Factors influencing the protective efficacy of a BCG homologous prime-boost vaccination regime against tuberculosis

Tuberculosis due to Mycobacterium spp. continues to represent a major threat to human and animal health, prompting the search for effective vaccines. We have previously reported a sequential prime and boost homologous vaccination regime, using live avirulent M. bovis bacille Calmette-Guerin (BCG) strain 1173P2, that can provide significant protection to red deer (Cervus elaphus) against virulent M. bovis infection. Here, we have investigated the influence of varying the time-periods during and following the vaccination regime on the subsequent outcome of disease following post-vaccination pathogen challenge. Deer vaccinated using the standard regime of BCG-prime (week 0) and BCG-boost (week 8) followed by M. bovis challenge (week 14–16) were highly protected, showing significant reductions in the incidence of M. bovis infection and tuberculous lesions, as well as reduced pathogen burdens in sentinel lymphatic tissues. Decreasing the time-period between primary and secondary immunisations from 8 to 4 weeks had no significant impact on the protective efficacy afforded by BCG vaccination, while increasing this period to 43 weeks largely ablated protection. Increasing the time-period between secondary immunisation and M. bovis challenge from 6 to 26 or 52 weeks also had no significant impact on protection, suggesting that an appropriately timed BCG prime-boost vaccination regime can establish long-lasting protective immunological memory in deer. Finally, increasing the time-period between virulent M. bovis challenge and the subsequent post-mortem examination of disease outcome indicated that – once vaccinated by the standard prime-boost regime – deer remain refractory to disease if challenged and maintained for up to 52 weeks, suggesting that vaccinated animals harbouring low numbers of virulent M. bovis organisms do not succumb to disease activation over time. These findings are discussed in relation to concurrent measurements of in vivo and ex vivo immunological markers of disease and protection, as well as the wider implications of a standardised vaccine regimen for practical use in animal health.