Joann D. Kuruc

Administration of vorinostat disrupts HIV-1 latency in patients on antiretroviral therapy

Despite antiretroviral therapy, proviral latency of human immunodeficiency virus type 1 (HIV-1) remains a principal obstacle to curing the infection. Inducing the expression of latent genomes within resting CD4+ T cells is the primary strategy to clear this reservoir. Although histone deacetylase inhibitors such as suberoylanilide hydroxamic acid (also known as vorinostat, VOR) can disrupt HIV-1 latency in vitro, the utility of this approach has never been directly proven in a translational clinical study of HIV-infected patients. Here we isolated the circulating resting CD4+ T cells of patients in whom viraemia was fully suppressed by antiretroviral therapy, and directly studied the effect of VOR on this latent reservoir. In each of eight patients, a single dose of VOR increased both biomarkers of cellular acetylation, and simultaneously induced an increase in HIV RNA expression in resting CD4+ cells (mean increase, 4.8-fold). This demonstrates that a molecular mechanism known to enforce HIV latency can be therapeutically targeted in humans, provides proof-of-concept for histone deacetylase inhibitors as a therapeutic class, and defines a precise approach to test novel strategies to attack and eradicate latent HIV infection directly.

HIV-1 expression within resting CD4+ T cells after multiple doses of vorinostat.

BACKGROUND A single dose of the histone deacetylase inhibitor vorinostat (VOR) up-regulates HIV RNA expression within resting CD4(+) T cells of treated, aviremic human immunodeficiency virus (HIV)-positive participants. The ability of multiple exposures to VOR to repeatedly disrupt latency has not been directly measured, to our knowledge. METHODS Five participants in whom resting CD4(+) T-cell-associated HIV RNA (rc-RNA) increased after a single dose of VOR agreed to receive daily VOR Monday through Wednesday for 8 weekly cycles. VOR serum levels, peripheral blood mononuclear cell histone acetylation, plasma HIV RNA single-copy assays, rc-RNA, total cellular HIV DNA, and quantitative viral outgrowth assays from resting CD4(+) T cells were assayed. RESULTS VOR was well tolerated, with exposures within expected parameters. However, rc-RNA measured after dose 11 (second dose of cycle 4) or dose 22 (second dose of cycle 8) increased significantly in only 3 of the 5 participants, and the magnitude of the rc-RNA increase was much reduced compared with that after a single dose. Changes in histone acetylation were blunted. Results of quantitative viral outgrowth and other assays were unchanged. CONCLUSIONS Although HIV latency is disrupted by an initial VOR dose, the effect of subsequent doses in this protocol was much reduced. We hypothesize that the global effect of VOR results in a refractory period of ≥ 24 hours. The optimal schedule for VOR administration is still to be defined.

Dual-Affinity Re-Targeting proteins direct T cell–mediated cytolysis of latently HIV-infected cells

Enhancement of HIV-specific immunity is likely required to eliminate latent HIV infection. Here, we have developed an immunotherapeutic modality aimed to improve T cell-mediated clearance of HIV-1-infected cells. Specifically, we employed Dual-Affinity Re-Targeting (DART) proteins, which are bispecific, antibody-based molecules that can bind 2 distinct cell-surface molecules simultaneously. We designed DARTs with a monovalent HIV-1 envelope-binding (Env-binding) arm that was derived from broadly binding, antibody-dependent cellular cytotoxicity-mediating antibodies known to bind to HIV-infected target cells coupled to a monovalent CD3 binding arm designed to engage cytolytic effector T cells (referred to as HIVxCD3 DARTs). Thus, these DARTs redirected polyclonal T cells to specifically engage with and kill Env-expressing cells, including CD4+ T cells infected with different HIV-1 subtypes, thereby obviating the requirement for HIV-specific immunity. Using lymphocytes from patients on suppressive antiretroviral therapy (ART), we demonstrated that DARTs mediate CD8+ T cell clearance of CD4+ T cells that are superinfected with the HIV-1 strain JR-CSF or infected with autologous reservoir viruses isolated from HIV-infected-patient resting CD4+ T cells. Moreover, DARTs mediated CD8+ T cell clearance of HIV from resting CD4+ T cell cultures following induction of latent virus expression. Combined with HIV latency reversing agents, HIVxCD3 DARTs have the potential to be effective immunotherapeutic agents to clear latent HIV-1 reservoirs in HIV-infected individuals.

Frequent detection of acute HIV infection in pregnant women.

Background:Universal prenatal HIV antibody testing, which does not detect acute HIV, is standard for pregnant women in the United States. Unrecognized HIV acquisition during pregnancy may result in higher rates of perinatal transmission. Objective:To determine the prevalence of acute (antibody-negative) HIV infection in pregnant women and to assess the potential for prompt initiation of antiretroviral therapy to prevent perinatal transmission. Methods:From 1 November 2002 to 30 April 2005, all publicly funded HIV testing sites participated in North Carolinas Screening and Tracing Active Transmission (STAT) Program, which retested all specimens that were HIV antibody negative for HIV RNA using specimen pooling. All patients with acute HIV infection were immediately traced for evaluation, confirmatory testing, counseling, and referral services. For this study, all pregnant women with acute HIV were immediately initiated onto antiretroviral therapy and followed prospectively for pregnancy outcomes. Results:During the study period, 443 women were HIV positive by antibody testing; 15 were HIV antibody negative but positive by RNA assay and of these five were pregnant at the time of testing. The pregnant women received antiretroviral drugs and delivered HIV-uninfected infants. Maternal testing records of all six HIV-infected infants born in North Carolina showed three mothers with chronic HIV infection and three HIV antibody negative at private prenatal testing facilities. Conclusions:In resource-rich settings, a substantial proportion of residual perinatal transmission may be from HIV acquisition during pregnancy. Standard antibody tests miss acute HIV infection and so algorithms that include pooled HIV RNA testing may improve its detection and represent a further opportunity to prevent perinatal transmission.

Investigating A Sexual Network of Black Men Who Have Sex with Men: Implications for Transmission and Prevention of HIV Infection in the United States

Background:HIV infections increased 48% among young Black men who have sex with men (MSM) in the United States between 2006 and 2009. Incomplete understanding of this trend undermines prevention strategy development. We investigated a sexual network to characterize the risk environment in which young Black MSM acquire HIV. Methods:Persons reported to the state after diagnosis of HIV or syphilis were included, along with sexual partners. We used network mapping alongside descriptive and bivariate statistics to characterize network connections. Generalized linear models assessed predictors of having untraceable sex partners. Results:The network included 398 individuals and 419 sexual relationships. Three-quarters were Black (n = 299); 92% were MSM. Median age at first network appearance was 26 years and decreased over time (P < 0.001). HIV prevalence was at least 29% (n = 117); serostatus was unknown for 47% of the network, either because they were untraceable (n = 150) or refused HIV testing (n = 39). One in 5 network members diagnosed with HIV had a subsequent incident sexually transmitted infection. In multivariable models, one-time encounters increased the risk of having an untraceable partner (risk ratio = 4.51, 95% CI: 2.27 to 8.97), whereas being acutely HIV infected at diagnosis reduced it (risk ratio = 0.27, 95% CI: 0.08 to 0.89). Conclusions:HIV prevalence in this sexual network of young Black MSM rivals that of sub-Saharan Africa, reflecting dramatically increased risk of acquiring HIV from the moment one entered the network. Prevention efforts for this population must consider the effect of sexual networks on HIV risk and find ways of leveraging network structure to reduce transmission.

Prevalence of transmitted antiretroviral drug resistance differs between acutely and chronically HIV-infected patients.

Abstract:The associations of acute HIV infection (AHI) and other predictors with transmitted drug resistance (TDR) prevalence were assessed in a cohort of HIV-infected, antiretroviral-naïve patients. AHI was defined as being seronegative with detectable HIV RNA. Binomial regression was used to estimate prevalence ratios and 95% confidence intervals for associations with TDR. Among 43 AHI patients, TDR prevalence was 20.9%, whereas prevalence was 8.6% among 677 chronically infected patients. AHI was associated with 1.9 times the prevalence of TDR (95% confidence intervals: 1.0 to 3.6) in multivariable analysis. AHI patients may represent a vanguard group that portends increasing TDR in the future.

Antiretroviral therapy initiated during acute HIV infection fails to prevent persistent T-cell activation.

Abstract:Initiation of antiretroviral therapy during acute HIV-1 infection may prevent persistent immune activation. We analyzed longitudinal CD38+HLA-DR+ CD8+ T-cell percentages in 31 acutely infected individuals who started early (median 43 days since infection) and successful antiretroviral therapy, and maintained viral suppression through 96 weeks. Pretherapy a median of 72.6% CD8+ T cells were CD38+HLA-DR+, and although this decreased to 15.6% by 96 weeks, it remained substantially higher than seronegative controls (median 8.9%, P = 0.008). Shorter time to suppression predicted lower activation at 96 weeks. These results support the hypothesis that very early events in HIV-1 pathogenesis may result in prolonged immune dysfunction.

CD4+CD8+ T Cells Represent a Significant Portion of the Anti-HIV T Cell Response to Acute HIV Infection

Previous studies have revealed that HIV-infected individuals possess circulating CD4+CD8+ double-positive (DP) T cells specific for HIV Ags. In the present study, we analyzed the proliferation and functional profile of circulating DP T cells from 30 acutely HIV-infected individuals and 10 chronically HIV-infected viral controllers. The acutely infected group had DP T cells that showed more proliferative capability and multifunctionality than did both their CD4+ and CD8+ T cells. DP T cells were found to exhibit greater proliferation and higher multifunctionality compared with CD4 T cells in the viral controller group. The DP T cell response represented 16% of the total anti-HIV proliferative response and >70% of the anti-HIV multifunctional response in the acutely infected subjects. Proliferating DP T cells of the acutely infected subjects responded to all HIV Ag pools with equal magnitude. Conversely, the multifunctional response was focused on the pool representing Nef, Rev, Tat, VPR, and VPU. Meanwhile, the controllers’ DP T cells focused on Gag and the Nef, Rev, Tat, VPR, and VPU pool for both their proliferative and multifunctional responses. Finally, we show that the presence of proliferating DP T cells following all HIV Ag stimulations is well correlated with proliferating CD4 T cells whereas multifunctionality appears to be largely independent of multifunctionality in other T cell compartments. Therefore, DP T cells represent a highly reactive cell population during acute HIV infection, which responds independently from the traditional T cell compartments.

No one's at home and they won't pick up the phone: using the Internet and text messaging to enhance partner services in North Carolina.

Background The Internet and mobile devices are increasingly used by men who have sex with men to find potential partners. Lack of partner information, besides e-mail addresses or user profiles, limits the ability to adequately perform partner notification by traditional means and test those at high risk. To streamline North Carolina Internet Partner Notification (IPN) services, University of North Carolina at Chapel Hill collaborated with the North Carolina Division of Public Health beginning in July 2011 to formalize state IPN and text messaging for partner notification (txtPN) policies and centralize notification practices by designating a single IPN/txtPN field coordinator within the University of North Carolina at Chapel Hill. Methods We compared the number of IPN and txtPN contacts initiated and their outcomes in July 1, 2011, to June 30, 2012, and compared with outcomes in January 1, 2010, to December 31, 2010, the year before the collaboration. Results Overall, 362 IPN contacts were initiated compared with 133 initiated in 2010. More than half (59.1%) were black; mean age was 28.8 years. Almost all were men who have sex with men (83.7%). Approximately two-thirds (n = 230; 63.5%) of contacts were successfully notified using centralized IPN. Seven new cases of HIV infection, 11 new cases of syphilis, and 19 known previous HIV-positive persons were identified. Text messaging for partner notification was used for 29 contacts who did not initially respond to traditional notification or IPN; 14 (48%) responded to txtPN in a median time of 57.5 minutes (interquartile range, 9–2708). Conclusions Centralization of IPN services augmented partner detection of new HIV and syphilis diagnoses. Text messaging for partner notification represents a potentially effective method for augmenting traditional partner services. In addition, IPN and txtPN allow identification of HIV-infected persons in need of linkage to care.

Efficacy of NNRTI-based antiretroviral therapy initiated during acute HIV infection.

Objective:Characterize responses to non-nucleoside reverse transcriptase inhibitor (NNRTI)-based antiretroviral treatment (ART) initiated during acute HIV infection (AHI). Design:This was a prospective, single-arm evaluation of once-daily, co-formulated emtricitabine/tenofovir/efavirenz initiated during AHI. Methods:The primary endpoint is the proportion of responders with HIV RNA less than 200 copies/ml by week 24. We examined time to viral suppression and CD8 cell activation in relation to baseline participant characteristics. We compared time to viral suppression and viral dynamics using linear mixed-effects models between acutely infected participants and chronically infected controls. Results:Between January 2005 and May 2009, 61 AHI participants were enrolled. Of participants whose enrollment date allowed 24 and 48 weeks of follow-up, 47 of 51 (92%) achieved viral suppression to less than 200 copies/ml by week 24, and 35 of 41 (85.4%) to less than 50 copies/ml by week 48. The median time from ART initiation to suppression below 50 copies/ml was 93 days (range 14–337). Higher HIV RNA levels at ART initiation (P = 0.02), but not time from estimated date of infection to ART initiation (P = 0.86), were associated with longer time to viral suppression. The median baseline frequency of activated CD8+CD38+HLA-DR+ T cells was 67% (range 40–95), and was not significantly associated with longer time to viral load suppression (P = 0.15). Viremia declined to less than 50 copies/ml more rapidly in AHI than chronically infected participants. Mixed-model analysis demonstrated similar phase I HIV RNA decay rates between acute and chronically infected participants, and more rapid viral decline in acutely infected participants in phase II. Conclusion:Once-daily emtricitabine/tenofovir/efavirenz initiated during AHI achieves rapid and sustained HIV suppression during this highly infectious period.