Tuberculosis (TB) remains a leading cause of morbidity and mortality world-wide, accounting for more than 2 million human lives every year. The overall goal of the TB VAC is to establish a joint academic-industrial consortium capable of taking tuberculosis vaccine candidates from the laboratory bench to phase I clinical trials.

TB is a leading infectious killer of adults in the world, causing about 2 million deaths a year, about 98% in developing countries. In these countries, the HIV pandemic has a strong impact on TB resulting in 1 million deaths per year. However, with the rise in drug-resistant strains of the disease, TB could once again become a major threat to Europe. Prevention through vaccination could be the most effective intervention, but as yet no vaccine effective against TB in the adult population is available.

In recent years several promising new vaccine strategies have been developed, particularly in the context of the FP5 project TB Vaccine Cluster. The FP6 TB VAC project aims to integrate the European efforts towards the development of novel tuberculosis vaccine candidates and forward these vaccines to small scale, Phase I human clinical trials in Europe and in Africa. Under the co-ordination of Animal Sciences Group in Lelystad, The Netherlands the project joins 30 leading European Institutions from 9 European countries and 2 African countries, including 2 major vaccine producers. A website for TB-VAC with a public and private section is available at www.tb-vac.org

The main goals of TB-VAC Integrated Project are:

  • The discovery and development of new promising tuberculosis vaccine candidates effective in the young adult population
  • The development of tests that predict vaccine efficacy in humans
  • The clinical evaluation of lead candidates in small initial trials in Europe and Africa
  • Capacity building in developing countries for clinical evaluation of vaccines
  • Liaise with other consortia (eg. MUVAPRED) to coordinate specific activities
  • Liaise with the European Developing Countries Clinical Trials Partnership (EDCTP) to enable further large clinical trials in African Countries.

TB-VAC consists of two tracks:

Track 1. Optimisation of existing vaccine candidates towards Phase I trials.

The strategic research focuses on goal-oriented research to optimize delivery, composition and evaluation of selected, promising candidate vaccines discovered during FP5 (subunit vaccines 72F, Hyb-1; modified vaccinia ankara (MVA) constructs expressing Ag85A), and new FP6 developed vaccines selected from track 2. In parallell, the strategic research component aims to identify correlates of protection and markers of TB disease that may facilitate (accelerated) monitoring of future clinical trials. The downstream development component will carry out GMP production of optimised vaccine candidates, establish pre-clinical files and carry out Phase I clinical trials in European and TB-endemic countries. According to the results, the Consortium will decide upon a strategy to exploit this portfolio.

A series of vaccine delivery and adjuvant combinations have been evaluated resulting in promising lead formulations for optimisation of effective and safe subunit vaccines. New models have been developed to monitor induction of memory T cells, an important tool to evaluate improved and prolongued memory necessary for a vaccine to be effective in adults (WP1). The lead vaccines from FP5 (72F, Hyb-1; MVAAg85A) have been further developed and tested in pre-clinical models (WP5), and optimised formulations have been produced under GMP conditions.

A DDA/TDB adjuvant combination (LipoVac) has been fully characterised and a stable GLP preparation is now available. Further improvement of this adjuvant through combination with other (viral) delivery systems or a series of recently identified mycobacterial immuno-modulators is underway, in addition to the study of their effects on antigen presentation, activation of presenting cells, induction of long-lived memory T cells, and polarising Th1/Th2 cells. A mycobacterial expression system has been developed allowing purification of subunit vaccines from a mycobacterial background to assess the effects of differential posttranslational modification of sub-unit vaccines on vaccine efficacy (WP1). Future work will focus on the most promising delivery systems and adjuvants currently discovered, and development of their use in a clinical setting. The safety, immunogenicity and efficacy of a range of subunit and live vaccines has been assessed in pre-clinical models (ao. Ag85A-expressing MVA booster vaccine candidate, new Mtb and BCG live attenuated vaccine candidates, and Hybrid-1 in IC31 and DDA/TDB adjuvants) to underpin their subsequent evaluation during clinical development. The specific effects of a range of vaccines in neonatal models as well as models pre-exposed to worm infection has been evaluated to assess their application in the neonate and in an endemic setting where the majority of individuals is exposed to helminths. Several candidate non protein antigens (sulfoglycolipids, PIMs, and phosphoantigens) have been evaluated in preclinical models to assess their applicability as sub-unit vaccine and/or their adjuvant capacities through the stimulation of CD1 restricted, gamma-delta or NK T cells. Based on these findings, a sulpho-glycolipid antigen has been selected for further optimisation and development as a vaccine candidate (WP5). Future work in WP5 will continue but focus on the preclinical models whereas the most promising non protein antigen work will be transferred to WP1. With the aim to identify and clinically apply correlate markers of protection or disease, a number of potential molecules associated with protection or disease have now been identified. These include the HBHA antigen, ESAT6, and several latency antigens that allow discrimination of latent from active TB. In addition, granulysin levels in serum have been shown to correlate with TB disease stages. New lipid components have been identified, as well as their various recognising T-cell populations, that are associated with differential presence in BCG vaccinated and latently infected individuals. DC-sign polymorphisms, or differential expression levels of DC-sign and small Ras GTP-ases (Rab 33A) in host cells associate with protection or with active disease. Various T cell populations with a differential expression of specific ligands were found to associate with either BCG vaccination, TB disease, or latent infection. Several of thee above are now being further tested in ongoing BCG trials to evaluate their potential for application in a clinical setting.

Downstream development activities

The production of new lots of candidate vaccines (MVA-Ag85A, Mtb72f H1-IC31) has been initiated as well as initial steps towards GMP production of a live improved recombinant BCG (Hly-BCG ). Four Product Development Teams (PDT) have been established to facilitate the move of each candidate vaccines or antigen combinations (H1-IC, MVA-Ag85A, Mtb72F, BCG-Hly) from discovery to development and clinical testing. Five PDT meetings were held to review and further define GMP processes, other regulatory issues and clinical development plans. From each of the candidate vaccines pre-clinical files are being centralised by collation of existing data from developers, assessment of physical procedures, immunological procedures, preparing of preliminary preclinical file, assessment and implementation of potential release criteria. Phase I/II clinical trials have been completed or initiated for MVA-Ag85A (In Oxford, UK and the Gambia; to assess vaccine safety and immunogenicity in individuals that were previously exposed to mycobacterial antigens, and for dose optimization), for Mtb72F (in Lausanne, Switzerland; in individuals previously exposed to BCG or infected with Mtb (previously treated)), or for H1-IC31(In Leiden, Netherlands; in PPD negative subjects). Further clinical trial sites are being prepared in AHRI, Ethiopia, and LEDANTEC, Senegal.

Track 2. Discovery of new vaccines.

This track will carry out innovative back-up research to identify and develop novel vaccines or vaccine antigens. We propose to optimise three selected live vaccine candidates that during FP5 showed improved efficacy as compared to BCG and show promise as vaccines in infants. Dormant or latent tuberculosis bacteria can persist for long periods in humans, and may be an important cause of tuberculosis in adults. We will focus on discovery of new vaccines based on antigens partiicularly produced in dormant or latent bacteria.

Discovery of new vaccines activities

New live vaccines (BCG ?Urea; LLO; BCG RD1 KO-mutant; PhoP/PhoR Mtb) have been constructed and evaluated in pre-clinical models. Strains without antibiotic markers are being constructed for application in a clinical setting. The promising findings with the BCG (?Urea; LLO) candidate resulted in it being selected to move for further production and clinical development. M.tb strains with mutations leading to different expression of PIM or mutated in genes for glycosyltransferase are also being constructed as new candidates. The down stream effects of Phop/PhoR mutations are being studied by lipidomic and proteomic approaches. An in vitro granuloma formation model based on beads coated with lipids has been established. Whereas TDM and LAM are only able to induce cellular recruitment during granuloma fomation, LM and PIMs induce both cellular recruitment and phagocyte differentiation.

Through transcriptome analyses of M. tuberculosis from lung lesions of patients suffering from active TB, several gene products were identified that were selectively expressed during active disease or latent infection. By comparative proteomics analyses of various TB-strains, antigens solely or differentially expressed by clinically relevant Beijing strains have been identified. Identified by other approaches, two promising new vaccine candidates (Rv3407 and heparin-binding hemagglutinin [HBHA]) showed considerable protective activity in experimental models using different prime-boost vaccination schemes. The HBHA antigen is now being considered for further clinical development in WP6. Cloning, expression and production of (recombinant) latency antigens and viral vectors encoding latency antigens has been completed, and recognition by human T cells as well as evaluation of their protective efficacy is being evaluated in preclinical models, including latency models.

 

PUBLICATIONS

Books

Kaufmann, Stefan H. E. / Rubin, Eric (eds.)
Handbook of Tuberculosis
Molecular Biology and Biochemistry
ISBN-10: 3-527-31886-0
ISBN-13: 978-3-527-31886-5

Kaufmann, Stefan H. E. / Britton, Warwick J. (eds.)
Handbook of Tuberculosis
Immunology and Cell Biology
ISBN-10: 3-527-31887-9
ISBN-13: 978-3-527-31887-2

Kaufmann, Stefan H. E. / van Helden, Paul (eds.)
Handbook of Tuberculosis
Clinics, Diagnostics, Therapy and Epidemiology
ISBN-10: 3-527-31888-7
ISBN-13: 978-3-527-31888-9

The three volumes can be ordered at http://eu.wiley.com/WileyCDA/WileyTitle/productCd-3527316833.html

Original Publications and Reviews

1. Rosenkrands I, Agger E M, W. Olsen A, S. Korsholm K, Swetman Andersen C, T. Jensen K, Andersen P. Cationic Liposomes Containing Mycobacterial Lipids: a New Powerful Th1 Adjuvant System. Infect. Immun. 2005, 73: 5817-5826.

2. Davidsen J, Rosenkrands I, Christensen D, Vangala A, Kirby D, Perrie Y, Agger E M, Andersen P. Characterization of cationic liposomes based on dimethyldioctadecylammonium and synthetic cord factor from M. tuberculosis (trehalose 6,6?-dibehenate)—A novel adjuvant inducing both strong CMI and antibody responses. Biochim Biophys Acta 2005, 1718:22-31.

3. Devilder M-C, Maillet S, Bouyge-Moreau I, Donnadieu E, Bonneville M, Scotet E Potentiation of Antigen-Stimulated Vg9Vd2 T Cell Cytokine Production by Immature Dendritic Cells (DC) and Reciprocal Effect on DC Maturation J.Immunol. 2006, 176:1386-93.

4. Ulrichs T, Lefmann M, Reich M, Morawietz L, Roth A, Brinkmann V, A. Kosmiadi G, Seiler P, Aichele P, Hahn H, Krenn V, B. Göbel U, H. E. Kaufmann S. Modified immunohistological staining allows detection of Ziehl-Neelsen-negative Mycobacterium tuberculosis organisms and their precise localization in human tissue. J.Pathol . 2005, 205: 633-40.

5. Ulrichs T, A. Kosmiadi G, Trusov V, Jör S, Pradl L, Titukhina M, Mishenko V, Gushina N, H. E. Kaufmann S. Human tuberculous granulomas induce peripheral lymphoid follicle-like structures to orchestrate local host defence in the lung. J. Pathol .2004, 204:217-28.

6. Jacobsen M, Repsilber D, Gutschmidt A, Neher A, Feldmann K, J. Mollenkopf H, Ziegler A, and H. E. Kaufmann S. Ras-Associated Small GTPase 33A, a Novel T Cell Factor, Is Down-Regulated in Patients with Tuberculosis. J.Inf. Dis. 2005,. 192:1211-8.

7. Mollenkopf H-J, Hahnke K , H. E. Kaufmann S. Transcriptional responses in mouse lungs induced by vaccination with Mycobacterium bovis BCG and infection with Mycobacterium tuberculosis. Microbes Infect. 2006, 8:136–144

8. H. E. Kaufmann S. Recent findings in immunology give tuberculosis vaccines a new boost . Trends. Immun ol. 2005, 26:660-7.

9. Rachman H, Strong M, Ulrichs T, Grode L, Schuchhardt J, Mollenkopf H, A. Kosmiadi G, Eisenberg D, H. E. Kaufmann S. Unique Transcriptome Signature of Mycobacterium tuberculosis in Pulmonary Tuberculosis. Infect. Immun. 2006, 74:1233-42

10. Repsilber D, Fink L, Jacobsen M, Blasing O, Ziegler A. Sample selection for microarray gene expression studies. Meth. Inf. Med. 2005;44:461-7.

11. Ulrichs T, A. Kosmiadi G, Jörg S, Pradl L, Titukhina M, Mishenko V, Gushina N, H. E. Kaufmann S. Differential Organization of the Local Immune Response in Patients with Active Cavitary Tuberculosis or with Nonprogressive Tuberculoma. J.Inf. Dis. 2005, 192:89-97.

12. Martin C. “The dream of a vaccine against tuberculosis; new vaccines improving or replacing BCG?” Eur Respir 2005 J 26: 162-167.

13. Brodin P, Majlessi L, Marsollier L, de Jonge MI, Bottai D, Demangel C, Hinds J, Neyrolles O, Butcher PD, Leclerc C, Cole ST, Brosch R. Dissection of ESAT-6 system 1 of Mycobacterium tuberculosis and impact on immunogenicity and virulence. Infect. Immun. 2006, 74: 88-98.

14. Tailleux L, Pham-Thi N, Bergeron-Lafaurie A, Herrmann J-L, Charles P, Schwart O, Scheinmann P, H. Lagrange P, de Blic J, Tazi A, Gicquel B, Neyrolles O. DC-SIGN Induction in Alveolar Macrophages Defines Privileged Target Host Cells for Mycobacteria in Patients with Tuberculosis. PLoS Med 2005, 2:1269-1279

15. Pitarque S, Herrmann J-L, Duteyrat J-L, Jackson M, R. Stewart G, Lecointre F, Payre B, Schwartz O, B. Young D, Marchal G, H. Lagrange P, Puzo G, Gicquel B, Nigou J, Neyrolles O. Deciphering the molecular bases of Mycobacterium tuberculosis binding to the lectin DC-SIGN reveals an underestimated complexity Biochem. J. 2005, 392:615-24.

16. B. Barreiro L, Neyrolles O, L. Babb C, Tailleux L, Quach H, McElreavey K, D. van Helden P, G. Hoal E, Gicquel B, Quintana-Murci L. Promoter Variation in the DC-SIGN–Encoding Gene CD209 Is Associated with Tuberculosis. PLoS Med 2006, 3: 203-235

17. Brodin P, de Jonge MI, Majlessi L, Leclerc C, Nilges M, Cole ST, Brosch R. Functional analysis of ESAT-6, the dominant T-cell antigen of Mycobacterium tuberculosis, reveals key residues involved in secretion, complex-formation, virulence and immunogenicity J. Biol. Chem. 2005, 280:33953-59.

18. Martin C, Williams A, Hernandez-Pando R, Cardona PJ, Gormley E, Bordat Y, Soto CY, Clark SO, Hatch GJ, Aguilar D, Ausina V, Gicquel B. The live Mycobacterium tuberculosis phoP mutant strain is more attenuated than BCG and confers protective immunity against tuberculosis in mice and guinea pigs. Vaccine. 2006 (on line)

19. Gonzalo Asensio J, Maia C, Ferrer NL, Barilone N, Laval F, Soto CY, Winter N, Daffe M, Gicquel B, Martin C, Jackson M. The virulence-associated two-component PhoP-PhoR system controls the biosynthesis of polyketide-derived lipids in Mycobacterium tuberculosis. J Biol Chem. 2006, 281:1313-6.

18. T. Temmerman S, Place S, Debrie A-S, Locht C, Mascart F. Effector functions of heparin-binding hemagglutinin-specific CD8+ T lymphocytes in latent human tuberculosis. J Infect Dis. 2005, 192:226-32.

19. Locht C, Hougardy J-M, Rouanet C, Place S, Mascart F. Heparin-binding hemagglutinin, from an extrapulmonary dissemination factor to a powerful diagnostic and protective antigen against tuberculosis. Tuberculosis. 2006. 27 Feb. Epub ahead of print.

20. Grode L, Seiler P, Baumann S, Hess J, Brinkmann V, Eddine AN, Mann P, Goosmann C, Bandermann S, Smith D, J. Bancroft G, Reyrat J-M, van Soolingen D, Raupach B, H.E. Kaufmann S. Increased vaccine efficacy against tuberculosis of recombinant Mycobacterium bovis bacille Calmette-Guérin mutants that secrete listeriolysin. J.Clin. Invest. 2005, 115:2472-2479.

21. Caccamo N, Meraviglia S, Ferlazzo V, Angelini D, Borsellino G, Poccia F, Battistini L, Dieli F, Salerno A. Differential requirements for antigen or homeostatic cytokines for proliferation and differentiation of human Vg9Vd2 naive, memory and effector T cell subsets. Eur. J.Immunol. 2005, 35:1764-72.

22. T. Kamath A, Fruth U, J. Brennan M, Dobbelaer R, Hubrechts P, Mei Ho M, E. Mayner R, Thole J, Walker K.B, Liu M, Lambert P-H. New live mycobacterial vaccines: the Geneva consensus on essential steps towards clinical development. Vaccine 2005, 23:3753-61

23. de la Salle H, Mariotti S, Angenieux C, Gilleron M, Garcia-Alles L-F, Malm D, Berg T, Paoletti S, Maître B, Mourey L, Salamero J, Cazenave J-P, Hanau D, Mori L, Puzo G,Gennaro De Libero M. Assistance of Microbial Glycolipid Antigen Processing by CD1e. Science 2005 :310: 1321 – 1324.