Development of a Mimotope Vaccine Targeting the Staphylococcus aureus Quorum Sensing Pathway

A major hurdle in vaccine development is the difficulty in identifying relevant target epitopes and then presenting them to the immune system in a context that mimics their native conformation. We have engineered novel virus-like-particle (VLP) technology that is able to display complex libraries of random peptide sequences on a surface-exposed loop in the coat protein without disruption of protein folding or VLP assembly. This technology allows us to use the same VLP particle for both affinity selection and immunization, integrating the power of epitope discovery and epitope mimicry of traditional phage display with the high immunogenicity of VLPs. Previously, we showed that using affinity selection with our VLP platform identifies linear epitopes of monoclonal antibodies and subsequent immunization generates the proper antibody response. To test if our technology could identify immunologic mimotopes, we used affinity selection on a monoclonal antibody (AP4-24H11) that recognizes the Staphylococcus aureus autoinducing peptide 4 (AIP4). AIP4 is a secreted eight amino acid, cyclized peptide produced from the S. aureus accessory gene regulator (agrIV) quorum-sensing operon. The agr system coordinates density dependent changes in gene expression, leading to the upregulation of a host of virulence factors, and passive transfer of AP4-24H11 protects against S. aureus agrIV-dependent pathogenicity. In this report, we identified a set of peptides displayed on VLPs that bound with high specificity to AP4-24H11. Importantly, similar to passive transfer with AP4-24H11, immunization with a subset of these VLPs protected against pathogenicity in a mouse model of S. aureus dermonecrosis. These data are proof of principle that by performing affinity selection on neutralizing antibodies, our VLP technology can identify peptide mimics of non-linear epitopes and that these mimotope based VLP vaccines provide protection against pathogens in relevant animal models.     

Epitope-Specific Anti-hCG Vaccines on a Virus Like Particle Platform

The possibility of a contraceptive vaccine targeting human chorionic gonadotropin has long been recognized, but never fully realized. Here we describe an epitope-specific approach based on immunogenic display of hCG-derived peptides on virus-like particles of RNA bacteriophage. A number of recombinant VLPs were constructed, each displaying a different hCG-derived peptide. Some were taken from the disordered C-terminal tail of the hormone, another came from an internal loop, and yet another was an epitope mimic produced by affinity-selection on an hCG-neutralizing antibody target. Immunization of mice with some VLPs yielded antisera that bound the hormone and neutralized hCG biological activity.     

Non-enzymatic recombination of RNA: Ligation in loops

Background

While the RNA world hypothesis is widely accepted, it is still far from complete: the existence of self-replicating ribozyme, consisting of potentially hundreds of nucleotides, is a core assumption for the majority of RNA world models. The appearance of such long RNA molecules under prebiotic conditions is not self-evident. Recombination seems to be a plausible way of creating RNA diversity, resulting in the appearance of functional RNAs, capable of self-replicating.

Effects of kaolin on the composition of generalist predator assemblages and parasitism of Choristoneura rosaceana (Lep., Tortricidae) in apple orchards

Abstract:  In three apple orchards, we tested how the hydrophilic kaolin clay particle film Surround WP affected the diversity of generalist arthropod predator assemblages in orchard foliage and the parasitism of the pest species Choristoneura rosaceana (Harris) (obliquebanded leafroller) (Lep., Tortricidae). In two orchards, kaolin was applied to orchard foliage once a week for 4 weeks, between mid-June and mid-July in 2004 and 2005. In the third orchard kaolin was applied to foliage twice over 2 weeks in June 2004. We quantified the proportion of larvae C. rosaceana parasitized, larval populations, and the relative abundance and assemblage composition of generalist predators (spiders and insects) in the orchards. Kaolin altered the species composition of the generalist predator assemblages and reduced the relative abundances of certain generalist predators, most notably jumping and crab spiders (Salticidae and Philodromidae), assassin bugs (Reduviidae), ants (Formicidae) and coccinellids (Coccinellidae). In contrast, the relative abundances of web-spinning spiders (Araneidae, Dictynidae, Theridiidae) were not affected. Kaolin did not affect the proportion of parasitized C. rosaceana larvae, which ranged from 24% to 47% in control and kaolin treatments, or the relative proportions of parasitoid taxa. The kaolin formulation did not affect the abundance of C. rosaceana larvae, but in one orchard, kaolin did reduce the abundance of the combined numbers of C. rosaceana and another tortricid pest, Argyrotaenia velutiana . Although kaolin does not affect parasitism of C. rosaceana , it significantly changes the composition of generalist predator assemblages in orchard foliage.     

Thermal biology and immersion tolerance of the Beringian pseudoscorpion <Emphasis Type="Italic">Wyochernes asiaticus</Emphasis>

Wyochernes asiaticus (Arachnida: Pseudoscorpiones: Chernetidae) is a pseudoscorpion distributed across Beringia, the areas of Yukon, Alaska and Siberia that remained unglaciated at the last glacial maximum. Along with low temperatures, its streamside habitat suggests that submergence during flood events is an important physiological challenge for this species. We collected W. asiaticus in midsummer from 66.8°N Yukon Territory, Canada, and measured thermal and immersion tolerance. Wyochernes asiaticus is freeze-avoidant, with a mean supercooling point of ?6.9 °C. It remains active at low temperatures (mean critical thermal minimum, CT_min, is ?3.6 °C) and has a critical thermal maximum (CT_max) of 37.8 °C, which is lower than other arachnids and consistent with its restriction to high latitudes. Fifty per cent of W. asiaticus individuals survived immersion in oxygen-depleted water for 17 days, suggesting that this species has high tolerance to immersion during flooding events. To our knowledge, these are the first data on the environmental physiology of any pseudoscorpion and a new addition to our understanding of the biology of polar microarthropods.