Disulfide linkage controls the affinity and stoichiometry of IgE Fcepsilon3-4 binding to FcepsilonRI.

IgE antibodies cause long-term sensitization of tissue mast cells and blood basophils toward allergen-induced cross-linking and triggering of allergic inflammation. This persistence of IgE binding is due to its uniquely high affinity for the receptor FcepsilonRI and in particular its slow rate of dissociation once bound. The binding interface consists of two subsites, one contributed by each Cepsilon3 domain of IgE Fc in a 1:1 complex.

A transcriptional regulatory element in the coding sequence of the human Bcl-2 gene.

We investigated the protein-binding sites in a DNAse I hypersensitive site associated with bcl-2 gene expression in human B cells. We mapped this hypersensitive site to the coding sequence of exon 2 of the bcl-2 gene in the bcl-2-expressing REH B-cell line. Electrophoretic mobility shift assays (EMSAs) with extracts from REH cells revealed three previously unrecognized B-Myb-binding sites in this sequence.

Transcription of Ig germline genes in single human B cells and the role of cytokines in isotype determination.

We have developed a critical test of the chromatin accessibility model of Ig isotype determination in which local unfolding of chromatin higher order structure (chromatin accessibility) in the region of specific germline genes in the H chain locus determines the Ab class to be expressed in the B cell. We show that multiple germline genes are constitutively transcribed in the majority of naive human B cells in a population. Thus, because chromatin in its higher order structure cannot be transcribed, the entire Ig H chain locus must be unfolded in naive B cells.

Ototoxic protection of sodium thiosulfate: daily vs constant infusion.

Objective: Compare the effect of a daily dose of sodium thiosulfate (STS) to continuous infusion of STS on Hartley albino guinea pig (HAGP) on cisplatin (CP)-induced sensorineural hearing loss. Study design Prospective randomized pilot study comparing a single daily dose of STS to a constant controlled infusion of STS given the middle ear space (MES) with an ototoxic regimen of CP. The hearing thresholds will be obtained by auditory brainstem response (ABR) 1 week before and after treatment.

Ibuprofen blocks changes in Na v 1.7 and 1.8 sodium channels associated with complete Freund's adjuvant-induced inflammation in rat.

Unlabelled: Although nerve growth factor plays a role in augmenting sodium channel expression in small dorsal root ganglion (DRG) cells, the cytochemical mediators responsible for enhanced expression in large DRG neurons are unknown. To narrow the search for mediators involved in the increased production of sodium channels in large DRG neurons, we examined the effect of cyclooxygenase inhibition on sodium channel production during inflammation. Thirty minutes before the subcutaneous injection of complete Freund's adjuvant (CFA), rats received ibuprofen (nonselective, cyclooxygenase inhibitor), NS-398 (selective, cyclooxygenase inhibitor), or vehicle.

Zeros of the partition function and pseudospinodals in long-range Ising models.

The relation between the zeros of the partition function and spinodal critical points in Ising models with long-range interactions is investigated. We find that the spinodal is associated with the zeros of the partition function in four-dimensional complex temperature/magnetic field space. The zeros approach the real temperature/magnetic field plane as the range of interaction increases.

PPAR gamma ligands, 15-deoxy-delta12,14-prostaglandin J2 and rosiglitazone regulate human cultured airway smooth muscle proliferation through different mechanisms.

The influence of two peroxisome proliferator-activated receptor gamma (PPARgamma) ligands, a thiazolidinedione, rosiglitazone (RG) and the prostaglandin D2 metabolite 15-deoxy-Delta(12,14)-prostaglandin J2 (15d-PGJ2) on the proliferation of human cultured airway smooth muscle (HASM) was examined. The increases in HASM cell number in response to basic fibroblast growth factor (bFGF, 300 pm) or thrombin (0.3 U ml-1) were significantly inhibited by either RG (1-10 microM) or 15d-PGJ2 (1-10 microM).

High precision measurement of the static dipole polarizability of cesium.

The cesium 6(2)S(1/2) scalar dipole polarizability alpha(0) has been determined from the time-of-flight of laser cooled and launched cesium atoms traveling through an electric field. We find alpha(0)=6.611+/-0.

Local somatic hypermutation and class switch recombination in the nasal mucosa of allergic rhinitis patients.

Immunoglobulin E is produced by nasal B cells in response to allergen. We have analyzed IgE V(H) region sequences expressed in the nasal mucosa of patients suffering from allergic rhinitis. V(H) region sequences were amplified by RT-PCR from IgE(+) B cells from nasal biopsies.

Cortical control mechanisms in volitional swallowing: the Bereitschaftspotential.

Objective: This research sought to identify a well-defined pre-motor potential, the Bereitschaftspotential (BP), as a manifestation of cortical contribution to the pre-motor planning of volitional swallowing.

Methods: EEG data were collected from 20 research participants during volitional execution of swallowing and finger movement tasks. A5 second pre-movement epoch for each task was triggered on EMG identification of movement onset.

Activity of human monocytes in IgE antibody-dependent surveillance and killing of ovarian tumor cells.

We have previously shown that a chimeric IgE antibody against the folic acid receptor (MOv18 IgE) inhibits tumor growth in a SCID mouse model of ovarian carcinoma. MOv18 IgE gave greater protection than the corresponding chimeric MOv18 IgG1. We have now confirmed these effects in a nude-mouse model of ovarian carcinoma and have demonstrated for the first time that human monocytes are active in IgE antibody-dependent cell-mediated cytotoxicity.

The biology of IGE and the basis of allergic disease.

Allergic individuals exposed to minute quantities of allergen experience an immediate response. Immediate hypersensitivity reflects the permanent sensitization of mucosal mast cells by allergen-specific IgE antibodies bound to their high-affinity receptors (FcepsilonRI). A combination of factors contributes to such long-lasting sensitization of the mast cells.

Knowledge and attitudes about the differences between emergency contraception and medical abortion among middle-class women and men of reproductive age in Mexico City.

Two reproductive technologies--emergency contraception and medical abortion--have the potential to reduce unintended pregnancy significantly in Latin America. Lack of knowledge and negative attitudes about the methods may limit their impact, however. Results from focus group discussions with middle-class men and women of reproductive age residing in Mexico City indicate that knowledge about emergency contraception and medical abortion is low.

Necessity of the stalk region for immunoglobulin E interaction with CD23.

Previously, a soluble mouse CD23 chimera, composed of an N-terminal trimeric isoleucine zipper motif (lz) followed by the entire extracellular region (amino acids 48-331) of CD23 (lz-CD2348-331), was prepared and exhibited strong binding to rodent immunoglobulin E (IgE). In the current study, we report the construction of a similar human chimeric protein (lz-huCD2345-321), as well as a series of murine chimeric lz-CD23 mutants with incremental portions of stalk deleted, to further investigate the role of the stalk region in mediating the CD23-IgE interaction. All chimeric proteins were designed such that the predicted heptad structure of the stalk was retained.

The crystal structure of IgE Fc reveals an asymmetrically bent conformation.

The distinguishing structural feature of immunoglobulin E (IgE), the antibody responsible for allergic hypersensitivity, is the C epsilon 2 domain pair that replaces the hinge region of IgG. The crystal structure of the IgE Fc (constant fragment) at a 2.6-A resolution has revealed these domains.

Mutagenesis within human FcepsilonRIalpha differentially affects human and murine IgE binding.

Soluble fragments of the alpha-chain of FcepsilonRI, the high-affinity receptor for IgE, compete with membrane-bound receptors for IgE and may thus provide a means to combat allergic responses. Mutagenesis within FcepsilonRIalpha is used in this study, in conjunction with the crystal structure of the FcepsilonRIalpha/IgE complex, to define the relative importance of specific residues within human FcepsilonRIalpha for IgE binding. We have also compared the effects of these mutants on binding to both human and mouse IgE, with a view to evaluating the mouse as an appropriate model for the analysis of future agents designed to mimic the human FcepsilonRIalpha and attenuate allergic disease.

Source generators of mismatch negativity to multiple deviant stimulus types.

The purpose of the present study was to investigate auditory stimulus feature processing and how neural generators might differ among the mismatch negativity (MMN) responses to intensity, frequency, and duration deviant stimuli. Data collected from 72 electrodes in twelve adult female subjects were analyzed. For each subject, peak amplitude and latency values at Fz were compared among responses to the three deviant stimulus types presented in individual conditions with a probability of 0.

Increases in allergen-specific IgE in BAL after segmental allergen challenge in atopic asthmatics.

IgE is important in both early and late allergic responses. Increases in the numbers of RNA transcripts coding for IgE have been observed in the bronchial mucosa of asthmatics and in the nasal mucosa of hay fever patients both during natural allergen exposure and after nasal allergen challenge, suggesting that IgE may be synthesized locally in the mucosa. In this study we have examined bronchoalveolar lavage (BAL) taken before and 24 h after bronchoscopic segmental allergen challenge from 18 atopic asthmatic patients, looking for evidence of increases in IgE protein.

Persistent IgE synthesis in the nasal mucosa of hay fever patients.

The location of IgE synthesis has been a longstanding controversy, with previous evidence favoring either the mucosa or lymphoid tissue in the region of allergen entry. The evidence for IgE synthesis in mucosal tissues has always been circumstantial. We have developed a novel explant culture system, using ELISA and radioactive amino acid incorporation, to measure de novo IgE protein synthesis in the nasal mucosa of hay fever patients.

Regulation of IgE production requires oligomerization of CD23.

Here we describe the production of a rabbit polyclonal Ab (RAS1) raised against the stalk of murine CD23. RAS1 inhibits release of CD23 from the surface of both M12 and B cells resulting in an increase of CD23 on the cell surface. Despite this increase, these cells are unable to bind IgE as determined by FACS.

IgEs targeted on tumor cells: therapeutic activity and potential in the design of tumor vaccines.

Surface-bound IgE play a central role in antiparasite immunity; to exploit IgE-driven immune mechanisms in tumor prevention and control, monoclonal IgEs of irrelevant specificity were loaded through biotin-avidin bridging onto tumor cells, either by systemic administration to tumor-bearing mice or pre-loading of tumor cells before inoculation. Here we show that systemic administration of biotinylated IgEs to mice bearing tumors pre-targeted with biotinylated antibodies and avidin significantly decreased tumor growth rate. In addition, as compared with IgG-loaded control cells, inoculation of suboptimal doses of IgE-loaded tumor cells suppressed tumor formation in a fraction of animals and induced protective host immunity by eliciting tumor-specific T-cell responses.

Endocytosis and recycling of the complex between CD23 and HLA-DR in human B cells.

The presentation of extremely low doses of antigen to T cells is enhanced by immunoglobulin E (IgE)-dependent antigen focusing to CD23, the low-affinity receptor for IgE, expressed on activated B cells. CD23 contains a C-type lectin domain in its extracellular sequence and a targeting signal for coated pits, required for endocytosis, in its cytoplasmic sequence. CD23 is non-covalently associated with the major histocompatibility complex class II antigen, human leucocyte antigen HLA-DR, on the surface of human B cells, but the fate of this complex following endocytosis is unknown.

IgE isotype determination: epsilon-germline gene transcription, DNA recombination and B-cell differentiation.

Immunoglobulin class switching is the process which determines whether a B-cell secretes antibodies of the IgM, IgG, IgA or IgE class (or isotype). IgE is the antibody that mediates the allergic response by sensitising mast cells to allergens at the mucosal barrier. Class switching proceeds by three successive steps, culminating in the synthesis and secretion of antibody: these are germline gene transcription, DNA recombination and B-cell differentiation.

The structure of the IgE Cepsilon2 domain and its role in stabilizing the complex with its high-affinity receptor FcepsilonRIalpha.

The stability of the complex between IgE and its high-affinity receptor, FcepsilonRI, on mast cells is a critical factor in the allergic response. The long half-life of the complex of IgE bound to this receptor in situ ( approximately 2 weeks, compared with only hours for the comparable IgG complex) contributes to the permanent sensitization of these cells and, hence, to the immediate response to allergens. Here we show that the second constant domain of IgE, Cepsilon2, which takes the place of the flexible hinge in IgG, contributes to this long half-life.