Biomedical subjects

P Wigley

Publications and source records attributed to P Wigley.

11 recordsLinked to original sources

PubMed2006

Cross-reactive cellular and humoral immune responses to Salmonella enterica serovars Typhimurium and Enteritidis are associated with protection to heterologous re-challenge.

Chickens infected with Salmonella enterica serovars Typhimurium (ST) and Enteritidis (SE) still represent a major source of human food poisoning via consumption of contaminated meat and eggs. Vaccination represents a sustainable approach to control Salmonella in the chicken and the serovar specificity of immunity has the potential to impact on the need for multivalent vaccines. The issue of cross-reactive immune responses and cross-serovar protection was examined in these experiments. Cellular and humoral immune responses were measured by antigen-specific ELISA and splenocyte proliferation assays during primary infections (with ST and SE) and during a second challenge with homologous or heterologous serovars. Primary infection with ST or SE induced strong lymphocyte proliferation and high levels of specific antibody (IgM, IgG and IgA) responses with substantial serovar cross-reactivity. The occurrence of high levels of splenocyte proliferation and strong antibody responses corresponded to the initiation of clearance with both ST and SE. Re-challenge of ST and SE infection-primed chickens with either serovar resulted in significant levels of protection (assessed by bacterial numbers and rate of clearance) with little difference between homologous or heterologous challenge schedules. Relatively low levels of antigen-specific splenocyte proliferation were detected during secondary infection, which may be caused by splenic T cells exiting to the gut. In contrast, the more rapid specific antibody responses (compared with primary infection controls) indicate the development of a secondary antigen-specific adaptive response. The substantial level of cross-protection between serovars and the level of antigenic cross-reactivity indicates the potential for single serovar live vaccines to protect against both group B and D salmonellae.

Animals↗

PubMed2004

Age at primary infection with Salmonella enterica serovar Typhimurium in the chicken influences persistence of infection and subsequent immunity to re-challenge.

Salmonella enterica remains one of the most important food-borne pathogens of humans and is often acquired through consumption of infected poultry meat or eggs. Control of Salmonella infections in chicken is therefore an important public health issue. Infection with S. enterica serovar Typhimurium results in a persistent enteric infection without clinical disease in chickens of more than 3 days of age, and represents a source for contamination of carcass at slaughter and entry into the human food chain. Data presented indicate a profound effect of age at initial exposure on the persistence of infection and a lesser effect on the development of effective immunity to re-challenge. The percentage of birds positive for Salmonella was high until 8-9 weeks of age, regardless of the age at which the birds were infected (1, 3 or 6 weeks). The birds infected at 3 and 6 weeks of age produced a more rapid and higher antibody response (IgY and IgA) than those infected at 1 week of age, but in all cases infection persisted for a considerable period despite the presence of high antibody levels. Following a re-challenge infection with S. Typhimurium, all three previously-infected groups had fewer bacteria in the gut, spleen and liver compared with age-matched birds receiving a parallel primary infection. However, the birds primary infected at 3 and 6 weeks of age cleared infection more rapidly than those infected at a younger age. Interestingly older-primed birds had higher specific T lymphocyte proliferative responses and specific circulating levels of IgY antibody at time of re-challenge. Although birds initially infected at 1 week of age and those that were previously uninfected produced a stronger antibody response following re-challenge, they were slower to clear Salmonella from the gut than the older-primed groups which expressed a stronger T lymphocyte response. The data presented indicate that clearance of Salmonella from the gut is age-dependent and we propose that this relates to the increased competence of the enteric T cell response. The findings that Salmonella persists beyond 8-9 weeks, irrespective of age at exposure, has implications for the broiler sector and indicates the need to remain Salmonella free throughout the rearing period. Moreover, the re-challenge data demonstrates that infection at a young age is less effective in producing protective immunity than in older chickens. This feature of the development of protective immunity needs to be considered when developing vaccines for the broiler sector of the poultry industry.

Age Factors↗

PubMed2004

Faecal shedding and intestinal colonization of Salmonella enterica in in-bred chickens: the effect of host-genetic background.

Considerable and reproducible differences were observed in the amount and duration of faecal excretion when in-bred lines of chickens were infected orally with S. enterica serovar Typhimurium at 6 weeks of age after being given a gut flora preparation when newly hatched. Similar but less pronounced results were observed with S. Enteritidis or S. Infantis. Differences in the viable numbers of the inoculated bacteria in caecal contents were detectable within 24 h of inoculation. No major differences were seen in Salmonella-specific serum IgA or IgG titres. Small differences were seen in the numbers of circulating heterophilic cells. Caecal contents taken from the more resistant lines immediately prior to challenge appeared to be no more inhibitory for Salmonella in vivo than contents taken from susceptible lines. The more resistant lines showed a slightly higher rate of intestinal flow, as indicated by the rate of production of faecal droppings, although there was no difference in the rate of emptying of the caeca. In an F1 generation resistance was dominant and not sex-linked. There was no MHC linkage or any association with SAL1, the gene implicated in resistance to systemic salmonellosis in chickens, or NRAMP1.

Age Factors↗

PubMed2004

Temporal dynamics of the cellular, humoral and cytokine responses in chickens during primary and secondary infection with Salmonella enterica serovar Typhimurium.

Salmonella enterica serovar Typhimurium (S. Typhimurium) infections cause systemic disease in the young chick, whereas in the older chicken the infection is mainly restricted to the intestine. Chickens infected orally with S. Typhimurium (F98) at 6 weeks of age and re-infected 10 weeks later were monitored for antibody production, T-cell proliferation and production of selected cytokines (interferon-gamma, interleukin-1beta and transforming growth factor-beta(4)). A strong coordinated antigen-specific humoral and cellular immune response was temporally linked to resolution of the primary infection. Enhanced levels of mRNA encoding the cytokines, interleukin-1beta, transforming growth factor-beta(4) and interferon-gamma were also evident during early phases of primary infection. Secondary infection was restricted to the intestine and of shorter duration than primary infection. Splenic immune responses were not further enhanced by secondary infection; indeed, antigen-specific proliferation was significantly reduced at 1 day after secondary infection, which may be interpreted as the trafficking of reactive T cells from the spleen to the gut.

Animals↗

PubMed2003

Contribution of proton-translocating proteins to the virulence of Salmonella enterica serovars Typhimurium, Gallinarum, and Dublin in chickens and mice.

We investigated the attenuating effects of a range of respiratory chain mutations in three Salmonella serovars which might be used in the development of live vaccines. We tested mutations in nuoG, cydA, cyoA, atpB, and atpH in three serovars of Salmonella enterica: Typhimurium, Dublin, and Gallinarum. All three serovars were assessed for attenuation in their relevant virulence assays of typhoid-like infections. Serovar Typhimurium was assessed in 1-day-old chickens and the mouse. Serovar Gallinarum 9 was assessed in 3-week-old chickens, and serovar Dublin was assessed in 6-week-old mice. Our data show variation in attenuation for the nuoG, cydA, and cyoA mutations within the different serovar-host combinations. However, mutations in atpB and atpH were highly attenuating for all three serovars in the various virulence assays. Further investigation of the mutations in the atp operon showed that the bacteria were less invasive in vivo, showing reduced in vitro survival within phagocytic cells and reduced acid tolerance. We present data showing that this reduced acid tolerance is due to an inability to adapt to conditions rather than a general sensitivity to reduced pH. The data support the targeting of respiratory components for the production of live vaccines and suggest that mutations in the atp operon provide suitable candidates for broad-spectrum attenuation of a range of Salmonella serovars.

Animals↗

PubMed2001

Salmonella enterica serovar Pullorum persists in splenic macrophages and in the reproductive tract during persistent, disease-free carriage in chickens.

Salmonella enterica serovar Pullorum is worldwide a poultry pathogen of considerable economic importance, particularly in those countries with a developing poultry industry. In addition to the characteristic high mortality rates among young chicks, one of the features of Salmonella serovar Pullorum infection is that it persists for long periods in convalescent chicks in the absence of clinical disease. This can lead to colonization of the reproductive tract of chickens and at sexual maturity can result in infected progeny through transovarian transmission to eggs. The sites of Salmonella serovar Pullorum persistence in convalescent birds are not known, and the mechanisms of persistence are not understood. Here we show that Salmonella serovar Pullorum can persist in both the spleen and the reproductive tract for over 40 weeks following experimental infection in chickens. During the period of sexual maturity, Salmonella serovar Pullorum colonized both the ovary and the oviduct of hens and led to 6% of laid eggs being infected by Salmonella serovar Pullorum. The colonization of several different sites of the reproductive tract suggests that Salmonella serovar Pullorum may employ more than one mechanism of egg infection. Persistence occurred despite a strong humoral response, suggesting an intracellular site of infection. By use of a Salmonella serovar Pullorum strain containing a plasmid stably expressing green fluorescent protein, we demonstrated that the main site of carriage in the spleen is within macrophages. This raises interesting questions about the biology of Salmonella serovar Pullorum, including why there is an increase in bacterial numbers when birds become sexually mature and in particular how Salmonella serovar Pullorum avoids clearance by macrophages and whether it modulates the immune system in other ways.

Animals↗

PubMed2001

Salmonella enterica serovar Gallinarum requires the Salmonella pathogenicity island 2 type III secretion system but not the Salmonella pathogenicity island 1 type III secretion system for virulence in chickens.

Salmonella enterica serovar Gallinarum is a host-specific serotype that causes the severe systemic disease fowl typhoid in domestic poultry and a narrow range of other avian species but rarely causes disease in mammalian hosts. Specificity of the disease is primarily at the level of the reticuloendothelial system, but few virulence factors have been described other than the requirement for an 85-kb virulence plasmid. In this work, by making functional mutations in the type III secretion systems (TTSS) encoded by Salmonella pathogenicity island 1 (SPI-1) and SPI-2, we investigated the role of these pathogenicity islands in interactions between Salmonella serovar Gallinarum and avian cells in vitro and the role of these pathogenicity islands in virulence in chickens. The SPI-1 mutant showed decreased invasiveness into avian cells in vitro but was unaffected in its ability to persist within chicken macrophages. In contrast the SPI-2 mutant was fully invasive in nonphagocytic cells but failed to persist in macrophages. In chicken infections the SPI-2 mutant was attenuated while the SPI-1 mutant showed full virulence. In oral infections the SPI-2 mutant was not observed in the spleen or liver, and following intravenous inoculation it was cleared rapidly from these sites. SPI-2 function is required by Salmonella serovar Gallinarum for virulence, primarily through promoting survival within macrophages allowing multiplication within the reticuloendothelial system, but this does not preclude the involvement of SPI-2 in uptake from the gut to the spleen and liver. SPI-1 appears to have little effect on virulence and survival of Salmonella serovar Gallinarum in the host.

Animals↗

PubMed2000

Multiple chromosomes in Burkholderia cepacia and B. gladioli and their distribution in clinical and environmental strains of B. cepacia.

Burkholderia cepacia is found in soils and waters, it can be used in biocontrol and bioremediation but is also a human pathogen. It is not yet clear what differentiates pathogenic from non-pathogenic strains of the organism. In this study the multiple replicon structure was investigated in 28 strains of B. cepacia by pulsed field gel electrophoresis. All strains examined, whether of clinical, environmental or plant pathogenic origin, were found to have two, three or four large (> 500 kbp) replicons. Many strains also contained small replicons. Clinical strains were more likely to have three or four large replicons than non-clinical strains. Multiple replicon structure was also demonstrated in B. gladioli and Alcaligenes eutrophus.

Burkholderia↗

PubMed2000

Susceptibility of multiresistant strains of Burkholderia cepacia to honey.

Twenty strains of Burkholderia cepacia, isolated principally from the sputum of cystic fibrosis patients, were tested for their susceptibility to eight antibiotics with a modified Kirby-Bauer Disc diffusion technique. All strains exhibited multiple but not identical patterns of antibiotic resistance. The sensitivity of all strains to honey was assessed with an agar dilution method. All strains exhibited susceptibility to concentrations of honey below 6% (v/v). This suggests that honey may have a potential role in the clinical management of B. cepacia infections.

Burkholderia Infections↗

PubMed1999

Genotypic and phenotypic relationships in Burkholderia cepacia isolated from cystic fibrosis patients and the environment.

Twenty-one strains of Burkholderia cepacia isolated from the environment, and 21 clinical strains isolated principally from sputum of cystic fibrosis (CF) patients, were characterized genotypically by macrorestriction analysis (genome fingerprinting) and PCR ribotyping, and phenotypically by susceptibility to antibiotics and the ability to macerate onion tissue. The plasmid content of the strains was also investigated. Environmental isolates showed a high degree of genetic variability, all strains differing from both one another and the CF isolates. The CF isolates were less variable, with common strains found in patients attending three geographically distinct CF centres. Phenotypic variation was found both within and between CF and environmental strains. Generally, CF isolates displayed higher levels of antibiotic resistance, while the ability to macerate onion tissue was more prevalent amongst environmental isolates. Plasmids were more frequently found in CF isolates, but were of similar size in both groups of strains. Such variability is not surprising in view of the existence of multiple genomovars within the B. cepacia complex.

Burkholderia cepacia↗

PubMed1994

Site-specific transgene insertion: an approach.

Methods to improve the production of transgenic animals are being developed. Conventional transgenesis, involving microinjection of DNA into fertilized eggs, has a number of limitations. These result from the inability to control both the site of transgene insertion and the number of gene copies inserted. The approach described seeks to overcome these problems and to allow single copy insertion of transgenes into a defined site in animal genomes. The method involves the use of embryonic stem cells, gene targeting and the FLP recombinase system.

Animals↗