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Abstract Blue stragglers are anomalously luminous core hydrogen-burning stars formed through mass-transfer in binary/triple systems and stellar collisions. Their physical and evolutionary properties are largely unknown and unconstrained. Here we analyze 320 high-resolution spectra of blue stragglers collected in eight galactic globular clusters with different structural characteristics and show evidence that the fraction of fast rotating blue stragglers (with rotational velocities larger than 40 km/s) increases for decreasing central density of the host system. This trend suggests that fast spinning blue stragglers prefer low-density environments and promises to open an unexplored route towards understanding the evolutionary processes of these stars. Since large rotation rates are expected in the early stages of both formation channels, our results provide direct evidence for recent blue straggler formation activity in low-density environments and put strong constraints on the timescale of the collisional blue straggler slow-down processes.

BACKGROUND HIV-1–infected CD4+ T cells contribute to latent reservoir persistence by proliferating while avoiding immune recognition. Integration features of intact proviruses in elite controllers (ECs) and people on long-term therapy suggest that proviruses in specific chromosomal locations can evade immune surveillance. However, direct evidence of this mechanism is missing.METHODS In this case report, we characterized integration sites and full genome sequences of expanded T cell clones in an EC before and after chemoradiation. We identified the cognate peptide of infected clones to investigate cell proliferation and virus production induced by T cell activation, and susceptibility to autologous CD8+ T cells.RESULTS The proviral landscape was dominated by 2 large clones with replication-competent proviruses integrated into zinc finger (ZNF) genes (ZNF470 and ZNF721) in locations previously associated with deeper latency. A third nearly intact provirus, with a stop codon in Pol, was integrated into an intergenic site. Upon stimulation with cognate Gag peptides, infected clones proliferated extensively and produced virus, but the provirus in ZNF721 was 200-fold less inducible. While autologous CD8+ T cells decreased the proliferation of cells carrying the intergenic provirus, they had no effect on cells with the provirus in the ZNF721 gene.CONCLUSIONS We provide direct evidence that upon activation of infected clones by cognate antigen, the lower inducibility of intact proviruses in ZNF genes can result in immune evasion and persistence.FUNDING Office of the NIH Director and National Institute of Dental & Craniofacial Research; NIAID, NIH; Johns Hopkins University Center for AIDS Research.

HIV-1 persists in a latent reservoir in resting CD4+ T cells despite antiretroviral therapy (ART). The reservoir decays slowly over the first 7 years of ART (t1/2 = 44 months). However, whether decay continues with long-term ART is unclear. Recent integration site studies indicate gradual selection against inducible, intact proviruses, raising speculation that decades of ART might allow treatment interruption without viral rebound. Therefore, we measured the reservoir in 42 people on long-term ART (mean 22 years) using a quantitative viral outgrowth assay. After 7 years of ART, there was no long-term decrease in the frequency of inducible, replication-competent proviruses but rather an increase with an estimated doubling time of 23 years. Another reservoir assay, the intact proviral DNA assay, confirmed that reservoir decay with t1/2 of 44 months did not continue with long-term ART. The lack of decay reflected proliferation of infected cells. Most inducible, replication-competent viruses (79.8%) had env sequences identical to those of other isolates from the same sample. Thus, although integration site analysis indicates changes in reservoir composition, the proliferation of CD4+ T cells counteracts decay, maintaining the frequency of inducible, replication-competent proviruses at roughly constant levels over the long term. These results reinforce the need for lifelong ART.

Background Antiretroviral therapy (ART) halts HIV-1 replication, decreasing viremia to below the detection limit of clinical assays. However, some individuals experience persistent nonsuppressible viremia (NSV) originating from CD4+ T cell clones carrying infectious proviruses. Defective proviruses represent over 90% of all proviruses persisting during ART and can express viral genes, but whether they can cause NSV and complicate ART management is unknown.Methods We undertook an in-depth characterization of proviruses causing NSV in 4 study participants with optimal adherence and no drug resistance. We investigated the impact of the observed defects on 5′-leader RNA properties, virus infectivity, and gene expression. Integration-site specific assays were used to track these proviruses over time and among cell subsets.Results Clones carrying proviruses with 5′-leader defects can cause persistent NSV up to approximately 103 copies/mL. These proviruses had small, often identical deletions or point mutations involving the major splicing donor (MSD) site and showed partially reduced RNA dimerization and nucleocapsid binding. Nevertheless, they were inducible and produced noninfectious virions containing viral RNA, but lacking envelope.Conclusion These findings show that proviruses with 5′-leader defects in CD4+ T cell clones can give rise to NSV, affecting clinical care. Sequencing of the 5′-leader can help in understanding failure to completely suppress viremia.Funding Office of the NIH Director and National Institute of Dental and Craniofacial Research, NIH; Howard Hughes Medical Institute; Johns Hopkins University Center for AIDS Research; National Institute for Allergy and Infectious Diseases (NIAID), NIH, to the PAVE, BEAT-HIV, and DARE Martin Delaney collaboratories.

Antiretroviral therapy (ART) effectively inhibits HIV-1 replication but is not curative due to the persistence of a latent viral reservoir in resting CD4+ T cells. This reservoir is a major barrier to cure. Sequencing studies have revealed that the population of proviruses persisting in ART-treated individuals is dominated by defective proviruses that cannot give rise to viral rebound due to fatal defects including large deletions and APOBEC3-mediated hypermutation. Near full genome sequencing (nFGS) of individual proviruses is used in reservoir assays to provide an estimate of the fraction of proviruses that are intact. nFGS methods rely on a long-distance outer PCR capturing most (~9 kb) of the genome, followed by nested inner PCRs. The outer PCR is carried out at limit dilution, and interpretation of the results is based on the assumption that all proviruses are quantitatively captured. Here, we evaluate nFGS methods using the intact proviral DNA assay (IPDA), a multiplex digital droplet PCR assay that quantitates intact and defective proviruses with single molecule sensitivity using only short, highly efficient amplicons. We analyzed proviral templates of known sequence to avoid the additional complication of sequence polymorphism. With the IPDA, we quantitated molecular yields at each step of nFGS methods. We demonstrate that nFGS methods are inefficient and miss ~70% of full-length proviruses due to amplification failure at the initial outer PCR step. In contrast, proviruses with large internal deletions encompassing 70% of the genome can be quantitatively amplified under the same conditions. Accurate measurement of the latent reservoir of HIV-1 is essential for evaluating the efficacy of cure strategies, and the bias against full length proviruses in nFGS methods must be considered.

High-resolution Hubble Space Telescope optical observations have been used to analyze the stellar population and the structure of the poorly investigated bulge globular cluster NGC 6316. We constructed the first high-resolution reddening map in the cluster direction, which allowed us to correct the evolutionary sequences in the color–magnitude diagram (CMD) for the effects of differential reddening. A comparison between the CMDs of NGC 6316 and 47 Tucanae revealed strikingly similar stellar populations, with the two systems basically sharing the same turnoff, subgiant branch, and horizontal branch morphologies, indicating comparable ages. The red giant branch in NGC 6316 appears slightly bluer than in 47 Tucanae, suggesting a lower metal content. This has been confirmed by the isochrone fitting of the observed CMD, which provided us with updated values of the cluster age, distance, average color excess, and metallicity. We estimated an absolute age of 13.1 ± 0.5 Gyr, consistent with the age of 47 Tucanae, an average color excess E ( B − V ) = 0.64 ± 0.01, and a true distance modulus ( m − M ) _0 = 15.27 ± 0.03 that sets the cluster distance at 11.3 kpc from the Sun. In addition, the photometric estimate of the cluster metallicity suggests [Fe/H] ≈ −0.9, which is ∼0.2 dex smaller than that of 47 Tucanae. We also determined the gravitational center and the density profile of the system from resolved stars. The latter is well reproduced by a King model. Our results confirm that NGC 6316 is another extremely old relic of the assembly history of the Galaxy.

Terzan 5 is a heavily obscured stellar system located in the inner Galaxy. It has been postulated to be a stellar relic, a bulge fossil fragment witnessing the complex history of the assembly of the Milky Way bulge. In this paper, we follow the chemical enrichment of a set of putative progenitors of Terzan 5 to assess whether the chemical properties of this cluster fit within a formation scenario in which it is the remnant of a primordial building block of the bulge. We can explain the metallicity distribution function and the runs of different element-to-iron abundance ratios as functions of [Fe/H] derived from optical-infrared spectroscopy of giant stars in Terzan 5 by assuming that the cluster experienced two major star formation bursts separated by a long quiescent phase. We further predict that the most metal-rich stars in Terzan 5 are moderately He-enhanced, and we predict a large spread of He abundances in the cluster, Y ≃ 0.26–0.335. We conclude that current observations fit within a formation scenario in which Terzan 5 originated from a pristine or slightly metal-enriched gas clump about one order of magnitude more massive than its present-day mass. Losses of gas and stars played a major role in shaping Terzan 5 the way we see it now. The iron content of the youngest stellar population is better explained if the white dwarfs that give rise to type Ia supernovae (the main Fe factories) sink toward the cluster center, rather than being stripped by the strong tidal forces exerted by the Milky Way in the outer regions.

We have used the “dynamical clock” to measure the level of dynamical evolution reached by three Galactic globular clusters (namely, NGC 3201, NGC 6316, and NGC 6440). This is an empirical method that quantifies the level of central segregation of blue straggler stars (BSSs) within the cluster half-mass radius by means of the ${A}_{\mathrm{rh}}^{+}$ parameter, defined as the area enclosed between the cumulative radial distribution of BSSs and that of a lighter population. The total sample with homogeneous determinations of ${A}_{\mathrm{rh}}^{+}$ currently includes 59 clusters: 52 old GCs in the Milky Way (including the three investigated here), five old clusters in the Large Magellanic Cloud, and two young systems in the Small Magellanic Cloud. The three objects studied here nicely nest into the correlation between ${A}_{\mathrm{rh}}^{+}$ and the central relaxation time defined by the previous sample, thus proving and consolidating the use of the dynamical clock as an excellent tracer of the stage of dynamical evolution of a star cluster in different galactic environments. Finally, we discuss the advantages of using the dynamical clock as an indicator of the dynamical ages of star clusters, compared to the present-day central relaxation time.

In the context of a project aimed at characterizing the dynamical evolution of old globular clusters in the Large Magellanic Cloud, we have secured deep HST/WFC3 images of the massive cluster NGC 1835. In the field of view of the acquired images, at a projected angular separation of approximately 2′ from the cluster, we detected the small stellar system KMK 88-10. The observations provided the deepest color–magnitude diagram ever obtained for this cluster, revealing that it hosts a young stellar population with an age of 600–1000 Myr. The cluster surface brightness profile is nicely reproduced by a King model with a core radius r _c = 4″ (0.97 pc), a half-mass radius r _hm = 12″ (2.9 pc), and a concentration parameter c ∼ 1.3 corresponding to a truncation radius r _t ∼ 81″ (19.5 pc). We also derived its integrated absolute magnitude ( M _V = −0.71) and total mass ( M ∼ 80–160 M _⊙ ). The most intriguing feature emerging from this analysis is that KMK 88-10 presents a structure elongated in the direction of NGC 1835, with an intracluster overdensity that suggests the presence of a tidal bridge between the two systems. If confirmed, this would be the first evidence of a tidal capture of a small star cluster by a massive globular.

In the context of the ESO-VLT Multi-Instrument Kinematic Survey of Galactic globular clusters, here we present the line-of-sight velocity dispersion profile of NGC 6440, a massive globular cluster located in the Galactic bulge. By combining the data acquired with four different spectrographs, we obtained the radial velocity of a sample of ∼1800 individual stars distributed over the entire cluster extension, from ∼0.″1 to 778″ from the center. Using a properly selected sample of member stars with the most reliable radial velocity measures, we derived the velocity dispersion profile up to 250″ from the center. The profile is well described by the same King model that best fits the projected star density distribution, with a constant inner plateau (at σ _0 ∼ 12 km s ^−1 ) and no evidence of a central cusp or other significant deviations. Our data allowed us to study the presence of rotation only in the innermost regions of the cluster ( r < 5″), revealing a well-defined pattern of ordered rotation with a position angle of the rotation axis of ∼132° ± 2° and an amplitude of ∼3 km s ^−1 (corresponding to V _rot / σ _0 ∼ 0.3). In addition, a flattening of the system qualitatively consistent with the rotation signal has been detected in the central region.

We report on the discovery of the companion star to the millisecond pulsar PSR J1835−3259B in the Galactic globular cluster NGC 6652. Taking advantage of deep photometric archival observations acquired through the Hubble Space Telescope in near-UV and optical bands, we identified a bright and blue object at a position compatible with that of the radio pulsar. The companion is located along the helium-core white dwarf cooling sequence, and the comparison with binary evolution models provides a mass of 0.17 ± 0.02 M _⊙ , a surface temperature of 11,500 ± 1900 K, and a very young cooling age of only 200 ± 100 Myr. The mass and the age of the companion are compatible with a progenitor star of about 0.87 M _⊙ , which started transferring mass to the primary during its evolution along the subgiant branch and stopped during the early red giant branch phase. Combining together the pulsar mass function and the companion mass, we found that this system is observed at an almost edge-on orbit and hosts a neutron star with a mass of 1.44 ± 0.06 M _⊙ , thus suggesting a highly nonconservative mass accretion phase. The young age of the WD companion is consistent with the scenario of a powerful, relatively young MSP indicated by the earlier detection of gamma-rays from this system.

In the context of a project aimed at characterizing the properties of star clusters in the Galactic bulge, here we present the determination of the internal kinematics and structure of the massive globular cluster NGC 6569. The kinematics has been studied by means of an unprecedented spectroscopic data set acquired in the context of the ESO-VLT Multi-Instrument Kinematic Survey of Galactic globular clusters, combining the observations from four different spectrographs. We measured the line-of-sight velocity of a sample of almost 1300 stars distributed between ∼0.″8 and 770″ from the cluster center. From a subsample of high-quality measures, we determined the velocity dispersion profile of the system over its entire radial extension (from ∼5″ to ∼200″ from the center), finding the characteristic behavior usually observed in globular clusters, with a constant inner plateau and a declining trend at larger radii. The projected density profile of the cluster has been obtained from resolved star counts, by combining high-resolution photometric data in the center, and the Gaia EDR3 catalog radially extended out to $\sim 20^{\prime} $ for a proper sampling of the Galactic field background. The two profiles are properly reproduced by the same King model, from which we estimated updated values of the central velocity dispersion, main structural parameters (such as the King concentration, the core, half mass, and tidal radii), total mass, and relaxation times. Our analysis also reveals a hint of ordered rotation in an intermediate region of the cluster (40″ < r < 90″, corresponding to 2 r _c < r < 4.5 r _c ), but additional data are required to properly assess this possibility.

Recently, slowly cooling white dwarfs (WDs) are a new class of WD that have been identified in two globular clusters (namely M13 and NGC 6752), showing a horizontal branch (HB) morphology with an extended blue tail. The cooling rate of these WDs is reduced by stable thermonuclear hydrogen burning in their residual envelope, and they are thought to originate by stars that populate the blue tail of the HB and then skip the asymptotic giant branch phase. Consistently, no evidence of such kind of WDs has been found in M3, a similar cluster with no blue extension of the HB. To further explore this phenomenon, we took advantage of deep photometric data acquired with the Hubble Space Telescope in the near-ultraviolet and investigated the bright portion of the WD cooling sequence in M5, another Galactic globular cluster with HB morphology similar to M3. The normalized WD luminosity function derived in M5 was found to be impressively similar to that observed in M3, in agreement with the fact that the stellar mass distribution along the HB of these two systems is almost identical. The comparison with theoretical predictions is consistent with the fact that the cooling sequence in this cluster is populated by canonical (fast cooling) WDs. Thus, the results presented in this paper provide further support to the scenario proposing a direct causal connection between the slow cooling WD phenomenon and the horizontal branch morphology of the host stellar cluster.

In the context of a project aimed at characterizing the properties of the so-called Bulge Fossil Fragments (the fossil remnants of the bulge formation epoch), here we present the first determination of the metallicity distribution of Liller 1. For a sample of 64 individual member stars we used ESO-MUSE spectra to measure the equivalent width of the Ca II triplet and then derive the iron abundance. To test the validity of the adopted calibration in the metal-rich regime, the procedure was first applied to three reference bulge globular clusters (NGC 6569, NGC 6440, and NGC 6528). In all the three cases, we found single-component iron distributions, with abundance values fully in agreement with those reported in the literature. The application of the same methodology to Liller 1 yielded, instead, a clear bimodal iron distribution, with a subsolar component at [Fe/H] = −0.48 dex ( σ = 0.22) and a supersolar component at [Fe/H] = +0.26 dex ( σ = 0.17). The latter is found to be significantly more centrally concentrated than the metal-poor population, as expected in a self-enrichment scenario and in agreement with that found in another bulge system, Terzan 5. The obtained metallicity distribution is astonishingly similar to that predicted by the reconstructed star formation history of Liller 1, which is characterized by three main bursts and a low, but constant, activity of star formation over the entire lifetime. These findings provide further support to the possibility that, similar to Terzan 5, Liller 1 is also a Bulge Fossil Fragment.

We used high-resolution spectra acquired with the Magellan Telescope to measure radial and rotational velocities of approximately 200 stars in the Galactic globular cluster NGC 3201. The surveyed sample includes blue straggler stars (BSSs) and reference stars in different evolutionary stages (main-sequence turnoff, subgiant, red giant, and asymptotic giant branches). The average radial velocity value (〈 V _r 〉 = 494.5 ± 0.5 km s ^−1 ) confirms a large systemic velocity for this cluster and was used to distinguish 33 residual field interlopers. The final sample of member stars has 67 BSSs and 114 reference stars. Similarly to what is found in other clusters, the totality of the reference stars has negligible rotation (< 20 km s ^−1 ), while the BSS rotational velocity distribution shows a long tail extending up to ∼200 km s ^−1 , with 19 BSSs (out of 67) spinning faster than 40 km s ^−1 . This sets the percentage of fast-rotating BSSs to ∼28%. Such a percentage is roughly comparable to that measured in other loose systems ( ω Centauri, M4, and M55) and significantly larger than that measured in high-density clusters (as 47 Tucanae, NGC 6397, NGC 6752, and M30). This evidence supports a scenario where recent BSS formation (mainly from the evolution of binary systems) is occurring in low-density environments. We also find that the BSS rotational velocity tends to decrease for decreasing luminosity and surface temperature, similarly to what is observed in main-sequence stars. Hence, further investigations are needed to understand the impact of BSS internal structure on the observed rotational velocities.

Abstract Despite antiretroviral therapy (ART) which halts HIV-1 replication and reduces plasma viral load to clinically undetectable levels, viral rebound inevitably occurs once ART is interrupted. HIV-1-infected cells can undergo clonal expansion, and these clonally expanded cells increase over time. Over 50% of latent reservoirs are maintained through clonal expansion. The clonally expanding HIV-1-infected cells, both in the blood and in the lymphoid tissues, contribute to viral rebound. The major drivers of clonal expansion of HIV-1-infected cells include antigen-driven proliferation, homeostatic proliferation and HIV-1 integration site-dependent proliferation. Here, we reviewed how viral, immunologic and genomic factors contribute to clonal expansion of HIV-1-infected cells, and how clonal expansion shapes the HIV-1 latent reservoir. Antigen-specific CD4+ T cells specific for different pathogens have different clonal expansion dynamics, depending on antigen exposure, cytokine profiles and exhaustion phenotypes. Homeostatic proliferation replenishes the HIV-1 latent reservoir without inducing viral expression and immune clearance. Integration site-dependent proliferation, a mechanism also deployed by other retroviruses, leads to slow but steady increase of HIV-1-infected cells harboring HIV-1 proviruses integrated in the same orientation at specific sites of certain cancer-related genes. Targeting clonally expanding HIV-1 latent reservoir without disrupting CD4+ T cell function is a top priority for HIV-1 eradication.

Abstract Antiretroviral therapy cannot cure HIV-1 infection due to the persistence of a small number of latently infected cells harboring replication-competent proviruses. Measuring persistent HIV-1 is challenging, as it consists of a mosaic population of defective and intact proviruses that can shift from a state of latency to active HIV-1 transcription. Due to this complexity, most of the current assays detect multiple categories of persistent HIV-1, leading to an overestimate of the true size of the latent reservoir. Here, we review the development of the viral outgrowth assay, the gold-standard quantification of replication-competent proviruses, and discuss the insights provided by full-length HIV-1 genome sequencing methods, which allowed us to unravel the composition of the proviral landscape. In this review, we provide a dissection of what defines HIV-1 persistence and we examine the unmet needs to measure the efficacy of interventions aimed at eliminating the HIV-1 reservoir.

Reactive oxygen species (ROS) are small oxygen-derived molecules that are used to control infections by phagocytic cells. In macrophages, the oxidative burst produced by the NOX2 NADPH-oxidase is essential to eradicate engulfed pathogens by both oxidative and non-oxidative killing. Indeed, while the superoxide anion (O2-) produced by NOX2, and the other ROS derived from its transformation, can directly target pathogens, ROS also contribute to activation of non-oxidative microbicidal effectors. The response of pathogens to the phagocytic oxidative burst includes the expression of different enzymes that target ROS to reduce their toxicity. Superoxide dismutases (SODs) are the primary scavengers of O2-, which is transformed into H2O2. In the Gram-negative Salmonella typhimurium, periplasmic SODCI has a major role in bacterial resistance to NOX-mediated oxidative stress. In Pseudomonas aeruginosa, the two periplasmic SODs, SODB, and SODM, appear to contribute to bacterial virulence in small-animal models. Furthermore, NOX2 oxidative stress is essential to restrict P. aeruginosa survival in macrophages early after infection. Here, we focused on the role of P. aeruginosa SODs in the counteracting of the lethal effects of the macrophage oxidative burst. Through this study of the survival of sod mutants in macrophages and the measurement of ROS in infected macrophages, we have identified a dual, antagonistic, role for SODB in P. aeruginosa survival. Indeed, the survival of the sodB mutants, but not of the sodM mutants, was greater than that of the wild-type (WT) bacteria early after infection, and sodB-infected macrophages showed higher levels of O2- and lower levels of H2O2. This suggests that SODB contributes to the production of lethal doses of H2O2 within the phagosome. However, later on following infection, the sodB mutants survived less that the WT bacteria, which highlights the pro-survival role of SODB. We have explained this defensive role through an investigation of the activation of autophagy, which was greater in the sodB-infected macrophages.

BackgroundHIV-1 proviruses can persist during ART in clonally-expanded populations of CD4+ T cells. To date, few examples of an expanded clones containing replication-competent proviruses exist, although it is suspected to be common. One such clone, denoted AMBI-1 (Maldarelli et al., 2014), was also a source of persistent viremia on ART, begging the question of how the AMBI-1 clone can survive despite infection with a replication-competent, actively-expressing provirus. We hypothesized that only a small fraction of cells within the AMBI-1 clone are activated to produce virus particles during cell division while the majority remain latent despite division, ensuring their survival. To address this question, we determined the fraction of HIV-1 proviruses within the AMBI-1 clone that expresses unspliced cell-associated RNA during ART and compared this fraction to 33 other infected T cell clones within the same individual.ResultsIn total, 34 different clones carrying either intact or defective proviruses in “Patient 1” from Maldarelli et al. (2014) were assessed. We found that 2.3% of cells within the AMBI-1 clone contained unspliced HIV-1 RNA. Highest levels of HIV-1 RNA were found in the effector memory (EM) T cell subset. The fraction of cells within clones that contained HIV-1 RNA was not different in clones with intact (median 2.3%) versus defective (median 3.5%) proviruses (p = 0.2). However, higher fractions and levels of RNA were found in cells with proviruses containing multiple drug resistance mutations, including those contributing to rebound viremia.ConclusionThese findings show that the vast majority of HIV-1 proviruses within expanded T cell clones, including intact proviruses, may be transcriptionally silent at any given time, implying that infected T cells may be able to be activated to proliferate without inducing the expression of the integrated provirus or, alternatelively, may be able to proliferate without cellular activation. The results of this study suggest that the long, presumed correlation between the level of cellular and proviral activation may not be accurate and, therefore, requires further investigation.

Combination antiretroviral therapy (cART) controls but does not eradicate HIV infection; HIV persistence is the principal obstacle to curing infections. The proportion of defective proviruses increases during cART, but the dynamics of this process are not well understood, and a quantitative analysis of how the proviral landscape is reshaped after cART is initiated is critical to understanding how HIV persists. Here, we studied longitudinal samples from HIV infected individuals undergoing long term cART using multiplexed Droplet Digital PCR (ddPCR) approaches to quantify the proportion of deleted proviruses in lymphocytes. In most individuals undergoing cART, HIV proviruses that contain gag are lost more quickly than those that lack gag. Increases in the fraction of gag-deleted proviruses occurred only after 1−2 years of therapy, suggesting that the immune system, and/or toxicity of viral re-activation helps to gradually shape the proviral landscape. After 10−15 years on therapy, there were as many as 3.5−5 times more proviruses in which gag was deleted or highly defective than those containing intact gag. We developed a provirus-specific ddPCR approach to quantify individual clones. Investigation of a clone of cells containing a deleted HIV provirus integrated in the HORMAD2 gene revealed that the cells underwent a massive expansion shortly after cART was initiated until the clone, which was primarily in effector memory cells, dominated the population of proviruses for over 6 years. The expansion of this HIV-infected clone had substantial effects on the overall proviral population.

This short paper presents the investigation of reaction mechanism induced by 6Li through a particle­γ coincidence measurement. The data have been taken from a 6Li+89Y experimentwhich is performed inINFN-LNL, Italy. In this experiment, the light charged particles are detected by a Si­ball, named EUCLIDES, and the γ rays are collected by a HPGe detector array, called GALILEO. In this contribution, scientific motivations, experimental details and some results, such as α­γ analysis, are presented.

Subtype A accounts for only 12% of HIV-1 infections worldwide but predominates in Russia and Former Soviet Union countries of Eastern Europe. After an early propagation via heterosexual contacts, this variant spread explosively among intravenous drug users. A distinct A1 variant predominates in Greece and Albania, which penetrated directly from Africa. Clade A1 accounts for 12.5% of non-B subtypes in Italy, being the most frequent after F1 subtype.Aim of this study was to investigate the circulation of A1 subtype in Italy and trace its origin and diffusion through phylogenetic and phylodynamic approaches.The phylogenetic analysis of 113 A1 pol sequences included in the Italian ARCA database, indicated that 71 patients (62.8%) clustered within 5 clades. A higher probability to be detected in clusters was found for patients from Eastern Europe and Italy (88.9% and 60.4%, respectively) compared to those from Africa (20%) (p < .001). Higher proportions of clustering sequences were found in intravenous drug users with respect to heterosexuals (85.7% vs. 59.3%, p = .056) and in women with respect to men (81.4% vs. 53.2%, p < .006). Subtype A1 dated phylogeny indicated an East African origin around 1961. Phylogeographical reconstruction highlighted 3 significant groups. One involved East European and some Italian variants, the second encompassed some Italian and African strains, the latter included the majority of viruses carried by African and Italian subjects and all viral sequences from Albania and Greece.Subtype A1 originated in Central Africa and spread among East European countries in 1982. It entered Italy through three introduction events: directly from East Africa, from Albania and Greece, and from the area encompassing Moldavia and Ukraine. As in previously documented A1 epidemics of East European countries, HIV-1 A1 subtype spread in Italy in part through intravenous drug users. However, Eastern European women contributed to the penetration of such variant, probably through sex work.

Investigation of the breakup and transfer effect of weakly bound nuclei on the fusion process has been an interesting research topic in the past several years. In comparison with radioactive ion beam (RIB), the beam intensities of stable weakly bound nuclei such as 6,7Li and 9Be, which have significant breakup probability, are orders of magnitude higher. Precise fusion measurements induced by these nuclei have already been performed. However, the conclusion of reaction dynamics was not clear and has contradiction. In order to have a proper understanding of the influence of breakup and transfer of weakly bound projectiles on the fusion process, the 6Li+89Y experiment with incident energies of 22 MeV and 34 MeV was performed on Galileo array in combination with Si-ball EUCLIDES at Legnaro National Laboratory (LNL) in Italy. Using the coincidence by the charged particles and γ-rays, the different reaction channels can be clearly identified.

Investigation of the breakup and transfer effect of weakly bound nuclei on the fusion process has been an interesting research topic in the past several years. However, owing to the low intensities of the presently available radioactive ion beam (RIB), it is difficult to clearly explore the reaction mechanisms of nuclear systems with unstable nuclei. In comparison with RIB, the beam intensities of stable weakly bound nuclei such as 6,7Li and 9Be, which have significant breakup probability, are orders of magnitude higher. Precise fusion measurements have already been performed with those stable weakly bound nuclei, and the effect of breakup of those nuclei on the fusion process has been extensively studied. Those nuclei indicated large production cross sections for particles other than the α + x breakup. The particles are originated from non-capture breakup (NCBU), incomplete fusion (ICF) and transfer processes. However, the conclusion of reaction dynamics was not clear and has the contradiction. In our previous experiments we have performed 6Li+96Zr and 154Sm at HI-13 Tandem accelerator of China Institute of Atomic Energy (CIAE) by using HPGe array. It is shown that there is a small complete fusion (CF) suppression on medium-mass target nucleus 96Zr different from about 35% suppression on heavier target nucleus 154Sm at near-barrier energies. It seems that the CF suppression factor depends on the charge of target nuclei. We also observed one neutron transfer process. However, the experimental data are scarce for medium-mass target nuclei. In order to have a proper understanding of the influence of breakup and transfer of weakly bound projectiles on the fusion process, we performed the 6Li+89Y experiment with incident energies of 22 MeV and 34 MeV on Galileo array in cooperation with Si-ball EUCLIDES at Legnaro National Laboratory (LNL) in Italy. Using particle-particle and particle-γ coincidences, the different reaction mechanisms can be clearly explored.

About 40% of the Italian HIV-1 epidemic due to non-B variants is sustained by F1 clade, which circulates at high prevalence in South America and Eastern Europe. Aim of this study was to define clade F1 origin, population dynamics and epidemiological networks through phylogenetic approaches. We analyzed pol sequences of 343 patients carrying F1 subtype stored in the ARCA database from 1998 to 2009. Citizenship of patients was as follows: 72.6% Italians, 9.3% South Americans and 7.3% Rumanians. Heterosexuals, Homo-bisexuals, Intravenous Drug Users accounted for 58.1%, 24.0% and 8.8% of patients, respectively. Phylogenetic analysis indicated that 70% of sequences clustered in 27 transmission networks. Two distinct groups were identified; the first clade, encompassing 56 sequences, included all Rumanian patients. The second group involved the remaining clusters and included 10 South American Homo-bisexuals in 9 distinct clusters. Heterosexual modality of infection was significantly associated with the probability to be detected in transmission networks. Heterosexuals were prevalent either among Italians (67.2%) or Rumanians (50%); by contrast, Homo-bisexuals accounted for 71.4% of South Americans. Among patients with resistant strains the proportion of clustering sequences was 57.1%, involving 14 clusters (51.8%). Resistance in clusters tended to be higher in South Americans (28.6%) compared to Italian (17.7%) and Rumanian patients (14.3%). A striking proportion of epidemiological networks could be identified in heterosexuals carrying F1 subtype residing in Italy. Italian Heterosexual males predominated within epidemiological clusters while foreign patients were mainly Heterosexual Rumanians, both males and females, and South American Homo-bisexuals. Tree topology suggested that F1 variant from South America gave rise to the Italian F1 epidemic through multiple introduction events. The contact tracing also revealed an unexpected burden of resistance in epidemiological clusters underlying the need of public interventions to limit the spread of non-B subtypes and transmitted drug resistance.

Clonal expansion of infected CD4+ T cells is a major mechanism of HIV-1 persistence and a barrier to achieving a cure. Potential causes are homeostatic proliferation, effects of HIV-1 integration, and interaction with antigens. Here, we show that it is possible to link antigen responsiveness, the full proviral sequence, the integration site, and the T cell receptor β-chain (TCRβ) sequence to examine the role of recurrent antigenic exposure in maintaining the HIV-1 reservoir. We isolated CMV- and Gag-responding CD4+ T cells from 10 treated individuals. Proviral populations in CMV-responding cells were dominated by large clones, including clones harboring replication-competent proviruses. TCRβ repertoires showed high clonality driven by converging adaptive responses. Although some proviruses were in genes linked to HIV-1 persistence (BACH2, STAT5B, MKL1), the proliferation of infected cells under antigenic stimulation occurred regardless of the site of integration. Paired TCRβ and integration site analysis showed that infection could occur early or late in the course of a clone's response to antigen and could generate infected cell populations too large to be explained solely by homeostatic proliferation. Together, these findings implicate antigen-driven clonal selection as a major factor in HIV-1 persistence, a finding that will be a difficult challenge to eradication efforts.