant sequence similarity to those of plant isoflavone reductase‐like proteins, which include a recently identified birch pollen allergen Bet v 5. Western blot analysis demonstrated that recombinant protein was recognized by cedar pollinosis patient IgE. In contrast to Bet v 5 being reported as a minor allergen, the recombinant protein exhibited 76% IgE binding frequency (19/25) against pollinosis patients.

Conclusion Here we identified the third member of Japanese cedar pollen allergen homologous to isoflavone reductase. Its high IgE‐binding frequency implicates that the isoflavone reductase homologue might be an additional major pollen allergen in C. japonica.
Suplatast tosilate inhibits thymus‐ and activation‐regulated chemokine production by antigen‐specific human Th2 cells
Authors: Oda N. , Minoguchi K. , Tanaka A. , Yokoe T. , Minoguchi H. , Matsuo H. , Nakashima M. , Tasaki T. , Adachi M.
Published: March 2003

Background Suplatast tosilate is an anti‐allergic agent that suppresses cytokine production by human Th2 cells.

Objective We investigated the effects of suplatast tosilate on the production of thymus‐ and activation‐regulated chemokine (TARC) by T cells from allergic patients with asthma.

Methods Purified protein derivative (PPD)‐specific Th1 cell lines and Dermatophagoides farinae (Der f)‐specific Th2 cell lines were established from nine patients with house dust mite‐allergic asthma. The effects of suplatast tosilate on mRNA expression of TARC and protein production of TARC from antigen‐specific Th1 or Th2 cell lines were investigated after stimulation with relevant antigens or phytohemagglutinin (PHA). In addition, the effects of IL‐4, IL‐10, and IFN‐γ on TARC production by Der f‐specific Th2 cell lines in the presence or absence of suplatast tosilate were studied.

Results Although PPD‐specific Th1 cell lines did not produce TARC after stimulation with PPD antigen or PHA, stimulation of Der f‐specific Th2 cell lines with Der f antigen or PHA increased production of TARC. Suplatast tosilate significantly and dose‐dependently inhibited production of TARC by Der f‐specific Th2 cell lines stimulated with either Der f antigen (76.5% inhibition at 100 µg/mL, P < 0.01) or PHA (81.9% inhibition at 100 µg/mL, P < 0.01). TARC production by Der f‐specific Th2 cell lines was significantly increased only by activation with IL‐4 but not with IL‐10 or IFN‐γ; this increase in TARC production was significantly inhibited by suplatast tosilate (97.5% inhibition at 100 µg/mL, P < 0.01).

Conclusion Suplatast tosilate inhibits TARC production by human Th2 cells. Therefore, this agent inhibits both Th2 cytokine and Th2 chemokine and may be a useful anti‐allergic agent.
Increased aeroallergen‐specific interleukin‐4‐producing T cells in asthmatic adults
Authors: Pala P. , Message S. D. , Johnston S. L. , Openshaw P. J. M.
Published: March 2003

Background Asthma, atopy and some forms of respiratory syncytial virus (RSV) disease are thought to be caused by T cells making IL‐4 (Th2 cells). However, not all patients with similar patterns of clinical disease have the same underlying pathogenesis and the ability to detect immunopathogenic T cells by examination of the peripheral blood remains in doubt. With the prospect of specific immunotherapy for diseases caused by T cell subsets, it is important to determine whether peripheral blood mononuclear cell (PBMC) reactivity can be used to establish the presence of immunopathogenic responses and therefore to predict therapeutic effects.

Objective To detect IL‐4 and IFN‐γ production as markers of Th1 and Th2 responses in the peripheral blood of atopic and asthmatic adults.

Methods PBMC from 22 adult asthmatics (18 of whom were atopic) and 21 non‐asthmatic volunteers (ten of whom were atopic) were stimulated with cat, birch and house dust mite allergens, human rhinovirus, RSV and recombinant chimaeric F/G protein from RSV in vitro. ELISPOT assays were used to enumerate cells producing IL‐4 and IFN‐γ.

Results Asthmatics had a sixfold increase in frequencies of IL‐4‐producing cells to cat and birch allergen (median values: 37 vs. 7 per million PBMC, P < 0.01 and 20 vs. 3 per million PBMC, P < 0.04, respectively) compared to non‐asthmatics. By contrast, non‐asthmatic atopics showed no specific increase in antigen‐specific IL‐4 responses and there was no evident correlation between skin prick test reactivity and ELISPOT results. Atopics had significantly more IFN‐ γ‐producing cells specific for FG than nonatopics. while IFN‐γ and IL‐4 responses to other antigens were not significantly different.

Conclusion Enhanced IL‐4 responses to non‐viral aeroallergens are seen in adults with asthma, while enhanced IFN‐γ responses to viral antigen FG were seen in atopics. In practical terms, ELISPOT assays for specific cytokines may provide a method that could be used to monitor antigen‐specific T cell responses in peripheral blood.
Comparison of four allergen‐sampling methods in conventional and allergy prevention classrooms
Authors: Karlsson A.‐S. , Renström A. , Hedrén M. , Larsson K.
Published: March 2003

Background Several allergen‐sampling methods are used to assess level of personal or indirect exposure to cat in homes, schools and other public buildings and working environments.

Objective To compare four different allergen‐sampling methods (dust collectors, Petri dishes, person‐carried pumps and intranasal samplers) by simultaneous sampling in classrooms and to compare the cat allergen levels between conventional classrooms and allergy prevention classrooms. Another aim was to relate the results to self‐reported frequency of allergy and asthma symptoms among the children, to their perception of the school environment.

Methods Among all compulsory schools (n = 257) in the Stockholm suburban area, 35 classrooms (five with implemented allergy prevention measures, seven with additional cleaning and 23 with normal cleaning routines) were chosen for allergen‐sampling. Dust collectors (two models), Petri dishes, person‐carried pumps and intranasal samplers were used simultaneously. All children (n = 829) received a self‐administered questionnaire which included questions about home and school environment, allergic disease, asthma symptoms and pet contact.

Results The correlation between sampling methods was generally poor. Furthermore, there was no significant difference in allergen levels between allergy prevention and allergen avoidance classes compared to conventional classes. Median levels were generally, but not significantly, lower in classes with few cat owners, compared to classes with many cat owners. Children in allergy prevention classes were more satisfied with the indoor air quality and cleaning than children attending classes with fewer or no allergy prevention measures (P < 0.0001). Nine per cent of all children reported allergic symptoms while at school.

Conclusion The lack of correlation between sampling methods used simultaneously demonstrates the difficulty in assessing allergen levels in schools and similar environments. The implemented intervention measures (allergy prevention/allergen avoidance) did not influence cat allergen levels at school.
Skin test with a timothy grass (Phleum pratense) pollen extract vs. IgE to a timothy extract vs. IgE to rPhl p 1, rPhl p 2, nPhl p 4, rPhl p 5, rPhl p 6, rPhl p 7, rPhl p 11, and rPhl p 12: epidemiological and diagnostic data
Authors: Mari A.
Published: March 2003

Background The diagnostic approach to grass pollen allergy is now possible by detecting specific IgE to its allergenic components.

Objective To compare the IgE reactivity to a timothy grass pollen extract with the IgE reactivity to eight allergenic components from the same source (Phl p 1, 2, 4, 5, 6, 7, 11, 12). Both were compared with the skin test reactivity to a timothy grass extract.

Methods A population survey was carried out by means of the skin test to identify grass‐allergic subjects, and to characterize them in terms of demographic and allergological parameters. Seven hundred and forty‐nine sera were available for IgE detection to a timothy extract, to the recombinant Phl p 1, 2, 5, 6, 7, 11, 12, and to native Phl p 4 and bromelain. Results were stratified by means of demographic and allergy parameters.

Results Ninety‐five per cent of the sera had detectable IgE to the timothy extract. Prevalence of IgE reactivity increased from 86.8% to 93.3% as the number of combined reactive molecules rose from 2 to 8. Adjusted prevalences for each allergen were: rPhl p 1 = 83%, rPhl p 2 = 55%, nPhl p 4 = 70%, rPhl p 5 = 50%, rPhl p 6 = 44%, rPhl p 7 = 7%, rPhl p11 = 43%, rPhl p 12 = 15%. Isolated reactivity to rPhl p 1 was 6%, whereas it was negligible for the remaining molecules. IgE reactivity prevalence and mean values differed when patients were stratified on the basis of their associated pollen reactivity and their skin test reactivity grade. No differences were found when age, symptom type and duration were considered. Up to eight‐fold higher IgE concentrations were found when the sum of IgE to molecules was compared with IgE to the extract. Testing for the IgE reactivity to the glycan of the native Phl p 4 allergen showed a possible interference with prevalence and value estimation. Higher prevalence values were found in previously immunotherapy‐treated patients.

Conclusions The use of a complete panel of grass allergenic molecules can mimic the current use of allergenic extracts, but new relevant information, such as individual pattern of reactivity, adjusted prevalence, correct specific IgE concentration, can be achieved only by means of discrete allergenic molecules.
HIV-induced IL-6/IL-10 dysregulation of CD4 cells is associated with defective B cell help and autoantibody formation against CD4 cells
Authors: Weimer R , Zipperle S , Daniel V , Zimmermann R , Schimpf K , Opelz G
Published: March 2003

To analyse CD4 cell cytokine secretion and helper/suppressor function at a clonal level we established 446 CD4+ T cell clones (TCC) in four healthy controls, three HIV− haemophilia patients, four CDC II,III and four CDC IV patients. Spontaneous TCC secretion of Th1 cytokines (IL-2, interferon-gamma (IFN-γ)) and Th2 cytokines (IL-4, IL-6, IL-10) was determined by ELISA. TCC helper and suppressor functions were tested in a pokeweed mitogen (PWM)-stimulated allogeneic co-culture system using a reverse haemolytic plaque assay for assessment of B cell responses. There was a significant association of TCC surface marker expression (Leu-8, CD45RA) with TCC IL-6 secretion in healthy controls (P < 0.01), HIV− patients (P 0.001) and CDC II,III patients (P 0.01) but not in CDC IV patients. Likewise, TCC expression of Leu-8 and CD45RA was significantly associated with TCC suppressor function in healthy controls (P 0.0005) but not in HIV-infected patients. A reduced TCC helper frequency (10% of TCC) and an enhanced TCC suppressor frequency (> 80% of TCC) were detected only in those HIV-infected patients who showed an excessively increased TCC IL-6 secretion (> 70% of TCC) together with a significantly diminished TCC IL-10 secretion (10% of TCC). CD4 cell autoantibodies also were found only in patients with this type of cytokine dysregulation. These data indicate that CD4 cell surface markers lose their functional relevance in HIV-infected patients. HIV-induced IL-6/IL-10 dysregulation of CD4+ T cells, i.e. the up-regulation of spontaneous IL-6 and down-regulation of spontaneous IL-10 secretion, appears to be involved in inducing CD4 helper defects and may promote autoantibody formation against CD4 cells.
Complement-regulatory protein expression and activation of complement cascade on erythrocytes from patients with rheumatoid arthritis (RA)
Authors: Arora M , Kumar A , Das S N , Srivastava L M
Published: March 2003

It has been previously reported that the expression of the complement receptor, CR1, on erythrocytes is reduced in patients with RA and that the reduced expression of CR1 is related to disease activity. In this study we investigate the role of other regulatory proteins, i.e. decay-accelerating factor (DAF) and CD59 (membrane inhibitor of reactive lysis), in the pathogenesis of RA by checking the expression of DAF and CD59 on erythrocytes of RA patients to establish whether reduced expression of DAF and CD59 on erythrocytes could be related to increased ability of erythrocytes to activate complement in RA. Flow cytometry was used to measure the expression of DAF and CD59 on erythrocytes from RA patients as well as the deposition of C3 fragments occurring in vivo or after in vitro complement activation. Significantly reduced expression of DAF and CD59 was observed on erythrocytes of RA patients. A significant inverse relationship was observed between DAF expression and in vitro complement activation, whereas no significant relationship between CD59 and complement activation was observed. Finally, we demonstrated an inverse relationship between CH50 activity and DAF expression. Thus, determination of DAF on erythrocytes can emerge as an additional tool in the assessment of extent of complement activation in RA.
Meningococcal disease and polymorphism of FcγRIIa (CD32) in late complement component-deficient individuals
Authors: Platonov A E , Kuijper E J , Vershinina I V , Shipulin G A , Westerdaal N , Fijen C A P , Van De Winkel J G J
Published: March 2003

Late complement component-deficient (LCCD) individuals lack plasma bactericidal activity and are highly susceptible to meningococcal disease. Phagocytosis plays a significant role in immune defence against meningococci and involves FcγRIIa (CD32) on leucocytes. Two allotypic forms are currently recognized: FcγRIIa-R131 and RIIa-H131. Neutrophils with the IIa-H/H131 allotype are more effective in phagocytosis than IIa-R/R131. We studied the distributions of IIa-R131 and IIa-H131 allotypes among 29 Russian LCCD patients who had suffered from recurrent episodes of meningococcal disease. The distribution of IIa-R/R131 to heterozygous IIa-R/H131 to homozygous IIa-H/H131 genotypes was 0.14:0.29:0.57 for LCCD patients who developed the first episode of disease before 10 years of age. The distribution was 0.21:0.64:0.14 for patients who experienced meningococcal disease above the age of 10 years (χ2 = 6, P < 0.05, odds ratio for IIa H/H131 versus R/R131 = 8). Meningococcal disease had a ‘grave’ course in 14 of 31 disease episodes in patients with IIa-R/R131 and IIa-R/H131 allotypes, in contrast to 1 of 18 episodes in patients with IIa-H/H131 allotype (χ2 = 7, P < 0.01, odds ratio = 14). We conclude that IIa-H/H131 individuals appear to have a higher acquired antibody-mediated phagocytosis-dependent resistance to meningococcal disease above the age of 10 years. Additionally, effective CD32-mediated phagocytosis may restrict the severity of meningococcal disease in LCCD patients with IIa-H/H131 phenotype.
Pentoxifylline decreasesin vivoandin vitrotumour necrosis factor-alpha (TNF-α) production in lepromatous leprosy patients with erythema nodosum leprosum (ENL)
Authors: Sampaio E P , Moraes M O , Nery J A C , Santos A R , Matos H C , Sarno E N
Published: March 2003

Increasing evidence has implicated TNF-α as a pivotal molecule involved in the systemic inflammatory manifestations of ENL, an acute inflammatory complication that may occur in the chronic course of leprosy. In the present study, the mechanism of action of pentoxifylline (PTX) as an alternative therapy for management of leprosy reactions has been evaluated. The effect of PTX on TNF-α production was examined in leprosy patients at the protein level and at the transcriptional level as well. Treatment of ENL patients with PTX (1200 mg daily) ameliorated the systemic symptoms and favoured the evolution of reactional leprosy lesions. Serum TNF-α was assayed before and during treatment with PTX in 15 patients. The increased TNF-α levels seen in the circulation during the reaction were dramatically reduced within 3–7 days of therapy. No significant effect on serum IL-6 was noted. In vitro TNF-α production was assayed upon culture stimulation with Mycobacterium leprae. A reduction of inducible TNF-α in peripheral blood mononuclear cells (PBMC) was seen after 1–2 weeks of in vivo administration of PTX. Furthermore, no effect of the drug on IL-10 secretion was detected in these cultures. A kinetic analysis of the expression of TNF-α and IL-6 mRNA at the site of leprosy lesion was performed in six reactional patients by semiquantitative reverse transcriptase-polymerase chain reaction (RT-PCR). The amount of TNF-α mRNA was increased in the tissue during ENL compared with before the reaction, and decreased thereafter following treatment for reaction (either PTX or thalidomide). These data suggest that PTX inhibits TNF-α production in ENL patients both in vivo and in vitro, and it may be useful in the treatment of leprosy patients undergoing ENL.
Essential pathogenic role for endogenous interferon-gamma (IFN-γ) during disease onset phase of murine experimental autoimmune orchitis. I.In vivostudies
Authors: Itoh M , Yano A , Xie Q , Iwahashi K , Takeuchi Y , Meroni P L , Nicoletti F
Published: March 2003

We previously found that immunization of CH3/He male mice with syngeneic testicular germ cells (TGC) without the aid of any adjuvants was sufficient to induce DTH to TGC and experimental autoimmune orchitis (EAO). To evaluate the role of endogenous IFN-γ in this model, C3H/He mice immunized subcutaneously with TGC on days 0 and 14 received a single injection of anti-murine IFN-γ MoAb on day 15, 20 or 25. On day 45, DTH to TGC was tested, testis specimens were collected for histological examination, and blood samples collected for IFN-γ measurement. The results showed that whilst MoAb treatment on day 15 or 25 did not influence DTH responses, EAO development, and appearance of IFN-γ in the circulation, treatment on day 20 significantly suppressed all of them. Thus, a single injection with anti-IFN-γ MoAb may successfully down-regulate testicular autoimmunity, provided that the treatment is given at an optimal time point during disease development.
Serum levels of soluble CD30 are increased in ulcerative colitis (UC) but not in Crohn's disease (CD)
Authors: Giacomelli R , Passacantando A , Parzanese I , Vernia P , Klidara N , Cucinelli F , Lattanzio R , Santori E , Cipriani P , Caprilli R , Tonietti G
Published: March 2003

Imbalance in Th1 and Th2 subsets and their derived cytokines seems to be involved in the immune abnormalities underlying UC and CD. CD30 is a member of the tumour necrosis factor/nerve growth receptor superfamily expressed on T cells producing Th2 cytokines and released as a soluble form. In this study high levels of soluble CD30 were found in sera of UC patients independently of disease activity. Furthermore, increased titres of soluble CD30 molecule were shown, in the same patients, by mitogen-stimulated cultures of peripheral blood mononuclear cells. Our data seem to indicate that an activation of Th2 immune response is involved in the pathogenesis of UC, but not of CD. Furthermore, this finding indicates that serum soluble CD30 measurement may be helpful for differentiating these two forms of inflammatory bowel disease.
Expression of low-density lipoprotein receptors in peripheral blood and tonsil B lymphocytes
Authors: De Sanctis J B , Blanca I , Rivera H , Bianco N E
Published: March 2003

B lymphocytes, purified from peripheral leucocytes from young normolipaemic humans, expressed and internalized low-density lipoprotein receptors (LDLR). The expression was assessed by a monoclonal anti-LDLR. The internalization of LDL was assessed by LDL labelled with 125I (125I-LDL) and 1,1′-dioctadecyl-3,3,3′,3′ tetramethyl-indocarboxycyanine perchlorate (LDL-DiI). The expression of LDLR, assessed by anti-LDLR, was: 38 ± 8% (n = 5) for fresh purified cells, 60 ± 10% (n = 12) for non-stimulated cells, 79 ± 5% (n = 10) for IL-2 (100 U/ml)-stimulated cells and 95 ± 5% (n = 8) for pokeweed mitogen (PWM) (1:200 dilution)-stimulated cells. The optimal concentrations of agonist were 100 U/ml of IL-2, and 1:200 dilution of PWM. IL-2 and PWM increased the internalization of LDL-DiI by 1.5-fold. The internalization of LDL-DiI was maximal at 60 μg of protein/ml (48 ± 8%). Scatchard analysis revealed a Kd of 3.2 ± 0.22 × 10−8 m and 2180 ± 190 binding sites in non-stimulated cells, a Kd of 7.73 ± 0.36 × 10−9 m and 12 500 ± 430 binding sites for IL-2 (100 U/ml)-stimulated cells, and a Kd of 7.2 ± 0.43 × 10−9 m and 13 250 ± 450 binding sites for PWM (1:200 dilution)-stimulated cells. Lineweaver–Burk analysis of LDL binding (LDL-DiI) revealed that the apparent Kd for non-stimulated cells was 1.3 ± 0.11 × 10−8 m, and 9.2 ± 0.2 × 10−9 m and 7.5 ± 0.25 × 10−9 m for IL-2- and PWM-stimulated cells, respectively. B lymphocytes from tonsils also showed a high expression of LDLR assessed with anti-LDLR (70 ± 6%). The high expression of LDLR and the avid internalization of LDL suggest that LDL may be important for B cell physiological responses.
P-selectin requirement for neutrophil accumulation and injury in the direct passive Arthus reaction
Authors: Santos L L , Huang X R , Berndt M C , Holdsworth S R
Published: March 2003

The aim of this study was to investigate the role of P-selectin in the accumulation of neutrophils in the direct passive Arthus reaction in rat skin. Direct passive Arthus dermal reaction was induced in male Sprague-Dawley (SD) rats by a single i.v. injection of rat anti-sheep globulin (SG) 1 h before i.d. injection of SG antigen. Anti-P-selectin or irrelevant control antibody was given 1 h before rat anti-SG injection. Complement depletion was also performed in a separate group by pretreatment with cobra venom factor (CVF). In all groups dermal swelling was assessed 4 h after antigen challenge. Four hours after antigen challenge, rats treated with control antibody developed skin swelling (2·29 ± 0·47 mm), prominent complement deposition and neutrophil accumulation. This response was associated with local up-regulation of endothelial P-selectin. Pre-treatment with anti-P-selectin antibody 1 h before passive Arthus induction prevented skin swelling (0·29 ± 0·06 mm, P < 0·05, cf with control antibody treatment), neutrophil accumulation and up-regulation of endothelial P-selectin despite complement deposition. CVF treatment prevented complement deposition, neutrophil accumulation and skin swelling (0·13 ± 0·07 mm, P < 0·05, cf with saline treatment). However, endothelial P-selectin expression was still present. Inhibition of skin swelling and neutrophil accumulation in direct passive Arthus by functional inhibition of P-selectin suggest a pivotal role for this adhesion molecule in this inflammatory process. These results also suggest that multiple steps are involved in the evolution of direct passive Arthus, including both P-selectin expression and complement activation. However, while complement activation is essential for neutrophil accumulation and expression of dermal injury, P-selectin up-regulation initiated by antibody/antigen deposition occurs independently of complement activation.
Single-cell analysis of lymphokine imbalance in asymptomatic HIV-1 infection: evidence for a major alteration within the CD8+ T cell subset
Authors: Sousa A E , Victorino R M M
Published: March 2003

In this study we investigated at single-cell level by flow cytometry the potential of T cell cytokine production in asymptomatic HIV-1-infected subjects with > 200 CD4 counts and possible correlation with T helper cell depletion and viral load. Mitogen-stimulated peripheral blood mononuclear cells from 32 HIV-1+ patients and 16 healthy subjects were intracytoplasmically stained for IL-2, interferon-gamma (IFN-γ), IL-4 or IL-10, and the frequency of cytokine-producing cells was assessed in total T cells, CD4, CD8 and CD45RO subsets as well as in CD69+ CD3+ gated lymphocytes. HIV-1+ patients, irrespective of their degree of CD4 depletion, exhibited a major increase in IFN-γ+ CD8 T cells, largely due to CD28− cells, as well as a decrease in the capacity of CD8 T cells to produce IL-2. Patients with > 500 CD4 counts showed a diminished frequency of IL-4 expression in CD4 T cells and a negative correlation was found between this parameter and the ex vivo CD4 counts in the 32 patients. Analysis of patients stratified according to viral load revealed a significantly higher proportion of IL-2-producing CD4 cells in the group with < 5000 RNA copies/ml. In short, using single-cell analysis and an antigen-presenting cell-independent stimulus, we have not been able to find any significant cytokine imbalances in the CD4 subset, suggesting that the well described T helper defects are not due to intrinsic alterations in the potential of CD4 T cells to produce cytokines. On the other hand, the major disturbances in the CD8 T lymphocytes agree with the marked activation and possible replicative senescence of CD8 T cells and emphasize the role of this subset in HIV immunopathogenesis.