20 research outputs found

    A Stochastic Model of Latently Infected Cell Reactivation and Viral Blip Generation in Treated HIV Patients

    Get PDF
    Motivated by viral persistence in HIV+ patients on long-term anti-retroviral treatment (ART), we present a stochastic model of HIV viral dynamics in the blood stream. We consider the hypothesis that the residual viremia in patients on ART can be explained principally by the activation of cells latently infected by HIV before the initiation of ART and that viral blips (clinically-observed short periods of detectable viral load) represent large deviations from the mean. We model the system as a continuous-time, multi-type branching process. Deriving equations for the probability generating function we use a novel numerical approach to extract the probability distributions for latent reservoir sizes and viral loads. We find that latent reservoir extinction-time distributions underscore the importance of considering reservoir dynamics beyond simply the half-life. We calculate blip amplitudes and frequencies by computing complete viral load probability distributions, and study the duration of viral blips via direct numerical simulation. We find that our model qualitatively reproduces short small-amplitude blips detected in clinical studies of treated HIV infection. Stochastic models of this type provide insight into treatment-outcome variability that cannot be found from deterministic models

    Predicting the outcomes of treatment to eradicate the latent reservoir for HIV-1

    Get PDF
    Massive research efforts are now underway to develop a cure for HIV infection, allowing patients to discontinue lifelong combination antiretroviral therapy (ART). New latency-reversing agents (LRAs) may be able to purge the persistent reservoir of latent virus in resting memory CD4+ T cells, but the degree of reservoir reduction needed for cure remains unknown. Here we use a stochastic model of infection dynamics to estimate the efficacy of LRA needed to prevent viral rebound after ART interruption. We incorporate clinical data to estimate population-level parameter distributions and outcomes. Our findings suggest that approximately 2,000-fold reductions are required to permit a majority of patients to interrupt ART for one year without rebound and that rebound may occur suddenly after multiple years. Greater than 10,000-fold reductions may be required to prevent rebound altogether. Our results predict large variation in rebound times following LRA therapy, which will complicate clinical management. This model provides benchmarks for moving LRAs from the lab to the clinic and can aid in the design and interpretation of clinical trials. These results also apply to other interventions to reduce the latent reservoir and can explain the observed return of viremia after months of apparent cure in recent bone marrow transplant recipients and an immediately-treated neonate.Comment: 8 pages main text (4 figures). In PNAS Early Edition http://www.pnas.org/content/early/2014/08/05/1406663111. Ancillary files: SI, 24 pages SI (7 figures). File .htm opens a browser-based application to calculate rebound times (see SI). Or, the .cdf file can be run with Mathematica. The most up-to-date version of the code is available at http://www.danielrosenbloom.com/reboundtimes

    Stochastic modelling of the eradication of the HIV-1 infection by stimulation of latently infected cells in patients under highly active anti-retroviral therapy

    Full text link
    HIV-1 infected patients are effectively treated with highly active anti-retroviral therapy (HAART). Whilst HAART is successful in keeping the disease at bay with average levels of viral load well below the detection threshold of standard clinical assays, it fails to completely eradicate the infection, which persists due to the emergence of a latent reservoir with a half-life time of years and is immune to HAART. This implies that life-long administration of HAART is, at the moment, necessary for HIV-1-infected patients, which is prone to drug resistance and cumulative side effects as well as imposing a considerable financial burden on developing countries, those more afflicted by HIV, and public health systems. The development of therapies which specifically aim at the removal of this latent reservoir has become a focus of much research. A proposal for such therapy consists of elevating the rate of activation of the latently infected cells: by transferring cells from the latently infected reservoir to the active infected compartment, more cells are exposed to the anti-retroviral drugs thus increasing their effectiveness. In this paper, we present a stochastic model of the dynamics of the HIV-1 infection and study the effect of the rate of latently infected cell activation on the average extinction time of the infection. By analysing the model by means of an asymptotic approximation using the semi-classical quasi steady state approximation (QSS), we ascertain that this therapy reduces the average life-time of the infection by many orders of magnitudes. We test the accuracy of our asymptotic results by means of direct simulation of the stochastic process using a hybrid multi-scale Monte Carlo scheme

    Determinants of the efficacy of HIV latency-reversing agents and implications for drug and treatment design

    Get PDF
    HIV eradication studies have focused on developing latency-reversing agents (LRAs). However, it is not understood how the rate of latent reservoir reduction is affected by different steps in the process of latency reversal. Furthermore, as current LRAs are host-directed, LRA treatment is likely to be intermittent to avoid host toxicities. Few careful studies of the serial effects of pulsatile LRA treatment have yet been done. This lack of clarity makes it difficult to evaluate the efficacy of candidate LRAs or predict long-term treatment outcomes. We constructed a mathematical model that describes the dynamics of latently infected cells under LRA treatment. Model analysis showed that, in addition to increasing the immune recognition and clearance of infected cells, the duration of HIV antigen expression (i.e., the period of vulnerability) plays an important role in determining the efficacy of LRAs, especially if effective clearance is achieved. Patients may benefit from pulsatile LRA exposures compared with continuous LRA exposures if the period of vulnerability is long and the clearance rate is high, both in the presence and absence of an LRA. Overall, the model framework serves as a useful tool to evaluate the efficacy and the rational design of LRAs and combination strategies

    Stochastic modelling of cellular populations: Effects of latency and feedbackl

    Get PDF
    [cat]L'objectiu principal d'aquesta tesi doctoral és l'estudi de l'efecte de les fluctuacions en poblacions acoblades en sistemes biològics, on cèl·lules en estat latent juguen un paper important. Intentant trobar el significat biològic de la dinàmica dels sistemes. Els punts específics que volem abordar i la organització de la tesi estan explicats a continuació. En el Capítol 2, estudiem el comportament de les poblacions de cèl·lules amb estructura jeràrquica des del punt de vista de les propietats d'estabilitat, En particular: - 1. Divisió simètrica contra asimètrica en el compartiment de les cèl·lules mare. Estudiem la robustesa de les poblacions amb estructura jeràrquica, depenent de si les cèl·lules mare es divideixen simètricament, asimètricament o de les dues maneres. Estudiem com la divisió simètrica afecta a l'estabilitat de la població, ja que això té una gran importància en la progressió del càncer. - 2. La competició entre dues poblacions amb diferents tipus de divisió de les cèl·lules mare. Això és crucial per trobar estratègies òptimes que maximitzin la robustesa (supervivència a llarg termini, resistència a invasions i habilitat per invadir) de poblacions amb estructura jeràrquica. - 3. La influència de paràmetres com son la duplicació i el ritme de mort de cèl·lules mare, el temps de vida mitjà de les cèl·lules completament diferenciades, la longitud de les cadenes de diferenciació i les fluctuacions al compartiment de les cèl·lules mare en la robustesa i arquitectura òptima de les cascades de diferenciació. En el Capítol 3 presentem un model homogeni de combinació de HAART amb teràpies d'activació de les cèl·lules latents del VIH-1 a la sang. Estem interessats en: - 1. L'efecte del ritme d'activació de les cèl·lules latents en el temps mitjà de vida de la infecció. En particular analitzem si les teràpies basades en incrementar aquest ritme són capaces de suprimir la infecció en un temps raonable. - 2. La importància de l'eficiència de les teràpies antiretrovirals, incloent els casos límit en que l'eficàcia és del 100%, en la quantitat de càrrega viral. - 3. La formulació d'una teoria asimptòtica basada en l'aproximació semi-clàssica amb aproximacions quasi estacionàries per descriure la dinàmica del procés. La precisió d'aquest mètode asimptòtic és comparat amb simulacions multi-scale proposades pel Cao et al. En el Capítol 4, estenem el model proposat pel Rong i el Perelson a un model no homogeni de la dinàmica del VIH-1 en el corrent sanguini, considerant que les cèl·lules i els virus no estan distribuïts de manera uniforme en la sang. Els punts específics que volem estudiar són: - 1. El mecanisme que fa que apareguin els episodis de virèmia per sobre els límits de detecció, coneguts com viral blips. En particular volem investigar si són producte de fluctuacions estocàstiques degudes a la inhomogenietat o un altre mecanisme ha de ser considerat. - 2. Si l'aparició dels viral blips està afectada pels procediments duts a terme en el laboratori, com el temps d'espera entre les extraccions i les observacions. - 3. Si la probabilitat, l'amplitud i la freqüència dels viral blips es veu afectada pels diferents possibles tipus de producció viral, és a dir, continua vs burst. En el Capítol 5 presentem i discutim els resultats obtinguts, i comparem, quan és possible, amb altres models o amb resultats experimentals, i discutim el treball que deixem pel futur. Els detalls relatius a qüestions metodològiques, això com una introducció a la modelització estocàstica fent servir equacions mestres es donen en els apèndixs. Per a aquells que no estan familiaritzats amb els models basats en equacions mestres, l'autor recomana llegir primer l'apèndix A que proporciona la base matemàtica per entendre el capítol 2. Els Apèndixs B, C i D juntament amb l'Apèndix A donen la base matemàtica necessària per seguir el capítol 3 i el capítol 4

    Intragenic elements support the transcription of defective HIV-1 proviruses

    Full text link
    Human immunodeficiency virus-1 (HIV-1) establishes a persistent proviral reservoir by integrating into the genome of infected host cells. Current antiretroviral treatments (ART) do not target this persistent population of proviruses which include latently infected cells that upon treatment interruption can be reactivated to contribute to HIV-1 rebound. Deep sequencing of persistent HIV-1 proviruses has revealed that greater than 90% of integrated HIV-1 genomes are defective and unable to produce infectious virions. We hypothesized that intragenic elements in the HIV genome support transcription of aberrant HIV-1 RNAs from defective proviruses that lack long terminal repeats (LTRs). Using an intact provirus detection assay, I observed that resting CD4+ T cells and monocyte-derived macrophages (MDMs) are biased towards generating defective HIV-1 proviruses. Multiplex reverse transcription droplet digital polymerase chain reaction (RT-ddPCR) identified env and nef transcripts which lacked 5’ untranslated regions (UTR) in acutely infected CD4+ T cells and MDMs indicating transcripts are generated that do not utilize the promoter within the LTR. 5’UTR-deficient env transcripts were also identified in a cohort of people living with HIV-1 (PLWH) on ART, suggesting that these aberrant RNAs are produced in vivo. Using 5’ rapid amplification of cDNA ends (RACE), I mapped the start site of these transcripts within the Env gene. This region bound several cellular transcription factors and functioned as a transcriptional regulatory element that could support transcription and translation of downstream HIV-1 RNAs. Transient expression of an HIV-1 5’UTR deletion construct in HEK293T cells demonstrated that HIV-1 transcripts and proteins are still produced when the 5’UTR is absent. These studies provide mechanistic insights into how defective HIV-1 proviruses are persistently expressed to potentially drive inflammation in PLWH

    Modulation of HIV-specific T cell responses during standard antiretroviral treatment and immunotherapy

    Full text link
    Seule une minorité des individus infectés par le virus de l’immunodéficience humaine (VIH) développe une réponse immunitaire capable de contrôler le virus. Chez la plupart des individus, on observe un échappement virologique et un épuisement des lymphocytes T CD8+ spécifiques du VIH. L’infection chronique non-traitée altère également les lymphocytes T CD4+ spécifiques du VIH caractérisé par l’expression accrue des récepteurs co-inhibiteurs et une signature des cellules auxiliaires T folliculaires (Tfh). La thérapie antirétrovirale (TAR) est très efficace pour supprimer durablement la charge virale dans le plasma. Néanmoins, elle ne permet pas une éradication complète du VIH car le virus persiste, intégré dans le génome des cellules réservoirs, desquelles le virus réapparaît lors de l’interruption de la thérapie. Cela démontre que l'immunité adaptive spécifiques du VIH n'est pas restaurée. Les anticorps neutralisants à large spectre (bNAbs) représentent une alternative potentielle à la TAR. En plus de la neutralisation du virus – et contrairement à la TAR – les bNAbs ne limitent pas la disponibilité de l'antigène et peuvent engager le système immunitaire. L'administration de bNAbs à des macaques rhésus induit des réponses immunitaires adaptatives associées à un contrôle prolongé de la virémie, mais cela n’a pas été établi chez l’Homme. Dans cette thèse, nous avons donc exploré la modulation des réponses des lymphocytes T spécifiques du VIH lors d'une TAR standard et d’une immunothérapie utilisant des bNAbs. Dans un premier objectif nous avons analysé la modulation persistante des réponses des lymphocytes T CD4+ spécifiques du VIH chez les individus sous TAR. Nous avons pu démontrer l'expansion persistante des Tfh spécifiques au VIH avec des caractéristiques phénotypiques et fonctionnelles les distinguant des Tfh spécifiques d’antigènes viraux comparatifs (cytomégalovirus, virus de l’hépatite B). Ces caractéristiques ont été induites au cours de l’infection chronique non-traitée, persistaient pendant la TAR et étaient associées au réservoir du VIH compétent pour la traduction. Ces données suggèrent qu’une stimulation antigénique persistante, malgré une TAR efficace, maintient des modifications immunologiques notamment au niveau des Tfh. Dans un second objectif, nous avons caractérisé les réponses T spécifiques du VIH à la suite d’un traitement utilisant des bNAbs et une interruption structurée de la TAR (IST). Des individus inclus dans une étude clinique de phase Ib ont reçu une perfusion d’une combinaison des bNAbs 10-1074 et 3BNC117 et ont démontré une suppression virale prolongée après l’IST. Chez ces participants, nous avons observé une augmentation des réponses immunitaires des lymphocytes T CD8+ et CD4+ spécifiques du VIH due à l'expansion des réponses immunitaires préexistantes et au développement de réponses ciblant de nouveaux épitopes. Cela suggère que la combinaison d’un traitement par bNAbs avec l’IST est associée au maintien de la charge virale plasmatique indétectable et à une intensification de la réponse immunitaire des lymphocytes T spécifiques du VIH. Nos travaux permettent une meilleure compréhension des réponses des lymphocytes T spécifiques du VIH au cours de la TAR et lors d’une immunothérapie. Ils peuvent contribuer au développement de stratégies thérapeutiques plus efficaces visant à contrôler la réplication virale sans la TAR.Only a small fraction of individuals infected with the human immunodeficiency virus (HIV) develops effective immune responses able to control the virus. In most individuals, the virus escapes the antiviral immune response and HIV-specific CD8+ T cell responses become exhausted. Untreated progressive HIV infection also leads to alterations in HIV-specific CD4+ T cells. This includes increased expression of co-inhibitory receptors and skewing towards a T follicular helper cell (Tfh) signature. Antiretroviral therapy (ART) is highly effective in controlling the HIV viral load at undetectable levels in the plasma. However, ART does not represent a cure as the virus integrates into the genome of infected cells from where the virus rebounds once ART is stopped. This demonstrates that the HIV-specific T cell immunity is not restored. However, the changes that are introduced during progressive infection and that are maintained after viral suppression with ART are poorly known. Broadly neutralizing antibodies (bNAbs) represent a potential alternative to ART. In addition to virus neutralization and unlike ART, bNAbs to do not limit HIV antigen availability and can engage the immune system. bNAb administration elicited adaptive immune responses that were associated with long-lasting viral control in a simian animal model but this has not been established in HIV-infected individuals. In this thesis, we therefore proceeded to study the modulation of HIV-specific T cell responses during standard ART and after an immunotherapeutic intervention using bNAbs. The first objective was to better understand persistent modulation of HIV-specific CD4+ T cell responses in ART-treated individuals. Our results demonstrated the persistent expansion of HIV-specific Tfh cell responses with multiple phenotypic and functional features that differed from Tfh cells specific for comparative viral antigens (cytomegalovirus, hepatitis B virus). These features were induced during chronic untreated HIV infection, persisted during ART and correlated with the translation-competent HIV reservoir. This suggests that persistent HIV antigen expression, despite effective ART, maintains these altered immunological features specifically for Tfh responses. For the second objective, we characterized changes in the HIV-specific CD8+ and CD4+ T cell immunity after bNAb treatment and analytical treatment interruption (ATI). For this, we used samples obtained from participants enrolled in a clinical phase Ib study that received combined infusion of bNAbs 10-1074 and 3BNC117 and demonstrated prolonged viral suppression after ATI. In these individuals, we detected an increase of HIV-specific CD8+ and CD4+ T cell responses during ART interruption when compared to baseline. Increased T cell responses were due to both expansion of pre-existing responses and the emergence of responses to new epitopes. In contrast, HIV-specific T cell responses remained unchanged in ART-treated individuals who did not receive bNAb infusions. This suggests that bNAb treatment and ATI is associated with increased HIV-specific T cell immunity while viral suppression is maintained. Together our results contribute to a better understanding of HIV-specific T cell responses during ART and immunotherapy treatment. Our findings may help to develop more effective HIV treatment strategies to improve the host’s immune system so that HIV can be controlled without the need for ART
    corecore