1. L’ INTRODUZIONE DI RALTEGRAVIR IN TERAPIA RIDUCE A LIVELLI NON DOSABILI LA VIRAL LOAD SEMINALE IN “SEMINAL SUPER SHEDDERS” HIV+ Implicazioni per la trasmissione e per le pratiche di fecondazione assistita Fiore J. R. Di Stefano M. Zoboli F. Faleo G. Fasano M. Santantonio T. A. Clinica di Malattie Infettive, Dipartimento di Medicina Clinica e Sperimentale, Università degli Studi di Foggia

2. Background discordant virologic responses and genotypic drug sensitivies between blood and semen observed in 7-15% of HIV infected men receiving HAART: SEMINAL SUPER-SHEDDERS Barroso PF et al. Effect of antiretroviral therapy on HIV shedding in semen. Ann Intern Med. 2000 Lafeuillade A et al. Differences in the detection of three HIV-1 protease inhibitors in non-blood compartments: clinical correlations. HIV Clin Trials. 2002 Leruez-Ville M et al. Decrease in HIV-1 seminal shedding in men receiving highly active antiretroviral therapy: an 18 month longitudinal study (ANRS EP012). AIDS. 2002 Lafeuillade A et al. HIV-1 RNA levels, resistance, and drug diffusion in semen versus blood in patients receiving a lopinavir-containing regimen. J Acquir Immune Defic Syndr. 2003 Barroso PF et al. Adherence to antiretroviral therapy and persistence of HIV RNA in semen. J Acquir Immune Defic Syndr. 2003 Bujan L et al. Factors of intermittent HIV-1 excretion in semen and efficiency of sperm processing in obtaining spermatozoa without HIV-1 genomes. AIDS. 2004 Lorello G et al. Discordance in HIV-1 viral loads and antiretroviral drug concentrations comparing semen and blood plasma. HIV Med. 2009 Seminal Plasma Viral Load Blood Plasma Viral Load >1 Taylor S. Sadiq T. Sabin C. Seminal super shedding of HIV: Implications for sexual transmission. Presented at the 10th Conference on Retroviruses and Opportunistic Infections; Boston, MA. Feb 10–14;2003

3. No seroconversion was reported in serodiscordant couples undergoing Assisted Reproduction programs Raltegravir and Maraviroc concentrations in semen are several fold-higher than those attained in blood and those required to inhibit viral replication in this compartment. Bujan L et al. Safety and efficacy of sperm washing in HIV-1-serodiscordant couples where the male is infected: results from the European CREAThE network. AIDS. 2007 Antoniou T et al. Pharmacokinetics of raltegravir in the semen of HIV-infected men. Antivir Ther. 2014

4. Materials 3 MEN with undetectable blood plasma viral load, but persistent seminal plasma viral load TDF+FTC and LPV/r TDF+FTC+EFV 2 1 RALTEGRAVIR toxicity Semen samples Baseline 1 month 2 months 4 months 6 months

5. Methods Semen was collected by masturbation without lubrificant after 48h of abstinence. Viral transport medium (2 ml of RMPI 1640 with 2 mMol L = glutamine and 10% fetal bovine serum (FBS), with the addition of 100 U/ml penicillin 100 µl/ml of streptomycin, and 200 U/ml of nystatin) was added to seminal samples at collection. Seminal plasma was separated from seminal cells by centrifugation at 700 x g for 12 minutes within 4 hours of collection and stored at -80°C and -150°C. In blood, HIV RNA was quantified by Artus HIV-1 RG RT-PCR kit QIAGEN.

6. SEMINAL PLASMA VIRAL LOAD #1#2#3 Baseline42500330002100 1 month 2 months 4 months 6 months undetectable Results At baseline all patients had undetectable blood plasma viral load

7. Discussion and Conclusions Raltegravir distribuites into and accumulates within seminal plasma by passive diffusion, by virtue of being slightly lipophilic and 83% bound to blood plasma proteins The small sample size is a limitation of our work, but these results suggest that Raltegravir can be used to reduce sexual transmission of HIV and improve the safety of Assisted Reproduction programs in serodifferent couples. Barau C, et al. High concentration of raltegravir in semen of HIV-infected men: results from a substudy of the EASIER- ANRS 138 trial. Antimicrob Agents Chemother. 2010 Carey D et al. A randomized study of pharmacokinetics, efficacy, and safety of 2 raltegravir plus atazanavir strategies in ART-treated adults. J Acquir Immune Defic Syndr. 2012

8. Immuno-virological aspects of genital fluids during viral infections Clinica di Malattie Infettive, Università degli Studi di Foggia Fiore J. R. Di Stefano M. Zoboli F. Faleo G. Ferrara S. Fasano M. Tartaglia A. Santantonio T. A. Istituto di Genetica, Università degli Studi di Foggia Margaglione M D’Andrea G. Clinica Ostetrica e Ginecologica, Università degli Studi di Ferrara Greco P. Microbiology & Tumorbiology Center, Karolinska Inst., Stockholm Chiodi F. Labor. Of Human Retrovirology, Institute Pasteur, Paris Menu E.