bluuebunny/crossref_metadata_2025 huggingface dataset:
Cross‐reactivity between Ficus benjamina latex and fig fruit in patients with clinical fig allergy
Authors: Focke M. , Hemmer W. , Wöhrl S. , Götz M. , Jarisch R.
Published: July 2003

Background  Anaphylactic reactions to fig fruits (Ficus carica) have been reported from subjects sensitized to Ficus benjamina (FB) latex allergens. Figs may also be involved in the latex–fruit syndrome.

Objective  To study the immunologic relationship between fig fruit, Ficus benjamina, natural rubber latex (Hevea brasiliensis), and other tropical fruits.

Methods  RAST inhibition and Western blotting with FB and fruit extracts was performed in five patients with oral allergy syndrome (OAS) or anaphylaxis after the ingestion of figs and one patient with symptoms from exposure to FB trees. Co‐sensitization to rubber latex and tropical fruits (kiwi, banana, avocado, papaya, pineapple, mulberry) was studied by skin testing.

Results  RAST to FB was inhibited >95% by FB extracts and 16–65% (mean 49%) by extracts from fresh fig. RAST to fig fruit was inhibited >95% by FB and fresh fig, 63–97% (mean 86%) by dried fig, and 0–84% (mean 35.5%) by kiwi fruit. FB and fig extracts lost most of their allergenicity when denatured by heat (95 °C) or reduced by dithiothreitol. Western blotting after non‐reducing gel electrophoresis revealed IgE binding to proteins of 22 and 28–34 kDa in FB latex; however, no corresponding allergens could be detected in fig extracts. Positive skin tests were obtained most often with kiwi fruit, papaya, and avocado. Sensitization to rubber latex could not be demonstrated in any of the patients. RAST to papain was positive in three of five patients.

Conclusions  Allergic reactions to fresh or dried figs can present as a consequence of primary sensitization to airborne FB allergens independent of sensitization to rubber latex allergens. Kiwi fruit, papaya, and avocado as well as pineapple and banana may be other fruits associated with sensitization to Ficus allergens.
Butterbur, a herbal remedy, attenuates adenosine monophosphate induced nasal responsiveness in seasonal allergic rhinitis
Authors: Lee D. K. C. , Carstairs I. J. , Haggart K. , Jackson C. M. , Currie G. P. , Lipworth B. J.
Published: July 2003

Background
Butterbur (BB) or Petasites hybridus, a herbal remedy, exhibits in vitro inhibition of cysteinyl leukotriene biosynthesis. However, no placebo‐controlled studies have been performed to evaluate the effectiveness of BB on objective outcomes such as nasal provocation testing in seasonal allergic rhinitis (SAR).

Methods
Twenty patients with grass‐pollen‐sensitized SAR were randomized in a double‐blind, cross‐over manner to receive for 2 weeks either BB 50 mg twice daily or placebo (PL) twice daily during the grass pollen season. Nasal adenosine monophosphate (AMP) challenge (the primary outcome) was administered as a single 400 mg/mL dose after each randomized treatment.

Results
Spontaneous recovery following AMP challenge (area under the response time profile curve as %.min±SEM) was significantly attenuated (P=0.028) with BB (584±289) compared to PL (1438±240); mean difference: 854 (95% CI 95–1614), and the maximum % peak nasal inspiratory flow reduction from baseline following AMP challenge was significantly blunted (P=0.036) with BB (30±4) compared to PL (43±5); mean difference: 13 (95% CI 1–25).

Conclusions
BB exhibited protection against AMP‐induced nasal responsiveness during the grass pollen season in sensitized patients. This is turn may explain its potential clinical efficacy in patients with SAR.
Quantitative measurement of airborne allergens from dust mites, dogs, and cats using an ion‐charging device
Authors: Custis N. J. , Woodfolk J. A. , Vaughan J. W. , Platts‐Mills T. A. E.
Published: July 2003

BackgroundIncreasing evidence suggests that children raised with an animal(s) in the house have a decreased risk of becoming sensitized. However, it is not clear whether this phenomenon is related to airborne exposure.

ObjectiveTo estimate airborne exposure to animal dander and dust mite allergens using a device that can sample large volumes of air silently.

MethodsThe device, which uses an ion‐charging technique to move air and to collect particles, was run at 1.7 m3/min for 24 h in 44 homes with and without animals. The allergen collected was measured by ELISA for Fel d 1, Can f 1, Der p 1, and Der f 1.

ResultsAirborne Fel d 1 was present in all homes with a cat (n=27). The quantities measured, i.e. 0.5–20 μg in 24 h, represent 0.01–0.3 μg Fel d 1 inhaled/day at normal breathing rates (20 L/h). Values for houseswithouta cat were 0.01–0.05 μg inhaled/day. Airborne Fel d 1 correlated significantly with floor Fel d 1 (r=0.58,P<0.001). Results for Can f 1 were similar in houses with a dog, but this allergen was only detected airborne in two houses without a dog. Neither Der p 1 nor Der f 1 (i.e. <0.01 μg) was detected, which represents  1 ng inhaled/day during normal domestic activity. During disturbance airborne mite was detected with both the ion‐charging device and a filter run in parallel. For cat and mite allergens there was a close correlation between the two techniques (r=0.84,P<0.001).

ConclusionExposure to cat or dog allergen airborne in homes with an animal can be up to 100 times higher than exposure to mite allergen. The results are in keeping with a model where immunological tolerance to animal dander allergens results from high exposure.
The major Platanus acerifolia pollen allergen Pla a 1 has sequence homology to invertase inhibitors
Authors: Asturias J. A. , Ibarrola I. , Eraso E. , Arilla M. C. , Martínez A.
Published: July 2003

Background
Sycamores or plane trees are an important source of airborne allergens in many cities of the United States and Western Europe. Pla a 1 has been described as a major allergen from Platanus acerifolia (London plane tree).

Objective
To clone and characterize the cDNA for Pla a 1 and to express the recombinant protein.

Methods
Pla a 1 was isolated by cationic exchange, gel filtration, and reverse‐phase chromato‐graphies. Pla a 1 cDNA was cloned by reverse transcription followed by polymerase chain reaction, using amino acid sequences from tryptic peptides of the allergen. The Pla a 1 encoding sequence has been subcloned into the pKN172 expression vector and expressed in Escherichia coli as a non‐fusion protein. Purified recombinant protein has been tested for its IgE‐binding capacity in immunoblot, immunoblot inhibition, and ELISA.

Results
Pla a 1 reacted with serum IgE from 35 of the 42 (83.3%) Platanus‐allergic patients studied and represented 60% of the total IgE‐binding capacity of the P. acerifolia pollen extract. The allergen displayed 43% sequence identity to a grape invertase inhibitor and showed a predicted secondary structure characteristic of all‐alpha proteins. Serological analysis revealed that both natural and recombinant forms of Pla a 1 displayed similar IgE‐binding capacity.

Conclusions
Pla a 1 belongs to a new class of allergens related to proteinaceous invertase inhibitors. Recombinant Pla a 1 binds IgE in vitro like its natural counterpart and, therefore, it can be useful for specific diagnosis and structural studies.
Fluticasone propionate and mometasone furoate have equivalent transcriptional potencies
Authors: Roumestan C. , Henriquet C. , Bousquet J. , Mathieu M.
Published: July 2003

Background
Glucocorticoids exert their anti‐inflammatory effects mainly through transrepression of the transcription factors activator protein‐1 (AP‐1) and nuclear factor‐kappa B (NF‐κB). Certain adverse effects of glucocorticoids are mediated through gene transactivation. Fluticasone propionate (FP) and mometasone furoate (MF) are the most recently developed topical glucocorticoids for the treatment of airway disorders. Their relative capacities to repress AP‐1 and NF‐κB activities are not known and comparison of their transactivation potencies has given unclear results.

Objective
To determine the relative transactivation and transrepression potencies of FP and MF.

Methods
Transactivation assays were performed in HeLa cells carrying a glucocorticoid‐inducible luciferase gene. To measure transrepressive potencies of FP and MF, A549 lung epithelial cells were transiently transfected with an AP‐1‐ or NF‐κB‐dependent luciferase gene. Using an immunoassay, we also evaluated the ability of MF and FP to inhibit the production of Regulated upon Activation, Normal T‐cell Expressed and Secreted (RANTES), a pro‐inflammatory cytokine, whose gene is controlled by AP‐1 and NF‐κB. Areas under the dose–response curve were calculated to determine relative potencies.

Results
FP and MF are equipotent for transactivation. Both molecules show globally the same potency to inhibit AP‐1 and NF‐κB activities and RANTES production. MF and FP have very significant transcriptional effects at 2×10−10 M, which is the peak concentration reached in the plasma after inhalation of high dosages. Indeed, they produce a 17‐fold induction of luciferase in the transactivation assay, and inhibit AP‐1 activity, NF‐κB activity and RANTES release by approximately 40%.

Conclusion
FP and MF have the same ability to trigger gene activation and also the same potency to inhibit AP‐1 and NF‐κB activities. Their strong transcriptional effects at 2×10−10 M suggest that these compounds act not only topically but also systemically, with the risk of provoking concomitant adverse effects at high dosages.
Reducing agents inhibit the contractile response of isolated guinea‐pig main bronchi
Authors: Casoni G. L. , Chitano P. , Pinamonti S. , Chicca M. , Ciaccia A. , Fabbri L. , Papi A.
Published: July 2003

Background
Oxidants are involved in many respiratory disorders, including asthma and chronic obstructive pulmonary diseases. Reduced glutathione (GSH), one of the most important antioxidant compounds against oxidant free radicals, is particularly abundant in the respiratory epithelial lining fluid, where its concentration is increased in inflammatory disorders.

Objective
We hypothesized that reducing agents may have a direct effect on airway smooth muscle. Therefore, we studied the effects of GSH on airway smooth muscle contractility in guinea‐pig main bronchi. In parallel, we evaluated superoxide anion generation associated with in vitro bronchial smooth muscle contraction.

Methods
Guinea‐pig main bronchi were mounted in organ baths filled with Krebs–Henseleit solution. Concentration–response curves to acetylcholine (Ach) (10−9–10−3 M), carbachol (10−9–10−4 M), or histamine (10−9–10−3 M) were performed in the presence or absence of either reduced or oxidized glutathione (GSSG) (10−5–10−3 M). We also evaluated the effects of GSH and GSSG on allergen‐induced contraction in main bronchi obtained from ovalbumin‐sensitized guinea‐pig. Superoxide dismutase (SOD)‐inhibited cytochrome c reduction kinetics was performed to evaluate superoxide anion (O2−) production during Ach‐induced contraction.

Results
Reduced but not oxidized glutathione significantly decreased smooth muscle contraction induced by Ach, carbachol, and histamine. Similarly, only the reduced form of glutathione attenuated the bronchoconstriction induced by allergen exposure in bronchi from sensitized animals. Finally, SOD‐inhibited cytochrome c reduction kinetics demonstrated increased O2− production following bronchial smooth muscle contraction. This production was not affected by epithelium removal.

Conclusion
Our findings show that GSH decreases bronchial smooth muscle contraction to different stimuli and that oxidant free radicals are produced during bronchial smooth muscle contraction. We suggest that oxidants are involved in the mechanisms of bronchoconstriction and that reducing agents could be a possible therapeutic option for airway obstruction sustained by bronchospasm.
An adult model of exclusive viral wheeze: inflammation in the upper and lower respiratory tracts
Authors: Mckean M. C. , Hewitt C. , Lambert P. C. , Myint St. , Silverman M.
Published: July 2003

Background
We have previously reported an experimental infection of young adults with a history of episodic and exclusive viral wheeze (EVW) using human coronavirus, in which 16 of 24 with EVW (15 atopic) and 11 of 19 healthy controls (seven atopic) developed a symptomatic cold with evidence of infection, but only those with EVW developed lower respiratory tract symptoms and increased airway responsiveness.

Objective
The aim of this study was to compare the EVW and control groups from this study for inflammatory changes occurring in the upper and lower respiratory tracts during the experimental infection, in particular, to determine whether eosinophil‐driven inflammation was associated with EVW.

Methods
Nasal lavage and induced sputum were collected prior to inoculation (day 0) and 2, 4 and 17 days later. Differential cell counts were performed and supernatant was assayed for IL‐8, IL‐5, IFN‐γ, and eosinophilic cationic protein (ECP).

Results
There was no difference between the two groups in any measurement at baseline. In both groups, during colds the volume of nasal secretion increased as did leucocyte counts in both upper and lower respiratory tracts. A modest increase in nasal neutrophil count was seen in both EVW and control groups with symptomatic colds on day 2 (median (quartile) difference from baseline 5.4 (0.0, 11.0) and 1.8 (−1.1, 2.2)×104/mL of secretions, respectively). The change in nasal neutrophil counts in all subjects correlated with nasal symptom scores. A significant relative increase in sputum differential neutrophil count was seen on day 4 in the EVW group with a cold but not in controls (mean difference (95% confidence interval) 20.4 (9.6, 31.1)% and 3.1 (−8.2, 14.5)%, respectively, P<0.01); however, this increase did not correlate with lower respiratory tract symptom scores. IL‐8 increased in both the upper and lower respiratory tracts in both EVW and control subjects with colds, the largest change being seen on day 4 in the sputum of those with EVW (mean difference from baseline (95% confidence interval) 2.5 (0.55–4.46) ng/mL). Only modest changes were seen in IFN‐γ and no changes were seen in IL‐5 or ECP. None of the results was influenced by the atopic status of the subjects in either group.

Conclusions
EVW wheeze is characterized by neutrophilic inflammation in both the upper and lower respiratory tracts without eosinophilia (even in atopic subjects). IL‐8 is likely to be an important chemokine in this process. Symptoms and airway responsiveness were correlated with change in neutrophils.
Mechanisms in cutaneous drug hypersensitivity reactions
Authors: Friedmann P. S. , Lee M.‐S. , Friedmann A. C. , Barnetson R. St C.
Published: July 2003

Up to 3% of all hospital admissions are due to adverse drug reactions (ADRs), and between 10% and 20% of hospital inpatients develop ADRs. Individual susceptibility to becoming ‘sensitized’ or allergic to a drug is thought to result from altered metabolic handling of the drug. Reactive intermediate compounds form haptens, bind to proteins and induce immune responses. Depending on whether the immune system generates antibodies or sensitized T cells, different clinical patterns of hypersensitivity may result. At present, both in vivo or in vitro tests to identify the culprit drug or to confirm the presence of hypersensitivity are not widely used because they are either not generally robust or not readily accessible. In vitro tests require the true immunogen/antigen to detect antibodies or sensitized T cells. As the metabolic basis underlying susceptibility to adverse drug reactions is elucidated, the resolution of immunological mechanisms and development of reliable tests will ensue. This will also become of great value for prediction of individuals at risk of becoming sensitized by a particular drug.
Localization and content of nerve growth factor in peripheral blood eosinophils of atopic dermatitis patients
Authors: Toyoda M. , Nakamura M. , Makino T. , Morohashi M.
Published: July 2003

Background  Atopic dermatitis (AD) is a chronic and relapsing eczematous skin disorder characterized by eosinophilia. Nerve growth factor (NGF) modulates the allergic response through interactions with immune‐inflammatory cells. Eosinophils have been reported to store NGF as a preformed mediator.

Objective  To gain further insight into the significance of eosinophils in association with NGF in the pathogenesis of AD, the localization of NGF within eosinophils and the difference of the eosinophil‐derived NGF content in the peripheral blood of normal volunteers vs. AD patients were investigated.

Methods  We examined the localization of NGF within human eosinophils using the post‐embedding immunoelectron microscopy and compared NGF content in freshly isolated eosinophil sonicates from the peripheral blood of 31 normal volunteers vs. 42 AD patients by immunoenzymatic assay. A possible correlation between the levels of NGF and major basic protein was also examined.

Results  Immunoelectron microscopic studies revealed that NGF was localized in the central core of normal eosinophil granules, where major basic protein is also present as a preformed mediator, in homogeneous granules and in intergranular ductal or vesicular structures adjacent to specific granules of eosinophils. NGF content in eosinophils was significantly increased in AD patients. Furthermore, there was a significant correlation between levels of NGF and major basic protein in eosinophils of AD patients.

Conclusions  Increased levels of NGF contained in eosinophils of the peripheral blood from AD patients, when released with other mediators such as basic proteins, could promote inflammation and local tissue damage.
Purification and characterization of mouse mast cell proteinase‐2 and the differential expression and release of mouse mast cell proteinase‐1 and ‐2 in vivo
Authors: Pemberton A. D. , Brown J. K. , Wright S. H. , Knight P. A. , McPhee M. L. , McEuen A. R. , Forse P. A. , Miller H. R.P.
Published: July 2003

Background
Gastrointestinal nematode infection is associated with mucosal mast cell (MMC) hyperplasia. In the mouse, this is accompanied by the release of substantial quantities of the chymase mouse mast cell proteinase‐1 (mMCP‐1) into the gut lumen and peripheral bloodstream. Expression of mMCP‐1 is largely restricted to intraepithelial MMC and is thought to play a role in the regulation of epithelial permeability. MMCs also express mouse mast cell proteinase‐2 (mMCP‐2), but less is known about the expression or biological function of this proteinase.

Objectives
(1) To purify and characterize mMCP‐2. (2) To compare the expression and release of mMCP‐2 and mMCP‐1 in vivo using specific antibodies.

Methods
Bone marrow‐derived mast cells (mBMMCs) were generated from mMCP‐1−/− BALB/c mice. mMCP‐2 was purified, characterized and used to generate rat and sheep polyclonal antibodies. The expression and systemic release of mMCP‐1 and ‐2 were compared in vivo by immunohistochemistry and ELISA.

Results
mMCP‐2 was successfully purified from mMCP‐1−/− mBMMC and its identity confirmed by N‐terminal amino acid sequencing. mMCP‐2 bound [3H]‐labelled DFP, indicating the presence of an active serine proteinase catalytic site, but showed little evidence of chymotryptic activity. MMC expressed comparable levels of mMCP‐1 and ‐2 in the jejunum but not in the gastric mucosa, where mMCP‐2 was more abundant. Expression of both proteinases increased substantially during primary Nippostrongylus brasiliensis infection and this was accompanied by a substantial increase in peripheral blood levels of mMCP‐1 (70 μg/mL on day 12). By contrast, mMCP‐2 was not detected in the serum of uninfected mice and only increased to approximately 25 ng/mL on day 12.

Conclusion
As in the case of mMCP‐1, mMCP‐2 expression is restricted to MMC. However, mMCP‐2 lacks chymase activity, is expressed at higher levels in gastric MMC and appears to be differentially released into the peripheral bloodstream.
Pituitary adenylate cyclase‐activating polypeptide, effects in the human nose
Authors: Kinhult J. , Adner M. , Uddman R. , Cardell L. O.
Published: July 2003

Background  Pituitary adenylate cyclase‐activating peptide (PACAP) is a neuropeptide with strong vaso‐ and bronchodilator capacity. There is recent evidence that PACAP decreases the release of proinflammatory cytokines and we have previously shown that PACAP inhibits neutrophil chemotaxis in vitro, but little is known about the effects of PACAP in human upper and lower airways.

Objective  To investigate the effects of PACAP in the human upper respiratory tract focusing on vasodilatation/nasal airway resistance (NAR), neutrophil recruitment, plasma extravasation and endogenous production of IL‐1‐related mediators.

Methods  Surgical specimens from five patients (aged 19–55 years), obtained in conjunction with nasal surgery, were used for immunohistochemical localization of PACAP in the nasal mucosa. In seven, healthy, non‐allergic, non‐smoking subjects (aged 19–45 years), NAR was measured with rhinomanometry. Nasal lavage was performed, before and after intranasal application of PACAP (200 μL of a 1 μm PACAP solution in each nasal cavity), with and without the addition of histamine. Cells, albumin and IL‐1‐related mediators were analysed in nasal lavage. In addition, the effects on pulse, blood pressure, ECG and pulmonary function were evaluated.

Results  In the nasal mucosa, PACAP‐like immunoreactive nerve fibres were seen close to blood vessels and seromucous glands. Application of PACAP in the nasal cavity increased NAR and augmented the increase in NAR induced by histamine. In addition, PACAP inhibited histamine‐induced recruitment of neutrophils, increased plasma leakage and reduced the level of IL‐1RA (an endogenously produced IL‐1 receptor antagonist) in nasal lavage. Cardiovascular and pulmonary parameters were not affected.

Conclusion  These results imply that PACAP is an important endogenous mediator in human upper airways, with a potential role as a regulator of vascular smooth muscle, secretion, plasma extravasation, neutrophil recruitment and cytokine activity.
Immunoglobulin E antibody reactivity to the major shrimp allergen, tropomyosin, in unexposed Orthodox Jews
Authors: Fernandes J. , Reshef A. , Patton L. , Ayuso R. , Reese G. , Lehrer S. B.
Published: July 2003

Background
Assessment of allergic (IgE antibody‐mediated) reactions to foods may become complicated by cross‐reactivity that can occur among certain food families and between foods and seemingly unrelated allergens.

Objective  The allergenic properties of tropomyosin (muscle‐derived protein) have been recently demonstrated in invertebrates such as cockroaches, dust mites, and shrimp. In view of a possible cross‐reactivity between food allergens and related allergens from animal sources, we designed a study to assess IgE antibody reactivity to the major shrimp allergen, Pen a 1, in an unexposed population of Orthodox Jews, who observe Kosher dietary laws that prohibit eating shellfish.

Methods
Nine subjects, who reacted positively by skin tests to shrimp (Penaeus setiferous), were selected for the study. Subjects (two females, seven males) ranged in age from 14 to 32 years (mean 20.4). All subjects were strictly observant of Jewish tradition and had no prior exposure to seafood (regarded as a non‐Kosher food). Serum was obtained from all the subjects and tested for IgE antibody reactivity to shrimp and dust mite.

Results
All subjects reported symptoms of perennial allergic rhinitis, five had history of asthma, atopic dermatitis, and/or sinusitis. All had positive skin prick tests to shrimp and house dust mite (HDM) (Dermatophagoides farinae, D. pteronyssinus, or both); 2/7 subjects were positive to cockroach mix (Blattella germanica and Periplaneta americana). Sera of 4/9 subjects demonstrated specific IgE antibodies by RAST to shrimp (7.0–20.0%), 3/9 to Pen a 1 (6.3–24.1%), and 3/9 to shrimp or Pen a 1 by immunoblot. IgE binding to Pen a 1 was inhibited with either mite or cockroach extracts as demonstrated by RAST and/or immunoblot inhibition analysis.

Conclusions
These studies indicate that IgE antibody reactivity to a major food allergen, shrimp, can occur in an unexposed population of individuals; some subjects allergic to HDM and/or cockroach show substantial IgE antibody reactivity to the major shrimp allergen Pen a 1 (tropomyosin). Based on inhibition with cockroach and/or dust mite extracts, this reactivity appears to be due to cross‐reacting tropomyosins.
Complement component C7 deficiency in a Spanish family
Authors: VÁZQUEZ-BERMÚDEZ M F , BARROSO S , WALTER K , ÁLVAREZ A J , ALARCÓN A , LÓPEZ-TRASCASA M , WICHMANN I , AGUILAR F , NÚÑEZ-ROLDÁN A , SÁNCHEZ B
Published: July 2003

Different genetic mutations have been described in complement component C7 deficiency, a molecular defect which is clinically associated with an increased susceptibility to neisserial recurrent infections, although some cases remain asymptomatic. In this work we report the genetic bases of C7 deficiency in one Spanish family. Exon-specific PCR and sequencing revealed a novel point mutation at nucleotide 615 (exon 6) leading to a stop codon (UGG to UGA) in the patient, his mother, and sister. This transversion causes the premature truncation of the C7 protein (W183X). Additionally, we detected a missense mutation at position 1135 (exon 9) located in the first nucleotide of the codon GGG (CGG), resulting in an amino acid change (G357R) in the patient, his father, as well as in his sister. This latter mutation had been previously described in individuals from Moroccan Sephardic Jewish ancestry. Since both heterozygous mutations were found in the patient as well as in his asymptomatic sister, we analyse other meningococcal defence mechanisms such as polymorphisms of the opsonin receptors on polymorphonuclear cells. Results showed that the patient and his sister bore identical combinations of FcγRIIA-H/R131 and FcγRIIIB-NA1/2 allotypes. Our results provide further evidence that the molecular pathogenesis of C7 deficiency as well as susceptibility to meningococcal disease are heterogeneous, since different families carry different molecular defects, although many of the C7 defects appear to be homogeneous in individuals from certain geographical areas. The missense mutation G357R would make an interesting topic of analysis with regard to meningococcal disease susceptibility in the Spanish population.
IgA class antineutrophil cytoplasmic antibodies in primary sclerosing cholangitis and autoimmune hepatitis
Authors: SCHWARZE C , TERJUNG B , LILIENWEISS P , BEUERS U , HERZOG V , SAUERBRUCH T , SPENGLER U
Published: July 2003

Antineutrophil cytoplasmic antibodies (ANCA) of IgG class have been described at high prevalence in autoimmune hepatitis (AIH) and primary sclerosing cholangitis (PSC). Data on IgA class ANCA in these diseases are limited. The aim of this study was to determine the prevalence and fluorescence patterns of IgA class ANCA in AIH and PSC and to examine a relationship between the presence of IgA ANCA and clinical characteristics in these patients. Sera from 35 patients with PSC (21 with concomitant inflammatory bowel disease), 40 patients with AIH and 10 healthy controls were studied. ANCA were detected on ethanol-fixed neutrophils using an indirect immunofluorescence technique. ANCA of the IgA class were found in 20% of sera from patients with PSC and in 50% of AIH sera. The majority of AIH patients with IgA class ANCA showed a ‘classical’ perinuclear staining pattern, whereas the ‘classical’ and ‘atypical’ perinuclear fluorescence patterns were distributed equally in PSC. In sera containing IgG and IgA class ANCA simultaneously, IgG class ANCA showed an ‘atypical’ pANCA fluorescence pattern whereas IgA class ANCA produced a ‘classical’ perinuclear staining. The presence of IgA class ANCA was not associated with disease-specific clinical characteristics. IgA class ANCA are more frequently detected in sera of patients with AIH than PSC. The diversity of fluorescence patterns points to different target antigens of IgA class ANCA with distinct subcellular localizations.
IL-16 in the airways of lung allograft recipients with acute rejection or obliterative bronchiolitis
Authors: LAAN M , LINDÉN A , RIISE G C
Published: July 2003

Acute rejection (AR) is the principal risk factor for obliterative bronchiolitis (OB), the major complication of lung transplantation. It is known that activated CD4+ T lymphocytes are involved in the development of AR and that interleukin (IL)-16 can inhibit the activity of CD4+ T lymphocytes. In this study, we evaluated whether the concentration of IL-16 in the airways is altered in AR or OB and, if so, how this IL-16 concentration relates to the number or activity of airway lymphocytes. The concentration of IL-16 protein was measured in bronchoalveolar lavage (BAL) fluid at three time-points in lung allograft recipients with either AR or OB and in matched controls using ELISA. The concentration of soluble IL-2 receptor (R) protein was measured in BAL fluid using ELISA as well, as an indicator of lymphocyte activity. The percentage of airway lymphocytes was evaluated by performing BAL differential cell counts. Lung allograft recipients with AR displayed lower IL-16 concentrations compared with matched control patients and this IL-16 concentration correlated negatively with the sIL-2R concentration, but it did not correlate with the percentage of lymphocytes in BAL fluid. In contrast, in BAL fluid from lung allograft recipients with OB, the IL-16 concentration was not altered compared with matched control patients and it did not correlate with the percentage of lymphocytes or with the sIL-2R concentration. These data are compatible with an increase in IL-16 playing a protective role against AR but not against OB and, hypothetically, this type of protective effect could be exerted via a down-regulation of the activity of T lymphocytes.
BPI–ANCA in transporter associated with antigen presentation (TAP) deficiency: possible role in susceptibility to Gram-negative bacterial infections
Authors: SCHULTZ H , SCHINKE S , WEISS J , CERUNDOLO V , GROSS W L , GADOLA S
Published: July 2003

Although HLA class I expression is diminished in patients with defects in the transporter associated with antigen presentation (TAP), recurrent Gram-negative bacterial lung infections are found from childhood onwards. As MHC class II-mediated responses are normal, other mechanisms that contribute to susceptibility to infections are presumed. The bactericidal/permeability-increasing protein (BPI) is a potent neutrophil antibiotic that neutralizes endotoxin efficiently. As antineutrophil cytoplasmic autoantibodies (ANCA) against BPI were found in the majority of cystic fibrosis patients and correlate with disease severity we examined the prevalence of BPI–ANCA and their contribution to susceptibility to bacterial infections in six TAP-deficient patients. Although only two patients showed ANCA in indirect immunofluorescence, BPI–ANCA occurred in five of six patients in ELISA. Purified IgG from BPI–ANCA-positive sera (five of six) inhibited the antimicrobial function of BPI in vitro. Epitope mapping revealed binding sites not only on the C-terminal but also on the antibiotic N-terminal portion of BPI, indicating that short linear BPI peptide fragments may be long-lived enough to become immunogens. In conclusion, BPI–ANCA are associated strongly with TAP deficiency. Inhibition of the antimicrobial BPI function by BPI–ANCA demonstrates a possible mechanism of how autoantibodies may contribute to increased susceptibility for pulmonary Gram-negative bacterial infections by diminished bacterial clearance.
Cytokine profile, HLA restriction and TCR sequence analysis of human CD4+ T clones specific for an immunodominant epitope of Mycobacterium tuberculosis 16-kDa protein
Authors: CACCAMO N , BARERA A , DI SANO C , MERAVIGLIA S , IVANYI J , HUDECZ F , BOSZE S , DIELI F , SALERNO A
Published: July 2003

The identification of immunodominant and universal mycobacterial peptides could be applied to vaccine design and have an employment as diagnostic reagents. In this paper we have investigated the fine specificity, clonal composition and HLA class II restriction of CD4+ T cell clones specific for an immunodominant epitope spanning amino acids 91–110 of the 16-kDa protein of Mycobacterium tuberculosis. Twenty-one of the tested 28 clones had a Th1 profile, while seven clones had a Th0 profile. None of the clones had a Th2 profile. While the TCR AV gene usage of the clones was heterogeneous, a dominant TCR BV2 gene family was used by 18 of the 28 clones. The CDR3 regions of BV2+ T cell clones showed variation in lengths, but a putative common motif R-L/V-G/S-Y/W-E/D was detected in 13 of the 18 clones. Moreover, the last two to three residues of the putative CDR3 loops, encoded by conserved BJ sequences, could also play a role in peptide recognition. Antibody blockade and fine restriction analysis using HLA-DR homozygous antigen-presenting cells established that 16 of 18 BV2+ peptide-specific clones were DR restricted and two clones were DR-DQ and DR-DP restricted. Additionally, five of the 18 TCRBV2+ clones recognized peptide 91–110 in association with both parental and diverse HLA-DR molecules, indicating their promiscuous recognition pattern. The ability of peptide 91–110 to bind a wide range of HLA-DR molecules, and to stimulate a Th1-type interferon (IFN)-γ response more readily, encourage the use of this peptide as a subunit vaccine component.
Non-enveloped HCV core protein as constitutive antigen of cold-precipitable immune complexes in type II mixed cryoglobulinaemia
Authors: SANSONNO D , LAULETTA G , NISI L , GATTI P , PESOLA F , PANSINI N , DAMMACCO F
Published: July 2003

Hepatitis C virus (HCV) infection has been detected in a large proportion of patients with mixed cryoglobulinaemia (MC). Circulating ‘free’ non-enveloped HCV core protein has been demonstrated in HCV-infected patients, and this suggests its possible involvement in the formation of cryoprecipitable immune complexes (ICs). Thirty-two anti-HCV, HCV RNA-positive patients with type II MC were evaluated. Non-enveloped HCV core protein, HCV RNA sequences, total IgM, rheumatoid factor (RF) activity, IgG and IgG subclasses, C3 and C4 fractions, C1q protein and C1q binding activity were assessed in both cryoprecipitates and supernatants. Non-enveloped HCV core protein was demonstrated in 30 of 32 (93·7%) type II MC patients. After separation of cold-precipitable material, the protein was removed completely from supernatant in 12 patients (40%), whereas it was enriched in the cryoprecipitates of the remaining 18. In addition, HCV RNA and IgM molecules with RF activity were concentrated selectively in the cryoprecipitates. Differential precipitation was found for both total IgG and IgG subclasses, as they were less represented in the cryoglobulins and no selective enrichment was noted. Immunological characterization of HCV core protein-containing cryoprecipitating ICs after chromatographic fractionation showed that the IgM monoclonal component had RF activity, whereas anti-HCV core reactivity was confined to the IgG fraction. C1q enrichment in addition to high avidity of ICs for C1q binding in the cryoprecipitates suggest that complement activation may occur through the C1q protein pathway. The present data demonstrate that non-enveloped HCV core protein is a constitutive component of cryoprecipitable ICs in type II MC patients.
Immune biology of macaque lymphocyte populations during mycobacterial infection
Authors: LAI X , SHEN Y , ZHOU D , SEHGAL P , SHEN L , SIMON M , QIU L , LETVIN N L , CHEN Z W
Published: July 2003

Immune responses of lymphocyte populations during early phases of mycobacterial infection and reinfection have not been well characterized in humans. A non-human primate model of Mycobacterium bovis bacille Calmette–Guerin (BCG) infection was employed to characterize optimally the immune responses of mycobacteria-specific T cells. Primary BCG infection induced biphasic immune responses, characterized by initial lymphocytopenia and subsequent expansion of CD4+, CD8+ and γδ T cell populations in the blood, lymph nodes and the pulmonary compartment. The potency of detectable T cell immune responses appears to be influenced by the timing and route of infection as well as challenge doses of BCG organisms. Systemic BCG infection introduced by intravenous challenge induced a dose-dependent expansion of circulating CD4+, CD8+ and γδ T cells whereas, in the pulmonary compartment, the systemic infection resulted in a predominant increase in numbers of γδ T cells. In contrast, pulmonary exposure to BCG through the bronchial route induced detectable expansions of CD4+, CD8+ and γδ T cell populations in only the lung but not in the blood. A rapid recall expansion of these T cell populations was seen in the macaques reinfected intravenously and bronchially with BCG. The expanded αβ and γδ T cell populations exhibited their antigen specificity for mycobacterial peptides and non-peptide phospholigands, respectively. Finally, the major expansion of T cells was associated with a resolution of active BCG infection and reinfection. The patterns and kinetics of CD4+, CD8+ and γδ T cell immune responses during BCG infection might contribute to characterizing immune protection against tuberculosis and testing new tuberculosis vaccines in primates.
Induction of mucosal tolerance with recombinant Hev b 1 and recombinant Hev b 3 for prevention of latex allergy in BALB/c mice
Authors: HUFNAGL K , WAGNER B , WINKLER B , BAIER K , HOCHREITER R , THALHAMER J , KRAFT D , SCHEINER O , BREITENEDER H , WIEDERMANN U
Published: July 2003

The prevalence of type I allergy to Hevea brasiliensis latex is particularly high among individuals with frequent exposure to latex products, such as health-care workers (HCW) and patients with spina bifida (SB). Treatment of latex allergy seems problematic as preventive measures, such as allergen avoidance, are not always possible and conventional immunotherapy with standardized latex extracts is not performed routinely. Thus, the aim of the present study was to establish a mouse model of latex allergy using two major latex allergens for HCWs and SB patients, Hev b 1 and Hev b 3, for sensitization. Prophylactic measures on the basis of mucosal tolerance induction with the recombinant allergens were tested in this model. Female BALB/c mice immunized intraperitoneally with recombinant (r)Hev b 1 or rHev b 3 displayed strong immune responses in vivo and in vitro. Intranasal treatment with rHev b 1 and rHev b 3 prior to sensitization led to reduced allergen-specific IgG1/IgE levels and significantly suppressed allergen-induced basophil degranulation. Moreover, lymphocyte proliferation and cytokine production (IL-4, IL-5, IFN-γ) in vitro were significantly suppressed after pretreatment with both allergens. Suppressive cytokines, such as interleukin (IL)-10 and transforming growth factor (TGF)-β, remained unchanged after the intranasal pretreatment, indicating mechanism of anergy rather than active immunosuppression. Taken together, these results suggest that mucosal tolerance induction with recombinant allergens could present a promising prevention strategy against latex allergy.
Evidence of expression of endotoxin receptors CD14, toll-like receptors TLR4 and TLR2 and associated molecule MD-2 and of sensitivity to endotoxin (LPS) in islet beta cells
Authors: VIVES-PI M , SOMOZA N , FERNÁNDEZ-ALVAREZ J , VARGAS F , CARO P , ALBA A , GOMIS R , LABETA M O , PUJOL-BORRELL R
Published: July 2003

CD14, a GPI-linked membrane protein, is a component of the lipopolysaccharide (LPS) receptor complex, one of the pattern-recognizing receptors (PRR) expressed by myeloid lineage cells. Here we report that CD14, the functionally linked toll-like receptor molecules, TLR2 and TLR4, and the associated molecule MD-2 are expressed in endocrine cells of the human pancreatic islets. CD14 expression in human pancreatic islets was determined by immunofluorescence staining of tissue sections and primary cultures, and confirmed by flow cytometry of dispersed normal islets and SV40-transformed islet cells (HP62). The latter cells synthesized and secreted CD14 in response to lipopolysaccharide (LPS) in a time- and dose-dependent manner. Reverse transcription polymerase chain reaction (RT-PCR)-Southern was positive for CD14, TLR2, TLR4 and MD-2 in human pancreas, purified islets and HP62 cells. In vitro experiments using rat islets (also positive for CD14 by RT-PCR) and HP62 cells showed that LPS regulates glucose-dependent insulin secretion and induces inflammatory cytokines [interleukin (IL)-1α, IL-6 and tumour necrosis factor (TNF)-α]. The functional expression of CD14 and associated molecules in islet β cells adds a new pathway that islet cells may follow to adjust their function to endotoxaemia situations and become vulnerable to the inflammatory events that occur during diabetogenic insulitis.
Normal human IgG prevents endothelial cell activation induced by TNFα and oxidized low-density lipoprotein atherogenic stimuli
Authors: RONDA N , BERNINI F , GIACOSA R , GATTI R , BALDINI N , BUZIO C , ORLANDINI G
Published: July 2003

Normal human immunoglobulin G (IgG) has anti-inflammatory and immuno-regulatory properties, which are exploited in the therapy of selected diseases. A putative mechanisms of action is the direct regulation of endothelial cell function by natural antiendothelial cell antibodies. Endothelium activation is a critical event in atherosclerosis. We have verified the ability of normal human IgG to modulate endothelial responses to the atherogenic stimuli tumour necrosis factor-α (TNFα) and oxidized low-density lipoproteins (oxLDL) in vitro. Confocal microscopy was used to visualize vascular cell adhesion molecule-1 (CD106) expression on endothelial cells, cytoplasmic free calcium ([Ca++]i) modifications and fluorescein-coupled oxLDL internalization. Cytokine secretion was measured by ELISA on cell supernatants. IgG prevented TNFα induced CD106 membrane expression and an increase in [Ca++]i, and inhibited the secretion of interleukin-6 (IL-6) and macrophage-colony-stimulating factor (M-CSF). IgG also inhibited CD106 expression induced by oxLDL and one pathway of their internalization, but were ineffective on oxLDL induced [Ca++]i rise and apoptosis. F(ab)′2 fragments from IgG, but not monoclonal IgG, reproduce IgG effects. These findings point to a regulatory role for specific antibodies included in circulating normal IgG towards proinflammatory responses of endothelial cells in atherogenesis and suggest possible development of new therapeutic strategies.
Relative influence of variables at multiple spatial scales on stream macroinvertebrates in the Northern Lakes and Forest ecoregion, U.S.A.
Authors: Weigel Brian M. , Wang Lizhu , Rasmussen Paul W. , Butcher Jason T. , Stewart Paul M. , Simon Thomas P. , Wiley Michael J.
Published: July 2003

We used 94 sites within the Northern Lakes and Forests ecoregion spanning Minnesota, Wisconsin and Michigan to identify environmental variables at the catchment, reach and riparian scales that influence stream macroinvertebrates. Redundancy analyses (RDA) found significantly influential variables within each scale and compared their relative importance in structuring macroinvertebrate assemblages.

Environmental variables included landcover, geology and groundwater delivery estimates at the catchment scale, water chemistry, channel morphology and stream habitat at the reach scale, and landcover influences at three distances perpendicular to the stream at the riparian scale. Macroinvertebrate responses were characterised with 22 assemblage attributes, and the relative abundance and presence/absence of 66 taxa.

Each scale defined macroinvertebrates along an erosional to depositional gradient. Wisconsin's macroinvertebrate index of biotic integrity, Ephemeroptera–Plecoptera–Trichoptera taxa and erosional taxa corresponded with forest streams, whereas organic pollution tolerant, Chironomidae and depositional taxa corresponded with wetland streams. Reach scale analyses defined the gradient similarly as dissolved oxygen and wide, shallow channels (erosional) opposed instream macrophytes and pool habitats (depositional). Riparian forests within 30 m of the stream coincided with an erosional assemblage and biotic integrity.

Next, we combined all significant environmental variables across scales to compare the relative influence of each spatial scale on macroinvertebrates. Partial RDA procedures described how much of the explained variance was attributable to each spatial scale and each interrelated scale combination.

Our results appeared consistent with the concept of hierarchical functioning of scale in which large‐scale variables restrict the potential for macroinvertebrate traits or taxa at smaller spatial scales. Catchment and reach variables were equally influential in defining assemblage attributes, whereas the reach scale was more influential in determining relative abundance and presence/absence.

Ultimately, comprehending the relative influence of catchment and reach scale properties in structuring stream biota will assist prioritising the scale at which to rehabilitate, manage and derive policies for stream ecosystem integrity.
Ecological strategies in the ancient asexual animal group Darwinulidae (Crustacea, Ostracoda)
Authors: Van Doninck Karine , Schön Isa , Maes Fre , De Bruyn Luc , Martens Koen
Published: July 2003

We investigated the relationship between geographical distribution and ecological tolerance within the ancient asexual family Darwinulidae. Distribution maps were compiled based on data from the literature, the Non‐marine Ostracod Distribution in Europe database and personal collections. Ecological tolerance was assessed experimentally by exposing individual ostracods to a combination of eight different salinities (range from 0 to 30 g L−1) and three different temperatures (10, 20 and 30 °C).

The type species of the family, Darwinula stevensoni, is ubiquitous and cosmopolitan; the two species Penthesilenula brasiliensis and Microdarwinula zimmeri also have an intercontinental distribution. Two other darwinulid species tested here (Vestalenula molopoensis and P. aotearoa) are known only from their type localities. The latter is also true for most extant darwinulids.

Darwinula stevensoni and P. brasiliensis had a broad salinity tolerance, tolerating distilled water and also salinity up to 25–30 g L−1, whereas the maximum salinity tolerance of V. molopoensis was 12 g L−1 and of P. aotearoa, 20 g L−1.

The results indicate that both ecological specialists and generalists, as well as intermediate forms, exist in the Darwinulidae and that taxa with the broadest ecological tolerance also have the widest distribution.
Ecosystem CO2 exchange and plant biomass in the littoral zone of a boreal eutrophic lake
Authors: Larmola T. , Alm J. , Juutinen S. , Martikainen P. J. , Silvola J.
Published: July 2003

In order to study the dynamics of primary production and decomposition in the lake littoral, an interface zone between the pelagial, the catchment and the atmosphere, we measured ecosystem/atmosphere carbon dioxide (CO2) exchange in the littoral zone of an eutrophic boreal lake in Finland during two open water periods (1998–1999). We reconstructed the seasonal net CO2 exchange and identified the key factors controlling CO2 dynamics. The seasonal net ecosystem exchange (NEE) was related to the amount of carbon accumulated in plant biomass.

In the continuously inundated zones, spatial and temporal variation in the density of aerial shoots controlled CO2 fluxes, but seasonal net exchange was in most cases close to zero. The lower flooded zone had a net CO2 uptake of 1.8–6.2 mol m−2 per open water period, but the upper flooded zone with the highest photosynthetic capacity and above‐ground plant biomass, had a net CO2 loss of 1.1–7.1 mol m−2 per open water period as a result of the high respiration rate. The excess of respiration can be explained by decomposition of organic matter produced on site in previous years or leached from the catchment.

Our results from the two study years suggest that changes in phenology and water level were the prime cause of the large interannual difference in NEE in the littoral zone. Thus, the littoral is a dynamic buffer and source for the load of allochthonous and autochthonous carbon to small lakes.