Dawn Nolt Lab
Background
Mycobacterium tuberculosis (Mtb), the bacterium responsible for tuberculosis (TB), infects 1/3 of the world's population and is a major cause of morbidity and mortality. The CD8+ T cell response is recognized as important in the coordinated immune response against Mtb.
Once phagocytosed by antigen-presenting cells (APCs), Mtb resides in a modified phagosome, a compartment normally associated with MHC II. There is increasing evidence that suggests the exciting possibility that the phagosome may participate in MHC I-mediated “cross-presentation” of Mtb antigens to CD8+ T cells. Our goal is to investigate the role of phagosomes in cross-presentation during Mtb infection.
Current projects
- Phenotyping the phagosome with relationship to dose and duration of Mtb infection of human dendritic cells
- Tracking the pathway of class I (particularly HLA-E, a non-classical MHC molecule) within human dendritic cells during Mtb infection
- Understanding the role of ubiquitination in defining the trafficking destination of class I
Techniques
- Flow cytometry and flow organellometry
- Immunofluorescent microscopy
- Cytokine analysis - ELISPOT
- Western blot analysis
- Extensive cell culturing and cloning
- Subcellular organelle separation
- Real-time RT PCR
- siRNA
- Transfections and transductions
Significance
Our long-term goal is to investigate the interaction of Mtb and CD8+ T cell immunity. Our studies will advance knowledge in key areas: 1) Mtb interactions in host cells and 2) modulation of immune function during Mtb infection. Since a future immunogenic vaccine against Mtb will likely need to incorporate CD8+ T cell responses, our findings will guide the design and development of future vaccines and therapeutics against Mtb.
