Enhanced immunisation and expression strategies using bacterial spores as heat-stable vaccine delivery vehicles

Spores of the gram positive bacterium Bacillus subtilis have been used as oral vaccine vehicles and have been shown to confer protection against tetanus in mice. In this work, we have used tetanus as a model with which to examine different antigen presentation strategies as well as dosing regimes using the TTFC antigen of Clostridium tetani. We show that display of the antigen on the spore surface produces a more pronounced, but less rapid, response than when the antigen is expressed in the germinating spore; though this response has a clear Th1 bias. When using the nasal dosing route ten times less spores were needed to produce the same level of protective antibodies using surface display of TTFC. Expression of the antigen within the germinating spore by contrast enables as little as three oral doses on consecutive days to generate protection with a noticeable Th2 bias. We have also shown that TTFC can be expressed using two routes, using display on the spore surface as well as in the germinating spore. This dual route produced the best responses in terms of magnitude and speed of neutralising response as well as a clear Th1 bias indicating the involvement of cellular as well as humoral responses.