2. Geographical Distribution http://www.who.int/emc-documents/surveillance/docs/whocdscsrisr2001.html 14 million cases (2 million new cases each year), distributed in 88 countries (350 million people at risk). cutaneous leishmaniasis visceral leishmaniasis

3. Clinical syndromes Cutaneous leishmaniasis – L. braziliensis, L. amazonensis, L. guyanensis L. major Diffuse leishmaniasis – L. amazonensis Mucocutaneous leishmaniasis – L. braziliensis Visceral leishmaniasis – L. chagasi, L. donovani New World species Old World species

4. Clinical picture AIDS (Persistence) Chemotherapy Gene regulation Virulence

5. Sacks, David L. (2001) Cellular Microbiology 3 (4), 189-196. Development in the insect vector Stage-regulated gene expression

6. 2.1 kb L S rRNA Characterisation of the meta 1 gene in L. major - 12 kDa, pI 9.7 - expressed in promastigotes, upregulated in metacyclics - localises to vacuoles close to the flagellar pocket - conserved in all Leishmania species

7. A BALC/c 46810121416 5 10 15 20 25 Lesion Index Weeks WT HE B Meta 1 overexpressor in vivo promastigotes amastigotes

8. 0 100 200 300 400 500 600 PBS pVXORF-1 pVX-mcp3 pVX-M1 pVX-mcp3- M1 Concentration (ng/mL) IgG1 IgG2a * * Meta 1 DNA Vaccine meta 1 is a strong Th2 inducer

9. L. major L. amaz. L. donovani META domain: Small domain family found in proteins of of unknown function. Some are secreted and implicated in motility in bacteria. Also occurs in Leishmania spp. as an essential gene. Over-expression in L.amazonensis increases virulence. A pair of cysteine residues show correlated conservation, suggesting that they form a disulphide bond. Taxa: cellular organisms cellular organisms

11. M1

12. 7,46 9,49 4,40 2,37 Probe B L S A 1,8 2,2 2,0 Kb Probe A L S A 2,37 Probe M2 L S A 1,35 0,24 Kb Probe K2 L S A Kb 1,8 2,0 Probe K1 L S A 2,1 Probe M1 L S A

14. 1 2 3 4 5 6 246810121416182022242628303234 contig Lm572 L. major (Sanger Centre) M1 CARD M2 AcyltransferaseHelicase ? Lariat-deb Pseudomonas ?

15. META2 META1NOD Acyltransf U5 VISTA UK Human Genome Mapping Project Resource Centre Funded by the MRC EMBOSS (European Molecular Biology Open Software) http://www.hgmp.mrc.ac.uk/

16. 0 20 40 60 80 100 1 3579 1113151719 -250 -200 -150 -100 -50 0 La gaps Lm gaps ORF UTR/IG Nt Similarity (%) Gaps calculated by Point Replacer v.2.2 (www.geocities.com/alvesjp/software)

17. Conserved synteny and linkage in Leishmania

18. Conserved synteny and linkage in trypanosomatids

19. B A 1 50 100 150 200 250 300 350 400 444 META M2a M2b M2c M2

20. A B C Pré-imune Anti- MBP Anti- M2/MBP

21. L a. WT L a. M2 L.a. M2/GFP Phase contrast DAPI  -Meta 2 e-GFP

23. A C BINIBIÇÃO

24. Conclusions Meta 1 and meta 2 genes- conserved in Leishmania, synteny with Trypanosoma Meta 1 related to virulence; Meta 2 ? Meta genes: several copies in Trypanosoma – evaluate function in T. brucei (RNAi) Meta 2 protein: Calcium? regulation of calpain activity? Meta locus: other stage-regulated genes - virulence ?

25. Camila S. Ramos Fernando A. L. Franco Amanda R. Franco Mariana Wajc Carlos H. Serezani Jenicer K. U. Yokoyama-Yasunaka Silvia R. B. Uliana Deborah F. Smith ICB - USP Imperial College University of London