Image1_Pre-clinical evaluation of antiproteases as potential candidates for HIV-1 pre-exposure prophylaxis.pdf

Previous studies on highly HIV-1-exposed, yet persistently seronegative women from the Punwami Sex Worker cohort in Kenya, have shed light on putative protective mechanisms, suggesting that mucosal immunological factors, such as antiproteases, could be mediating resistance to HIV-1 transmission in the female reproductive tract. Nine protease inhibitors were selected for this study: serpin B4, serpin A1, serpin A3, serpin C1, cystatin A, cystatin B, serpin B13, serpin B1 and α-2-macroglobulin-like-protein 1. We assessed in a pilot study, the activity of these antiproteases with cellular assays and an ex vivo HIV-1 challenge model of human ecto-cervical tissue explants. Preliminary findings with both models, cellular and tissue explants, established an order of inhibitory potency for the mucosal proteins as candidates for pre-exposure prophylaxis when mimicking pre-coital use. Combination of all antiproteases considered in this study was more active than any of the individual mucosal proteins. Furthermore, the migration of cells out of ecto-cervical explants was blocked indicating potential prevention of viral dissemination following amplification of the founder population. These findings constitute the base for further development of these mucosal protease inhibitors for prevention strategies.</p

Proteins found to be significantly overabundant (A) and under abundant (B) in the cationic protein-depleted cervicovaginal secretions of HIV-exposed seronegative sex workers (<3 years) compared to HIV seronegative low-risk controls.

<p>^a General functions are based on each protein’s gene ontology obtained from the UniProt website.</p><p>^b Statistical significance was deduced via Student’s T-test, p < 0.05.</p><p>* denotes a known association with HIV-1.</p><p>Proteins found to be significantly overabundant (A) and under abundant (B) in the cationic protein-depleted cervicovaginal secretions of HIV-exposed seronegative sex workers (<3 years) compared to HIV seronegative low-risk controls.</p

Proteins found to be significantly overabundant (A) and under abundant (B) in the cationic protein-depleted cervicovaginal secretions of HIV-positive sex workers compared to HIV seronegative low-risk controls.

<p>^a General functions are based on each protein’s gene ontology obtained from the UniProt website.</p><p>^b Statistical significance was deduced via Student’s T-test, p < 0.05.</p><p>Proteins found to be significantly overabundant (A) and under abundant (B) in the cationic protein-depleted cervicovaginal secretions of HIV-positive sex workers compared to HIV seronegative low-risk controls.</p

Proteins found to be significantly overabundant (A) and under abundant (B) in the cationic protein-depleted cervicovaginal secretions of HIV-exposed seronegative sex workers (>3 years) compared to HIV-seronegative low-risk controls.

<p>^a General functions are based on each protein’s gene ontology obtained from the UniProt website.</p><p>^b Statistical significance was deduced via Student’s T-test, p < 0.05.</p><p>* denotes a known association with HIV-1.</p><p>Proteins found to be significantly overabundant (A) and under abundant (B) in the cationic protein-depleted cervicovaginal secretions of HIV-exposed seronegative sex workers (>3 years) compared to HIV-seronegative low-risk controls.</p

HIV neutralizing activity in unprocessed and cationic protein-depleted cervicovaginal secretions.

<p>The HIV neutralizing activity of unprocessed cervicovaginal secretions (CVS) and cationic protein-depleted CVS, respectively, was assessed in pooled samples from the study groups. Green bars: HIV seronegative low-risk women (Low risk) [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0130404#pone.0130404.ref025" target="_blank">25</a>]; Yellow bars: HIV seronegative women with less than 3 years of reported active sex work (HESN<3yr); Blue bars: HIV seronegative women with more than 3 years of reported active sex work (HESN>3yr); Red bars: HIV seropositive sex working women (HIV positive). The results shown are from two to three representative experiments using duplicate wells (median values ±SEM).</p

Biological processes of differentially abundant proteins between female sex worker and low-risk groups.

<p><b>A:</b> HIV-exposed seronegative female sex workers for a time greater than three years <b>B:</b> HIV-exposed seronegative female sex workers for a time less than three years <b>C:</b> HIV-seropositive female sex workers (DAVID Bioinformatics Resources 6.7, UniProt) (Blue bars represent overabundant proteins and red bars represent under abundant proteins compared to the low-risk, uninfected control group).</p

Combined effect of age and allele phenotype on disease outcome using logistic regression analysis comparing asymptomatic (AS), symptomatic (S), and neuroinvasive disease (ND) outcome groups.

<p>Combined effect of age and allele phenotype on disease outcome using logistic regression analysis comparing asymptomatic (AS), symptomatic (S), and neuroinvasive disease (ND) outcome groups.</p

Demographics of the BSRI and McMaster cohorts.

<p>Demographics of the BSRI and McMaster cohorts.</p

Combinations of “susceptible” (S) and “protective” (P) alleles.

<p>The histogram displays the percentage of subjects with the phenotype indicated below the X axis in the three WNV⁺ groups: asymptomatic (AS), symptomatic (S), and having a neuroinvasive disease (ND). ** <i>P</i><0.01 and * <i>P</i><0.05.</p

Pair-wise analysis of HLA Class I phenotype frequencies^* in asymptomatic (AS), symptomatic (S), and neuroinvasive disease (ND) groups of WNV⁺ subjects.

<p>*Phenotype frequencies >5% are listed.</p>!<p>n represents the number of subjects with a specific allele phenotype. Alleles in bold showed an uncorrected <i>P</i><0.05 followed by a corrected <i>Pc</i> (<i>P/Pc</i>).</p