bluuebunny/crossref_metadata_2025 huggingface dataset:
The Role of the Invariant Chain in Mucosal Immunity
Authors: Barrera Carlos A. , Almanza Ruben J. , Ogra Pearay L. , Reyes Victor E.
Published: April 2003

The invariant chain (Ii) due to its intimate association with major histocompatibility complex (MHC) α and β chains is a determining element in the development of immune responses. Ii plays a major role in the assembly, the intracellular transport and peptide selection by class II MHC. A segment of Ii designated as CLIP (class II–associated Ii peptide) binds into the antigen binding site of class II MHC molecules until class II MHC reach intracellular compartments that contain peptides from internalized antigens. This association limits the self endogenous peptides that can bind to class II MHC molecules. The removal of CLIP from class II MHC catalyzes the binding of antigenic peptides and their subsequent cell surface expression. An isoform of Ii, known as chondroitin sulfate–modified Ii (IiCS), that is surface–expressed enhances T cell activation while acting as a coreceptor for CD44. The expression of class II MHC molecules by mucosal epithelial cells has generated interest in the role that these cells may have in mucosal immunity. Since in classical antigen–presenting cells (APC) the biology of class II MHC is regulated by Ii, it is necessary to bring into perspective the known functions of Ii in conventional APC to understand the role that Ii may play in mucosal epithelial cells as potential regulators of local immune responses.
Neoallergens in Heated Soybean Hull
Authors: Codina Rosa , Oehling Jr. Albert G. , Lockey Richard F.
Published: April 2003

Background: During the process of harvest, transport and storage, microbial and mold contamination can raise the temperature of soybeans to 75°C or higher. The purposes of this study were (1) to evaluate the allergenicity of fresh and stored soybean hulls and (2) to ascertain whether heat alters the allergenicity of stored soybean hulls. Methods: Allergen extracts were prepared from (1) stored soybean hulls, (2) fresh soybean hulls and (3) stored soybean hulls heated to 37°C (E1), 55°C (E2) and 80°C (E3) or kept at room temperature (E4) for 16 h. Individual serum from 68 soybean asthmatic (SA) subjects, 30 nonallergic subjects and two serum pools made from 4 SA sera and 4 sera from asthmatics not sensitive to soybean were studied. All sera and serum pools were assayed for content of specific IgE (radioallergosorbent test) and IgG4 (ELISA). The following additional studies were done for extracts E1–E4: (1) SDS–PAGE, (2) SDS–PAGE/Western blot for specific IgE and IgG4 using both serum pools, and (3) study of the effects of heat on inhibiting activity of the extracts prepared from stored soybean hulls using the pool of SA sera. Results: Test results demonstrated a reduced binding of specific IgE and IgG4 to fresh soybean hull extract compared to stored soybean hull extract, and an increased binding for heated extracts (E1–E3) compared to unheated ones (E4). Moreover, there was an increase in potency for IgE and IgG4 bindings for the heated (E1–E3) compared to unheated (E4) extract, as measured by the amount of protein to produce 50% inhibition. Several protein bands with a molecular weight (MW) higher than 20 kD were absent from the SDS–PAGE for E3 but were present in E1, E2 and E4, and a new protein band (MW 15.3 kD) appeared for E3 only. Two new protein bands, with MWs of 15.3 and 10 kD, which bind specific IgE, were present on Western blot and one of the 3 main soybean hull allergens, probably Gly m 2, disappeared in E3. IgG4 Western blot showed similar results, but only the 10 kD protein band was present. Conclusion: The results demonstrate that soybean hull allergenicity is affected by heat, and suggest that the heat generated during storage and transport of soybeans could generate 2 new allergen determinants or increases in epitope exposure as a result of conformational changes. The significance of these new IgE and IgG4 binding proteins has yet to be determined.
IgE Binding Capacity of Synthetic and Recombinant Peptides of the Major Storage Mite (Lepidoglyphus destructor) Allergen, Lep d 2
Authors: Elfman Lotta H.M. , Whitley Paul , Schmidt Margit , van Hage-Hamsten Marianne
Published: April 2003

Background: Lepidoglyphus destructor is an important non–pyroglyphid mite species in Europe and a dominant allergen in farming environments. The major allergen of L. destructor, Lep d 2, is a protein of 13.2 kD that is recognised by about 90% of sera RAST positive to this mite species. Methods: The cDNA of two isoallergens of the Lep d 2 has previously been sequenced and the protein expressed in different protein expression systems. In order to map the B–cell epitopes, the full length protein and the truncated forms of the protein have been expressed in Escherichia coli as glutathione–S–transferase (GST) fusion proteins. Recombinant Lep d 2 fragments and synthetic overlapping 15 mer peptides spanning Lep d 2 were probed with sera from patients allergic to storage mite. Results: The full–length (125 amino acids) GST fusion protein reacted strongly with patient IgE in Western blots and dot blots. Synthetic peptides failed to react with IgE antibodies from mite–allergic patients and the truncated fusion proteins displayed weak IgE–binding capacity. Conclusion: We conclude that there are no dominant linear IgE–binding epitopes in Lep d 2. Recombinant or synthetic Lep d 2 fragments may, however, be further evaluated as hypoallergenic candidate molecules for specific immunotherapy.
Hypersensitivity of Bronchial Asthmatics to Cockroach in Taiwan
Authors: Tsai Jaw-Ji , Kao Mau-Han , Wu Chii-Huei
Published: April 2003

This study was performed to test the hypersensitivity of asthmatics to American and German cockroaches, which are both common in Taiwan. A total of 236 asthmatic patients received skin prick test using allergen extracts from both American and German cockroaches, and 596 sera from asthmatic patients were analyzed for their specific IgE against German cockroach extract. The results of skin test showed that 39.4 and 36.4% asthmatic patients were hypersensitive to American and German cockroaches. Fifteen among 236 patients were only allergic to American cockroaches and 8 were only allergic to German cockroaches. Using the Pharmacia CAP system, 36% of the sera were found to contain the specific IgE to German cockroach extract. Eighty–nine sera positive for German cockroach extract were then tested for their reactivity to American cockroach extract using the fluoroallergosorbent test (FAST). Sixty among 89 (68%) of their sera contained American cockroach–specific IgE. The correlation coefficient between both parameters was r = 0.45. Immunoblot and immunoblot inhibition studies were performed to analyze the IgE–binding components and the cross–reactivity between American and German cockroaches. The results showed that there are different IgE–binding components between American and German cockroaches. Sera containing specific IgE to both species of cockroach were absorbed with both species of cockroach extracts. The specific IgE to German cockroaches can be absorbed by American cockroach extract in all selected sera and the specific IgE to American cockroaches can only partially be absorbed by German cockroaches. The nonabsorbed allergens in American cockroaches had molecular weights of 33 and 50 kD. In conclusion, one–third of the asthmatic population tested was allergic to cockroaches. Although most cockroach–hypersensitive patients were allergic to both American and German cockroaches, more asthmatic patients were allergic to American cockroaches in Taiwan. The use of non–crossreacting allergen in detecting American cockroach–specific IgE might be important not only for the diagnosis and treatment of cockroach hypersensitivity in asthmatics but also for the differentiation between German and American cockroach hypersensitivity.
The Biological Function of Allergens: Relevant for the Induction of Allergic Diseases?
Authors: Bufe Albrecht
Published: April 2003

During the last 10 years an increasing number of allergens were identified and biochemically as well as immunologically characterized. More and more information on the primary structure of these molecules, revealed by recombinant technologies, became available. Furthermore for several allergens the biological functions could be elucidated. Most recently it was shown that beside IgE–binding capacity functional properties such as enzyme activity may contribute to the allergenicity and the induction of allergic inflammation. These findings could stimulate novel concepts in allergy prevention and therapy. This review summarizes recent data in this area.
Quantitative Assessment of Immediate Cutaneous Hypersensitivity in a Model of Genetic Predisposition to Atopy
Authors: Daser Angelika , Bätjer Nina , Kölsch Uwe , Kötz Karsten , Schmeling Heinrike , Schou Carsten , Renz> Harald
Published: April 2003

Genetic predisposition and environmental factors modulate the expression of allergic phenotypes. The frequent allergic phenotype 'immediate cutaneous hypersensitivity' was established in mice as a model for atopy. Genetic dissection of this trait requires a robust procedure to assess the allergic phenotype. To this end, different mouse strains were immunized with birch pollen extract. Immediate cutaneous hypersensitivity reactions were induced through intradermal allergen exposure. Wheel formation was quantitated and expressed as a hypersensitivity score according to the bonitur method. This procedure identified A/J and C57BL/6 mice as high– and low–responder strains, respectively. Crosses of A/J and C57BL/6 mice should allow the characterization of mendelian factors responsible for the two extreme phenotypes identified here.
Serum Levels of Soluble Interleukin–2 Receptor at Acute Asthma Exacerbation: Relationship with Severity of Exacerbation and Bronchodilator Response
Authors: Kim Jin Tack , Kim Chang Keun , Koh Young Yull
Published: April 2003

Background: T lymphocytes play a central role in the regulation of airway inflammation in asthma, and T cell activation appears to be a characteristic feature of acute asthma. It is not clear, however, whether this is proportional to the severity of acute asthma and is directly related to airway inflammation relevant to airflow obstruction during acute asthma. It is presumed that the extent to which bronchoconstriction or inflammation contributes to airflow obstruction in acute asthma may determine responsiveness to bronchodilator therapy. Methods: Fifty asthmatic children who visited the emergency room due to acute exacerbation were studied. Serum levels of soluble interleukin–2 receptor (sIL–2R), a marker of T cell activation, were measured at the time of acute exacerbation and of clinical remission. At acute exacerbation, FEV1 was assessed before and after the administration of aerosolized salbutamol. Results: The mean (±SD) serum level of sIL–2R at acute exacerbation (854±248 U/ml) was significantly higher (p<0.01) than at clinical remission (676±211 U/ml). It correlated positively with the severity of exacerbation (r = 0.47, p<0.01), but showed no significant relationship with bronchodilator response (r = –0.17, p = 0.20). Conclusion: A higher serum level of sIL–2R at acute exacerbation was associated with more severe exacerbation but not with lower bronchodilator response. These findings suggest that in the context of acute asthma, T cell activation is proportional to disease activity, but its relationship to airway inflammation relevant to the genesis of airflow obstruction remains obscure.
Effect of Interleukin–3, Interleukin 5 and Hyaluronic Acid on Cultured Eosinophils Derived from Human Umbilical Cord Blood Mononuclear Cells
Authors: Ohashi Hiroshi , Takei Masao , Ide Youichi , Ishii Hiromi , Kita Hirohito , Gleich Gerald J. , Ishikawa Masaharu , Fukamachi Hiromi
Published: April 2003

Background: Several studies have shown that cultured eosinophils can be generated from human umbilical cord blood mononuclear cells (UCMC) in the presence of interleukin (IL)–3 and IL–5 in vitro. Other reports have indicated that cellular adhesion to hyaluronic acid (HA) enhances the proliferation of cultured eosinophils derived from CD34+ cells purified from UCMC. The aim of this study was to obtain large numbers of mature eosinophils from UCMC using IL–3, IL–5 and HA, and to investigate their functions. Methods: We examined several combinations of IL–3 and IL–5 and their effect on eosinophil development from UCMC in HA–coated on non–coated flasks. We also examined whether cultured eosinophils degranulated eosinophil–derived neurotoxin (EDN) induced by secretory immunoglobulin A conjugated to sepharose beads (sIgA–beads) and responded to eotaxin. Results: Culture with HA–coated flasks for 35 days (in the presence of IL–3 and IL–5, with IL–3 omitted after day 14 of culture) caused a 11.2–fold augmentation in the proliferation of UCMC. On day 35 of the culture, 98% of cultured cells were eosinophils judging from May–Grünwald and Giemsa staining and transmission electron micrographs. The EDN content of the cultured eosinophils on day 35 was 156 ng/105 cells. Cultured eosinophils degranulated EDN induced by sIgA–beads and responded to eotaxin by chemotaxis and intracellular Ca2+ mobilization. Conclusion: We found a useful culture system to obtain large numbers of eosinophils derived from UCMC, which may facilitate the investigation of eosinophil function, since there was no significant difference in response to sIgA–beads and eotaxin between cultured and peripheral eosinophils.
Regulation of IL–5 and IL–5 Receptor Expression in the Bone Marrow of Allergic Asthmatics
Authors: Denburg Judah A. , Sehmi Roma , Upham John , Wood Lorna , Gauvreau Gail , O’Byrne Paul
Published: April 2003

Background: Following consistent demonstrations of the clinical relevance of fluctuations in eosinophil–basophil (Eo–B) progenitors in the blood of patients with a variety of allergic airway disorders, we have turned our attention recently to hemopoietic events occurring in the bone marrow of allergic asthmatic subjects, utilizing a model of airway allergen challenge. Methods: Flow–cytometric analyses of CD34/45+ progenitors for coexpression of surface α–receptor subunits for IL–3, IL–5 and GM–CSF, as well as in situ hybridization and in situ PCR methodologies to detect mRNA for IL–5 and GM–CSF in developing Eo–B in colony and liquid culture assays were employed before and after in vivo allergen challenge. Results: An early, specific upregulation of IL–5R α expression on CD34/45 progenitors was observed after allergen challenge, concomitant with the development of the late–phase asthmatic response. Protein and mRNA for both GM–CSF and IL–5 were expressed in a time–dependent manner ex vivo, in developing (β 7–integrin–positive), colony–derived Eo–B after allergen challenge in vivo. Both retinoic acid and corticosteroids were able to downregulate IL–3– and IL–5–induced expression of IL–5R on cord–blood–derived as well as HL–60 cloned Eo–B progenitors. Conclusion: These studies indicate the critical involvement of IL–5 and IL–5R in the induction of Eo–B differentiation and eosinophilic airway inflammation in allergic asthmatics, and point to these events as potential targets for long–term therapy of atopic disease.
Linked Suppression in Peripheral T Cell Tolerance to the House Dust Mite Derived Allergen Der p 1
Authors: Hoyne Gerard F. , Dallman Margaret J. , Lamb Jonathan R.
Published: April 2003

Background: Peripheral tolerance is required to maintain balance within the immune system. A feature of peripheral tolerance is linked suppression, in which tolerance induced to a single T cell epitope inhibits the response to all epitopes in the same protein. It is suggested that this phenomenon is mediated by regulatory T cells through either the activity of immunopressive cytokines or direct cell contact. In previous experiments we failed to detect inhibitory cytokines when T cells from mice rendered tolerant by intranasal delivery of the immunodominant peptide of Der p 1 (p 1, 110–131) were restimulated with peptide in vitro. Therefore, the aim of this study was to determine if cognate interactions between T cells mediated by Notch/Delta signaling induce and maintain peripheral T cell tolerance. Methods: Using in situ hybridization and viral mediated gene transfer, the expression and function of Delta1 were investigated in a murine model of T cell tolerance to Der p 1 in vivo. Results: Delta1 expression is increased on peripheral T cells during the induction of tolerance with high–dose peptide delivered intranasally and when tolerant animals are rechallenged under immunogenic conditions. Peptide p 1, 110–131–specific CD4+ T cells transfected with Delta1 inhibited the response of antigen–primed T cells and induced linked suppression. Conclusions: High–dose peptide delivered intranasally induces transient expression of Delta 1 on inhibitory CD4+ T cells. Ligation of the Notch1 receptor on neighbouring T cells by Delta1+ regulatory T cells inhibits clonal expansion of the former and mediates linked suppression.
Neural Hyperresponsiveness and Nerve Growth Factor in Allergic Rhinitis
Authors: Sanico A.M. , Koliatsos V.E. , Stanisz A.M. , Bienenstock J. , Togias A.
Published: April 2003

Background: In allergic rhinitis, symptoms are triggered not only by allergens but also by environmental irritants. Hereinafter we address the hypothesis that this is reflective of increased responsiveness of the neural apparatus which, in turn, may be attributable to upregulation of nerve growth factor (NGF) in this disease. Methods: We compared subjects with active allergic rhinitis and healthy volunteers in terms of sensitivity and/or magnitude of three nerve–mediated responses, namely (1) the sneezing reflex induced by histamine, (2) the central or nasonasal reflex depicted by contralateral secretions induced by unilateral nasal challenge with capsaicin, and (3) the axonal reflex depicted by plasma extravasation upon capsaicin challenge. We have also measured NGF levels in nasal lavage fluids at baseline and with allergen provocation in rhinitis and healthy subjects. Results: Compared to healthy individuals, subjects with active allergic rhinitis were found to have (1) significantly greater sensitivity and reactivity of the sneezing reflex, (2) significantly greater secretory responsiveness to sensory nerve stimulation, and (3) significantly greater plasma extravasation indicated by albumin leakage following capsaicin nasal challenge. We also found that subjects with active allergic rhinitis have significantly greater baseline levels of NGF in nasal lavage fluids compared to their healthy counterparts, and that these levels can be increased by allergen nasal provocation. Conclusion: The responsiveness of the neural apparatus of the nose is significantly greater in patients with active allergic rhinitis. The increased presence of NGF in the nasal mucosa of these patients supports the hypothesis that this neurotrophin may be implicated in neural hyperresponsiveness.
IgE–Mediated Reactions to Autoantigens in Allergic Diseases
Authors: Appenzeller Ulrich , Meyer Christina , Menz Günter , Blaser Kurt , Crameri Reto
Published: April 2003

Background: There is increasing evidence that IgE–mediated reactivity against autoantigens could play a role in the pathology of severe chronic atopic diseases. Methods: Human mitochondrial managenese superoxide dismutase (MnSOD), acidic ribosomal P2 protein and cyclophilins were cloned by PCR, expressed as inclusion body proteins in Escherichia coli, purified by Ni2+–chelate affinity chromatography, refolded in vitro and used for further experiments. Results: Human MnSOD, P2 ribosomal protein and cyclophilins respectively, show sequence identity and similarity of >50% and 70% to the corresponding allergens of Aspergillus fumigatus. The human proteins were able to bind IgE from sera of individuals sensitized to the mould proteins and to elicit strong skin reactions of the immediate type in these individuals. Conclusions: These observations provide evidence for autoreactivity to human proteins in allergic persons chronically sensitized to environmental allergens from A. fumigatus which share a high degree of sequence homology to the corresponding human proteins.
Cloning of the Patatin–Like Latex Allergen Hev b 7, Its Expression in the Yeast Pichia pastoris and Its Immunological Characterization
Authors: Breiteneder Heimo , Sowka Slawomir , Wagner Stefan , Krebitz Monika , Hafner Christine , Kinaciyan Tamar , Yeang Hong Yeet , Scheiner Otto
Published: April 2003

The 43–kD latex allergen Hev b 7 was purified from the latex of Hevea brasiliensis and identified by N–terminal and internal peptide sequences as highly homologous to patatins. Patatins are storage proteins encoded by a multigene family found in plants such as potato and tomato. We have obtained a cDNA clone coding for a cytoplasmic form of Hev b 7. The recombinant protein was expressed in the methylotrophic yeast Pichia pastoris at 10 mg/l culture supernatant. Both natural Hev b 7 and rHev b 7 were recognized by IgE in 11% of the latex–allergic patients. rHev b 7 inhibited binding to its counterpart in natural rubber latex extracts. Purified rHev b 7 used at concentrations of 10 μg/ml in skin prick tests produced wheal–and–flare reactions of sizes equal to those produced by nHev b 7. Furthermore, we were able to show that rHev b 7 possessed esterase activity. A plant expression system for the production of larger quantities of recombinant latex allergens as an alternative to the preparation from H. brasiliensis sap is discussed.
IL–10 Induces a Th2 Cell Tolerance in Allergic Asthma
Authors: Adachi Mitsuru , Oda Naruhito , Kokubu Fumio , Minoguchi Kenji
Published: April 2003

Background: Interleukin (IL)–10 induces a long–term antigen–specific anergy in human CD4+ T cells. Methods: Peripheral blood mononuclear cells (PBMCs) were isolated from house dust mite (Dermatophagoides farinae, Der f)–sensitized asthmatic patients. PBMCs were stimulated with Der f antigen for 72 h immediately after purification or after 48 h of resting culture with medium, and IL–10 and IL–5 in the culture supernatant were measured. PBMCs were also stimulated with Der f antigen for 7 days either immediately after purification or after 48 h of resting culture, after which cells were collected. Secondary proliferative responses of these cells to stimulation for 3 days with Der f antigen and mitomycin C–treated PBMCs as antigen–presenting cells or with phorbol myristate acetate (PMA) plus calcium ionophore were investigated. Results: Stimulation of PBMCs with Der f antigen immediately after purification significantly increased the proliferative response and IL–5 production. Stimulation of PBMCs with Der f antigen after resting culture with medium alone for 48 h significantly decreased IL–5 production and markedly increased IL–10 production. Although activation of cells with Der f antigen immediately after purification significantly increased secondary proliferative responses, stimulation after 48 h of resting culture failed to increase secondary proliferative responses. However, proliferation recovered when cells were activated with PMA plus calcium ionophore. Conclusion: These results suggest that antigen–specific Th2 cells are anergized by IL–10 and that Th2 cell tolerance may suppress eosinophilic inflammation in allergic asthma.
Environmental Risk Factors for Respiratory and Skin Atopy: Results from Epidemiological Studies in Former East and West Germany
Authors: Ring J. , Krämer U. , Schäfer T. , Abeck D. , Vieluf D. , Behrendt H.
Published: April 2003

The fall of the Berlin wall in 1989 offered the unique opportunity to compare populations of a similar genetic and geographic background which had been living under quite different environmental exposure conditions for over 40 years. Since 1990 comparative epidemiological studies were performed between various regions in former East and West Germany with yearly questionnaires and 3 years’ physical, dermatological, allergological and exposure examination in a total of about 30,000 preschool children. There were striking differences between the various German regions with higher prevalence rates of respiratory atopy (hay fever, asthma) and atopic sensitization (prick test, RAST) in West Germany, while atopic eczema was significantly higher in East Germany (17.5 vs. 11.4% in West Germany). Total serum IgE levels were markedly higher in children in the east of Germany, similarly to the prevalence of parasitic infestation (questionnaire data as well as positive ascaris RAST). In multivariate logistic regression analysis the following factors were significantly associated with atopic eczema: animal contact (odds ratio, OR 2.9), animal furs in the bedroom (OR 2.2), use of gas without ventilation (OR 1.7) and living near road with heavy traffic (OR 1.7). Furthermore, socioeconomic factors measured as parental educational status (university vs. elementary school) were significantly associated (OR 2.3) with atopy. Respiratory tract infections and irritant responses decreased together with decreasing SO2 and suspended particulate air pollution in East Germany from 1991 to 1997. It is concluded that environmental factors from the physical, chemical, biological, and psychological environment (characteristic of a ‘modern’ or ‘western’ society) do influence the development of atopic sensitization and disease.
Bacterial DNA and CpG–Containing Oligodeoxynucleotides Activate Cutaneous Dendritic Cells and Induce IL–12 Production: Implications for the Augmentation of Th1 Responses
Authors: Jakob Thilo , Walker Patricia S. , Krieg Arthur M. , von Stebut Esther , Udey Mark C. , Vogel Jonathan C.
Published: April 2003

Background: Unmethylated CpG sequences in bacterial DNA act as adjuvants selectively inducing Th1 predominant immune responses during genetic vaccination or when used in conjunction with protein Ag. The precise mechanism of this adjuvant effect is unknown. Because dendritic cells (DC) are thought to be crucially involved in T cell priming and Th1/Th2 education during vaccination via skin, we characterized the effects of bacterial DNA and CpG–containing oligodeoxynucleotides (CpG ODN) on cutaneous DC. Methods and Results: Stimulation with CpG ODN 1826 (6 μg/ml) induced activation of immature Langerhans cell (LC)–like DC as determined by an increased expression of MHC class II and costimulatory molecules, loss of E–cadherin–mediated adhesion and increased ability to stimulate allogeneic T cells. Composition–matched control ODN 1911 lacking CpG sequences at equal concentrations was without effect. In comparison to LPS and ODN 1911, CpG ODN 1826 selectively stimulated DC to release large amounts of IL–12 (p40) and little IL–6 or TNF–α within 18 h and detectable levels of IL–12 p70 within 72 h. Stimulation with Escherichia coli DNA, but not calf thymus DNA, similarly induced DC maturation and IL–12 p40 production. Injection of CpG ODN into murine dermis induced enhanced expression of MHC class II and CD86 by LC in the overlying epidermis and intracytoplasmic IL–12 p40 accumulation in a subpopulation of activated LC. Conclusion: Bacterial DNA and CpG ODN stimulate DC in vitro and in vivo and may preferentially elicit Th1–predominant immune responses because they can activate and mobilize DC, inducing them to produce IL–12.
Serologic and Clinical Response to Treatment of Systemic Vasculitis and Associated Autoimmune Disease with Intravenous Immunoglobulin
Authors: Levy Yair , Sherer Yaniv , George Jacob , Langevitz Pnina , Ahmed Alaa , Bar-Dayan Yaron , Fabbrizzi Fabrizio , Terryberry Jeff , Peter James , Shoenfeld Yehuda
Published: April 2003

Background: Autoimmune vasculitides cannot always be controlled by steroids and immunosuppressive drugs. Intravenous immunoglobulin (IVIg) treatment was found beneficial in several vasculitides including systemic and organ–specific diseases. In this article we tested whether the beneficial clinical response of IVIg treatment in vasculitides was accompanied by a decrease in vasculitis–associated autoantibody levels. Methods: Ten patients diagnosed as having vasculitis were treated with high–dose (2 g/kg) IVIg monthly, in a 5–day schedule. In all the patients, other therapeutic measures failed to control disease progression prior to IVIg treatment. Each patient received between 1 and 6 treatment courses. All patients were evaluated for the levels of 5 autoantibodies (Abs) related to vasculitis before and after each treatment course. Results: In 6 out of the 10 patients, a beneficial clinical response followed IVIg treatment. Moreover, no treatment–related adverse effects were observed in any of the patients. Anti–myeloperoxidase antibodies and cytoplasmic–antineutrophil cytoplasmic antibodies levels decreased concomitantly with the clinical improvement observed in the patients with Churg–Strauss vasculitis and Wegener’s granulomatosis, respectively. Levels of cytoplasmic–antineutrophil cytoplasmic antibodies (ANCA) with specificity for bacteridial/permeability–increasing protein and human lysosomal–associated membrane protein increased after each treatment course, but returned to normal values before the following one. Conclusions: When other therapeutic measures, such as immunosuppressive therapy, fails to control disease manifestations in patients with vasculitides, IVIg is a possible effective intervention method with a high response rate. IVIg probably exerted its effects on disease progression via different mechanisms. Among these mechanisms, a decrease in relevant Ab levels is often found (probably by anti–idiotypes in IVIg), and thus ANCA levels are expected to associate with disease activity.
Selective Cloning of Peanut Allergens, Including Profilin and 2S Albumins, by Phage Display Technology
Authors: Kleber-Janke Tamara , Crameri Reto , Appenzeller Ulrich , Schlaak Max , Becker Wolf-Meinhard
Published: April 2003

Background: Peanut kernels contain many allergens able to elicit IgE–mediated type 1 allergic reactions in sensitized individuals. Sera from sensitized patients recognize variable patterns of IgE–binding proteins. The identification of the IgE–binding proteins of peanut extract would faciliate improvement of diagnostic and immunotherapeutic approaches as well as development of sensitive test systems for the detection of hidden peanut allergens present as additives in various industrial food products and the investigation of their stability during processing of food products. Methods: We applied the pJuFo cloning system based on the phage surface display of functional cDNA expression products to clone cDNAs encoding peanut allergens. Sera (n = 40) of peanut–allergic individuals were selected according to case history, radioallergosorbent test and immunoblot analysis to demonstrate IgE binding towards the newly identified recombinant allergens. Results: In addition to the known allergens Ara h 1 and Ara h 2 we were able to identify four allergens with estimated molecular weights of 36, 16, 14.5 and 14 kDa. Three of them formally termed Ara h 4, Ara h6 and Ara h 7 show significant sequence similarities to the family of seed storage proteins and the fourth (Ara h 5) corresponds to the well–known plant allergen profilin. Immunoblotting of the six expressed recombinant allergens with 40 patients sera shows 14 individual recognition patterns and the following frequency of specific IgE binding: Ara h 1 was recognized by 65%, Ara h 2 by 85%, Ara h 4 by 53%, Ara h 5 by 13%, Ara h 6 by 38% and Ara h 7 by 43% of the selected sera. Conclusions: All of the selected peanut–positive sera can detect at least one of the six identified recombinant allergens which can be used to establish individual patients’ reactivity profiles. A comparison of these profiles with the clinical data will possibly allow a further insight into the relationship between clinical severity of the symptoms and specific IgE levels towards the six peanut allergens.
Time Course of Pharmacological Modulation of Peak Eosinophilic Airway Inflammation after Mite Challenge in Guinea Pigs: A Therapeutic Approach
Authors: Hsiue Tzuen-Ren , Lei Huan-Yao , Hsieh Ai-Ling , Chang Han-Yu , Chen Chang-Wen
Published: April 2003

Background: It is well known that eosinophilic airway inflammation develops after allergen challenge in sensitized humans and animals. However, the detailed time course of suppression of early eosinophilic airway inflammation by pharmacological agents given just after challenge has not been discussed. Therefore, we aimed to evaluate the time course relationship of the suppression of peak eosinophilia by anti–cytokines and pharmacological agents given several hours after the aerosol challenge by a therapeutic approach. Methods: We used crude mite extract as an allergen to create a sensitization and inhalation challenge, and performed bronchoalveolar lavages (BAL) after the inhalation challenge to observe the degree of eosinophilic airway inflammation in guinea pigs. Various anti–cytokines (anti–IL–3 and anti–IL–5) and pharmacological agents (dexamethasone, theophylline, and roxithromycin) were given within several hours after the acute aeorosol challenge to evaluate the suppressive effect on peak eosinophilia in BAL fluid, which occurred 24 h after the challenge. Results: Our results show that anti–IL–5 and dexamethasone, given within 4 and 8 h after the inhalation challenge, respectively, inhibit the acute allergen–induced peak eosinophilia in BAL fluid. However, anti–IL–3, theophylline, and roxithromycin had no effect on peak eosinophilic airway inflammation after challenge. Conclusion: These observations suggest that several hours are needed to complete the process of cytokine–induced recruitment of eosinophils from the blood to the airways after acute allergen challenge. This may be the optimal time to administer anti–cytokines and dexamethasone to attenuate the subsequent eosinophilic airway inflammation after acute allergen–induced asthmatic attacks.
Natural Rubber Latex Allergy Is Not a Cause of Sudden Infant Death
Authors: Gutgesell Carsten , Seubert Sigrid , Saternus Klaus-Steffen , Fuchs Thomas
Published: April 2003

Background: The causes for sudden infant death (SID) remain unclear. As infants can become sensitized to NRL allergens by pacifiers and latex mattresses, we tried to establish whether there is a relationship between SID and natural rubber latex (NRL) allergy. Methods: We determined NRL–specific IgE concentrations in 112 unselected cases of SID by the CAP–FEIA method. Results: NRL–specific IgE could be detected only in 1 sample (0.64 kU/l; CAP class 1). Conclusions: We conclude that NRL allergy is not a cause of SID.
Recent Advances in Mastocytosis Research
Authors: Valent Peter , Escribano Luis , Parwaresch Reza M. , Schemmel Verena , Schwartz Lawrence B. , Sotlar Karl , Sperr Wolfgang R. , Horny Hans-Peter
Published: April 2003

The term mastocytosis denotes a heterogenous group of disorders characterized by abnormal growth and accumulation of mast cells in one or more organs. Cutaneous and systemic variants of the disease have been described. Mast cell disorders have also been categorized according to other aspects, such as family history, age, course of disease, or presence of a concomitant myeloid neoplasm. However, so far, generally accepted disease criteria are missing. Recently, a number of diagnostic (disease–related) markers have been identified in mastocytosis research. These include the mast cell enzyme tryptase, CD2, and mast cell growth factor receptor c–kit (CD117). Several gain–of–function–mutations in the kinase domain of c–kit appear to occur in mastocytosis supporting the clonal (neoplastic) nature of the disease. Also, certain point mutations appear to be associated with distinct variants of mastocytosis, i.e. Asp–816→Val with a subset of sporadic persistent (systemic) mastocytosis (mostly adults), and Gly–839→Lys with (a subset of) typical pediatric (mostly cutaneous) mastocytosis. Another potential indicator of mast cell neoplasm is the T–/NK–cell–associated marker CD2. This antigen (LFA–2) is abnormally expressed on neoplastic mast cells in cases of systemic mastocytosis or mast cell leukemia, but not found on normal mast cells. The mast cell enzyme tryptase is increasingly used as a serum– and immunohistochemical marker to estimate the actual spread of disease (burden of neoplastic mast cells). The clinical significance of novel mastocytosis markers is currently under investigation. First results indicate that they may be useful to define reliable criteria for the delineation of the disease.
HgCl2–Induced Human Lymphocyte Activation in vitro: A Superantigenic Mechanism?
Authors: Loftenius Annika , Ekstrand Jan , Möller Erna
Published: April 2003

Mercuric chloride (HgCl2) has been proposed to be a mitogen for human blood lymphocytes in vitro. In our previous study, we demonstrated that HgCl2 preferentially stimulates the CD4+ T cell subset to blast transformation and DNA synthesis and that the reaction is dependent on CD14+ accessory cells. In order to characterise the responding cells further and to elucidate the mechanism of T cell activation, the T cell receptor (TCR) Vβ chain expression of the blast–transformed cells was analysed by monoclonal antibodies and flow cytometry. The 22 TCR–Vβ–specific antibodies used were found to react with 55–80% of the naive CD4+ and CD8+ blood T cells from the different donors. Six to 18% of the lymphocytes, mainly CD4+ T cells, were blast transformed after addition of HgCl2. The distribution of the lymphoblasts carrying certain TCR Vβ chains were skewed, and 15–40% expressed the TCR Vβ2 chain. Furthermore, if cells were pretreated for 5 days with HgCl2, whereafter recombinant interleukin–2 in fresh medium was added, the TCR Vβ7+ T cell subset was also stimulated to blast transformation. The superantigen staphyloccal enterotoxin B, as a control, induced blast transformation in 10–26% of the lymphocytes, mainly CD4+ T cells, which were, as expected, positive for Vβ3, Vβ12, Vβ14 or Vβ17. We conclude that HgCl2 has characteristics of a superantigen, activating the human lymphocytes in a Vβ–chain–selective manner in vitro.
Clinical Significance of IgE–Binding Activity to Enzymatic Digests of Ovomucoid in the Diagnosis and the Prediction of the Outgrowing of Egg White Hypersensitivity
Authors: Urisu Atsuo , Yamada Kazue , Tokuda Reiko , Ando Hitoshi , Wada Eiko , Kondo Yasuto , Morita Yutaka
Published: April 2003

Background: We frequently encounter subjects without overt symptoms despite high IgE antibodies to egg white and its components. The measurements of these antibodies are not necessarily efficient for the diagnosis or the prediction of the outcome of egg allergy in children. Methods: Specific IgE antibodies to egg white and its components, including ovomucoid, ovalbumin, ovotransferrin and lysozyme, were measured by direct RAST assays. IgE–binding activity to ovomucoid degraded by pepsin, trypsin and chymotrypsin was examined by RAST inhibition. Thirty subjects were divided into two groups with positive (n=18; mean age ± SD = 42 ±25 months) and negative (n=12; mean age ± SD = 48 ±31 months) oral challenge tests with egg white antigens. The individuals with positive results to the first challenge tests were given the second provocation tests at mean intervals of 32 months. IgE–binding activity of the sera collected on the first challenge to these ovomucoid fragments was compared between subjects with positive and negative reactions to the follow–up challenge tests. Results: There were no significant differences in IgE antibody titers to egg white and its components between the positive and negative groups at the first and the second challenge tests. IgE–binding activity to ovomucoid digests after treatments with pepsin (p = 0.000008) and trypsin (p=0.037), except chymotrypsin (p=0.062), were significantly higher in subjects with positive challenge tests than in those with negative results. The difference was most remarkable in the IgE–binding to pepsin digests; the average concentrations (mean – SD and mean + SD) needed for 50% RAST inhibition in the positive group and in the negative group were 2.6 μg/ml (0.3 and 25) and 94.2 μg/ml (24.7 and 358.7), respectively. A significant difference was still observed in the inhibition tests using filtrates of pepsin digests with a membrane with MW 10,000 (p=0.014) and 3,000 (p=0.042) of cutoff. The concentration (mean= 0.8, mean – SD=0.2, mean + SD=3.4; μg/ml) of pepsin–treated ovomucoid required for 50% RAST inhibition in the subjects with positive second challenge results was significantly (p=0.033) lower than that (mean=6.8, mean–SD=0.6, mean + SD=73.9) of the negative group. Conclusion: IgE–binding activity to pepsin–digested ovomucoid was of diagnostic value to distinguish the challenge–positive subjects from the negative subjects. Subjects with high IgE–binding activity to pepsin–treated ovomucoid are unlikely to outgrow egg white allergy.
Distinct Serum Cytokine Levels in Drug– and Measles–Induced Exanthema
Authors: Hari Yvonne , Urwyler Adrian , Hurni Michael , Yawalkar Nikhil , Dahinden Clemens , Wendland Thomas , Braathen Lasse R. , Matter Lucas , Pichler Werner J.
Published: April 2003

Background: Macular or maculopapular skin reactions are frequent events in drug allergy as well as in viral infections. Clinically, the differentiation may be difficult in the absence of a clear relationship to drug intake or failure to detect virus–specific antibodies of the IgM class. Studies on drug–specific T cell lines and T cell clones isolated from drug–allergic patients have suggested that these cells may represent a significant source of IL–5. On the other hand, viral infections are frequently associated with elevated IFN–γ levels. Objective: Determination of serum–cytokine levels to differentiate between drug– and virally induced skin eruptions. Patients: 18 patients suffering from acute drug allergy and 19 patients with acute measles, rubella or parvovirus infection. Measurements: Cytokine–ELISA (IL–5, IL–4 and IFN–γ) of sera collected during acute drug allergy or during acute measles, rubella or parvovirus infection. Results: In 12/18 patients with drug allergy, IL–5 and/or IL–4 were elevated. A significant correlation (rSpearman = 0.84) between IL–5 serum levels and eosinophil counts in the blood was found. No correlation was detected between IL–4 and blood eosinophilia or between IL–4 and IL–5 levels. After remission, IL–5 and IL–4 decreased to undetectable levels. IFN–γ on the other hand was not measurable in patients with drug allergy while elevated IFN–γ serum levels were detected in 17/19 patients with measles, rubella or parvovirus infection; 2 patients with acute virus infection had elevated IL–5, and/or IL–4 and IFN–γ levels. Conclusion: These data underline the distinct pathogenesis of these morphologically similar exanthemas and suggest that the combined analysis of eosinophilia in the blood, IL–4 and IFN–γ might help in differentiating skin eruptions.
Vascular–Endothelial Cadherin (CD144)– but Not PECAM–1 (CD31)–Based Cell–to–Cell Contacts Convey the Maintenance of a Quiescent Endothelial Monolayer
Authors: Halama Thomas , Staffler Günther , Hoch Susanne , Stockinger Hannes , Wolff Klaus , Petzelbauer Peter
Published: April 2003

Background: In vivo, all blood vessels are lined by a single layer of flattened noncycling endothelial cells. We tested the hypothesis that the maintenance of such a quiescent endothelial monolayer depends on homotypic contacts between not yet defined growth–inhibitory molecules located at interendothelial junctions. Methods: ECV304 cells, which lack endogenous vascular endothelial cadherin (VE cadherin) or CD31 expression, were transfected with cDNA encoding for the respective proteins or with the empty vector. Results: In VE cadherin transfectants, β–catenin was targeted to junctional regions and the F–actin–based cytoskeleton formed parallel bundles reaching from one cell border to the other. In contrast, in CD31 transfectants and in empty vector cells, β–catenin was dispersed throughout the cytoplasm, and F–actin formed short, plump and criss–cross bundles. On a two–dimensional plastic matrix, both, VE cadherin and CD31 transfectants formed clusters of polygonal cells, whereas in three–dimensional gels, only VE cadherin cells were able to form tubes. Empty vector cells grew in a fibroblast–like pattern and neither formed clusters nor tubes. Most importantly, whereas CD31 and empty vector cells grew on top of each other, formed polylayers and maintained cycling even after reaching confluence, VE cadherin cells strictly maintained a single layer of flattened cells and the numbers of cycling cells dramatically dropped after reaching a continuous monolayer. Conclusion: The insertion of VE cadherin into ECV304 cells produces a cell type which mimics endothelial growth characteristics seen in vivo.
Allergy to Mammalian Proteins: At the Borderline between Foreign and Self?
Authors: Spitzauer S.
Published: April 2003

Atopic individuals are characterized by their genetically determined tendency to mount IgE antibody responses against otherwise harmless antigens. Many of the environmental allergens (e. g., plant–, insect–, mould–derived allergens) are recognized as foreign intruders by the immune system. However, a substantial proportion of atopic individuals mount IgE responses to allergens of mammalian origin and, as recently reported, certain of them even react with human proteins (i. e., autoallergens). Through the application of molecular biological techniques for allergen characterization, the molecular nature and biological function of an increasing number of mammalian allergens have been revealed. This review provides a summary of well characterized mammalian allergens derived from hair/dander, saliva, serum and urine of animals (cat, dog, rodents, cow, horse) as well as of some recently characterized autoallergens. In addition, mammalian allergens were grouped in protein families with similar sequences and/or biological functions. The importance of cross–reactivities is exemplified for albumins which represent abundant mammalian serum proteins and highly cross–reactive allergens. Finally, possible advantages of using recombinant animal allergens for novel forms of component–resolved diagnosis and treatment of allergies to mammalian proteins are described.
Identification of Sunflower Seed IgE-Binding Proteins
Authors: Kelly Jack D. , Hlywka Jason J. , Hefle Susan L.
Published: April 2003

Background: Sunflower seed can cause severe anaphylactic reactions in some susceptible individuals. It is conceivable that the 2S sunflower seed protein is an allergen based on its high degree of homology (34%) with the allergenic mature 2S albumin protein of the Brazil nut. The first step in determining the allergenicity of sunflower seed proteins is to identify IgE-binding proteins. Methods: Sera from sunflower seed-sensitive individuals were evaluated by radioallergosorbent test (RAST), isoelectric focusing (IEF) and sodium dodecyl sulfate-polyacrylamide gel electrophoresis immunoblotting with sunflower seed proteins. Results: Positive RAST scores (>2) were observed in 3 individuals and immunoblotting demonstrated IgE-binding to 2–7 distinct proteins ranging in size from 10 to 50 kD. Two out of 3 sera recognized two proteins between 16 and 17 kD. The lower molecular weight protein (16 kD) approximates to the prepo region of the precursor methionine-rich 2S albumin protein found in sunflower seed (SFA-8/SSA). IEF followed by immunoblotting demonstrated several IgE-binding proteins, including two proteins with isoelectric points of 5.97 and 5.3, respectively, which are consistent with the mature and immature forms of the SFA-8/SSA region. Conclusions: Sunflower seed contains several IgE-binding proteins, including regions of the high-methionine 2S albumin SFA-8/SSA.