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Malaria and Dengue A mosquito-based approach

Malaria and Dengue A mosquito-based approach. Design anti-pathogen genes. Mosquito Population. Culex quinquefasciatus. West Nile fever. Ae. aegypti. An. darlingi. An. aquasalis. Malaria. Ae. aegypti Ae. albopictus. Dengue fever. Anti-pathogen genes. Recombinant antibodies

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Malaria and Dengue A mosquito-based approach

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  1. Malaria and Dengue A mosquito-based approach

  2. Design anti-pathogen genes • Mosquito Population

  3. Culex quinquefasciatus West Nile fever

  4. Ae. aegypti An. darlingi An. aquasalis Malaria

  5. Ae. aegypti Ae. albopictus Dengue fever

  6. Anti-pathogen genes • Recombinant antibodies • (scFv - single chain antibody) • Active peptides • Protein domains

  7. Recombinant antibodies • (scFv - single chain antibody) 1A10-2 1B7 – E protein 3H5 9A3D-8 2H2 – prM

  8. VH VL VL his L VH L 1A10-2-scFv Sindbis expression system

  9. cDNA RArm LArm 3xP3 EGFP SV40 VG-Promoter 5’UTR 3’UTR pBac[3xP3-EGFP, Vg-effector]

  10. scFv-Transgenic 21 Sporozoites/gland 1 0 2 5 3 10 99.8% 4 31 5 34 6 40 7 41 8 61 9 4,800 10 6,000 11 7,500 12 10,000 13 10,500 14 14,000 15 14,500 16 14,500 Plasmodium gallinaceum and Aedes aegypti

  11. Promoter 3’UTR 5’UTR Transgene Effectors Molecules

  12. Aedes aegypti fat body transcriptome 50.82% 30.05% 5.16% 7.97%

  13. Macro array ♀ ♂ 130 transcripts

  14. Proteomic approach on hemolymph proteins ♀ BF Sucrose 24 hs OV 48 hs 72 hs

  15. 17 11 12 13 15 14 6 18 7 20 8 10 16 19 9 1 2 3 4 5

  16. Pg1 Non-infected P.gal infected

  17. Mosquitoes Population

  18. José Eduardo Bracco e Dr. Maria Anice Sallun 3 1 23 5 9 7 2 14 Uganda, Singapore, Cambodia and Tahiti Senegal 13 18 The haplotype network for mitochondrial ND4 from Aedes aegypti populations from Brazil. Each line in the network represents a single mutational change. White circle represents a theoretical interior node. 17 15 20 22

  19. Ae. albopictus Ae. aegypti Specie-specific markers

  20. Dr. José Eduardo Levi • IMT - USP Dr. Margareth L. Capurro ICB – USP Bianca Burini Fernanda Rosa – MT - FAPESP Ricardo V. Araújo – DD - CNPq Fabiana M. Feitosa – DD - FAPESP JoãoV. Camargo e Silva – DD - CAPES Daniela G. Rodrigues – MT - CNPq Ceres M. de Miranda – MT - CNPq Márcia Sperança – PosDoc - FAPEMA José Eduardo Bracco – DT- (SUCEN) Alexandre Moura – technicalsupport • Dr. Anthony A. James • UCI • Dr. Ken Olson • Colorado State • Dr. Maria Anice Sallun • FSP - USP • Dr. Tania Bijovsky • ICB - USP Supported by FAPESP

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