Low dose BCG vaccination in mice : immune responses and implications for tuberculosis control
Gebreyohannes, Tadele Kiros
The outcome of an infection is often determined by the qualitative nature of the immune response generated against the infectious agent. Various intracellular pathogens, including those that cause leprosy, tuberculosis, leishmaniasis, and most probably malaria and AIDS appear to require a predominant cell-mediated, Th1, response for effective containment, whereas the generation of a mixed Th1/Th2 or predominantly Th2 response is associated with progressive disease. Therefore, any attempt to develop universally efficacious vaccination against these pathogens must generate an immunological imprint that ensures a strong and stable cell-mediated response upon natural infection with the relevant pathogen. We report here critical tests of a strategy designed to achieve such an imprint using Bacille-Calmette-Guérin (BCG) vaccine. BCG vaccine is an attenuated form of M. bovis, the causative agent of tuberculosis in cattle, and is the most widely used vaccine in humans. However, considerable uncertainty still surrounds its efficacy against tuberculosis both in man and animals. As the protective dose is not known, BCG has been given at the maximum tolerable dose. However, recent studies in mice and other animals have shown that the dose of an antigen can be a critical factor in determining the type of immune response generated. I tested the general hypothesis that low dose vaccination would preferentially induce cell-mediated immune response and generate a Th1 imprint that can protect the host against intracellular pathogens in the particular case of mycobacteria. To this end, both adult and newborn mice were vaccinated with different doses of BCG or saline and cell-mediated and humoral immune responses were assessed at different times post-vaccination. Several weeks after vaccination, mice from each group were challenged with a dose of BCG that induces a mixed Th1/Th2 response in naïve mice, and the T-cell and antibody responses were assessed using ELISPOT and ELISA assays, respectively. The splenic bacterial burden was also determined using colony formation on agar plates. Our results show that the class of immunity induced by BCG depends on the dose employed for vaccination, independent of the route of administration and the age and strain of mice used. In all cases, lower doses induce an exclusive cell-mediated, Th1, response with no antibody production, while higher doses induce either a mixed Th1/Th2 response or a predominantly Th2, humoral response, with higher titers of both IgG1 and IgG2a antibodies. Following intravenous high dose BCG challenge, all mice in the vaccinated groups developed a Th1 response associated with a more efficient clearance of BCG from the spleen. The greatest clearance of mycobacteria was generated following vaccination with lower doses, as low as 33 cfu of BCG. In addition, our findings demonstrate that newborn mice are not inherently biased towards generating Th2 responses, but they can generate Th1 responses and Th1 imprints if appropriate vaccination protocols (dose, route and time) are employed. Furthermore, subcutaneous exposure of young mice to environmental mycobacteria can induce a mixed Th1/Th2 response that can abrogate the potential to generate Th1 responses and Th1 imprints upon vaccination with low doses of BCG vaccine. Low dose neonatal BCG vaccination can circumvent the interference caused by “impingement” of environmental mycobacteria on the immune system. Therefore, our observations strongly support a neonatal low dose BCG vaccination strategy to provide universally efficacious protection against infections by pathogenic mycobacteria.
DegreeDoctor of Philosophy (Ph.D.)
DepartmentMicrobiology and Immunology
ProgramMicrobiology and Immunology
SupervisorBretscher, Peter A.
CommitteeHoward, S. Peter; Havele, Calliopi; Griebel, Philip J.; Tabel, Henry