bluuebunny/crossref_metadata_2025 huggingface dataset:
Autocrine Regulation of Eotaxin in Normal Human Bronchial Epithelial Cells
Authors: Saito Hiroshi , Shimizu Hisako , Akiyama Kazuo
Published: April 2003

Normal human bronchial epithelial cells (NHBECs) spontaneously synthesize and store chemokines including eotaxin, RANTES, or MCP-3. We also observed the constitutive expression of mRNA specific for CC chemokine receptor-3 (CCR3) in these cells. The stimulation of these cells by eotaxin (1–100 ng/ml) dose-dependently increased eotaxin production. In addition, RANTES or eotaxin-2 but not MCP-3 at the higher concentrations (100 ng/ml), also induced slight eotaxin production. Eotaxin elicited the expression of eotaxin mRNA at 6 h after stimulation. Antibody against eotaxin or CCR3, and actinomycin D inhibited these effects of eotaxin. These antibodies also significantly inhibited the ability of eotaxin to produce further eotaxin. These findings strongly suggest that eotaxin which originates from inflammatory cells or cells resident in the airway of bronchial asthmatics can induce further production of eotaxin and participate in expansion of allergic inflammation.
Inhibition of IgE-Dependent Histamine Release from Human Peripheral Blood Basophils by Humanized Fab Fragments That Recognize the Membrane Proximal Domain of the Human FcεRI α-Chain
Authors: Takai Toshiro , Yuuki Toshifumi , Ra Chisei
Published: April 2003

Background: Inhibition of the interaction between IgE and the α-chain of FcΕRI (FcΕRIα) is a straightforward strategy to develop therapeutic reagents for IgE-mediated allergic diseases. Objective: The purpose of this study is the humanization of CRA2 and/or CRA4, mouse anti-human FcΕRIα monoclonal antibodies (mAbs) which recognize the IgE-binding membrane proximal immunoglobulin-like domain of FcΕRIα. Methods: The two mAbs were humanized by CDR grafting onto human V region frameworks encoded by human germline V and J genes. The activities of the recombinant antibodies to bind FcΕRIα and inhibit IgE binding to FcΕRIα were analyzed by flow cytometry and ELISA. Human peripheral blood basophils were pretreated with the Fab fragments of the humanized CRA2 and stimulated with IgE and an anti-IgE polyclonal antibody. The released histamine was measured. Results: The humanized CRA2 had almost the same activities of binding and inhibition of IgE binding to FcΕRIα as the original mouse CRA2. Although the FcΕRI-binding activity was maintained following humanization of the CRA4 light chain V region, it was lost by the humanization of the CRA4 heavy chain V region. Pretreatment of human peripheral blood basophils with the Fab fragments of the humanized CRA2 inhibited their subsequent degranulation activated by cross-linking of the FcΕRI. Conclusion: In the humanized CRA2, all amino acid residues except CDR are replaced with the residues encoded by human germline genes. The humanization of CRA2 might be an important step in the development of immunotherapy to manipulate the IgE network in which mast cells, basophils, and various types of FcΕRIα expressing cells are involved.
Suppressive Effect of Topically Applied CX-659S, a Novel Diaminouracil Derivative, on the Contact Hypersensitivity Reaction in Various Animal Models
Authors: Goto Yuso , Inoue Yoshifumi , Tsuchiya Masami , Isobe Masakazu , Ueno Takamasa , Uchi Hiroshi , Furue Masutaka , Hayashi Hideya
Published: April 2003

Background: The T-cell-mediated contact hypersensitivity reaction (CHR) is thought to be involved in the pathogenesis of clinical cutaneous disorders including atopic dermatitis. A novel diaminouracil derivative, CX-659S, has been reported to have an inhibitory activity against picryl chloride (PC)-induced CHR when administered either orally or percutaneously. The inhibitory effect of topical CX-659S was assessed in three CHR models in the present study. In addition, to elucidate the mechanism of action of this compound, we examined the effect of CX-659S on the expression of messenger RNAs for proinflammatory cytokines after elicitation in PC models. Methods: For the in vivo evaluation of the efficacy of CX-659S, we used PC- or oxazolone-induced CHR in mice and 2,4-dinitrochlorobenzene (DNCB)-induced CHR in guinea pigs. CX-659S was topically applied immediately after the hapten challenge in each model. To assess the effect on gene expression of cytokines, we used the reverse transcriptase-polymerase chain reaction (RT-PCR), a semiquantitative technique with specific primers. Results: Topical CX-659S dose-dependently inhibited ear swelling at 24 h after the challenge in the two mouse models. This inhibitory effect was histologically confirmed in the PC model. Topically applied CX-659S also inhibited erythema and edema formation 24 h after challenge in the guinea pig model. CX-659S inhibited the expression of mRNA for proinflammatory cytokines IL-1β and TNF-α in vivo. Conclusions: Topically applied CX-659S showed significant inhibitory activities against CHR models both in mice and in guinea pigs. Inhibition profiles of CX-659S toward mRNA expression for proinflammatory cytokines corroborated these findings. CX-659S thus could be a useful therapeutic agent for allergic cutaneous disorders such as allergic contact dermatitis and atopic dermatitis.
A Perspective: Regulation of IgE Receptor-Mediated Mast Cell Responses by a LAT-Organized Plasma Membrane-Localized Signaling Complex
Authors: Rivera Juan , Arudchandran Ramachandran , Gonzalez-Espinosa Claudia , Manetz Timothy Scott , Xirasagar Sandhya
Published: April 2003

Background: To understand how the high-affinity IgE receptor (FcΕRI) communicates with downstream effectors, we focused on exploring the functional importance of the FcΕRI-mediated formation and localization of a signaling complex that contains the hematopoietic cell-specific scaffolding protein linker for activation of T cells (LAT) and the guanine nucleotide exchange factor Vav1. Methods: Using the mast cell line RBL-2H3, we explored the localization of these proteins by confocal microscopy and cell fractionation. Additionally, the mechanism of function and the importance of LAT and Vav1 to mast cells was studied in genetically disrupted mice and in mast cells derived from their bone marrow. Results: We found that LAT, Vav1 and the adapter molecule SLP-76 associated in detergent-resistant microdomains (lipid rafts) found in the plasma membrane upon FcΕRI stimulation. In the absence of LAT, mast cells showed a remarkable loss of the secretory response and reduced cytokine responses. Vav1 deficiency also affected secretion, although not to the extent of LAT deficiency, and inhibited IL-2 and IFN-γ production. LAT- and Vav1-deficient mice showed reduced blood histamine levels after a systemic anaphylaxis challenge as compared to their normal counterparts. Conclusions: The results demonstrate that LAT is a central mediator in IgE receptor signaling by regulating multiple signaling pathways that affect mast cell degranulation and cytokine production. Vav1, a component of this LAT-containing signaling complex, regulates a specific subset of these responses.
Further Characterization of FcγRII and FcγRIII Expression by Cultured Human Mast Cells
Authors: Okayama Yoshimichi , Hagaman David D. , Woolhiser Michael , Metcalfe Dean D.
Published: April 2003

Background: We have reported that resting human mast cells exhibit minimal expression for FcγRI, and that interferon-γ will upregulate this expression. The expression of FcγRII and FcγRIII by human mast cells remains to be fully examined. Methods: To investigate FcγRII and FcγRIII expression, we determined mRNA and protein expression of FcγRII and FcγRIII in human peripheral blood CD34+ derived cultured mast cells by RT-PCR and flow cytometry. The expression of FcγRII and FcγRIII in intact and permeabilized mast cells was also compared. We measured histamine release to monitor mast cell degranulation following cross-linking of FcγRII. Results: We found by RT-PCR that resting human mast cells exhibit mRNA for FcγRIIA, FcγRIIb1, FcγRIIb2 and FcγRIII but not FcγRIIC. FACS analysis of Fcγ receptors in intact versus permeabilized mast cells showed expression of FcγRII to be 42.2 ± 3.9% and this was unchanged by permeabilization. FcγRIII protein expression was minimal and this was also unchanged by permeabilization. Aggregation of FcγRII on human mast cells led to no significant degranulation as evidenced by histamine release. Conclusions: In addition to FcγRI expression, human mast cells express FcγRIIA, FcγRIIb1, FcγRIIb2 and FcγRIII mRNA, and significant surface expression of FcγRII. Aggregation of FcγRII on cultured human mast cells in this model was not followed by histamine release.
Mechanism of IL-10-Induced T Cell Inactivation in Allergic Inflammation and Normal Response to Allergens
Authors: Akdis Cezmi  A. , Joss Andrea , Akdis Mübeccel , Blaser Kurt
Published: April 2003

Background: Induction of specific unresponsiveness (tolerance/anergy) in peripheral T cells and recovery by cytokines from the tissue microenvironment represent two key steps in specific immunotherapy (SIT) with whole allergen or antigenic T cell peptides. Methods: Antigen-specific T cell responses and molecular mechanisms of T cell inactivation were investigated during conventional SIT, T cell epitope peptide immunotherapy and natural exposure to bee venom in allergic and hyperimmune individuals. Results: T cell unresponsiveness, initiated by autocrine action of IL-10, is characterized by suppressed proliferative and cytokine responses. The unresponsive T cells can be reactivated by different cytokines that may mimic the microenvironmental cytokine influence. IL-10 initiates peripheral tolerance by blocking the CD28 costimulatory signal in T cells. Coprecipitation experiments reveal that upon stimulation CD28 and IL-10 receptor are physically associated in T cells. Accordingly, IL-10 binding to its receptor inhibits CD28 tyrosine phosphorylation, the initial step of the CD28 signaling pathway. This leads to inhibition of phosphatidylinositol 3-kinase p85 binding to CD28. IL-10 only affects T cells that receive a stimulation with low numbers of triggered T cell receptors and that require costimulatory signals by CD28. Conclusion: These data demonstrate the pivotal role of autocrine IL-10 and the interaction of its receptor with CD28 in the induction of T cell tolerance as an immunoregulatory mechanism controlling antigen-specific T cell responses.
T Cytotoxic 1 and T Cytotoxic 2 CD8 T Cells Both Inhibit IgE Responses
Authors: Thomas Matthew J. , MacAry Paul A. , Noble Alistair , Askenase Philip W. , Kemeny David M.
Published: April 2003

Background: It is well recognized that CD8 T cells inhibit IgE responses. In this study, we investigated the mechanism of CD8 T cell-mediated IgE suppression by comparing the capacity of T cytotoxic 1 (Tc1) and T cytotoxic 2 (Tc2) CD8 T cells to inhibit IgE responses to ovalbumin (OVA). Methods: Tc1 and Tc2 CD8 T cells were generated from OVA257–264-specific Vβ5.2 T cell receptor (TcR) transgenic mice by stimulation with anti-CD3 and anti-CD28 under Tc1 and Tc2 polarizing conditions. Tc1 and Tc2 Vβ5.2 TcR CD8 T cells (106) were adoptively transferred to syngeneic mice, and following immunization with 100 µg of OVA/alum, serum IgE antibodies were measured by passive cutaneous anaphylaxis and expressed as the highest dilution that gave a detectable skin response. Results: Both Tc1 and Tc2 CD8 T cells from OT-I mice inhibited IgE. Conclusion: Both Tc1 and Tc2 CD8 T cells promote Th1 immunity and inhibit IgE responses. This process appears to be independent of CD8 T cell-derived IFN-γ, as both Tc2 (IFN-γ–) and Tc1 (IFN-γ+) CD8 T cells inhibited IgE.
Allergic Sensitization and Allergen Exposure during Pregnancy Favor the Development of Atopy in the Neonate
Authors: Herz Udo , Joachim Ricarda , Ahrens Birgit , Scheffold Alexander , Radbruch Andreas , Renz Harald
Published: April 2003

Background: Several studies have considered that the in utero environment plays an important role in the onset of the allergic phenotype. We assessed whether allergic sensitization and allergen exposure during pregnancy favor the postnatal onset of allergy in the neonate. Methods: BALB/c mice were sensitized to ovalbumin (OVA) before mating followed by allergen aerosol exposure during pregnancy. T and B cell responses in offspring were followed up until day 60 postpartum. At the age of 4 weeks offspring were exposed to a heterologous antigen, β-lactoglobulin (BLG). Results: Pregnant mice developed immediate hypersensitivity responses and Th-2/ Th-0 immunity following allergen aerosol exposure. At birth, T cells from offspring of nonsensitized BALB/c mice were characterized by an impaired IFN-γ production, which was lowered even further in offspring of OVA-sensitized BALB/c mice. Offspring of OVA-sensitized BALB/c mice responded with immediate-type cutaneous hypersensitivity reactions to OVA which could be related to the pre- and postnatal transfer of maternal OVA-specific IgG1 antibodies. After exposure to BLG, offspring of OVA-sensitized BALB/c mice developed an accelerated Th-2-driven immune response compared to offspring from nonsensitized BALB/c mice as indicated by enhanced anti-BLG IgG1 antibody production and increased numbers of positive immediate-type cutaneous hypersensitivity reactions to BLG. Conclusion: Our data suggest that Th-2/Th-0 immunity present during pregnancy has a decisive impact on shaping the Th-1/Th-2 T cell profile in response to postnatal allergen exposure.
Regulation of IL-5 Receptor on Eosinophil Progenitors in Allergic Inflammation: Role of Retinoic Acid
Authors: Denburg Judah A. , Sehmi Roma , Upham John
Published: April 2003

We and others have shown that IL-5 plays a central role in eosinophil and basophil differentiation, exerting its effects through the IL-5 receptor (IL-5R). Little is currently known concerning regulation of IL-5Rα gene transcription in the context of commitment of hemopoietic progenitor cells to the eosinophil and basophil lineages; recent studies have indicated that IL-5 itself can regulate IL-5Rα expression on mature eosinophils. We now provide evidence to indicate that IL-5 can upregulate IL-5Rα on bone marrow CD34+ progenitors in vitro, as we have demonstrated in vivo in atopic asthmatics. Given that all-trans retinoic acid (ATRA) is known to modulate some granulopoiesis, causing neutrophilic differentiation, we examined the effects of ATRA on eosinophil/basophil differentiation and IL-5Rα expression. In cultures of normal human bone marrow, ATRA selectively suppressed eosinophil/basophil differentiation. Similarly, ATRA inhibited eosinophil/basophil differentiation of cord blood CD34+ cells, while neutrophil differentiation proceeded without impediment. Most importantly, these effects of ATRA on CD34+ cells were associated with selective, dose-dependent inhibition of membrane-bound IL-5Rα, upregulation of soluble IL-5Rα transcription, but no change in GM-CSF receptor expression. These findings indicate that retinoids can differentially regulate membrane and soluble isoforms of IL-5Rα, and that these effects have functional consequences in vitro on eosinophil and basophil differentiation. ATRA may be of therapeutic benefit in allergic inflammatory disorders in which eosinophil differentiation and membrane-bound IL-5R are upregulated.
Antigen-Induced Airway Migration of a T Helper 2 Clone
Authors: Kaminuma Osamu , Fujimura Hisako , Fushimi Keiko , Ogawa Koji , Nakata Aya , Kikkawa Hideo , Okudaira Hirokazu , Mori Akio
Published: April 2003

Background: The crucial role of T helper (Th) 2 cells in allergic eosinophilic inflammation has been established. Methods: To determine the dynamics of Th2 cells in airway eosinophilic inflammation, ovalbumin-reactive murine Th2 clones were infused into unprimed mice. Results: The infused T cells were activated and migrated into the bronchial submucosa upon provocation with the relevant inhaled antigen. The T cell migration was followed by massive eosinophilic infiltration, edema, swelling of epithelial cells in the lung and a significant increase in bronchial responsiveness to acetylcholine. Conclusion: Clonal Th2 cells have the potential to accumulate and induce eosinophilic inflammation in the lung upon antigenic stimulation.
Pathogenesis of Drug-Induced Exanthema
Authors: Yawalkar N. , Pichler W.J.
Published: April 2003

Background: In vitro data derived from drug-specific T cell clones have revealed that heterogeneous T cell subsets with distinct phenotypes (CD4+ > CD8+) and cell functions (strong IL-5 production, cytotoxic potential) are generated. The aim of this study was to elaborate the relevance of these findings in vivo. Methods: Skin biopsy specimens from drug-induced maculopapular exanthema and normal skin were analyzed for CD4, CD8, CD25, HLA-DR, CD54, perforin, granzyme B, IL-5 and IFN-γ using immunohistochemistry. Results: The majority of infiltrating lymphocytes in maculopapular drug eruptions were CD4+. Both CD4+ and CD8+ T cells expressed perforin and granzyme B and were partly located at the dermoepidermal junction and in the epidermis. In addition, strong immunoreactivity for IL-5 and moderate immunoreactivity for IFN-γ were observed in the mononuclear cell infiltrate. Conclusions: Our data indicate that skin infiltrating T cells with a cytotoxic potential and the ability to produce IL-5 and IFN-γ may contribute to the damage of keratinocytes and the activation of eosinophils, which are typical features of drug-induced maculopapular exanthema.
Mucosal Tolerance Induction with Hypoallergenic Molecules in a Murine Model of Allergic Asthma
Authors: Wiedermann U. , Herz U. , Vrtala S. , Neuhaus-Steinmetz U. , Renz H. , Ebner C. , Valenta R. , Kraft D.
Published: April 2003

Type I allergy, frequently elicited by airborne allergens, has constantly increased within recent years. Birch pollen and its major allergen Bet v 1 represent a major source of type I allergens. By genetic engineering hypoallergenic Bet v 1 fragments were produced, which lost the IgE binding capacity but retained the T cell epitopes. We have established a murine model of aerosol sensitization to birch pollen and its major allergen Bet v 1, leading to type I allergic immune responses and airway hyperresponsiveness. In the present study we demonstrate that mucosal administration of recombinant Bet v 1 prior to sensitization led to allergen-specific suppression of B and T cell responses in vivo and in vitro, reduction of eosinophilic infiltration in the lungs and inhibition of airway hyperresponsiveness. Intranasal pretreatment with the nonanaphylactic fragments of Bet v 1 prevented allergic immune responses and airway inflammation to the same degree as the pretreatment with the complete molecule. We conclude from our studies that mucosal tolerance induction with hypoallergenic molecules could provide a safe and convenient treatment strategy against type I allergies.
FcεRI-Mediated Activation of Transcription Factors in Antigen-Presenting Cells
Authors: Kraft Stefan , Bieber Thomas
Published: April 2003

Professional antigen-presenting cells (APC) such as monocytes and dendritic cells (DC) bearing high-affinity IgE receptors (FcΕRI) efficiently present IgE-bound antigens to T cells. FcΕRI expression is upregulated on APC from atopic donors, especially in inflamed tissues. These data suggest a pathophysiological concept of an IgE-mediated delayed-type hypersensitivity reaction in atopic diseases. However, FcΕRI ligation also leads to the synthesis of proinflammatory cytokines and other molecules involved in inflammatory reactions. The investigation of transcription factors mediating these effects has only recently commenced. In general, members of the NF-ĸB family are known to regulate APC function and differentiation, with the RelB subunit being especially important in DC generation. In addition, Ikaros and PU.1 have also been shown to be essential factors for DC differentiation, whereas Oct-2 is upregulated by differentiation towards macrophages. Recently, FcΕRI has been demonstrated to induce NF-ĸB activation via IĸB-α serine phosphorylation and degradation in monocytes and DC. Inhibitors of NF-ĸB activation such as N-acetylcysteine or N-tosyl-L-phenylalanine chloromethyl ketone can suppress FcΕRI-induced TNF-α and MCP-1 release. Interestingly, in human epidermal Langerhans’ cells (LC), NF-ĸB activation can only be observed when large amounts of FcΕRI are present. In addition, the composition of NF-ĸB complexes differs between monocytes, monocyte-derived DC and LC, suggesting a cell type-specific regulation. Moreover, the transcription factor NFAT is induced upon FcΕRI ligation in human APC. The elucidation of further transcription factors involved in FcΕRI signaling in APC should contribute to the employment of new inhibition strategies for the treatment of atopic and other inflammatory diseases.
Dual TCR-Expressing T Lymphocytes in Health and Disease
Authors: Hinz Thomas , Weidmann Eckhart , Kabelitz Dieter
Published: April 2003

The authors briefly review recent experimental advances in elucidating the role of dual T cell receptor (TCR)-expressing lymphocytes in the developement of diseases with special emphasis on autoimmunity. Moreover, they summarize present knowledge about these cells concerning their proportion among peripheral blood mononuclear cells, their functionality, and their impact on allorecognition and memory both in humans and in mice. Finally, they describe disease-associated clonal expansions of dual TCR-expressing cells in humans, most of which have been observed in peripheral T cell malignancies. Other cases occurred in inflammatory bowel disease and in HIV infection. They propose that expression of two distinct TCR on malignant T lymphocytes might be much higher than is suggested by the few cases described so far, and that their presence might impinge on therapeutic immunization strategies which make use of the TCR itself as a target.
High-Level Expression in Mammalian Cells of Recombinant House Dust Mite Allergen ProDer p 1 with Optimized Codon Usage
Authors: Massaer Marc , Mazzu Pasqualina , Haumont Michèle , Magi Mauro , Daminet Véronique , Bollen Alex , Jacquet Alain
Published: April 2003

Background: The major house dust mite allergen Der p 1 is associated with allergic diseases such as asthma. Production of recombinant Der p 1 was previously attempted, but with limited success. The present study describes the expression of recombinant (rec) ProDer p 1, a recombinant precursor form of Der p 1, in CHO cells. Methods: As optimization of the codon usage may allow successful overexpression of protein in mammalian cells, a synthetic gene encoding ProDer p 1 was designed on the basis of the codon usage frequently found in highly expressed human genes. Gene synthesis was accomplished from a set of 14 mutually priming overlapping oligonucleotides and after two runs of polymerase chain reaction. Results: COS cells transiently transfected with the synthetic ProDer p 1 gene produced up to 5–10 times as much ProDer p 1 compared with the expression level obtained after transfection with the authentic gene. To stably express the recombinant allergen, CHO-K1 cells were transfected with the ProDer p 1 synthetic gene, and one amplified recombinant clone produced up to 30 mg of recProDer p 1 per liter in the culture medium before purification. recProDer p 1 was secreted as an enzymatically inactive single-chain molecule presenting three glycosylated immunoreactive forms of 41, 38 and 36 kD. When examined with respect to direct binding, recProDer p 1 and natural Der p 1 displayed very similar IgE reactivities. However, IgE inhibition and histamine release assays showed a much higher reactivity to natural Der p 1 compared to recProDer p 1. Conclusions: These data indicated that codon optimization represents an attractive strategy for high-level production of allergen in mammalian cells.
Identification of a Hevea brasiliensis Latex Manganese Superoxide Dismutase (Hev b 10) as a Cross-Reactive Allergen
Authors: Wagner Stefan , Sowka Slawomir , Mayer Christina , Crameri Reto , Focke Margit , Kurup Viswanath P. , Scheiner Otto , Breiteneder Heimo
Published: April 2003

Background: Cross-reactive allergens play an increasingly important role in latex allergy in complicating both the diagnosis and time course of allergic symptoms. Manganese superoxide dismutase (MnSOD), a ubiquitous protein of prokaryotic and eukaryotic organisms, was described as a cross-reactive allergen in Aspergillus fumigatus. Little information is available on the importance of this pan-allergen in Hevea brasiliensis latex. The aim of this study was to clone and express MnSOD from H. brasiliensis latex, and to obtain the soluble and immunologically active recombinant allergen for diagnosis of latex allergy and to investigate possible cross-reactivities with the structurally related A. fumigatus and human MnSODs. Methods: A complementary DNA coding for Hevea latex MnSOD was amplified by PCR. The recombinant protein was produced in Escherichia coli with an N-terminal hexahistidyl tag. Enzymatic activity of the recombinant protein was determined using an enzyme assay for SODs. IgE immunoblotting and IgE inhibition assays were performed to characterize the recombinant allergen and its cross-reactivity. Results: A Hevea latex MnSOD consisting of 206 amino acid residues was cloned and expressed in E. coli. The allergen was designated Hev b 10. The recombinant protein was enzymatically active, indicating the correct folding of the protein. In immunoblots, latex- as well as A. fumigatus-allergic patients revealed IgE binding to recombinant (r)Hev b 10. Cross-reactivity to Asp f 6, the MnSOD from A. fumigatus, and human MnSOD was determined by inhibition of IgE binding to these MnSODs by rHev b 10. Conclusions: Hev b 10 is a new cross-reactive allergen of H. brasiliensis which belongs to the ‘latex-mold’ group of latex allergens. Furthermore, it is a candidate for primary sensitization in patients allergic to the pan-allergen MnSOD.
Characterization of High-Molecular-Mass Allergens in Oilseed Rape Pollen
Authors: Chardin H. , Mayer C. , Sénéchal H. , Tepfer M. , Desvaux F.-X. , Peltre G.
Published: April 2003

Background:Oilseed rape pollen allergies have been previously described as the result of cross-sensitization with various pollens. Recently, several proteins have been identified as oilseed rape allergens. The aim of the present work was the characterization of oilseed rape pollen allergens by two-dimensional (2-D) gel analysis and amino acid microsequencing. Methods: Water extractable proteins from oilseed rape pollen were separated by isoelectrofocusing and then transferred onto a nitrocellulose sheet. Twenty-one human sera from pollen- or mustard-allergic individuals were screened for their reactivity to oilseed rape proteins. Eleven sera possessed IgE which recognized oilseed rape pollen proteins and one serum was selected for further 2-D characterization and amino acid microsequencing of the allergens. Results: The results showed that three molecules from oilseed rape pollen were identified as oilseed rape allergens which have not yet been described. These three proteins were molecules of 70 kD with a pI >8, 40 kD with a pI around 10 and 80 kD with a pI around 5. These proteins displayed identities with the berberine bridge protein, a receptor-like protein kinase and the cobalamin-independent methionine synthetase from Arabidopsis thaliana, respectively. The genes encoding the putative Arabidopsis molecules are located on chromosome 1 (berberine bridge protein) and chromosomes 3 and 4 (receptor-like protein kinases). Conclusion: These results show that certain high-molecular-mass proteins from oilseed rape pollen are allergens.
Urticaria and Rhinitis to Shrubs of Ficus benjamina and Breadfruit in a Banana-Allergic Road Worker: Evidence for a Cross-Sensitization between Moracea, Banana and Latex
Authors: De Greef J.M. , Lieutier-Colas F. , Bessot J.C. , Vérot A. , Gallerand A.M. , Pauli G. , de Blay F.
Published: April 2003

Background: We report the case of a road worker with a food allergy to banana, who developed urticaria and rhinitis when cutting shrubs of Ficus benjamina and breadfruit. He did not develop an allergy to latex of Hevea brasiliensis.Results: Sensitization to latex of F. benjamina, H. brasiliensis, breadfruit and banana was demonstrated using skin tests and specific IgE measurements. RAST inhibitions procedures showed that specific IgE to breadfruit latex cross-reacted more strongly with latex of H. brasiliensis and banana than with latex of F. benjamina with the same extract. Conclusion: Given the wide distribution of Moracea trees in tropical regions, sensitization to latex of H. brasiliensis and banana could be a consequence of sensitization to Moracea members; F. benjamina does not seem to be the only Moracea responsible for cross-allergy with latex and fruit. Consequently, it seems interesting to test other members of the Moracea family in patients sensitized to latex of H. brasiliensis and banana. Sensitization to breadfruit could be a risk factor for sensitization to latex of H. brasiliensis.
Effect of Low-Dose Cyclosporin A Microemulsion on Disease Severity, Interleukin-6, Interleukin-8 and Tumor Necrosis Factor Alpha Production in Severe Pediatric Atopic Dermatitis
Authors: Bunikowski Rita , Gerhold Kerstin , Bräutigam Matthias , Hamelmann Eckard , Renz Harald , Wahn Ulrich
Published: April 2003

Background: The release of cytokines [interleukin-6 (IL-6), IL-8 and tumor necrosis factor-α (TNF-α)] by skin cells is involved in the pathogenesis of atopic dermatitis (AD). Objective: To evaluate the effect of low-dose cyclosporin A (CyA) on clinical symptoms and cytokine secretion in severe pediatric AD. Methods: Ten children with severe AD (SCORAD index >50) were treated for 8 weeks with CyA. The initial dose of 2.5 mg/kg/day was titrated to a maximum of 5 mg/kg/day until a SCORAD reduction of ≧35% was achieved (‘treatment response’). After stopping CyA all patients entered a 4-week follow-up period. Cytokine secretion (IL-6, IL-8 and TNF-α) from patients’ PBMC was assessed by ELISA before and after CyA treatment and was compared with 18 healthy nonatopic controls. Only the data of patients, who responded to CyA and did not experience a relapse during the follow-up period, were evaluated for this paper. Results: Seven patients responded to CyA without relapse during the follow-up period. The median SCORAD index in these patients improved from 71 at baseline to 22 after CyA treatment (p < 0.001). AD patients’ PBMC produced more IL-6, IL-8 and TNF-α than PBMC of controls. Suppression of IL-6 (p < 0.05) and IL-8 (p < 0.05) production was observed after CyA treatment. TNF-α levels were unchanged by CyA in all patients. Conclusions: The reduction in severity of pediatric AD with CyA is associated with decreased production of IL-6 and IL-8, but not TNF-α by PBMC.
Allergic and Immunologic Parameters in Patients with Fanconi`s Anemia
Authors: Roxo Jr. Pérsio , Arruda L. Karla , Nagao Aparecida T. , Carneiro-Sampaio Magda M.S. , Ferriani Virginia P.L.
Published: April 2003

Background: Fanconi`s anemia (FA) is a rare recessive chromosomal instability disorder, characterized by progressive bone marrow failure and congenital defects. Patients with FA present with recurrent infections, particularly those of the respiratory tract. Objective: The aim of the present study was to evaluate whether patients with FA have altered antibody-mediated immune responses. Methods: A group of 12 patients with FA, 5–32 years old (6 males) was studied. Serum levels of IgG, IgM, IgA and IgG subclasses, isohemagglutinin titers and specific IgG antibodies to poliovirus and measles were determined using standard methods. Immediate skin tests to common inhalant allergens were performed, and total and specific serum IgE was quantitated using a fluoroenzymatic assay (Uni-CAP, Pharmacia). Antipneumococcal antibodies were measured by ELISA before and 4–8 weeks after immunization with pneumococcal vaccine (Pneumo 23, Pasteur Mérieux Connaught). Responses to serotypes 1, 3, 5, 6B, 9V and 14, which are the most prevalent in our country, were studied. Results: Ten patients had elevated IgE levels in sera, and 7 of them had detectable specific IgE and positive immediate skin tests. An inadequate response to pneumococcal vaccination was found in 2 of the 12 patients. Isohemagglutinin titers and levels of IgG, IgM, IgA and IgG subclasses and antipoliovirus and antimeasles antibodies were within the normal limits for age in all patients. Two patients had undetectable IgG4 levels (below 5 mg/dl). Conclusions: The results indicate that a proportion of patients with FA (2/12) in our study had inadequate responses to pneumococcal vaccination. No other significant abnormalities of the immune system were found in these patients.
Dexamethasone Does Not Modulate Eosinophil Adhesion to Endothelial Cells
Authors: Sutani Akihisa , Nagata Makoto , Yamamoto Hideaki , Sakurai Manami , Sakamoto Yoshio
Published: April 2003

Background: Glucocorticoids are representative agents in the anti-inflammatory treatment of allergic airway diseases. Although they reduce the number of tissue eosinophils, its exact mechanism has not been completely established. Objective: The present study was undertaken to evaluate whether dexamethasone modulates the adhesive property of eosinophils. The effect on the expression of endothelial adhesion molecules was also evaluated. Methods: Blood eosinophils from healthy subjects were incubated in the presence or absence of 0.1–1 µM of dexamethasone for 6 h at 37°C. Eosinophil chemotaxis and adhesion to paraformaldehyde-fixed human pulmonary vascular endothelial cells (HPMEC) were then examined. The effects of dexamethasone on the expression of VCAM-1 and ICAM-1 on HPMEC were examined by cell ELISA. Results: 1 µM dexamethasone significantly inhibited the chemotactic response of eosinophils to 1 µM formyl-methionyl-leucyl-phenylalanine (FMLP). On the other hand, dexamethasone did not modify either the spontaneous or FMLP-stimulated adhesion of eosinophils to resting HPMEC. Similarly, dexamethasone did not alter either the spontaneous adhesion of eosinophils to or the expression of VCAM-1 or ICAM-1 on HPMEC stimulated with IL-4 + TNF-α. Conclusion: Dexamethasone does not modify the adhesive property of eosinophils or the expression of adhesion molecules on endothelial cells. Thus, it is unlikely that glucocorticoids directly modulate the adhesive interaction between eosinophils and endothelial cells.
Regulation of Chemokine Receptor Expression in Eosinophils
Authors: Nagase Hiroyuki , Miyamasu Misato , Yamaguchi Masao , Fujisawa Takao , Kawasaki Hiroshi , Ohta Ken , Yamamoto Kazuhiko , Morita Yutaka , Hirai Koichi
Published: April 2003

Signals via chemokine receptors play an important role in the accumulation of eosinophils at allergic inflammatory sites. Eosinophils constitutively express CC chemokine receptor 3 (CCR3) and, to a lesser extent, CCR1. CCR3 is mainly responsible for migration of resting eosinophils, and its specific ligand, eotaxin, represents the most potent chemoattractant for eosinophils. Some reports also suggest the expression of CXC chemokine receptor 1 (CXCR1) and/or CXCR2 in eosinophils. In addition, we recently reported the functional expression of CXCR4. The ligand of CXCR4, stromal cell-derived factor-1 (SDF-1), was able to induce a strong migratory response comparable to that by eotaxin. In contrast to the CCR3/eotaxin system which is mainly regulated at the level of ligand production, the CXCR4/SDF-1 system is regulated at the level of receptor expression. CXCR4 expression was completely attenuated by IL-4 and IL-5 and upregulated by IFN-γ and dexamethasone, while CCR3 expression was only marginally affected. The balance between the biological effects of these chemokine systems may affect the distribution and migration of eosinophils.
Mouse Models for the Genetic Dissection of Atopy
Authors: Heinzmann Andrea , Daser Angelika
Published: April 2003

Atopic disorders are complex traits with a significant contribution of heritability. Several mouse models for atopic phenotypes such as allergen-induced and intrinsic asthma, atopic dermatitis and immediate cutaneous hypersensitivity have been used to systematically dissect the genetics of these diseases. Many of the chromosomal regions identified in genome-wide screens colocalize with linkage regions of autoimmune phenotypes suggesting common regulatory genes in combination (or addition) to disease-specific genes. Moreover, many of these regions correspond to regions found in the human system, thus strengthening the likelihood of a gene contributing to the phenotype. The role of individual candidate genes has been vigorously pursued in genetically modified mice by destruction or overexpression of the candidate gene. The complex interactions of multiple contributing genes may further be elucidated in randomly mutated mouse strains. Thus mouse models can contribute largely to the elucidation of disease genes.
Lipid A Analogue, ONO-4007, Inhibits IgE Response and Antigen-Induced Eosinophilic Recruitment into Airways in BALB/c Mice
Authors: Iio Jun , Katamura Kenji , Takeda Hiroshi , Ohmura Kayo , Yasumi Takahiro , Meguro Taka-aki , Ohshima Yusei , Nakahata Tatsutoshi
Published: April 2003

Background: Since antigen-specific IgE and eosinophils are major inducing factors of allergic inflammation of the airways, both factors are therapeutic targets of asthma. We investigated the effects of ONO-4007, a nontoxic lipid A analogue, on antigen-specific antibody response and the recruitment of eosinophils into airways in murine systems. Methods: BALB/c mice were injected ONO-4007 intraperitoneally during sensitization with ovalbumin (OVA) and aluminium hydroxide to determine its effects on the antigen-specific antibody response. ONO-4007 was also injected intravenously during either systemic sensitization and inhalation with OVA, or sensitization or inhalation alone to determine its effects on antigen-induced airway inflammation. In vitro effects of ONO-4007 on the functional differentiation of naive CD4+ T cells were investigated by culturing naive CD4+ T cells derived from DO11.10 mice and OVA-pulsed dendritic cells (CDCs) with ONO-4007. Results: ONO-4007 inhibited antigen-specific IgE and IgG1, but not IgG2a responses. ONO-4007 decreased the recruitment of eosinophils and the levels of IL-5 in bronchoalveolar lavage fluid, not only when it was injected during systemic sensitization and inhalation with OVA, but also during inhalation alone. ONO-4007 inhibited the differentiation of IL-4- and IL-13-producing CD4+ T cells in vitro, which was partly mediated by DCs. Conclusions: ONO-4007 inhibited antigen-specific IgE and IgG1 responses and antigen-induced eosinophil recruitment into the airways in BALB/c mice. These effects were mediated, at least partly, by the modulation of DCs, although there may also be other mechanisms.
Overcoming the Problem of Cytomegalovirus Infection after Organ Transplantation: Calling for Heracles?
Authors: van Son Willem J. , de Maar Eltjo F. , van der Bij Wim , van den Berg Arie P. , Verschuuren Erik A.M. , The T. Hauw
Published: April 2003

Although diagnosis of CMV infections and treatment of CMV disease with effetive antiviral drugs have become much easier, the persistent problem of CMV infection after solid-organ transplantation still requires solid knowledge of the pathophysiology of its clinical manifestations in order to minimize the impact of CMV infections in the future. The complex symptomatology of CMV infection after solid-organ transplantation is reviewed as well as some of the new theories attempting to explain the myriad of symptoms seen after transplantation.
Clinical Significance of Cytomegalovirus-Specific T Helper Responses and Cytokine Production in Lung Transplant Recipients
Authors: Zeevi Adriana , Spichty Kathy , Banas Richard , Cai Jane , Donnenberg Vera S. , Donnenberg Albert D. , Ahmed Mamun , Dauber James , Iacono Aldo , Keenan Robert , Griffith Barthley
Published: April 2003

Cytomegalovirus (CMV) disease continues to be a major problem for lung transplant recipients. In CMV-seropositive individuals, we detected two types of CMV-specific responses: a self-restricted response stimulated by soluble CMV antigen (sCMV-Ag) and a non-self-restricted response induced by CMV-infected cells (cCMV-Ag). Lung transplant recipients who develop the CMV-specific self-restricted T helper response have a low risk of recurrent CMV disease. In contrast, during CMV disease, lung transplant recipients exhibit only the non-self-restricted T helper responses. We characterized the T cell activation and the kinetics of cytokine production of sorted CD4+ and CD8+ T cells from PBLs of CMV seropositive donors. The two types of CMV antigens induced cytokine production in both T cell subsets. We also performed competitive RT-PCR for Granzyme B (GB) in BAL cells of lung transplant recipients prior to, during and following CMV disease. CMV disease was associated with increase in GB gene expression when was accompanied by acute cellular rejection while it remained low in patients with CMV disease that did not have a complicated course. In summary, CMV-activated T cells within the allograft may produce inflammatory cytokines and effector molecules that may promote allograft rejection.
Viral Dynamics during Active Cytomegalovirus Infection and Pathology
Authors: Emery Vincent C.
Published: April 2003

The central role that cytomegalovirus (CMV) load plays in its pathogenesis is being unravelled. In AIDS patients with active CMV replication, many months prior to the development of CMV disease, elevated CMV load in the blood and urine are significantly associated with an increased risk of disease progression. In addition, elevated load in blood is associated with an increased risk of death. Intervention with ganciclovir acts to rapidly inhibit CMV replication in vivo and has allowed estimates of the clearance/replication rate of CMV to be performed. These data indicate that CMV replicates dynamically in the human host with a doubling time of approximately 1 day. This knowledge has been used to determine the relative contribution of initial viral load and rate of change of viral load as predictors of CMV disease in organ transplant recipients. The data show that both these parameters have prognostic value in multivariate models and should allow the development of novel patient management strategies.
Optimization Strategies for DNA Vaccines
Authors: Weeratna Risini D. , McCluskie Michael J. , Comanita Lacrimiora , Wu Tong , Davis Heather L.
Published: April 2003

DNA immunization is a relatively new vaccination strategy that involves the direct introduction into the host of plasmid DNA encoding the desired antigen. The DNA enters host cells and results in immune responses following in vivo expression of the antigen. Although DNA-based immunization works well in animal models for the induction of both humoral and cell-mediated immune responses, its success in humans has been limited. This paper discusses different approaches that have attempted to optimize DNA vaccines, and presents results evaluating some of these approaches in mice.
Clinical Efficacy of Micronized Purified Flavonoid Fraction: An Overview
Authors: Struckmann Jan R.
Published: April 2003

Flavonoids are widely used for the treatment of chronic venous insufficiency (CVI). In clinical trials, micronized purified flavonoid fraction (MPFF, 450 mg diosmin plus 50 mg hesperidin, Daflon 500 mg) has demonstrated its activity in CVI by improving venous tone and vein elasticity assessed by plethysmography. Randomized, double-blind, placebo-controlled clinical studies have shown an improvement in signs and symptoms related to CVI and a decrease in leg circumferences at the levels of the ankle and calf. The effect of MPFF on microcirculatory parameters suspected of participating in the pathophysiological process of venous ulceration has been investigated in patients. These include hemorheological parameters and transcutaneous oxygen tension measurements, which were shown to improve after treatment with MPFF. Finally, a randomized double-blind, placebo-controlled clinical study has shown that MPFF treatment for 2 months, in addition to standard compression therapy, accelerates leg ulcer healing in patients with ulcers ≤10 cm.
The CytoscanTM Model E-II, a New Reflectance Microscope for Intravital Microscopy: Comparison with the Standard Fluorescence Method
Authors: Harris A.G. , Sinitsina I. , Messmer K.
Published: April 2003

The CytoscanTM Model E-II (Cytometrics Inc., Philadelphia, Pa., USA) is a newly developed instrument which functions as an intravital microscope and is small and easily portable. Through the use of orthogonal polarization spectral (OPS) imaging, the Cytoscan Model E-II delivers images of the microcirculation which are comparable to those achieved with intravital fluorescence videomicroscopy (IFM), but without the use of fluorescent dyes. The purpose of this study was to validate the Cytoscan Model E-II instrument against IFM. The experiments were carried out on striated muscle in the dorsal skinfold chamber of the awake Syrian hamster. The following parameters were measured in identical regions of interest in the same animal under baseline conditions and 0.5 and 2 h after a 4-hour period of pressure-induced ischemia: arteriolar diameter, venular diameter and venular red blood cell velocity. Bland-Altman plots showed good agreement between the two techniques for venular red blood cell velocity. As expected, arteriolar and venular diameters as measured by the Cytoscan were on average 5 µm smaller than the values from IFM, since the Cytoscan measures the red blood cell column width and IFM measures luminal diameter. Thus, OPS imaging can be used to make valid measurements of microvascular diameter and red blood cell velocity in tissues.
Enhanced Cutaneous Lymphatic Network in the Forearms of Women with Postmastectomy Oedema
Authors: Mellor R.H. , Stanton A.W.B. , Azarbod P. , Sherman M.D. , Levick J.R. , Mortimer P.S.
Published: April 2003

Postmastectomy oedema (PMO) of the arm is a common aftermath of axillary lymphatic damage during treatment for breast cancer. The aim of the present study was to quantify the forearm dermal lymphatic capillaries in order to determine whether they exhibit adaptive responses to PMO. Both forearms were examined by fluorescence microlymphography in 16 patients with oedema following treatment for breast cancer (mean swelling 25 ± 4%) and 19 patients treated for breast cancer but without oedema. Delineated lymphatic networks were analysed stereologically. The main findings were: (1) lymphatic density at any specified distance from the injection site was greater in the swollen arm than the control arm (p < 0.01, t test); (2) taking into account the increased skin area, the total length of lymphatic capillaries in a 1-cm annulus of skin was 676 ± 56 cm (swollen), compared with 385 ± 30 cm (control) (p < 0.001, t test); (3) fluorescent marker was transported over a greater distance before draining deep in the swollen arm (2.74 ± 0.33 cm) than in the control arm (1.59 ± 0.24 cm) (p = 0.02); (4) there was no evidence of lymphatic dilatation in the swollen arm, and (5) in breast cancer patients without swelling, the arm on the side of radiotherapy/surgery (otherwise referred to as the unswollen arm) showed none of the above changes, indicating that the changes are linked to the oedema rather than being universal responses to breast cancer or its treatment. It is concluded that microlymphatic changes occur in the swollen arm, namely a local superficial rerouting of lymph drainage and either lymphangiogenesis and/or increased recruitment of dormant lymphatic vessels. Since blood capillary angiogenesis occurs in the swollen arms, and lymphangiogenesis occurs in experimental lymphoedema, there is a precedent for proposing lymphangiogenesis in PMO. An increased number of functional vessels would help to maintain the ratio of local tissue drainage capacity to filtration capacity.