bluuebunny/crossref_metadata_2025 huggingface dataset:
Characterization of Api g 1.0201, a New Member of the Api g 1 Family of Celery Allergens
Authors: Hoffmann-Sommergruber Karin , Ferris Rosemary , Pec Martina , Radauer Christian , O’Riordain Gabriel , Laimer da Camara Machado Margit , Scheiner Otto , Breiteneder Heimo
Published: April 2003

Background: The association of pollinosis with allergy to plant foods occurs in up to 70% of tree pollen-allergic patients. In recent years, some of the relevant cross-reacting proteins have been characterized at the molecular and immunological level. Api g 1 has been identified as the celery homologue of the major birch pollen allergen, Bet v 1. Although a number of Bet v 1 isoforms have been characterized from birch pollen, little is known about isoforms of food allergens and their allergenic features. Methods: Api g 1.0201, an isoform of Api g 1, was isolated from a cDNA library, cloned and sequenced. The cDNA was expressed in Escherichia coli and the purified recombinant protein was tested in immunoblots. Results: Api g 1.0201 displays 72% sequence similarity to the previously identified Api g 1.0101 and consists of 159 amino acid residues. The sequence of Api g 1.0201 has five additional amino acid residues at the carboxy-terminus as compared to Api g 1.0101. Purified recombinant Api g 1.0201 is recognized by IgE from the sera of celery-allergic patients, as well as by the murine monoclonal anti-Bet v 1 antibody. In general, this isoform displays a weaker IgE-binding capacity than Api g 1.0101, as concluded from immunoblotting experiments. Results from inhibition assays revealed that IgE-binding to Api g 1.0201 is only slightly reduced by preincubation with either purified recombinant Api g 1.0101 or purified recombinant Bet v 1a. Total inhibition was only achieved when using purified natural Bet v 1. Conclusions: At present, little is known about the IgE-binding capacity of isoforms of Bet v 1 homologues of food allergens. Identification and characterization of such isoforms may help to contribute to a better understanding of food allergy and the observed cross-reactivity to pollen allergy.
Effects of Perilla Seed Oil Supplementation on Leukotriene Generation by Leucocytes in Patients with Asthma Associated with Lipometabolism
Authors: Okamoto Makoto , Mitsunobu Fumihiro , Ashida Kozo , Mifune Takashi , Hosaki Yasuhiro , Tsugeno Hirofumi , Harada Seishi , Tanizaki Yoshiro , Kataoka Mikio , Niiya Kenji , Harada Mine
Published: April 2003

Background: Dietary sources of α-linolenic acid, such as perilla seed oil, may have the capacity to inhibit the generation of leukotrienes (LTs) by leucocytes in patients with asthma, as has been reported with the consumption of other long-chain n-3 fatty acids. Methods: The factors affecting the suppression of leukotriene (LT) C4 generation by leucocytes were examined by comparing the clinical features of patients with asthma who had been given dietary perilla seed oil (n-3 fatty acids). Group A consisted of patients in whom the leucocyte generation of LTC4 was suppressed by dietary perilla seed oil. Group B consisted of those in whom LTC4 generation was not suppressed. Results: LTC4 generation by leucocytes decreased significantly in group A after 2 (p < 0.05) and 4 weeks (p < 0.05); conversely, it increased significantly in group B after 4 weeks (p < 0.05). The two study groups differed significantly in terms of LTC4 generation by leucocytes after 4 weeks of dietary supplementation (p < 0.05). Ventilatory parameters such as peak expiratory flow (PEF), forced vital capacity (FVC) and forced expiratory volume in 1 s (FEV1) increased significantly after 4 weeks of dietary supplementation in group A (p < 0.05). Values of PEF, FVC, FEV1 and maximum expiratory flow at 25% of the forced vital capacity (v̇25) differed significantly between groups A and B prior to dietary supplementation. Serum levels of total cholesterol, low-density lipoprotein (LDL) cholesterol and phospholipid were significantly decreased by dietary supplementation in group A after 4 weeks. Serum levels of total cholesterol, triglyceride, high-density lipoprotein cholesterol, LDL cholesterol and phospholipid differed significantly between the two study groups prior to dietary supplementation. Serum levels of triglyceride and LDL cholesterol differed significantly between the two study groups after 4 weeks of dietary supplementation. Conclusions: Dietary supplementation with perilla seed oil in selected patients with asthma suppresses the generation of LTC4 and is associated with clinical features such as respiratory function and lipometabolism.
Transmission of IgA and IgG Monoclonal Antibodies to Mucosal Fluids following Intranasal or Parenteral Delivery
Authors: Falero-Diaz Gustavo , Challacombe Stephen , Rahman Durdana , Mistry Mukesh , Douce Gill , Dougan Gordon , Acosta Armando , Ivanyi Juraj
Published: April 2003

Background: The efficacy by which passive antibodies can reach the lungs could be important for the outcome of immunotherapy of respiratory pulmonary infections. We examined how transmission to a number of mucosal sites is affected by the route of inoculation. Methods: Transmission of newly raised IgA class Mabs against mycobacterial surface antigens to saliva, lung or vaginal lavage, bile and serum of BALB/c mice was compared with existing IgG Mabs. ELISA was used for testing body fluids obtained 1–24 h after intranasal or intravenous inoculation and 1–7 days following back-pack tumour growth of hybridomas. Results: Intranasal inoculation resulted in a rapid rise and high levels of both IgA and IgG class Mabs in lung lavage. In contrast, following intravenous Mab injection or back-pack tumour growth of hybridoma cells, effective lung transmission was observed for the IgG1 and IgG2b MAbs, but not for the IgA Mabs. The secretory component was acquired by the transmitted IgA MAbs in the mucosal fluids, but not in the serum. Nevertheless, the time course of mucosal IgA antibody levels was similar to that of the tested IgG Mabs. Furthermore, the relative proportion of transmission to saliva and bile varied between individual Mabs indicating a role of tissue-specific, immunoglobulin class-unrelated mechanisms. Conclusions: Intranasal, rather than parenteral inoculation of mice is required for the efficient delivery of IgA antibodies against respiratory pulmonary pathogens. Interestingly, IgA-secretory component complexing of intranasally applied Mabs did not significantly influence their persistence in the lungs.
The Adjuvant Effect of Particles – Importance of Genetic Background and Pre-Sensitisation
Authors: Granum Berit , Gaarder Per Ivar , Eikeset Åse , Stensby Berit A. , Løvik Martinus
Published: April 2003

Background: We have previously reported that simple and well-characterised particles, such as polystyrene particles (PSP), have an IgE adjuvant effect in mice. The purpose of this study was to explore the importance of genetic background concerning the adjuvant effect of PSP in different strains of mice. Methods: Inbred NIH/Ola, BALB/c and C3H/HeJ mice were given two intraperitoneal injections with either PSP plus OVA or OVA alone, and then an intraperitoneal challenge with OVA alone. NIH/Ola mice were also pre-sensitised to develop a weak or strong IgE response to OVA, and then given an intraperitoneal challenge with PSP plus OVA or OVA alone. Serum levels of total and allergen-specific IgE and IgG2a were measured. Results: PSP had a specific IgE and IgG2a adjuvant effect in NIH/Ola mice but not in C3H/HeJ and BALB/c mice. Weakly pre-sensitised NIH/Ola mice showed the same response pattern as the naive NIH/Ola mice. In contrast, strongly pre-sensitised mice showed an antibody response pattern similar to that of high-responder BALB/c mice. Conclusions:  Our results indicate that the allergen responder status, either genetic or induced, is of importance for the adjuvant effect from particles. The IgE and IgG2a adjuvant effect may depend on the genetically determined susceptibility of an individual to be influenced by exposure to the adjuvant. Therefore, the allergy-enhancing effect from particle pollution may differ between individuals.
Immunoblot Analysis of IgE-Binding Antigens in Paprika and Tomato Pollen
Authors: van Toorenenbergen A.W. , Waanders J. , Gerth van Wijk R. , Vermeulen A.M.
Published: April 2003

Background: The high incidence of occupational allergy in horticulture has only recently been recognized. We determined IgE against pollen and fruit from paprika and tomato plants in sera from 3 greenhouse workers and in 3 sera from food-allergic patients. Methods: Proteins in extracts of paprika and tomato pollen were incubated with patients’ sera after covalent coupling of these proteins to agarose beads, or in immunoblots. Results: IgE against paprika pollen, but no IgE against tomato pollen, was found in serum from 2 greenhouse workers who worked with paprika plants only. IgE binding of these 2 sera to agarose-bound paprika pollen extract could be inhibited by paprika pollen but not by tomato pollen extract. A greenhouse worker, who cultivated tomato plants, had IgE against both tomato and paprika pollen. IgE binding of this serum to agarose-bound paprika pollen extract could be inhibited by both paprika pollen and tomato pollen extract. Three food-allergic patients also had IgE against tomato and paprika pollen. IgE from 2 food-allergic patients recognized IgE-binding structures in paprika or tomato pollen that were also present in fruit from the corresponding plant. In contrast, no substantial cross-reactivity was observed between paprika pollen and fruit towards IgE from 3 greenhouse workers. In 4 of 5 sera that were positive in the paprika pollen immunoblot major IgE binding to allergens of about 30 and 64 kD occurred. Conclusion: The presence of IgE against paprika or tomato pollen is not restricted to workers in horticulture; IgE against these pollen can also be present in food-allergic patients who have serum IgE against paprika and/or tomato fruit.
Involvement of Eosinophils in the Early-Phase Allergic Reaction in a Guinea Pig Rhinitis Model
Authors: Imai Norio , Miyahara Atsushi , Yamazaki Yuji , Homma Reiko , Ino Yoshitaka , Kurumi Masateru
Published: April 2003

Background: Eosinophils are found in the nasal lavage fluid (NLF) and nasal biopsies of patients with allergic rhinitis after a nasal antigen challenge, and associated not only with a late-phase allergic reaction (LPR) but also an early phase allergic reaction (EPR). Numerous studies have been carried out to clarify the participation of eosinophils in LPR or airway hyperresponsiveness. However, there has been no published report describing in detail the role of eosinophils during EPR. To better understand the involvement of eosinophils in EPR, we studied the effects of repeated antigen challenges on nasal airway responsiveness and eosinophilic inflammation in EPR using a guinea pig rhinitis model. Methods: Nasal airway responsiveness was measured as the nasal airway resistance (NAR) after nasal antigen provocation. Eosinophilic inflammation during EPR was assessed by nasal lavage and histopathological examination using two groups of animals: those in group 1 were subjected to a sensitization pretreatment only, and those in group 2 were subjected to a pretreatment of sensitization followed by repeated nasal challenges. Results: Repeated antigen challenges induced nasal hyperresponsiveness as indicated by a decrease in the antigen provocation dose and a significant increase in NAR. Furthermore, significant increases in eosinophil counts, eosinophil peroxidase (EPO) activity and protein content in NLF during EPR were observed following antigen provocation in group 2. There were significant correlations between the levels of these parameters, and albumin was the most prevalent of the proteins in NLF. Histopathological examination showed that the degree of eosinophil infiltration into the lamina propria of the nasal mucosa of the animals in group 2 was significantly and apparently higher than in group 1. Particularly, epithelial disruption and mucosal edema were significantly elevated after antigen provocation in group 2. Conclusions: These results suggest that chronic eosinophil accumulation is induced by repeated antigen challenges in the nasal tissue, and that once antigen provocation occurs, eosinophils in the tissue are activated and responsible for the amplification of EPR such as vascular permeability and mucosal edema.
Autoantibody Reactivity in a Case of Schnitzler’s Syndrome: Evidence for a Th1-Like Response and Detection of IgG2 Anti-FcεRIα Antibodies
Authors: Sperr Wolfgang R. , Natter Susanne , Baghestanian Mehrdad , Smolen Josef , Wolff Klaus , Binder Bernd R. , Müller Michael R. , Lechner Klaus , Valenta Rudolf , Valent Peter
Published: April 2003

Schnitzler’s syndrome is a rare disease characterized by chronic urticaria, monoclonal IgM, and clinical and laboratory signs of inflammation. In a subset of patients, the urticarial lesions cause pruritus. However, the pathophysiology of the disease and the biochemical basis of urticaria are not known. We describe a female patient with Schnitzler’s syndrome suffering from chronic urticaria associated with pruritus. The patient’s serum was found to contain IgG antibodies recognizing cellular components of the microvasculature. In particular, IgG3 antibodies directed against proteins (14–100 kD) expressed in cultured dermal microvascular endothelial cells and mast cells, were found by immunoblotting. Moreover, IgG2 antibodies specific for the α-chain of the FcεRI were detectable. However, the autoantibodies did not mediate histamine release in mast cells or basophils. In patients with IgM paraproteinemia who did not have Schnitzler`s syndrome, antibodies against endothelial/mast cells or FcεRI were not detectable. In summary, we describe subclass-specific IgG reactivity against microvascular endothelial cells and mast cells indicating Th1 autoimmunity in a patient with Schnitzler’s syndrome. Whether such autoantibodies are recurrently produced in patients with Schnitzler’s syndrome and play a role in the pathophysiology of the disease remains to be determined.
Etiopathogenesis of Systemic Lupus Erythematosus
Authors: Herrmann Martin , Winkler Thomas , Gaipl Udo , Lorenz Hanns-Martin , Geiler Thomas , Kalden Joachim R.
Published: April 2003

Systemic lupus erythematosus is an autoimmune disease of unknown etiology. Research efforts of the last few years have mainly focused on basic molecular and cellular pathogenetic processes of the disease. Consequently, this paper reviews the etiopathogenetic hallmarks, such as impaired amount and presentation of nuclear antigens, production of antinuclear antibodies by T-cell-dependent B cell stimulation and organ damage by anti-dsDNA antibodies or immune complexes that are discussed at the present time. In summary, the hypothesis of a dysregulation of apoptotic cell clearance is strongly supported and broadly discussed.
Use of Immunoglobulin Variable-Region Genes by Normal Subjects and Patients with Systemic Lupus Erythematosus
Authors: Hansen A. , Dörner T. , Lipsky P.E.
Published: April 2003

Antibodies to specific autoantigens are serological hallmarks of systemic autoimmune diseases. These autoantibodies are thought to represent a consequence of immune dysregulation in these conditions, and, in part, have been shown to be involved in their pathologic consequences. However, the mechanisms that lead to the production of autoantibodies are still unknown. The observation that certain autoantibodies are frequently encoded by a limited number of immunoglobulin (Ig) variable-region gene segments suggested that a bias in the development of the Ig repertoire might play a role in the tendency to develop autoimmunity. Whether the use of these individual gene segments is random or different in normal subjects and patients with systemic autoimmune disorders remains a matter of controversy. New approaches for the analysis of variable-region genes from unstimulated individual human B cells employing the single-cell polymerase chain reaction have provided new insights in the B cell repertoire of both normal subjects and patients with systemic autoimmune diseases. Using this approach, the analysis of nonproductive and productive Ig variable-region gene rearrangements made it possible to distinguish molecular processes, as manifested in the nonproductive repertoire, from subsequent selection influences. An initial study in a patient with systemic lupus erythematosus has led to the hypothesis that the molecular generation of the B cell repertoire is similar in patients and normal subjects but subsequent influences and, most notably, extensive mutations and receptor editing differ significantly in shaping the peripheral IgV gene use by persons with autoimmune diseases.
Antiphospholipid Syndrome
Authors: Gromnica-Ihle Erika , Schössler W.
Published: April 2003

Antiphospholipid syndrome (APS) is a disease characterized by venous and arterial thromboses or spontaneous abortions and the repeated detection of antiphospholipid antibodies (aPL). APS may be associated with another autoimmune disease (secondary APS), particularly systemic lupus erythematosus (SLE), or unrelated to an underlying disease (primary APS). APS affects almost all organs. In addition to the clinical criteria, lupus anticoagulant testing and immunological aPL determinations are required to establish the diagnosis of APS.
Diagnostic and Prognostic Relevance of Autoantibodies in Uranium Miners
Authors: Conrad Karsten , Mehlhorn Jürgen
Published: April 2003

Uranium miners exposed to silica dust have a higher risk of developing systemic sclerosis (SSc) and systemic lupus erythematosus (SLE). Sera of 1976 former uranium miners were analysed for autoantibodies typical of connective tissue disease. The frequency of some of these antibodies (anti-centromere, -topoisomerase I, -nucleolar, -dsDNA, -Ro/SSA, -La-SSB and U1-RNP antibodies) was significantly higher compared to a gender- and age-matched control group and was associated with the intensity of exposure as well as with clinical symptoms of SSc or SLE. It was also shown that SSc-associated autoantibodies may serve as an early indicator of disease development. Some differences in the autoantibody production between silica-dust-associated and idiopathic SLE/SSc were observed that might be caused by environmental factors in the population of uranium miners.
Cultivar-Specific IgE-Epitopes in Date (Phoenix dactylifera L.) Fruit Allergy
Authors: Kwaasi A.A.A. , Harfi H.A. , Parhar R.S. , Collison K.S. , Al-Sedairy S.T. , Al-Mohanna F.A.
Published: April 2003

Background: Date fruits are allergenic and standardized extracts are required for diagnosis and therapy of this allergy. Since there are several cultivars of dates, this study was carried out to assess the allergenicity of different cultivars in order to select suitable source material for standardization. Methods: The protein profiles of 18 of the most commonly sold varieties were compared by SDS-PAGE and their relative allergenicity assessed by SPT and IgE-based ELISA and immunoblotting. Thirty-two date fruit-sensitive patients were skin tested with a pooled extract from all the cultivars. Six of the patients with high SPT results (≥3+) who volunteered were further tested with the 18 cultivars and their sera used in ELISA and immunoblotting. Results: Six of the cultivars gave high SPT-positive reactions in ≥4 of patients. Five of these high SPT-reactive cultivars gave high IgE ELISA scores (≥0.58) but individual cultivars varied in their number of IgE immunoblot bands. Cultivar-specific IgE-binding patterns indicated that only certain cultivars bound IgE at molecular weights of ≤14.3 and 27–33 kDa whilst all cultivars bound to a 54–58 kDa doublet. Cultivars that bind to the ≤14.3 and 27–33 kDa bands appeared to form the majority of the high SPT-reactive cultivars. When individual sera of 24 of the 32 SPT-positive patients were used in IgE immunoblots with the pooled cultivar extract, all sera bound IgE at ≤14.3 and 27–33 kDa and about 60% of sera bound to a 54–58 kDa doublet bands. Conclusions: These results indicate that allergenicity of date fruits is a cultivar-specific phenomenon. Sixty to 100% of sera from date fruit-allergic patients bind IgE to three major allergens of ≤14.3, 27–33 and 54–58 kDa. Five of the cultivars that evoke high SPT reactions, high IgE ELISA scores and bind IgE to the major allergens, can be selected for the preparation of ‘in-house’ allergen extracts and for allergen standardization.
Early DNA Synthesis and Cytokine Expression in the Nickel Activation of Peripheral Blood Mononuclear Cells in Nickel-Allergic Subjects
Authors: Falsafi-Amin Hale , Lundeberg Lena , Bakhiet Moiz , Nordlind Klas
Published: April 2003

Background and Objective: Nickel sulphate stimulates the proliferation of lymphocytes in nickel-allergic subjects. However, nickel-induced stimulation of lymphocytes from control persons without clinical symptoms of nickel allergy has also been reported. The aim of the present study was to correlate T cell activity, measured by DNA synthesis and the expression of Th1 [interleukin (IL)-2, tumour necrosis factor (TNF)-β and interferon (IFN)-γ] and Th2 (IL-4) cytokines, in short-term (up to 72 h) culture of nickel-stimulated peripheral blood mononuclear cells (PBMC) from nickel-allergic patients compared to control subjects. Methods: DNA synthesis was measured by the incorporation of [methyl-3H]thymidine. The production of IL-2, TNF-β, IFN-γ and IL-4 was measured in the supernatants of the cultures by ELISA. In situ hybridization for mRNA was performed using oligonucleotide probes for IL-4, IFN-γ and TNF-β in cell smears. Results: Already after 24 h and proceeding through the remaining culture period, there was a statistically significant (p < 0.001) difference in the concentrations of IL-2 between patients and controls. There was a significant (p < 0.01) difference in DNA synthesis (stimulation index) between the patients and control subjects at 72 h, and also at the same time a difference in the concentrations of TNF-β (p < 0.05) and IL-4 (p < 0.01). In the in situ hybridization study, TNF-β was found to be the only one of the studied cytokines that differed between the nickel-allergic and control subjects, this difference being most evident at 72 h (p < 0.01). Conclusions: Our results indicate a difference between nickel-allergic and non-allergic subjects in the synthesis of DNA and production of cytokines when PBMC are stimulated by nickel sulphate, and IL-2 may be regarded as a critical and early-occurring cytokine
A Soybean G2 Glycinin Allergen
Authors: Helm Ricki M. , Cockrell Gael , Connaughton Cathie , Sampson Hugh A. , Bannon Gary A. , Beilinson Vadim , Livingstone Donald , Nielsen Niels C. , Burks A. Wesley
Published: April 2003

Background: Multiple allergens have been documented in soybean extracts. IgE from individuals allergic to soybeans, but not to peanut, was shown by immunoblot analysis to bind to proteins with a molecular weight of approximately 21 kD. These findings suggested that unique proteins in soybeans might be responsible for soybean allergic reactivity. The objective of the present study was to identify unique proteins in soybean extracts that bind to specific IgE from soybean-sensitive individuals, and to characterize the allergen using physicochemical methods and IgE binding. Methods: Two-dimensional and preparative SDS-PAGE/IgE immunoblot analysis was used to identify a 22-kD soybean-specific allergen from crude soybean extracts. N-terminal sequence analysis was used to determine the identification of the protein binding IgE from soybean-sensitive individuals. Results: IgE immunoblot and amino acid sequence analysis identified the 22-kD protein as a member of the G2 glycinin soybean protein family. Further investigation revealed that the IgEs reacted with basic chains from each member of the glycinin family of soybean storage proteins. Conclusions: Each of the subunits from glycinin, the storage protein that is the most prevalent component of soybean, are major allergens.
Characterisation of Horse Dander Allergen Glycoproteins Using Amino Acid and Glycan Structure Analyses
Authors: Bulone Vincent , Rademaker Geert Jan , Pergantis Spiros , Krogstad-Johnsen Tonje , Smestad-Paulsen Berit , Thomas-Oates Jane
Published: April 2003

Separation of horse dander allergens using two-dimensional PAGE resulted in the identification of 16 proteins that react with allergic patient sera. A sensitive method has been developed for analysing the structures of the glycan chains of individual glycoprotein allergens transferred to blots following two-dimensional PAGE, and has allowed the structural identification of the glycan chains of the most abundant isoforms of Equ c 1, a glycosylated horse dander major allergen. The method involves separation of the allergens by two-dimensional PAGE, transfer to polyvinylidene difluoride membranes, release of the glycan chains using peptide N-glycosidase F, permethylation and mass spectrometric analysis of the derivatised glycans. The amino acid compositions of the 16 horse dander allergens separated by two-dimensional PAGE have been determined, allowing the identification of the various isoforms of Equ c 1. These results also confirmed that the two non-glycosylated major allergens, Equ c 2.0101 and Equ c 2.0102, belong to the lipocalin family, and support the idea that these two allergens are most probably isoforms of the same protein. The glycan structures identified using the mass spectrometric method are common biantennary and triantennary glycan chains. These carbohydrate moieties may have a role in the binding of IgE; however, it is more likely that the overall glycoprotein structure involving both the glycan and protein moieties, rather than the structure of the glycan chains alone, is responsible for eliciting allergic responses.
Identification of a Sequential B-Cell Epitope on Major Allergen (Cry j 1) of Japanese Cedar (Cryptomeria japonica) Pollen in Mice
Authors: Tamura Yukiko , Sasaki Reiko , Inouye Sakae , Kawaguchi Junko , Serizawa Nobufusa , Toda Masako , Takemori Toshitada , Sakaguchi Masahiro
Published: April 2003

Background: Japanese cedar (Cryptomeria japonica; CJ) pollinosis is one of the most common allergic diseases in Japan. B cell epitopes on Cry j 1, a major allergen of CJ pollen, have been analyzed by the specific monoclonal antibodies to Cry j 1, and most of these epitopes may be conformational, but no previous report has addressed the analysis of sequential epitope mapping with synthetic peptides. The main purpose of the present study is to identify IgE and IgG B cell epitopes on Cry j 1 by using a synthetic peptide approach in mice. Methods: We synthesized 35 overlapping peptides that cover the entire length of Cry j 1 and examined whether mouse IgE and IgG antibodies produced by immunization with Cry j 1 reacted to the Cry j 1 peptides. Results and Conclusion: We found that mouse IgE and IgG antibodies reacted strongly to Cry j 1 peptide No. 15 (141GVEPVHPQDGDALTLRTATN160), though those antibodies did not react with other peptides. IgE and IgG antibody binding to peptide No. 15 was completely inhibited by Cry j 1 and the peptide. To determine the minimum epitope in peptide No. 15, we conducted an ELISA inhibition test. IgE and IgG antibody binding to peptide No. 15 was inhibited by smaller peptides of this peptide. We found the core of the epitope to be 145VHPQDGDA152.
Monoclonal Antibodies against Der s 1, a Major Allergen of Dermatophagoides siboney
Authors: Sewer Minerva , Uyema Keiko , Labrada Mayrel , González Maritza , Coca Marcos
Published: April 2003

Six stable clones secreting murine monoclonal antibodies (Mabs) against Der s 1 were obtained. The binding of Mabs showed cross-reactivity with Dermatophagoides farinae, as determined by enzyme-linked immunosorbent assay (ELISA). In a Western blot assay, antibodies reacted with a 24-kD protein considered to represent the major allergen Der s 1. The repertoire of antigenic sites on Der s 1 was studied using a panel of Mabs. Epitope specificity of the Mabs was determined by both competitive inhibition and sandwich ELISA assays. The results defined six different, non-overlapping and non-repeated antigenic sites on the allergen molecule. Der s 1 allergen from Dermatophagoides siboney extracts was purified by Mab affinity chromatography, this procedure gave 43% recovery of >90% pure allergen. The purified allergen had capacity to bind specific human IgE and demonstrated an allergenic activity of up to 77% of total D. siboney extract. An Mab-ELISA was developed using Mabs directed against different epitopes on Der s 1. This assay could detect up to 1 ng/ml of Der s 1 and Der f 1 in allergen preparations.
MMP-2- and MMP-9-Linked Gelatinolytic Activity in the Sputum from Patients with Asthma and Chronic Obstructive Pulmonary Disease
Authors: Cataldo D. , Munaut C. , Noël A. , Frankenne F. , Bartsch P. , Foidart J.M. , Louis R.
Published: April 2003

Background: The course of asthma and chronic obstructive pulmonary disease (COPD) is associated with bronchial morphological changes. Metalloproteinases are thought to play a role in these structural changes. Methods: We studied the gelatinolytic activity present in the induced sputum from 20 patients with asthma, 20 with COPD and 19 healthy controls. The assessment of gelatinolytic activity was performed by quantitative zymography, and gelatinolytic species were identified by Western blot analysis. Tissue inhibitor of metalloproteinase-1 (TIMP-1) was detected by reverse zymography and ELISA. Results: From zymography, we found significantly higher gelatinolytic activity linked to pro-matrix metalloproteinase-9 (pro-MMP-9) in the sputum from asthmatics (p < 0.0001) and COPD patients (p < 0.0001) compared to the control group. Furthermore, the activated form of MMP-9 (85 kD) was found in the sputum from 60% of asthmatics and 85% of COPD patients, but was absent in that of control subjects (p < 0.0001). Importantly, although less frequently detectable than pro-MMP-9, pro- MMP-2 (72 kD) was found more frequently in asthmatics (50%) than in control subjects (5%) (p < 0.005). We also described two unusual gelatinolytic species of 45 and 120 kD and showed that they derived from MMP-9 according to their ability to bind gelatin and anti-MMP-9 antibody. Levels of TIMP-1 were higher in asthmatics (p < 0.05) and COPD patients (p < 0.05) than in controls. Conclusion: Asthmatics and COPD patients display an increased gelatinolytic activity linked to MMP-2 and MMP-9 and higher levels of TIMP-1 in their sputum.
Double Sensitization to Honeybee and Wasp Venom: Immunotherapy with One or with Both Venoms?
Authors: Straumann F. , Bucher C. , Wüthrich B.
Published: April 2003

Background: Double sensitization to honeybee (Apis mellifera) and wasp venom (Vespula spp.) as determined by skin test and measurement of specific IgE is common in hymenoptera sting allergy. Double-sensitized patients have either distinct antibodies for each venom or cross-reacting antibodies that recognize similar or identical epitopes in both venoms. Unfortunately, patients often fail to identify the stinging insect which makes it difficult to distinguish cross-reactors from non cross-reactors. However, for economic reasons as well as for the benefit of the patients, it would be useful to identify complete cross-reactors. Methods: In this study we investigated 24 double-sensitized patients who were candidates for venom immunotherapy. Homologous and heterologous FEIA inhibition was carried out with honeybee (Apis mellifera) and wasp venom (Vespula spp.) preparations from two different providers. The inhibitor concentrations were ranging from 0 to 100 μg protein/ml. Results: Sera of 4 patients were completely cross-reacting for one venom (3 honeybee, 1 wasp), 8 patients were partially cross-reacting and 10 patients were not cross-reacting. Two patients were excluded from the study due to insufficient homologous inhibition. Data from specific IgE measurements, skin test, and clinical history were not useful for the identification of cross-reacting patients. Conclusion: FEIA inhibition is easy to perform and useful for the identification of patients with complete cross-reactivity. In these patients immunotherapy might be restricted to one venom which is beneficial for the patient and cost-effective.
Truncated Dipeptidyl Peptidase IV Is a Potent Anti-Adhesion and Anti-Metastasis Peptide for Rat Breast Cancer Cells
Authors: Abdel-Ghany Mossaad , Cheng Hung-Chi , Levine Roy A. , Pauli Bendicht U.
Published: April 2003

A novel adhesion receptor/ligand pair was shown recently to mediate lung vascular arrest and metastasis of rat breast cancer cells. The interacting adhesion molecules are endothelial dipeptidyl peptidase IV (DPP IV) and tumor cell surface-associated, polymeric fibronectin (FN). A truncated DPP IV (DPP IV(31–767): amino acids 31–767) in which the FN-binding site is preserved is shown here to mask the breast cancer cell surface-associated FN complexes, causing a dose-dependent inhibition of adhesion to endothelial DPP IV and impeding lung colony formation by approximately 80%. Since surface accumulation of FN is chiefly occurring during dissemination in the blood and since many cancer cell types have surface receptors by which they may initiate FN accumulation on their surfaces, the present anti-metastatic treatment modality may extend its efficacy farther than appreciated by this study.
Invasion Strategies of the Oral Pathogen Porphyromonas gingivalis: Implications for Cardiovascular Disease
Authors: Deshpande Rajashri G. , Khan Mahfuz , Genco Caroline Attardo
Published: April 2003

Microorganisms have evolved a variety of mechanisms designed to evade detection and/or destruction by the host. Many pathogens evade host defenses by invading cells, thus providing the bacterium with an environment free of competing microorganisms. Adherence and invasion are active processes in which microorganisms often use host proteins and enzymes to gain entry into the cell, thus stimulating their own uptake. The investigation of invasion by the periopathogen Porphyromonas gingivalis is in its infancy in comparison with that of the enteric pathogens. However, recent studies with P. gingivalis have revealed that these organisms have developed invasion strategies and mechanisms similar to those of the enteric pathogens for both epithelial and endothelial cells. The study and elucidation of the mechanisms by which microorganisms such as P. gingivalis persist in chronic infection will provide valuable insight into the pathogenesis of P. gingivalis-mediated periodontal disease. The ability to multiply in and to activate endothelial cells may be one of the pathogenic mechanisms exerted by P. gingivalis that may explain the recently described association between this organism and cardiovascular disease.
Varicella-Zoster Virus Infections in Pregnancy
Authors: Sauerbrei A.
Published: April 2003

Chickenpox is an uncommon disease in women during pregnancy. If, however, maternal varicella infects the fetus, intrauterine death or severe diseases (congenital or fetal varicella syndrome) may ensue depending on the time of maternal infection. On the basis of our own clinical observations and virological testing we report on 31 women with varicella during pregnancy and their infants. Five neonates born to mothers with varicella between the 8th and 21st week showed clinical signs of the congenital varicella syndrome. Varicella-zoster virus DNA could be detected in different tissues, including lung, liver, skin and adrenal glands. As a consequence of maternal chickenpox within 12 days before delivery, 6 newborn babies developed neonatal varicella. Measures for diagnosis, treatment and prophylaxis are discussed.
Epidemiology of Hepatitis C Virus in Japan
Authors: Moriya Takashi , Koyama Tomiko , Tanaka Junko , Mishiro Shunji , Yoshizawa Hiroshi
Published: April 2003

Blood screening for hepatitis C virus (HCV) antibodies at Japanese Red Cross blood centers has indicated an age-dependent prevalence of HCV infection in the general population of Japan: the older the age, the higher the prevalence. The same pattern was corroborated by the sentinel study conducted in an HCV-endemic area. The high prevalence of HCV in the elderly is most likely due to a spread of HCV infection during the turmoil period just after World War II: in particular by illicit intravenous amphetamine abuse. Fortunately, however, since the rate of newly acquired infection is currently too low to influence HCV prevalence in each age group, the total number of HCV carriers (estimated to be 691,852 for the age group 20–64 years) would decrease spontaneously with generation takeover to ∼50% of the current number during the decade to come.
Stretch-Induced Collagen Synthesis in Cultured Smooth Muscle Cells from Rabbit Aortic Media and a Possible Involvement of Angiotensin II and Transforming Growth Factor-β
Authors: Li Qing , Muragaki Yasuteru , Hatamura Ikuji , Ueno Hikaru , Ooshima Akira
Published: April 2003

Mechanical strain reportedly stimulates the synthesis of collagen in vascular smooth muscle cells (SMCs). The present study was designed to investigate a possible involvement of angiotensin II (Ang II) and transforming growth factor (TGF)-β in stretch-induced collagen synthesis of cultured SMCs derived from the rabbit aortic media. SMCs were cyclically stretched at a rate of 10% elongation and 30 cycles/min for 24 h using the Flexercell® strain unit (Flexcell International Corp., McKeesport, Pa.). A two-fold increase in collagen synthesis and a concurrent increase in total protein synthesis were noted in stretched SMCs. Concentration of immunoreactive Ang II in the conditioned medium was elevated under the mechanical strain. Stretch-induced collagen and total protein synthesis were inhibited by either a selective antagonist to Ang II (saralasin), an angiotensin I-converting enzyme inhibitor (captopril) or an antisense oligonucleotide for angiotensinogen mRNA. An elevated secretion of TGF-β, both active and latent forms, was found in the medium of stretched SMCs. Saralasin inhibited the stretch-induced secretion of TGF-β from SMCs. Stretch-induced collagen and total protein synthesis was further inhibited by either an anti-TGF-β1 neutralizing antibody or an adenovirus-mediated transfer of a truncated TGF-β type II receptor. Elevated expression of collagen α1(III) chain and TGF-β1 mRNAs, and its reversal by saralasin were also demonstrated in stretched SMCs. Results indicate that the stretch-induced collagen and total protein synthesis appears to be mediated via an autocrine-paracrine mechanism of Ang II and TGF-β released from SMCs.
Galectin 1 Modulates Attachment, Spreading and Migration of Cultured Vascular Smooth Muscle Cells via Interactions with Cellular Receptors and Components ofExtracellu lar Matrix
Authors: Moiseeva Elena P. , Spring Elizabeth L. , Baron Julia H. , de Bono DavidP.
Published: April 2003

Galectin 1 (Gal-1), a lactose-binding lectin, is a component of vascular extracellular matrix and secreted by human vascular smooth muscle cells (SMCs). The purpose of this study was to investigate a possible role of Gal-1 in controlling adhesion and migration of cultured human vascular SMCs. Gal-1 co-localised with laminin and cellular fibronectin in extracellular matrix (ECM) secreted by cultured human vascular SMCs. Recombinant glutathione S-transferase (GST)-Gal-1 fusion protein bound to laminin and cellular fibronectin in ELISA. GST-Gal-1 inhibited SMC attachment to laminin via interactions with both SMCs and laminin. GST-Gal-1 inhibited SMC spreading on plastic or on laminin, but not on cellular fibronectin. GST-Gal-1 modulated SMC migration on laminin and inhibited migration on cellular fibronectin. GST-Gal-1 bound to several 35S-labelled proteins in SMC extracts including laminin and α1β1 integrin, identified by depletion of SMC protein extracts with respective antibodies. We conclude that Gal-1 is able to modulate SMC attachment, spreading and migration via interactions with ECM proteins and α1β1 integrin.
Relaxation in Different-Sized Rat BloodVessels Mediated by Endothelium-Derived Hyperpolarizing Factor: Importance of Processes Mediating Precontractions
Authors: Tomioka Hiroshi , Hattori Yuichi , Fukao Mitsuhiro , Sato Atsushi , Liu Ming-Yue , Sakuma Ichiro , Kitabatake Akira , Kanno Morio
Published: April 2003

To clarify the mechanisms involved in relaxations mediated by endothelium-derived hyperpolarizing factor (EDHF), acetylcholine (ACh)-induced endothelium-dependent relaxations and hyperpolarizations were examined in the rat aorta, the main branch of the mesenteric artery (MBMA) and the first branch of the mesenteric aftery (FBMA). In the presence of 100 μM NG-nitro-L-arginine (L-NNA) and 10 μM indomethacin, ACh (1 nM to 100 μM) produced no relaxation in the phenylephrine-precontracted aorta. The L-NNA-resistant relaxations by ACh in MBMA precontracted with phenylephrine were eliminated in the presence of 1 μM nifedipine where contractions were independent of L-type Ca2+ channel activation. In FBMA precontracted with phenylephrine, the L-NNA-resistant relaxations were only partially inhibited by nifedipine. When vessels had been contracted with 300 nM phorbol-12,13-dibutyrate in the presence of nifedipine, ACh-induced L-NNA-resistant relaxations were observed in FBMA only. Pinacidil produced relaxations in all different-sized blood vessels, although sensitivity was inversely related to vessel size. The extent of the ACh hyperpolarizing responses was much smaller than that by pinacidil in the aorta. The membrane potential changes by ACh and pinacidil were almost the same in FBMA. These results indicate that the contribution of EDHF to endothelium-dependent relaxations increases as the vessel size decreases. This may be partly explained by precontractile processes dependent on Ca2+ entry through L-type Ca2+ channels, because Ca2+ channel deactivation seems to be involved as a major mechanism of EDHF-mediated vasorelaxations. However, EDHF may also generate vasorelaxations by an additional mechanism, probably a reduced Ca2+ sensitivity of contractile elements, as proposed for ATP-sensitive K+ channel openers.
Microvascular Endothelial Cells Differ in Their Basal and Tumour Necrosis Factor-α-Regulated Expression of Adhesion Molecules and Cytokines
Authors: Lehmann I. , Brylla E. , Sittig D. , Spanel-Borowski K. , Aust G.
Published: April 2003

We recently located a rare cytokeratin-positive (CK+) type of microvascular endothelial cell (MVEC) in the corpus luteum and aorta. Bovine corpus luteum MVEC are known to be involved in the cyclic accumulation of eosinophils and macrophages. Since leukocyte migration is specifically mediated by adhesion molecules and the release of cytokines, we compared the expression of these factors in basal and TNF-α-stimulated CK+ MVEC and in common cytokeratin-negative (CK–) MVEC in order to obtain an initial insight into the functional capacities of CK+ MVEC. CK– MVEC revealed significantly higher basal RANTES mRNA expression than CK+ MVEC, and TNF- α up-regulated RANTES mRNA in both types of MVEC. Only resting and stimulated CK– MVEC expressed granulocyte-macrophage colony-stimulating factor mRNA. Both MVEC types expressed monocyte colony-stimulating factor mRNA, but remained negative for eotaxin and interleukin (IL)-5 mRNA even after stimulation. Resting CK+ MVEC were positive for CD29, CD31, CD49a and CD49e, but expressed most of these antigens at a significantly lower density than did CK– MVEC. In contrast to CK– MVEC, CK+ MVEC failed to express CD49b or MHC class II. The activation of CK+ MVEC with TNF-α induced the expression of CD62P, but not of CD49b or MHC class II. In summary, phenotypically variable MVEC derived from the microvascular bed of one organ differ in their TNF-α-regulated expression of cytokine mRNA and adhesion molecules. Morphological heterogeneity is related to a particular specialisation of functional MVEC.
Effects of Chronic Nitric Oxide Synthase Inhibition on Renal Function and Histology in Polycythemic Rats
Authors: Kawata Tetsuya , Hashimoto Seiji , Koike Takao
Published: April 2003

Augmented endogenous nitric oxide (NO) production may ameliorate derangement of renal functions or glomerular damage in polycythemia. To investigate this possibility, we examined the effect of NO synthase inhibition with Nω-nitro-L-arginine methyl ester (L-NAME; 50 mg/dl in drinking water) on renal functions and histology in heminephrectomized Sprague-Dawley rats treated for 4 weeks with recombinant human erythropoietin (rh-EP; 500 IU/kg on alternate days). L-NAME elevated the blood pressure which was aggravated by concomitant rh-EP and was ameliorated by treatment with a nonpeptide angiotensin type 1 receptor blocker (CV116; 60 mg/kg in chow). The hematocrit level was prominently increased by rh-EP. The glomerular filtration rate was impaired by L-NAME alone, but was maintained by concomitant administration of rh-EP or CV116. Micropuncture experiments revealed that the glomerular capillary pressure was similarly elevated by L-NAME alone or in combination with rh-EP. L-NAME significantly, although not prominently, aggravated glomerular sclerosis observed with rh-EP alone, and concomitant CV116 ameliorated the glomerular damage. These results suggest that, in polycythemia, enhanced NO production buffers the glomerular damage, and the balance between NO and angiotensin II may play an important role in maintaining renal function and glomerular structure.
Angiotensin-Converting Enzyme Gene Polymorphism Determines the Antiproteinuric and Systemic Hemodynamic Effect of Enalapril in Patients with Proteinuric Renal Disease
Authors: Haas M. , Yilmaz N. , Schmidt A. , Neyer U. , Arneitz K. , Stummvoll H.K. , Wallner M. , Auinger M. , Arias I. , Schneider B. , Mayer G.
Published: April 2003

Angiotensin-converting enzyme (ACE) inhibitors are known to reduce blood pressure and proteinuria in a variety of different glomerular diseases. Nonetheless, a marked interindividual difference in the efficacy of these agents exists. The activity of the ACE and therefore of the renin-angiotensin-aldosterone system (RAAS) has been shown to be under genetic influence. Patients with a deletion genotype at the intron 16 of the ACE gene have been shown to exhibit higher activity of plasmatic ACE when compared to patients with the insertion genotype. We therefore studied prospectively the hemodynamic and antiproteinuric effect of a 6-month therapy with enalapril in patients with biopsy-proven proteinuric glomerular diseases and the DD (n = 10) and ID/II (n = 26) genotype. Although patients with the DD genotype received a slightly higher dose of enalapril, blood pressure and proteinuria did not change significantly. However, both were significantly reduced in the II/ID group after 10 weeks and 6 months of therapy. Creatinine clearance decreased steadily in DD patients. In II/ID patients, creatinine clearance was reduced significantly after 10 weeks of therapy but increased again thereafter and the value at 6 months was again comparable to the one obtained in the DD patients. We conclude from our study that the ACE genotype influences the blood pressure-lowering and antiproteinuric effect of enalapril in patients with proteinuric glomerular disease.