Wellcome Trust Research Career Development Fellowship: A personal perspective Samantha Sampson Department of Microbiology Centre for Molecular Microbiology and Infection

Career History Cape Town, South Africa: BSc (Biochemistry & Chemistry) BSc Hons, MSc & PhD (Medical Biochemistry) Mycobacterium tuberculosis strain diversity

Career History Boston, USA: Post-doc Novel TB vaccine efficacy studies

Career History London, UK: Research Fellow Host-pathogen interactions: PE/PPE proteins

WT RCDF Opportunity for postdoctoral scientists to become independent research scientists and to undertake research of high quality Eligibility: 3-6 years research experience from date of PhD degree intellectual contributions to published research demonstrate potential to carry out independent research

WT RCDF Application Procedure: Preliminary application form (25/9/08) Full application (to appropriate funding committee) Panel Interview

My research Worldwide TB incidence

TB: Disease Burden 1/3 rd of world’s population infected with Mycobacterium tuberculosis 8 million incident cases 2 million deaths annually

PE/PPE proteins PPE-MPTR subgroup (up to ~3720 aa) (GG A / N GG A / N ) n PE-PGRS subgroup (up to >1500 aa) PE NXGXGNXG PPE PPE-SVP subgroup (up to ~470 aa) GXXSVPXXWPPE PE “PE-only” subgroup (~110 aa) Subgroup with unique C-terminal region (up to ~560 aa) Subgroup with unique C-terminal region (up to ~600 aa) PE PPE

The proposal Title: The PE and PPE proteins of Mycobacterium tuberculosis and Mycobacterium bovis: exploring their potential as variable antigens Hypothesis: PE and PPE genes of M. tuberculosis and M. bovis encode variable antigens

The proposal Aims: (i) Establish whether the PE and PPE genes are differentially expressed under different in vitro growth conditions and during host infection. (ii) Determine whether PE and PPE epitopes are differentially recognised by the host immune system over the course of an infection. (iii) Elucidate mechanisms of PE and PPE gene regulation.

Kinetics of PE/PPE immune responses >PE35 MEKMSHDPIAADIGTQVSDNALHGVTAGSTALTSVTGLVPAGADEVSAQAATAFTSEGIQLLASNASA... MEKMSHDPIAADIGTQVSDN IAADIGTQVSDNALHGVTAG VSDNALHGVTAGSTALTSVT VTAGSTALTSVTGLVPAGAD TSVTGLVPAGADEVSAQAAT AGADEVSAQAATAFTSEGIQ QAATAFTSEGIQLLASNASA ºC, 5% CO 2, 48 hours Store -80 ºC until use

PE/PPE regulation Labeled probe (0.8 ng Rv1195 upstream region) Unlabeled probe Protein (100 ng, rRv0485-His 6 ) DNA-protein complex Unbound, labelled DNA

Why was my application successful? WT Remit: “… research across all biomedical science … to produce knowledge that can be used to protect and improve human and animal health.” Preliminary data Sponsor Institution Track record Preparation Right place at the right time!

Independent funding: Pros Independence Limited teaching obligations (if you choose) Technical assistance Opportunity to develop own research interests Develop management skills (time, budget, personnel)

Independent funding: Cons Independence Finding a balance Measurable outputs required

Advice Be flexible (personally and professionally) Be pro-active Have a Plan B Research your options D O WHAT YOU ENJOY !!

Acknowledgments Prof. Rob Warren (PhD Supervisor) Prof. Barry Bloom (Post-doc PI) Prof. Douglas Young (RCDF Sponsor) Rachael Goldstone (RA)