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Bioimpedance analysis is a label-free and easy approach to obtain information on cellular barrier integrity and cell viability more broadly. In this work, we introduce a small, low-cost, portable in vitro impedance measurement system for studies where a shadow-free exposure of the cells is a requirement. It can be controlled by a user-friendly web interface and can perform measurements automated and autonomously at short intervals. The system can be integrated into an existing IoT network for remote monitoring and indepth analyses. A single-board computer (SBC) serves as the central unit, to control, analyze, store and forward the measurement data from the single-chip impedance analyzer. Various materials and manufacturing methods were used to produce a purpose-built lid on top of a modified 24-well microtiter plate in a “do it yourself” fashion. Furthermore, three different sensor designs were developed utilizing anodic aluminum oxide (AAO) membranes and gold-plated electrodes. Preliminary tests with potassium chloride (KCl) showed first promising results.

Bioimpedance analysis is a label-free and easy approach to obtain information on cellular barrier integrity and cell viability more broadly. In this work, we introduce a small, low-cost, portable in vitro impedance measurement system for studies where a shadow-free exposure of the cells is a requirement. It can be controlled by a user-friendly web interface and can perform measurements automated and autonomously at short intervals. The system can be integrated into an existing IoT network for remote monitoring and indepth analyses. A single-board computer (SBC) serves as the central unit, to control, analyze, store and forward the measurement data from the single-chip impedance analyzer. Various materials and manufacturing methods were used to produce a purpose-built lid on top of a modified 24-well microtiter plate in a “do it yourself” fashion. Furthermore, three different sensor designs were developed utilizing anodic aluminum oxide (AAO) membranes and gold-plated electrodes. Preliminary tests with potassium chloride (KCl) showed first promising results.

Since the beginning of this century, allergen immunotherapy has been widely used to treat allergic disorders. In addition to the long-term clinical efficacy of this therapy, there are immediate beneficial effects as observed in rush immunotherapy for which there is no clear mechanism. We investigated the direct impact of an Ag on its specific IgE in terms of IgE measurability in immunoassays and subsequent binding of IgE to the high-affinity receptor for IgE. As a model we used a chimeric IgE specific for NIP that exhibits similar biologic properties as serum or myeloma IgE. To mimic particulate and soluble allergens we coupled 15 NIP molecules to BSA. Using this "artificial allergen" we could show that the presence of the Ag reduced the IgE measurability in immune assays. Furthermore, IgE binding to the Fc epsilon RI was 60% inhibited in the presence of the Ag shown with an optical biosensor that monitors molecular interactions. In normal basophils passively sensitized with IgE that was preincubated with the Ag, no sulfidoleukotriene release could be induced. Rush immunotherapy may invoke a similar phenomenon, resulting in the short-term alteration of symptoms by blocking or substantially reducing binding of IgE to its high-affinity receptor. Thus, our result may explain some of the short-term beneficial effects observed in rush immunotherapy.

The population dynamics of macromolecules that reproduce by means of template-directed ligation of two fragments are shown to be represented by a replicator equation with a special non-linear response function. This result is obtained through detailed consideration of the mechanism of ligation autocatalysis, In contrast to treatments which involve simplification to a parabolic growth law and the expectation of global coexistence of all species, we find that strong selection can take place in such systems, even when there is slow uncatalysed synthesis of replicators. Also, systems of this type are subject to invasion by new species that have a selective advantage. An expression is derived for the survival threshold in terms of species parameters and it is shown that this threshold depends on the total concentration of all species in the system. For a plausible distribution of species parameters, the number of surviving species coexisting above the threshold increases monotonically with increasing concentration. Illustrative numerical simulations are presented.

We have previously described a mouse monoclonal anti-human IgE antibody (BSW17) capable of recognizing receptor-bound IgE without inducing mediator release from human basophils or mast cells. Moreover, immune complexes of IgE and BSW17 are not able to bind to the IgE receptor. An initial attempt to map the precise epitope recognized by this mAb by using Fcε-derived peptides of variable length was unsuccessful. However, by screening random peptide phage display libraries we isolated circular nona- and octapeptides specifically recognized by BSW17. These constrained peptides mimic at least a part of a conformational epitope and are thus called mimotopes. These mimotopes, either phage displayed or synthetically synthesized, did not react with any other anti-human IgE antibody tested, but efficiently inhibited the binding of human IgE to BSW17 only. The use of Rhodol-Green-labeled free cyclic peptide proved that these interactions were not carrier dependent. Immunization of rabbits with phage clones displaying the specific peptides on the surface induced an anti-human IgE response specific for the epitope of BSW17. Therefore, we conclude that such mimotopes or mimotope-derived peptides might be used for vaccination to induce in vivo a beneficial anti-IgE response as a novel immunotherapy.

Natural antibodies provide an early defense mechanism against pathogens, show a frequent self-reactivity, and are present throughout life. Two questions concern the physiological control of self-reactivity and the pathogenetic link to autoimmune disease. Here we propose a concept of conditional autoimmunity involving natural antibodies against the alpha chain of the high-affinity receptor for IgE (Fc(epsilon)RIalpha ). Like other natural antibodies, anti-Fc(epsilon)RIalpha antibodies are found in sera of healthy donors. We now report the first human recombinant anti-Fc(epsilon)RIalpha autoantibodies isolated by repertoire cloning from a human tonsillar IgM library. These high-affinity antibodies recognize Fc(epsilon)RIalpha on cells and trigger histamine release from freshly isolated blood basophils. However, the latter effect requires IgE removal from the Fc(epsilon)RI. The same conditional histamine release is seen when using sera from individual normal donors and affinity-purified anti-Fc(epsilon)RIalpha antibodies isolated from multidonor therapeutic IgG preparations. We propose that such anti-Fc(epsilon)RIalpha antibodies can become pathogenic and that this is dependent on the state of occupancy of the Fc(epsilon)RIalpha by its natural ligand IgE. We suggest that an imbalance between Fc(epsilon)RIalpha occupancy and natural anti-Fc(epsilon)RIalpha antibodies may be implicated in the pathogenesis of autoimmune urticaria.