Brazilian Ministry of Health Oswaldo Cruz Foundation Oswaldo Cruz Institute

1. Brazilian Ministry of Health Oswaldo Cruz Foundation Oswaldo Cruz Institute Brazilian Experiences in Collaborative and Preventive Multicentric Studies – FIOCRUZ Mariza G. Morgado Lab. AIDS & Molecular Immunology HIV Vaccine Research: Ethical and Regulatory Issues PN-DST/AIDS – Brazilian Ministry of Health Brasília – Out 2006

2. WHO-Global Programme on AIDS Network for HIV Isolation and characterization • Objective: To obtain information on genetic and antigenic variation of HIV and its possible implications for vaccine development • Primary laboratories – Brazil, Rwanda, Thailand and Uganda • Secondary International laboratories for: • Standardized virus isolation – repository • Genetic subtyping – PCR gag fingerprint, HMA, V3 loop and env DNA sequencing • Biological and immunological characterization - peptide binding EIA, CD4/gp120 blocking antibodies, in vitro replication characteristics and cross-neutralization assays

3. Brazilian Network of HIV Isolation and Characterization (1993) Objective: To establish a Brazilian surveillance program to monitor the genetic and antigenic variation of HIV and its possible implications for vaccine development Major strategies • Identification of potential participants; • Organization of local seminars for protocol definition and implementation; • Submission to WHO and PN-DST/AIDS funding; • Organization of local “International Workshop Trainings”; • In-country and international personnel trainings; • Elaboration and conduction of scientific projects

4. 40 35 30 25 For. 20 Braz. 15 10 5 0 92-93 94-95 96-97 98-99 00-01 02-03 04-05 Medline available scientific publications 1992-2005 No. of papers (N=106) (N=113) (N=194) (N=120) (N=189) (N=242) (N=285) Source: PubMed – Key words: HIV and Brazil * Papers including HMA, DNA sequencing, RFLP, Neutralizing antibodies, peptide ELISA, ELISPOT, CMI

5. Molecular epidemiology of HIV-1 in Brazil B, F, B/F B, F, B/F B, F, B/F, C B, F, B/F, C, D, A,CRF02_AG, CRF28_BF, CRF_29BF C, B, B/C, F, B/F, A Updated 2006

6. Clinical Research - Brazilian HPTN Protocols NICHD/HPTN 040 PHASE III RAMDOMIZED TRIAL OF THE SAFETY AND EFFICACY OF THREE NEONATAL ANTIRETROVIRAL REGIMENS FOR PREVENTION OF INTRAPARTUM HIV-1 TRANSMISSION Belo Horizonte, MG Rio de Janeiro, RJ São Paulo,SP HPTN 052 A RANDOMIZED TRIAL TO EVALUATE THE EFFECTIVENESS OF HIV PRIMARY CARE VERSUS HIV PRIMARY CARE PLUS ANTIRETROVIRAL THERAPY TO PREVENT THE SEXUAL TRANSMISSION OF HIV-1 IN SERODISCORDANT COUPLES Porto Alegre, RS NICHD/HPTN 040 NICHD/HPTN 040/HPTN052

7. HIV-1 subtype infection among pregnant women from Rio de Janeiro, RJ, and Porto Alegre, RS 3% 10% 87% RJ N=3,689 1.35% of seropositive women (49) * RJ (n=39) BB 3% 11% 25% FF (env & pol) CC * BC BF CBF 61% PoA N=1,524 6,3% of eropositive women (99) PoA (n=64) Morgado et al.2004 CROI

8. Major requirements for project participation • Good Laboratory Practices • Biosafety rules • Ethical Issues • Regulatory Issues

9. NICHD-HPTN040 – Strategy shipping, storage and testing of biological samples UFSP São Paulo, SP HSE and HGNI Rio de Janeiro, RJ Results Plasma, blood cell pellets and cryopreseved mononuclear cells Plasma, blood cell pellets and cryopreseved mononuclear cells FIOCRUZ, RJ Study Coordination Data Manegement Central Laboratory Central Laboratory Transport of clinical specimens Transport of clinical specimens ldo Cruz Foundation) Repository RNA and DNA assays UFMG Belo Horizonte, MG HC, HF, HSC Porto Alegre, RS Plasma, blood cell pellets and cryopreseved mononuclear cells Plasma, blood cell pellets and cryopreseved mononuclear cells Results

10. Hospital dos Servidores Hosp. Geral de Nova Iguacu Results Sample processing Sample storage LDMS CD4 counts Sample processing Sample storage LDMS FIOCRUZ, RJ Sample processing Sample storage/LDMS RNA assay CD4 counts Genotyping* Hematol./Biochem/Immunol/Microbiol Shipping & Training ldo Cruz Foundation) Clinical specimens Clinical specimens • *Reference of Genotyping also for the following centers: • Hospital Conceição, RS • IMPACTA, Lima, Peru HPTN052 – Strategy shipping, storage and testing of biological samples Johns Hopkins Central Lab

11. Laboratory procedures INMETRO NIT-DICLA 083 / ISO-IEC 15189 • SOPs • Internal and external controls • Data register • Data storage and control • Results

12. Sample processing and storage in biosafety conditions Biological repository maintenance • Cryopreserved mononuclear cells • Plasma • Sera • Blood cell pellets * RESOLUÇÃO N o 347, DE 13 DE JANEIRO DE 2005

13. National and international laboratory certifications • UKNEQAS (England)– Immunophenotyping • Virology Quality Assessment Program (USA) – DNA PCR; RT-PCR; Genotyping • Immunology Quality Assessment Program – PBMC cryopreservation • College of American Pathologists (USA) – CBC, serology, etc • Brazilian AIDS Program – immunophenotyping, VL and Genotyping

14. Major ethical and regulatory issues • Timeline to get the protocols approved. • Concerns about the storage of biological samples for further studies. • Timeline to prepare the documents (IL) and have them accepted for drug and reagent reception • Timeline to prepare the documents and have them accepted for importation and/or customs clearance of biological panels for laboratory proficiency tests. • Constant modifications in documentation required for shipping and/or clearance of drugs, reagents, and biological materials, including proficiency panels.