1. www.aids2014.org HIV-Exposed Seronegative Men who have Sex with Men overexpress potential antiviral antiproteases in their rectal mucosa Laura Romas (MSc. Student) Burgener Lab, University of Manitoba, Canada

2. www.aids2014.org GALT Why is the rectal mucosa so susceptible to HIV infection? 4. Mucosal Fluid? 1. Thin epithelial layer 2. Many HIV target cells 3. Easily accessible lymphatic system

3. www.aids2014.org Natural Immunity in HIV Exposed Seronegative (HESN) Individuals HIV Exposed Seronegative HIV + Partner HESN MSM from the Venhälsan Cohort (est. 2007): –Gay Men`s Health Clinic in Stockholm, Sweden. –Serodiscordant couples >5yrs –Men have HIV neutralizing IgA within their mucosal fluid, but no HIV antibodies in serum (Hasselrot et al. AIDS. 2009)

4. www.aids2014.org Using Proteomics to Identify Correlates of Protection P24 Peptides Mucosal Fluid HESN vs. Control (n=23) (n=20) ID/ QuantTargets Target B. A.

5. www.aids2014.org HESN MSM over-express several immune proteins in their mucosal fluid * 80% power (≥2 fold difference), p<0.05, Benjimani-Hochberg corrected Intelectin 2 α-1-acid glycoprotein 1 Antiprotease 1 S100A7 Antiprotease 2 Antirpotease S100A9 S100A8 α -1-acid glycoprotein 1 Myeloperoxidase Ig Heavy chain Immunoglobulin Immunoglboulin Immunoglobulin Tetraspanin 8

6. www.aids2014.org Antiproteases in Immunity AP 1 (Log 2 Rel. Abund.) AP 2 (Log 2 Rel. Abund.) **p<0.001

7. www.aids2014.org Epidemiological Correlations Antiprotease expression does not correlate with: –Vial exposure (viral load of HIV+ partner) –Mucosal response to HIV (HIV-neutralizing IgA) –Seminal/microbial exposure from unprotected intercourse (Frequency of unprotected sex)

8. www.aids2014.org AP1 inhibits infection in PBMCs C. D. B.A. P>0.01 P<0.01 P>0.01

9. www.aids2014.org Conclusions HESN MSM overexpress antiproteases AP expression does not correlate with exposure AP1 show antiviral properties in vitro –PBMC culture –Maintains cell viability Future Directions: –Follow up in colorectal explants (underway) –Antiviral mechanism Application: –Increase knowledge of HIV pathogenesis through the rectum –Guide future microbicide research

10. www.aids2014.org Acknowledgements Mentors Adam Burgener and Blake Ball; Kevin Coombs and Emmanuel Ho Carolina Herrerra (Imperial College of London, Shattock Lab) University of Manitoba, HIV Research Group, Canada Lindsay Aboud, Melissa Herman, Sue Ramdahin, Kenzie Birse, Max Abou, Derek Stein, Michelle Perner, Jen Butler and Liane Arcinas National Microbiology Laboratory, Canada Frank Plummer; Garrett Westmacott, Stuart McCorrister and Patrick Chong Karolinska Institutet, Sweden Kristinal Broliden, Klara Hasselrot, A. Tjerlund, P. Levinson, L. Persson, P. Saberg, G. Bratt, E. Sandstrom MSM / HESN Participants of the Gay Men’s Health Clinic, Stockholm, SWD.

11. www.aids2014.org AP1 inhibits HIV infection in TZM-bl cells B. C. D. A.