Zarmina Durrani

Differences in immunogenicity and protection in mice and guinea pigs following intranasal immunization with Helicobacter pylori outer membrane antigens

Mice and guinea pigs were intranasally immunized with either recombinant lipoprotein 20 or Helicobacter pylori outer membrane vesicles (OMV). Cholera toxin was used as mucosal adjuvant. In mice, both vaccines elicited systemic and local IgG responses, which correlated with significantly lower levels of H. pylori colonization. In contrast, only OMV proved immunogenic in guinea pigs, with the development of both systemic and local immune responses. These antibodies did not, however, correlate with protection in these animals, which suggests that vaccine formulation is as important as choice of antigen in the development of an H. pylori vaccine.

Investigation of the biological relevance of Helicobacter pylori cagE locus diversity, presence of CagA tyrosine phosphorylation motifs and vacuolating cytotoxin genotype on IL-8 induction in gastric epithelial cells

Isolates of Helicobacter pylori from dyspeptic patients in England and South Africa were tested for ability to induce interleukin-8 (IL-8) in gastric cells. All isolates were cagA-positive, which was used as a marker for the presence of the cag pathogenicity island. The aims were to determine if activities were related to diversity within cagE (HP0544), a locus encoding a key component in the Type IV secretion system, and if disease severity might be linked to a combination of strain features. We found that isolates were heterogeneous in ability to induce IL-8 activity with the 23 positive isolates (59%) showing activities ranging from 260 to 3200 pg ml(-1). The cagE locus was detected in most isolates and RFLP analysis of a 1.52-kb internal fragment showed interstrain diversity with 12 combined (MboI/NlaIII) types. Most cagE genotypes were not associated with IL-8 induction, however two genotypes were found only in IL-8-inducing strains and one genotype was associated with lack of IL-8 induction. IL-8 activity was not associated with either the number or composition of cagA tyrosine phosphorylation motifs and vacA m-type. Although we found a weak association between cagE type and the ability to induce IL-8, our results imply that gastric cell factors or bacterial factors other than vacA, cagA and cagE are involved in the induction of IL-8 and the development of severe gastric disease.

Orogastric vaccination of guinea pigs with Helicobacter pylori sonicate and a high dose of cholera toxin lowers the burden of infection

Guinea pigs were vaccinated orogastrically with Helicobacter pylori cell sonicate (CS) and 10 microg or 100 microg cholera toxin (CT) or CT only. Nai;ve animals were used as a control. In both experiments, vaccination primed the local IgG and IgA response, irrespective of the CT dose. After challenge, only the group of animals immunised with CS and 100 microg CT had a significantly lower number of H. pylori in the antral region of the stomach, but vaccination did not prevent H. pylori infection. This protective effect was not associated with a switch in IgG subclass, which remained predominantly IgG2. The levels of specific antibodies in serum and the gastric mucosa which were similar to naive unprotected animals. In conclusion, the ability of mucosal adjuvants such as CT to induce a protective immune response may be host dependent and findings in the Helicobacter-mouse model should be interpreted with caution.

A Study of PA DNA/Dendron Nanoparticles for Genetic Immunisation Against Anthrax

Protective antigen (PA), the binding subunit of toxins produced by Bacillus anthracis is singularly the most important antigen required for specific immunity to anthrax disease. We used cationic poly-lysine dendrons to develop a genetic anthrax vaccine. Plasmid CMV/ER PA83 which encoded full length PA 83 was complexed with dendrons to form dendriplexes. Two types of dendron were used: C0 and C18.These were mixed with DNA to form dendriplexes, of ap- proximately 80 nm in size, which were tested for immunogenicity. A/J and BALB/c mice were vaccinated with dendri- plexes containing 1� g and 50 � g plasmid DNA per dose over a period of 6 weeks. Immunisation with naked PA DNA did not induce an antibody response even after secondary boosting, whereas both dendriplexes produced strong anti-PA anti- body response. This response was dose dependent. We conclude that dendriplexes show superior immunogenicity com- pared to naked PA DNA in both mouse strains and that C18 dendriplexes with 50 �g plasmid DNA are most efficacious. However, the elicited antibodies did not neutralise lethal toxin in vitro. Therefore further work is required to improve these preparations in order to elicit functional antibodies.