1. Highly-exposed HIV seronegative persons (HESN) had higher levels of inflammatory and immune activation-associated serum biomarkers than lower risk HIV seroconverters (LRSC) O Martínez-Maza, R Detels, L Magpantay, J Phair, JH Bream, CR Rinaldo and LP Jacobson Multicenter AIDS Cohort Study (MACS) no conflict of interest Hypothesis: HESN will display lower levels of immune/inflammatory molecules than LRSN Figure 1. Role of inflammation in HIV infection. Pro-inflammatory molecules can enhance HIV infection and replication in newly infected cells. Molecules that were assessed in this study are represented in bold face. microbial translocation (endotoxin, TLR-L) Figure 1. Role of inflammation in HIV infection. Pro-inflammatory molecules can enhance HIV infection and replication in newly infected cells. Molecules that were assessed in this study are represented in bold face. microbial translocation (endotoxin, TLR-L) Inflammation and immune activation can enhance initial infection with HIV. The objective of this study was to define serum levels of cytokines and biomarkers of inflammation in participants in the MACS who were at high risk for acquiring HIV infection but remained HIV seronegative (HESN), compared to those at lower risk who did (LRSC), or did not (LRSN), subsequently undergo HIV seroconversion.

2. Figure 2. Selection of HESC, LRSN, and LRSC groups from the MACS cohort. HESN-SC are those who were in the initial HESN group and went on to sero-convert prior to 2009. HESN-SC were not included in the analyses shown here.

3. Serum levels of inflammatory cytokines and biomarkers were measured using multiplexed assays (Luminex, R&D Systems) in archival specimens from HESN, LRSC, and LRSN, collected during the period of highest exposure (mid 1980s), prior to seroconversion: Figure 1. Role of inflammation in HIV infection. Pro-inflammatory molecules can enhance HIV infection and replication in newly infected cells. Molecules that were assessed in this study are represented in bold face. microbial translocation (endotoxin, TLR-L) Figure 1. Role of inflammation in HIV infection. Pro-inflammatory molecules can enhance HIV infection and replication in newly infected cells. Molecules that were assessed in this study are represented in bold face. microbial translocation (endotoxin, TLR-L)

4. Table 2: Serum levels - cytokines and biomarkers of inflammation Results and Conclusions HESN displayed significantly higher serum levels of sTNFR2 than LRSC HESN had significantly higher levels of sCD14, sTNFR2, and IL8, and lower levels of IFNγ, than LRSN The magnitude of these differences was generally small This is consistent with the conclusion that there were modestly elevated levels of inflammation in the HESN, compared to the LRSC and the LRSN These findings do not support our initial working hypothesis that HESN would have lower inflammatory biomarker levels than LRSC Results and Conclusions HESN displayed significantly higher serum levels of sTNFR2 than LRSC HESN had significantly higher levels of sCD14, sTNFR2, and IL8, and lower levels of IFNγ, than LRSN The magnitude of these differences was generally small This is consistent with the conclusion that there were modestly elevated levels of inflammation in the HESN, compared to the LRSC and the LRSN These findings do not support our initial working hypothesis that HESN would have lower inflammatory biomarker levels than LRSC HESN displayed significantly higher serum levels of sTNFR2 than LRSC HESN had significantly higher levels of sCD14, sTNFR2, and IL8, and lower IFNγ, than LRSN The magnitude of these differences was small This is consistent with the conclusion that the HESN men may display modestly elevated levels of inflammation, compared to the LRSC and the LRSN These findings do not support our initial working hypothesis that HESN have lower inflammatory biomarker levels than LRSC