Searching for line active molecules on biphasic lipid monolayers.

In membranes with phase coexistence, line tension appears as an important parameter for the determination of the amount of domains, as well as their size and their shape, thus defining the membrane texture. Different molecules have been proposed as "linactants" (i.e. molecules that reduce the line tension, thereby modulating the membrane texture). In this work, we explore the efficiency of different molecules as linactants in monolayers with two coexisting phases of different thicknesses. We tested the linactant ability of a molecule with chains of different saturation degrees, another molecule with different chain lengths and a bulky molecule. In this way, we show in the same system the effect of molecules with chains of different rigidities, with an intrinsic thickness mismatch and with a bulky moiety, thereby analyzing different hypotheses of how a molecule may change the line tension in a monolayer system. Both lipids with different hydrocarbon chains did not act as linactants, while only one of the bulky molecules tested decreased the line tension in the monolayer studied. We conclude that there are no universal rules for the structure of a molecule that enable us to predict that it will behave as a linactant and thus, designing linactants appears to be a difficult task and a challenge for future studies. Furthermore, in regard to the membrane texture, there was no direct influence of the line tension in the distribution of domain sizes.

Basophil expansion protects against invasive pneumococcal disease in mice.

BACKGROUND: Protein-based vaccination using pneumococcal proteins is a promising approach for efficient vaccines against Streptococcus pneumoniae. Basophils play an important role in enhancing memory immune responses to intact proteins. We examined the impact of increased basophil pool sizes on humoral memory responses to pneumococcal surface protein A (PspA). METHODS: Basophil pool sizes in blood, spleen, and bone marrow were increased by either interleukin 3 (IL-3) treatment or by adoptive basophil transfer before secondary PspA immunization. Subsequently, PspA-specific antibody titers and resistance of mice against invasive pneumococcal disease (IPD) was determined. RESULTS: Mice treated with IL-3, which increased basophil pool sizes, and mice receiving a single basophil transfusion responded with significantly higher PspA-specific antibody titers after immunization with PspA. Importantly, however, just a single transfusion of flow-sorted basophils into mice before secondary immunization with PspA significantly protected mice from lethal IPD. Moreover, concomitant blockade of inhibitory FcγRIIB on transfused basophils further substantially increased basophil-mediated protection against IPD in mice. CONCLUSIONS: This is the first study to find that a single transfusion of basophils is sufficient to boost protein-based memory responses against pneumococcal protein antigens, thereby providing significant protection against IPD in mice.

FMS-like tyrosine kinase 3 ligand treatment of mice aggravates acute lung injury in response to Streptococcus pneumoniae: role of pneumolysin.

FMS-like tyrosine kinase-3 ligand (Flt3L) is a dendritic cell (DC) growth and differentiation factor with potential in antitumor therapies and antibacterial immunization strategies. However, the effect of systemic Flt3L treatment on lung-protective immunity against bacterial infection is incompletely defined. Here, we examined the impact of deficient (in Flt3L knockout [KO] mice), normal (in wild-type [WT] mice), or increased Flt3L availability (in WT mice pretreated with Flt3L for 3, 5, or 7 days) on lung DC subset profiles and lung-protective immunity against the major lung-tropic pathogen, Streptococcus pneumoniae. Although in Flt3L-deficient mice the numbers of DCs positive for CD11b (CD11b(pos) DCs) and for CD103 (CD103(pos) DCs) were diminished, lung permeability, a marker of injury, was unaltered in response to S. pneumoniae. In contrast, WT mice pretreated with Flt3L particularly responded with increased numbers of CD11b(pos) DCs and with less pronounced numbers of CD103(pos) DCs and impaired bacterial clearance and with increased lung permeability following S. pneumoniae challenge. Notably, infection of Flt3L-pretreated mice with S. pneumoniae lacking the pore-forming toxin, pneumolysin (PLY), resulted in substantially less lung CD11b(pos) DCs activation and reduced lung permeability. Collectively, this study establishes that Flt3L treatment enhances the accumulation of proinflammatory activated lung CD11b(pos) DCs which contribute to acute lung injury in response to PLY released by S. pneumoniae.

Molecular determinants for the line tension of coexisting liquid phases in monolayers.

The line tension (λ) in biphasic membranes has been determined in monolayers and bilayers using a variety of techniques. In this work we present a novel approach to the determination of λ in monolayers with liquid/liquid phase coexistence, overcoming several of the drawbacks of current techniques. Using our method, we determined the line tension of liquid/liquid phases in binary mixtures of different lipids and a molecule similar to cholesterol but less oxidizable. We analyzed the effect of the hydrocarbon chain length and the polar head-group of the non-sterol lipid and found the latter to exert much more influence than the former. The presence of PE led to high λ values, PG to low values and PS and PC to intermediate values. The line tension showed a strong correlation with the critical packing parameter of the phospholipid. The spontaneous curvature displayed by the phases constituted by a particular lipid appears to be an important parameter for determining the line tension in mixed films.

Evaluation of invertebrate infection models for pathogenic corynebacteria.

For several pathogenic bacteria, model systems for host-pathogen interactions were developed, which provide the possibility of quick and cost-effective high throughput screening of mutant bacteria for genes involved in pathogenesis. A number of different model systems, including amoeba, nematodes, insects, and fish, have been introduced, and it was observed that different bacteria respond in different ways to putative surrogate hosts, and distinct model systems might be more or less suitable for a certain pathogen. The aim of this study was to develop a suitable invertebrate model for the human and animal pathogens Corynebacterium diphtheriae, Corynebacterium pseudotuberculosis, and Corynebacterium ulcerans. The results obtained in this study indicate that Acanthamoeba polyphaga is not optimal as surrogate host, while both Caenorhabtitis elegans and Galleria larvae seem to offer tractable models for rapid assessment of virulence between strains. Caenorhabtitis elegans gives more differentiated results and might be the best model system for pathogenic corynebacteria, given the tractability of bacteria and the range of mutant nematodes available to investigate the host response in combination with bacterial virulence. Nevertheless, Galleria will also be useful in respect to innate immune responses to pathogens because insects offer a more complex cell-based innate immune system compared with the simple innate immune system of C. elegans.