- To clone novel mycobacterial antigen genes by direct T cell expression cloning using M. tuberculosis specific human T cell lines and clones.
- To prioritize recombinant M. tuberculosis antigens based upon their T cell recognition by a panel of PPD positive healthy donors and by patients with active tuberculosis.
- To prioritize recombinant M. tuberculosis antigens based upon their ability alone or in combination, to prime T cell responses in mice and guinea pigs.
- To evaluate the vaccine potential of recombinant antigens in mice, guinea pigs, and monkey models of tuberculosis, emphasizing adjuvants and delivery systems suitable for use in humans.
- To develop a vaccine for a class C bioterrorism agent, multi-drug resistant TB (MDR-TB). GLP data including immunogenicity, safety, protection, and therapeutic cure of existing M. tuberculosis infections, using pre-clinical grade material, will be generated.
Capabilities
Antigen identification and validation
Development of large scale protein purification, and the characterization of proteins for vaccine development.
Antigen characterization using human, non-human primate and rodent blood products
Activities within this area include 1) the conception, design and development of vaccine candidate antigens for TB 2) production, laboratory analysis and release of recombinant mycobacterial antigens 3) basic research to establish mechanisms of inducing long-lasting protective immunity
Immunology
Our group has expertise in using advanced multicolor flow cytometry to identify and characterize T-cell subsets and cytokines, and to determine their role in tuberculosis pathogenesis. We evaluate the heterogeneity of T-cells, key differences in the role of T cells in the progression of tuberculosis disease, and the broad spectrum of potential functions that T cells can generate in response to vaccination or infection. We apply our experience and knowledge of human immunology and flow cytometry to better understand immune responses to vaccines in humans and animal models of tuberculosis with the hope of identifying surrogates of disease progression and correlates of protective immunity.
In vivo models
We perform immunogenicity and efficiacy testing of experimental vaccine antigen and adjuvant candidates for TB vaccines using standardized animal models.
Adjuvant development
IDRI has dedicated BL3 facilities for in vitro and in vivo work.
Microbiology
The group has extensive experience in handling Mycobacterium cultures, and can perform determination of colony-forming units (CFU) from infected organs in-house.
Key skills and expertise
The TB Vaccine Development Research group has extensive experience with Mycobacterium tuberculosis:
- Culture and handling
- Microbiology
- Molecular biology (gene cloning, protein expression & purification)
- Development of vaccine and diagnostic antigens
- In vivo models of TB
Publications
1. Keeler, E., M.D. Perkins, P. Small, C. Hanson, S.G. Reed, J. Cunningham, J.E. Aledort, L. Hillborne, M.E. Rafael, F. Girosi, and C. Dye. 2006. Reducing the global burden of tuberculosis: The contribution of improved diagnostics. Nature Nov 23;444 Suppl 1:49-57.
2. Tsenova, L., R. Harbacheuski, A.L. Moreira, E. Ellison, W. Dalemans, M.R. Alderson, B. Mathema, S.G. Reed, Y.A. Skeiky, and G. Kaplan. 2006. Evaluation of the Mtb72F polyprotein vaccine in a rabbit model of tuberculous meningitis. Infect. Immun. 74:2392-2401.
3. Mukherjee S., S.S. Kashino, Y. Zhang, N. Daifalla, V. Rodrigues, Jr., S.G. Reed, and A. Campos-Neto. 2005. Cloning of the gene encoding a protective Mycobacterium tuberculosis secreted protein detected in vivo during the initial phases of the infectious process. J. Immunol. 175: 5298–5305.
4. Irwin, S.M., A.A. Izzo, S.W. Dow, Y.A. Skeiky, S.G. Reed, M.R. Alderson, and I.M. Orme. 2005. Tracking antigen-specific CD8 T lymphocytes in the lungs of mice vaccinated with the Mtb72F polyprotein. Infect. Immun. 73:5809-5816.
5. Skeiky, Y.A., M.R. Alderson, P.J. Ovendale, Y. Lobet, W. Dalemans, I.M. Orme, S.G. Reed, and A. Campos-Neto. 2005 Protection of mice and guinea pigs against tuberculosis induced by immunization with a single Mycobacterium tuberculosis recombinant antigen, MTB41. Vaccine 23:3937-3745.
6. Reece, S.T., N. Stride, P. Ovendale, S.G. Reed, and A. Campos-Neto. 2005. Skin test performed with highly purified Mycobacterium tuberculosis recombinant protein triggers tuberculin shock in infected guinea pigs. Infect. Immun. 73:3301-3306.
7. Brandt, L., Y.A. Skeiky, M.R. Alderson, Y. Lobet, W. Dalemans, O.C. Turner, R.J. Basaraba, A.A. Izzo, T.M. Lasco, P.L. Chapman, S.G. Reed, and I.M. Orme. 2004. The protective effect of the Mycobacterium bovis BCG vaccine is increased by coadministration with the Mycobacterium tuberculosis 72-kilodalton fusion polyprotein Mtb72F in M. tuberculosis-infected guinea pigs. Infect. Immun. 72:6622-6632.
8. Skeiky, Y.A., M.R. Alderson, P.J. Ovendale, J.A. Guderian, L. Brandt, D.C. Dillon, A. Campos-Neto, Y. Lobet, W. Dalemans, I.M. Orme, and S.G. Reed. 2004. Differential immune responses and protective efficacy induced by components of a tuberculosis polyprotein vaccine, Mtb72F, delivered as naked DNA or recombinant protein. J. Immunol. 172:7618-7628.
9. Mukherjee, S., N. Daifalla, Y. Zhang, J. Douglass, L. Brooks, T. Vedvick, R. Houghton, S.G. Reed, and A. Campos-Neto. 2004. Potential serological use of a recombinant protein that is a replica of a Mycobacterium tuberculosis protein found in the urine of infected mice. Clin. Diagn. Lab. Immunol. 11:280-286.
10. Liu, C., E. Flamoe, H.J. Chen, D. Carter, S.G. Reed, and A. Campos-Neto. 2004. Expression and purification of immunologically reactive DPPD, a recombinant Mycobacterium tuberculosis skin test antigen, using Mycobacterium smegmatis and Escherichia coli host cells Can. J. Microbiol. 50:97-105.
11. Reed, S.G., M.R. Alderson, W. Dalemans, Y. Lobet, and Y.A. Skeiky. 2003. Prospects for a better vaccine against tuberculosis. Tuberculosis (Edinb).83:213-219.
12. Campos-Neto, A., V. Rodrigues-Júnior, D.B. Pedral-Sampaio, E.M. Netto, P.J. Ovendale, R.N. Coler, Y.A.W. Skeiky, R. Badaró and S.G. Reed. 2001. Evaluation of DPPD, a single recombinant Mycobacterium tuberculosis protein, as an alternative antigen for the Mantoux test. Tuberculosis 81:353-358.
13. Lodes, M.J., D.C. Dillon, R. Mohamath, C.H. Day, D.R. Benson, L.D. Reynolds, P. McNeill, Y.A.W. Skeiky, R. Badaro, D.H. Persing, S.G. Reed, and R.L. Houghton. 2001. Serological expression cloning and immunological evaluation of MTB48, a novel Mycobacterium tuberculosis antigen. J. Clin. Microbiol. 39:2485-2493.
14. Coler, R.N., A. Campos-Neto, P. Ovendale, F.H. Day, S.P. Fling, L. Zhu, N. Serbina, J.L. Flynn, S.G. Reed, and M.R. Alderson. 2001. Vaccination with the T cell antigen Mtb 8.4 protects against challenge with Mycobacterium tuberculosis. J. Immunol. 166:6227-6235.
15. Lewinsohn, D.M., L. Zhu, V.J. Madison, D.C. Dillon, S.P. Fling, S.G. Reed, K.H. Grabstein, and M.R. Alderson. 2001. Classically restricted human CD8(+) T lymphocytes derived from Mycobacterium tuberculosis-infected cells: definition of antigenic specificity. J. Immunol. 166:439-446.
16. Lewinsohn, D.M., A.L. Briden, S.G. Reed, K.H. Grabstein, and M.R. Alderson. 2000. Mycobacterium tuberculosis-reactive CD8+ T lymphocytes: the relative contribution of classical versus nonclassical HLA restriction.J. Immunol. 165:925-930.
17. Dillon, D.C., M.R. Alderson, C.H. Day, T. Bement, A. Campos-Neto, Y.A.W. Skeiky, T. Vedvick, R. Badaro, S.G. Reed, and R.L. Houghton. 2000. Molecular and immunological characterization of Mycobacterium tuberculosis CFP-10, an immunodiagnostic antigen missing in Mycobacterium bovis BCG.J. Clin. Microbiol. 38:3285-3290.
18. Coler, R.N., Y.A.W. Skeiky, P.J. Ovendale, T.S. Vedvick, L. Gervassi, J. Guderian, S. Jen, S.G. Reed, and A. Campos-Neto. 2000. Cloning of a Mycobacterium tuberculosis gene encoding a PPD protein that elicits strong tuberculosis specific delayed type hypersensitivity. J. Infect. Dis. 182:224-233.
19. Hendrickson, R.C., J.F. Douglass, L.D. Reynolds, P.D. McNeill, D. Carter, S.G. Reed, and R.L. Houghton. 2000. Mass Spectrometric identification of Mtb81: a novel serological marker for tuberculosis. J. Clin. Microbiol. 38:2354-2361.
20. Skeiky, Y.A.W., P.J. Ovendale, S. Jen, M.R. Alderson, D.C. Dillon, S. Smith, C.B. Wilson, I.M. Orme, S.G. Reed, and A. Campos-Neto. 2000. T cell expression cloning of a Mycobacterium tuberculosis gene encoding a protective antigen associated with the early control of infection. J. Immunol. 165:7140-7149.
21. Alderson, M.R., T. Bement, C.H. Day, L. Zhu, D. Molesh, Y.A.W. Skeiky, R. Coler, D.M. Lewinsohn, S.G. Reed, and D.C. Dillon. 2000. Expression cloning of an immunodominant family of Mycobacterium tuberculosis antigens using human CD4+ T cells. J. Exp. Med. 191:551-559.
22. Skeiky, Y.A.W., M.J. Lodes, J.A. Guderian, R. Mohamath, T. Bement, M.R. Alderson, and S.G. Reed. 1999. Cloning, expression, and immunological evaluation of two putative secreted serine protease antigens of Mycobacterium tuberculosis. Infect. and Immun. 67:3998-4007.
23. Dillon, D., M.R. Alderson, C.H. Day, D. Lewinsohn , R. Coler, T. Bement, A. Campos-Neto, Y.A.W. Skeiky, I.M. Orme, S. Steen, W. Dalemans, R. Badaro, and S.G. Reed. 1999. Molecular characterization and human T cell responses to a member of a novel Mycobacterium tuberculosis Mtb39 gene family. Infect. and Immun. 67:2941-2950.
24. Coler R.N., Y.A.W. Skeiky, T. Vedvick, T. Bement, P. Ovendale, A. Campos-Neto, M.R. Alderson, and S.G. Reed. 1998. Molecular cloning and immunological reactivity of a novel low-molecular-mass antigen of Mycobacterium tuberculosis. J. Immunol. 161:2356-2364.
25. Webb, J.R., T.S. Vedvick, M.R. Alderson, J.A. Guderian, P.J. Ovendale, S.M. Johnson, S.G. Reed, and Y.A.W. Skeiky. 1998. Molecular cloning, expression and immunogenicity of MTB12, a novel low molecular weight secreted antigen of Mycobacterium tuberculosis. Infect. and Immun. 66:4208-4214.
26. Lewinsohn, D.M., M.R. Alderson, A.L. Briden, S.R. Riddell, S.G. Reed, and K.H. Grabstein. 1998. Characterization of human CD8+ T cells reactive with Mycobacterium tuberculosis-infected antigen presenting cells. J. Exp. Med. 187:1633-1640.