Kuala Lumpur, Malaysia, 30 June - 3 July 2013 Protein engineering of the lectin BanLec reduces its mitogenic activity and prevents vaginal HIV transmission Michael Swanson University of North Carolina

Kuala Lumpur, Malaysia, 30 June - 3 July 2013 BanLec binds to high- mannose on gp120 Broad, potent activity anti-HIV activity Microbicide Candidate Can its stimulatory activity be reduced? – Substitution of amino acids in binding sites Engineering BanLec to Reduce its Mitogenic and Stimulatory Activity TFV 3.2 µM Strategy Outline cDNA for BanLec Introduce amino substitutions Express in E. coli Purify variants Assess stimulatory activity

Kuala Lumpur, Malaysia, 30 June - 3 July 2013 Topically-applied BanLec H84T Prevents Vaginal HIV Transmission Apply 75 µg BanLec H84T Topically Expose Vaginally with HIV-1 JR-CSF 10 minutes later Measure Viral Load p=0.0359

Kuala Lumpur, Malaysia, 30 June - 3 July 2013 Conclusions BanLec H84T has reduced mitogenicity and is effective ant preventing vaginal HIV transmission in vivo BanLec H84T should be investigated as a potential microbicide. Engineering of mitogenic, anti-HIV lectins could lead to more microbicide candidates

Kuala Lumpur, Malaysia, 30 June - 3 July 2013 Acknowledgements University of Leuven, Rega Institute for Medical Research -Dominique Schols -Dana Huskens University of Michigan -David Markovitz -Dan Bodreaux University of North Carolina -J. Victor Garcia NIH Grants: AI100775