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Peter J Hudson

Publications and source records attributed to Peter J Hudson.

At least 19 recordsLinked to original sources
PubMed2006

Latest technologies for the enhancement of antibody affinity.

High affinity antibodies are crucial both for the discovery and validation of biomarkers for human health and disease and as clinical diagnostic and therapeutic reagents. This review describes some of the latest technologies for the design, mutation and selection of high affinity antibodies that provide a paradigm for molecular evolution of a far wider range of proteins including enzymes. Strategies include both in vivo and in vitro methods and embrace the latest concepts for antibody display and selection. Specifically, affinity enhancement can be tailored to the target-binding surface, typically the complementary determining region (CDR) loops in antibodies, whereas enhanced stability, expression or catalytic properties can be affected by selected changes to the core protein scaffold. Together, these technologies provide a rapid and powerful strategy to drive the next generation of protein-based reagents for numerous clinical, environmental and agribusiness applications.

Animals↗
PubMed2006

Is a healthy ecosystem one that is rich in parasites?

Historically, the role of parasites in ecosystem functioning has been considered trivial because a cursory examination reveals that their relative biomass is low compared with that of other trophic groups. However there is increasing evidence that parasite-mediated effects could be significant: they shape host population dynamics, alter interspecific competition, influence energy flow and appear to be important drivers of biodiversity. Indeed they influence a range of ecosystem functions and have a major effect on the structure of some food webs. Here, we consider the bottom-up and top-down processes of how parasitism influences ecosystem functioning and show that there is evidence that parasites are important for biodiversity and production; thus, we consider a healthy system to be one that is rich in parasite species.

Animals↗
PubMed2006

Engineering of the Escherichia coli Im7 immunity protein as a loop display scaffold.

Protein scaffolds derived from non-immunoglobulin sources are increasingly being adapted and engineered to provide unique binding molecules with a diverse range of targeting specificities. The ColE7 immunity protein (Im7) from Escherichia coli is potentially one such molecule, as it combines the advantages of (i) small size, (ii) stability conferred by a conserved four anti-parallel alpha-helical framework and (iii) availability of variable surface loops evolved to inactivate members of the DNase family of bacterial toxins, forming one of the tightest known protein-protein interactions. Here we describe initial cloning and protein expression of Im7 and its cognate partner the 15 kDa DNase domain of the colicin E7. Both proteins were produced efficiently in E.coli, and their in vitro binding interactions were validated using ELISA and biosensor. In order to assess the capacity of the Im7 protein to accommodate extensive loop region modifications, we performed extensive molecular modelling and constructed a series of loop graft variants, based on transfer of the extended CDR3 loop from the IgG1b12 antibody, which targets the gp120 antigen from HIV-1. Loop grafting in various configurations resulted in chimeric proteins exhibiting retention of the underlying framework conformation, as measured using far-UV circular dichroism spectroscopy. Importantly, there was low but measurable transfer of antigen-specific affinity. Finally, to validate Im7 as a selectable scaffold for the generation of molecular libraries, we displayed Im7 as a gene 3 fusion protein on the surface of fd bacteriophages, the most common library display format. The fusion was successfully detected using an anti-Im7 rabbit polyclonal antibody, and the recombinant phage specifically recognized the immobilized DNase. Thus, Im7 scaffold is an ideal protein display scaffold for the future generation and for the selection of libraries of novel binding proteins.

Amino Acid Sequence↗
PubMed2006

Testing the role of parasites in driving the cyclic population dynamics of a gamebird.

The role of parasites in regulating populations has been the subject of debate. We tested whether parasites caused population cycles in red grouse by manipulating parasite intensities in four, paired 1 km(2) study areas during cyclic population declines over 4 years. Parasite reductions led to (1) larger grouse broods, (2) higher population densities in both autumn and spring, (3) reduced autumn population declines in one of two regions, and (4) reduced spring declines, but only in the first year. We infer that a single trophic interaction between a parasite and its host does not explain cyclic dynamics in spring breeding density in this species, although it contributed to the start of a cyclic decline. Another process was operating to drive the populations down. Together with our other results these findings emphasize that both trophic and intrinsic processes may act within populations to cause unstable dynamics.

Animals↗
PubMed2006

Localized deer absence leads to tick amplification.

Deer support high tick intensities, perpetuating tick populations, but they do not support tick-borne pathogen transmission, so are dilution hosts. We test the hypothesis that absence of deer (loss of a dilution host) will result in either an increase or a reduction in tick density, and that the outcome is scale dependent. We use a complementary methodological approach starting with meta-analysis, followed up by a field experiment. Meta-analysis indicated that larger deer exclosures reduce questing (host-seeking) tick density, but as the exclosure becomes smaller (<2.5 ha) the questing tick density is increased (amplified). To determine the consequences for tick-borne pathogen transmission we carried out a field experiment, comparing the intensity of ticks that fed on hosts competent for tickborne pathogen transmission (rodents) in two small (<1 ha) deer exclosures and their replicated controls. Intensity of larval ticks on rodents was not significantly different between treatments, but nymph intensity, the tick stage responsible for tick-borne encephalitis (TBE) transmission, was higher in deer exclosures. TBE seropositive rodents were found in a deer exclosure but not in the controls. We propose that localized absence of deer (loss of a dilution host) increases tick feeding on rodents, leading to the potential for tick-borne disease hotspots.

Animals↗
PubMed2005

Interactions between intrinsic and extrinsic mechanisms in a cyclic species: testosterone increases parasite infection in red grouse.

Field studies of mechanisms involved in population regulation have tended to focus on the roles of either intrinsic or extrinsic factors, but these are rarely mutually exclusive and their interactions can be crucial in determining dynamics. Experiments on red grouse Lagopus lagopus scoticus have shown that population instability can be caused both by the effects of a parasitic nematode, Trichostrongylus tenuis, on host production or by changes in testosterone influencing aggressive behaviour and recruitment. We experimentally tested for an interaction between testosterone and T. tenuis in free-living male grouse. A total of 123 grouse were caught in autumn, treated with an anthelmintic to remove parasites, and then given either testosterone or empty, control, implants. After one month grouse were re-infected with a standard dose of parasites. We show that males with increased testosterone levels had greater parasite intensities than controls after one year. We discuss possible physiological and behavioural mechanisms linking testosterone and increased parasite intensity, and the implications for our understanding of complex, unstable population dynamics.

Aggression↗
PubMed2005

Separating behavioral and physiological mechanisms in testosterone-mediated trade-offs.

Testosterone often mediates trade-offs between reproduction and other life-history traits, which are usually investigated using testosterone implants. However, this approach does not distinguish between the physiological and behavioral effects of testosterone. We studied a wild game bird, the red grouse Lagopus lagopus scoticus, and took a new approach to investigate mechanisms linking elevated testosterone to increased parasite intensity. We caught males in autumn, removed their parasites, implanted them with the antiandrogen flutamide in combination with an aromatase inhibitor (FA males) or with empty implants (control males), and challenged them with parasites. The FA treatment increased testosterone concentration and physiological stress, but without enhancing testosterone-dependent behaviors, because testosterone receptors were blocked. FA males ended up with more parasites than the control males the following autumn, an effect similar to that of a testosterone treatment reported elsewhere. However, and unlike the testosterone treatment, the FA treatment did not affect home range, pairing, or breeding success. The results supported a physiological mechanism (increased susceptibility) linking elevated testosterone and increased parasite intensity. The FA treatment provided a new way of investigating testosterone-mediated trade-offs whereby testosterone concentration was increased while the effects on behavior were blocked, resulting in physiological costs without phenotypic benefits.

Albumins↗
PubMed2005

Parasites and climate synchronize red grouse populations.

There is circumstantial evidence that correlated climatic conditions can drive animal populations into synchronous fluctuations in abundance. However, it is unclear whether climate directly affects the survival and fecundity of individuals, or indirectly, by influencing food and natural enemies. Here we propose that climate affects trophic interactions and could be an important mechanism for synchronizing spatially distributed populations. We show that in specific years the size of red grouse populations in northern England either increases or decreases in synchrony. In these years, widespread and correlated climatic conditions during May and July affect populations regionally and influence the density-dependent transmission of the gastrointestinal nematode Trichostrongylus tenuis, a parasite that reduces grouse fecundity. This in turn forces grouse populations into synchrony. We conclude that specific climatic events may lead to outbreaks of infectious diseases or pests that may cause dramatic, synchronized changes in the abundance of their hosts.

Animals↗
PubMed2005

Prevalence, intensity and aggregation of intestinal parasites in mountain hares and their potential impact on population dynamics.

Mountain hare Lepus timidus populations show unstable dynamics and since hares carry a significant helminth infection and host-parasite interactions are known to be destabilising, they have been proposed as a possible causal mechanism for the observed instability. We assessed the prevalence, intensity of infection and aggregation of the helminth parasites Graphidium strigosum and Trichostrongylus retortaeformis recovered from 589 mountain hares culled from 30 Scottish sporting estates in 1999 and 2000. Graphidium strigosum showed low prevalence and intensity of infection and was highly aggregated. In contrast, T. retortaeformis showed high prevalence and intensity of infection and a low degree of aggregation. Differences in body condition of the hares were best explained by a model including sex and month of collection and interaction terms for sex-month and intensity of infection of T. retortaeformis-month. The low degree of aggregation of T. retortaeformis and the significant negative effect of intensity of infection on body condition are in accordance with the hypothesis that the host-parasite interaction is the causative destabilising mechanism for mountain hare dynamics.

Animals↗
PubMed2005

Engineered antibody fragments and the rise of single domains.

With 18 monoclonal antibody (mAb) products currently on the market and more than 100 in clinical trials, it is clear that engineered antibodies have come of age as biopharmaceuticals. In fact, by 2008, engineered antibodies are predicted to account for >30% of all revenues in the biotechnology market. Smaller recombinant antibody fragments (for example, classic monovalent antibody fragments (Fab, scFv)) and engineered variants (diabodies, triabodies, minibodies and single-domain antibodies) are now emerging as credible alternatives. These fragments retain the targeting specificity of whole mAbs but can be produced more economically and possess other unique and superior properties for a range of diagnostic and therapeutic applications. Antibody fragments have been forged into multivalent and multi-specific reagents, linked to therapeutic payloads (such as radionuclides, toxins, enzymes, liposomes and viruses) and engineered for enhanced therapeutic efficacy. Recently, single antibody domains have been engineered and selected as targeting reagents against hitherto immunosilent cavities in enzymes, receptors and infectious agents. Single-domain antibodies are anticipated to significantly expand the repertoire of antibody-based reagents against the vast range of novel biomarkers being discovered through proteomics. As this review aims to show, there is tremendous potential for all antibody fragments either as robust diagnostic reagents (for example in biosensors), or as nonimmunogenic in vivo biopharmaceuticals with superior biodistribution and blood clearance properties.

Animals↗
PubMed2004

Ticks need not bite their red grouse hosts to infect them with louping ill virus.

For pathogens transmitted by biting vectors, one of the fundamental assumptions is often that vector bites are the sole or main route of host infection. Here, we demonstrate experimentally a transmission route whereby hosts (red grouse, Lagopus lagopus scoticus) became infected with a member of the tick-borne encephalitis virus complex, louping ill virus, after eating the infected tick vector. Furthermore, we estimated from field observations that this mode of infection could account for 73-98% of all virus infections in wild red grouse in their first season. This has potential implications for the understanding of other biting vector-borne pathogens where hosts may ingest vectors through foraging or grooming.

Animals↗
PubMed2004

Competition and mutualism among the gut helminths of a mammalian host.

Most animal species are infected with multiple parasite species; however, the role of interspecific parasite interactions in influencing parasite dynamics and shaping parasite communities has been unclear. Although laboratory studies have found evidence of cross-immunity, immunosuppression and competition, analyses of hosts in the field have generally concluded that parasite communities are little more than random assemblages. Here we present evidence of consistent interspecific interactions in a natural mammalian system, revealed through the analysis of parasite intensity data collected from a free-ranging rabbit (Oryctolagus cuniculus) population, sampled monthly for a period of 23 yr. The wild rabbit plays host to a diverse gut helminth community that reflects the communities seen in other economically important domestic herbivores. These findings suggest that parasite interactions could have profound implications for the dynamics of parasite communities. The efficacy of parasite control programmes could be jeopardized if such interactions are not taken into account. In contrast, a clear understanding of such interactions may provide the basis for the development of more environmentally acceptable methods of parasite control.

Animals↗
PubMed2004

Selection and affinity maturation of IgNAR variable domains targeting Plasmodium falciparum AMA1.

The new antigen receptor (IgNAR) is an antibody unique to sharks and consists of a disulphide-bonded dimer of two protein chains, each containing a single variable and five constant domains. The individual variable (V(NAR)) domains bind antigen independently, and are candidates for the smallest antibody-based immune recognition units. We have previously produced a library of V(NAR) domains with extensive variability in the CDR1 and CDR3 loops displayed on the surface of bacteriophage. Now, to test the efficacy of this library, and further explore the dynamics of V(NAR) antigen binding we have performed selection experiments against an infectious disease target, the malarial Apical Membrane Antigen-1 (AMA1) from Plasmodium falciparum. Two related V(NAR) clones were selected, characterized by long (16- and 18-residue) CDR3 loops. These recombinant V(NAR)s could be harvested at yields approaching 5mg/L of monomeric protein from the E. coli periplasm, and bound AMA1 with nanomolar affinities (K(D)= approximately 2 x 10(-7) M). One clone, designated 12Y-2, was affinity-matured by error prone PCR, resulting in several variants with mutations mapping to the CDR1 and CDR3 loops. The best of these variants showed approximately 10-fold enhanced affinity over 12Y-2 and was Plasmodium falciparum strain-specific. Importantly, we demonstrated that this monovalent V(NAR) co-localized with rabbit anti-AMA1 antisera on the surface of malarial parasites and thus may have utility in diagnostic applications.

Amino Acid Sequence↗
PubMed2003

Structure of an influenza neuraminidase-diabody complex by electron cryomicroscopy and image analysis.

The structure of a complex between a bivalent diabody and its antigen, influenza neuraminidase, has been determined by electron cryomicroscopy of single particles and image analysis. A three-dimensional reconstruction has been interpreted in terms of high-resolution X-ray models of the component proteins. The complex consists of two neuraminidase tetramers cross-linked by four diabodies with 422 point symmetry. The structure and symmetry of the complex is determined uniquely by packing constraints consistent with the maximum possible number of diabody cross-links. Diabodies may provide a useful approach to the structure determination of small proteins by incorporating the proteins into large symmetric complexes followed by single-particle electron microscopy.

Cryoelectron Microscopy↗
PubMed2003

Transmission dynamics of a trematode parasite: exposure, acquired resistance and parasite aggregation.

This paper examines the relative importance of exposure and susceptibility to the infection of rainbow trout ( Oncorhynchus mykiss) with the trematode parasite Diplostomum spathaceum under natural conditions. A total of 93 individually marked, similarly aged fish were introduced into three cages at regular time intervals and the intensity of infection in individuals recorded by counting parasites in live fish using ophthalmic techniques. Fish introduced into the cages became infected faster than fish that were already in the cages, indicating that fish developed resistance to infection after repeated exposure. Fish kept in the cages experienced similar levels of exposure and the distribution of parasites between these fish was not significantly different from a random distribution. In contrast, parasites from 16 Finnish wild roach populations were highly aggregated. The differences between the caged fish and the wild fish indicate that the aggregated distribution in wild fish might be determined by variations in exposure rather than variations in susceptibility between fish. This is one of the few studies to demonstrate the development of resistance in fish against the parasite under natural conditions, and to attempt to separate exposure and susceptibility as causative agents of parasite aggregation.

Animals↗
PubMed2003

Thresholds for disease persistence in models for tick-borne infections including non-viraemic transmission, extended feeding and tick aggregation.

Lyme disease and Tick-Borne Encephalitis (TBE) are two emergent tick-borne diseases transmitted by the widely distributed European tick Ixodes ricinus. The life cycle of the vector and the number of hosts involved requires the development of complex models which consider different routes of pathogen transmission including those occurring between ticks that co-feed on the same host. Hence, we consider here a general model for tick-borne infections. We assumed ticks feed on two types of host species, one competent for viraemic transmission of infection, the second incompetent but included a third transmission route through non-viraemic transmission between ticks co-feeding on the same host. Since a blood meal lasts for several days these routes could lead to interesting nonlinearities in transmission rates, which may have important effects.We derive an explicit formula for the threshold for disease persistence in the case of viraemic transmission, also for the case of viraemic and non-viraemic transmission. From this formula, the effect of parameters on the persistence of infection can be determined. When only viraemic transmission occurs, we confirm that, while the density of the competent host has always a positive effect on infection persistence, the density of the incompetent host may have either a positive effect, by amplifying tick population, or a negative ("dilution") effect, by wasting tick bites on an incompetent host. With non-viraemic transmission, the "dilution" effect becomes less relevant. On the other hand, if the nonlinearity due to extended feeding is included, the dilution effect always occurs, but often at unrealistically high host densities. Finally, we incorporated the effects of tick aggregation on the hosts and correlation of tick stages and found that both had an important effect on infection persistence, if non-viraemic transmission occurred.

Animals↗