Dynamic and regulation of LILRB2/MHC-I inhibitory axis during HIV/SIV infection

Les cellules dendritiques classiques (cDC) jouent un rôle crucial dans l’efficacité des réponses immunitaires précoces conduisant au contrôle ou à la persistance virale. A cet égard, il a été montré que l’infection par le VIH induit des dysfonctions des cDC caractérisées par une inhibition de leur capacité à stimuler les cellules T et associées à la progression de la maladie. Parmi les mécanismes moléculaires impliqués dans ces dysfonctions, des études in vitro ont mis en évidence le rôle du récepteur inhibiteur LILRB2. Néanmoins, la dynamique d’expression de LILRB2 ainsi que son rôle dès les premiers stades de l'infection restent à démontrer. Chez des patients en primo-infection VIH-1, nous observons une augmentation de l’expression de LILRB2 et de ses ligands HLA-I à la surface des cDC. Par ailleurs, la cinétique d’expression de LILRB2 et CMH-I au cours de l’infection de macaques cynomolgus par le SIVmac251 montre une augmentation transitoire de l'expression de LILRB2 et du CMH-I sur les cDC du sang et des ganglions lymphatiques dès les premiers jours de l’infection. Parmi les mécanismes qui pourraient être impliqués dans la régulation de l’expression de LILRB2, nos résultats indiquent que la réplication du VIH-1, l'activation de voies TLR7/8 ainsi que la présence d’IL-10 et d’IFN-I induisent une forte expression de LILRB2. Enfin, cette expression exacerbée de LILRB2 sur les cDC semble être spécifique à l'infection par le VIH/SIV. En effet, l’infection de macaques cynomolgus par le virus chikungunya, qui est caractérisée par une réponse immunitaire antivirale robuste aboutissant à un contrôle de la virémie, est associée à une expression diminuée de LILRB2 sur les cDC dès les premiers jours de l’infection. L’ensemble de nos données suggèrent un rôle majeur de l’axe inhibiteur LILRB2/MHC-I dans les mécanismes de dérégulations des cDC qui pourrait participerait à l’inefficacité des réponses immunitaires adaptatives et à la persistance du VIH/SIV.Conventional dendritic cells (cDCs) play a crucial role in setting up early immune responses leading to viral control or persistence. In this regard, it has been shown that HIV-1 infection induces cDC dysfunctions characterized by inhibitions in their ability to stimulate T-cells and associated with disease progression. In vitro studies have shown the implication of LILRB2 inhibitory receptor in cDC dysfunctions. However, the dynamic of LILRB2 expression and its role in the early stages of infection are yet to be characterized. In primary HIV-1 infected patients, we observe an increased expression of LILRB2 and its ligands, HLA-I, on the surface of cDCs. Kinetics of LILRB2 and MHC-I expressions during SIV infection of Cynomolgus macaques shows a transient increase in LILRB2 and MHC-I expressions on blood and lymph node cDCs during the first days of infection. We also show that HIV replication, activation of TLR7/8 pathways, and presence of IL-10 and IFN-I drive upregulated expression of LILRB2. Finally, this strong induced LILIRB2 expression seems specific to HIV/SIV infections. Indeed, chikungunya virus infection of cynomolgus macaques, which characterized by a robust antiviral immune response leading to viral control, is associated with decreased expression of LILRB2 on cDCs in the first days of infection. Taken together, our data suggest a major role of the LILRB2/HLA-I inhibitory axis, mediating cDC dysfunctions and thus contributing to inefficient adaptive immune responses and viral persistence

Etude de la dynamique de l’axe inhibiteur LILRB2/CMH-I et de sa régulation au cours de l’infection par le VIH/SIV

Conventional dendritic cells (cDCs) play a crucial role in setting up early immune responses leading to viral control or persistence. In this regard, it has been shown that HIV-1 infection induces cDC dysfunctions characterized by inhibitions in their ability to stimulate T-cells and associated with disease progression. In vitro studies have shown the implication of LILRB2 inhibitory receptor in cDC dysfunctions. However, the dynamic of LILRB2 expression and its role in the early stages of infection are yet to be characterized. In primary HIV-1 infected patients, we observe an increased expression of LILRB2 and its ligands, HLA-I, on the surface of cDCs. Kinetics of LILRB2 and MHC-I expressions during SIV infection of Cynomolgus macaques shows a transient increase in LILRB2 and MHC-I expressions on blood and lymph node cDCs during the first days of infection. We also show that HIV replication, activation of TLR7/8 pathways, and presence of IL-10 and IFN-I drive upregulated expression of LILRB2. Finally, this strong induced LILIRB2 expression seems specific to HIV/SIV infections. Indeed, chikungunya virus infection of cynomolgus macaques, which characterized by a robust antiviral immune response leading to viral control, is associated with decreased expression of LILRB2 on cDCs in the first days of infection. Taken together, our data suggest a major role of the LILRB2/HLA-I inhibitory axis, mediating cDC dysfunctions and thus contributing to inefficient adaptive immune responses and viral persistence.Les cellules dendritiques classiques (cDC) jouent un rôle crucial dans l’efficacité des réponses immunitaires précoces conduisant au contrôle ou à la persistance virale. A cet égard, il a été montré que l’infection par le VIH induit des dysfonctions des cDC caractérisées par une inhibition de leur capacité à stimuler les cellules T et associées à la progression de la maladie. Parmi les mécanismes moléculaires impliqués dans ces dysfonctions, des études in vitro ont mis en évidence le rôle du récepteur inhibiteur LILRB2. Néanmoins, la dynamique d’expression de LILRB2 ainsi que son rôle dès les premiers stades de l'infection restent à démontrer. Chez des patients en primo-infection VIH-1, nous observons une augmentation de l’expression de LILRB2 et de ses ligands HLA-I à la surface des cDC. Par ailleurs, la cinétique d’expression de LILRB2 et CMH-I au cours de l’infection de macaques cynomolgus par le SIVmac251 montre une augmentation transitoire de l'expression de LILRB2 et du CMH-I sur les cDC du sang et des ganglions lymphatiques dès les premiers jours de l’infection. Parmi les mécanismes qui pourraient être impliqués dans la régulation de l’expression de LILRB2, nos résultats indiquent que la réplication du VIH-1, l'activation de voies TLR7/8 ainsi que la présence d’IL-10 et d’IFN-I induisent une forte expression de LILRB2. Enfin, cette expression exacerbée de LILRB2 sur les cDC semble être spécifique à l'infection par le VIH/SIV. En effet, l’infection de macaques cynomolgus par le virus chikungunya, qui est caractérisée par une réponse immunitaire antivirale robuste aboutissant à un contrôle de la virémie, est associée à une expression diminuée de LILRB2 sur les cDC dès les premiers jours de l’infection. L’ensemble de nos données suggèrent un rôle majeur de l’axe inhibiteur LILRB2/MHC-I dans les mécanismes de dérégulations des cDC qui pourrait participerait à l’inefficacité des réponses immunitaires adaptatives et à la persistance du VIH/SIV

Using Machine Learning in WSNs for Performance Prediction MAC Layer

To monitor environments, Wireless Sensor Networks (WSNs) are used for collecting data in divers domains such as smart factories, smart buildings, etc. In such environments, different medium access control (MAC) protocols are available to sensor nodes for wireless communications and are of a paramount importance to enhance the network performance. Proposed MAC layer protocols for WSNs are generally designed to achieve a good performance in packet reception rate. Once chosen, the MAC protocol is used and remains the same throughout the network lifetime even if its performance decreases over time. In this paper, we adopt supervised machine learning techniques to predict the performance of CSMA/CA MAC protocol based on the packet reception rate. Our approach consists of three steps: experiments for data collection, offline modeling and performance evaluation. Our analysis shows that XGBoost prediction model is the better supervised machine learning technique to enhance network performance at the MAC layer level. In addition, we use SHAP method to explain predictions

ETV3 and ETV6 enable monocyte differentiation into dendritic cells by repressing macrophage fate commitment

International audienceIn inflamed tissues, monocytes differentiate into macrophages (mo-Macs) or dendritic cells (mo-DCs). In chronic nonresolving inflammation, mo-DCs are major drivers of pathogenic events. Manipulating monocyte differentiation would therefore be an attractive therapeutic strategy. However, how the balance of mo-DC versus mo-Mac fate commitment is regulated is not clear. In the present study, we show that the transcriptional repressors ETV3 and ETV6 control human monocyte differentiation into mo-DCs. ETV3 and ETV6 inhibit interferon (IFN)-stimulated genes; however, their action on monocyte differentiation is independent of IFN signaling. Instead, we find that ETV3 and ETV6 directly repress mo-Mac development by controlling MAFB expression. Mice deficient for Etv6 in monocytes have spontaneous expression of IFN-stimulated genes, confirming that Etv6 regulates IFN responses in vivo. Furthermore, these mice have impaired mo-DC differentiation during inflammation and reduced pathology in an experimental autoimmune encephalomyelitis model. These findings provide information about the molecular control of monocyte fate decision and identify ETV6 as a therapeutic target to redirect monocyte differentiation in inflammatory disorders

Functional specialization of short-lived and long-lived macrophage subsets in human tonsils

International audienceMacrophages play a central role in tissue homeostasis and host defense. However, the properties of human macrophages in non-diseased tissues remain poorly understood. Here, we characterized human tonsil macrophages and identified three subsets with distinct phenotype, transcriptome, life cycle, and function. CD36hi macrophages were related to monocytes, while CD36lo macrophages showed features of embryonic origin and CD36int macrophages had a mixed profile. scRNA-seq on non-human primate tonsils showed that monocyte recruitment did not pre-exist an immune challenge. Functionally, CD36hi macrophages were specialized for stimulating T follicular helper cells, by producing Activin A. Combining reconstruction of ligand-receptor interactions and functional assays, we identified stromal cell-derived TNF-α as an inducer of Activin A secretion. However, only CD36hi macrophages were primed for Activin A expression, via the activity of IRF1. Our results provide insight into the heterogeneity of human lymphoid organ macrophages and show that tonsil CD36hi macrophage specialization is the result of both intrinsic features and interaction with stromal cells

Mass Cytometry Analysis Reveals Complex Cell-State Modifications of Blood Myeloid Cells During HIV Infection

International audienceDendritic cells (DC), which are involved in orchestrating early immune responses against pathogens, are dysregulated in their function by HIV infection. This dysregulation likely contributes to tip the balance toward viral persistence. Different DC subpopulations, including classical (cDCs) and plasmacytoid (pDCs) dendritic cells, are subjected to concomitant inflammatory and immunoregulatory events during HIV infection, which hampers the precise characterization of their regulation through classical approaches. Here, we carried out mass cytometry analysis of blood samples from early HIV-infected patients that were longitudinally collected before and after 1 year of effective combination antiretroviral therapy (cART). Blood samples from HIV controller patients who naturally control the infection were also included. Our data revealed that plasma HIV RNA level was positively associated with a loss of cDC and pDC subpopulations that display high expression of LILR immunomodulatory receptors. Conversely, specific monocyte populations co-expressing high levels of HLA-I, 3 immunomodulatory receptors, CD64, LILRA2, and LILRB4, and the restriction factor CD317 (also known as BST2/Tetherin), were more abundant in early HIV-infection. Finally, our analysis revealed that the blood of HIV controller patients contained in a higher abundance a particular subtype of CD1c+ cDCs, characterized by elevated co-expression of CD32b inhibitory receptor and HLA-DR antigen-presentation molecules. Overall, this study unravels the modifications induced in DC and monocyte subpopulations in different HIV+ conditions, and provides a better comprehension of the immune regulation/dysregulation mechanisms induced during this viral infection

Mass Cytometry Analysis Reveals the Landscape and Dynamics of CD32a+ CD4+ T Cells From Early HIV Infection to Effective cART

International audienceCD32a has been proposed as a specific marker of latently HIV-infected CD4 + T cells. However, CD32a was recently found to be expressed on CD4 + T cells of healthy donors, leading to controversy on the relevance of this marker in HIV persistence. Here, we used mass cytometry to characterize the landscape and variation in the abundance of CD32a + CD4 + T cells during HIV infection. To this end, we analyzed CD32a + CD4 + T cells in primary HIV infection before and after effective combination antiretroviral therapy (cART) and in healthy donors. We found that CD32a + CD4 + T cells include heterogeneous subsets that are differentially affected by HIV infection. Our analysis revealed that naive (N), central memory (CM), and effector/memory (Eff/Mem) CD32a + CD4 + T-cell clusters that co-express LILRA2-and CD64-activating receptors were more abundant in primary HIV infection and cART stages. Conversely, LILRA2 − CD32a + CD4 + T-cell clusters of either the TN, TCM, or TEff/Mem phenotype were more abundant in healthy individuals. Finally, an activated CD32a + CD4 + TEff/Mem cell cluster co-expressing LILRA2, CD57, and NKG2C was more abundant in all HIV stages, particularly during primary HIV infection. Overall, our data show that multiple abundance modifications of CD32a + CD4 + T-cell subsets occur in the early phase of HIV infection, and some of which are conserved after effective cART. Our study brings a better comprehension of the relationship between CD32a expression and CD4 + T cells during HIV infection

image_5_Mass Cytometry Analysis Reveals the Landscape and Dynamics of CD32a+ CD4+ T Cells From Early HIV Infection to Effective cART.PDF

<p>CD32a has been proposed as a specific marker of latently HIV-infected CD4⁺ T cells. However, CD32a was recently found to be expressed on CD4⁺ T cells of healthy donors, leading to controversy on the relevance of this marker in HIV persistence. Here, we used mass cytometry to characterize the landscape and variation in the abundance of CD32a⁺ CD4⁺ T cells during HIV infection. To this end, we analyzed CD32a⁺ CD4⁺ T cells in primary HIV infection before and after effective combination antiretroviral therapy (cART) and in healthy donors. We found that CD32a⁺ CD4⁺ T cells include heterogeneous subsets that are differentially affected by HIV infection. Our analysis revealed that naive (_N), central memory (_CM), and effector/memory (_Eff/Mem) CD32a⁺ CD4⁺ T-cell clusters that co-express LILRA2- and CD64-activating receptors were more abundant in primary HIV infection and cART stages. Conversely, LILRA2⁻ CD32a⁺ CD4⁺ T-cell clusters of either the T_N, T_CM, or T_Eff/Mem phenotype were more abundant in healthy individuals. Finally, an activated CD32a⁺ CD4⁺ T_Eff/Mem cell cluster co-expressing LILRA2, CD57, and NKG2C was more abundant in all HIV stages, particularly during primary HIV infection. Overall, our data show that multiple abundance modifications of CD32a⁺ CD4⁺ T-cell subsets occur in the early phase of HIV infection, and some of which are conserved after effective cART. Our study brings a better comprehension of the relationship between CD32a expression and CD4⁺ T cells during HIV infection.</p

image_2_Mass Cytometry Analysis Reveals the Landscape and Dynamics of CD32a+ CD4+ T Cells From Early HIV Infection to Effective cART.PDF

<p>CD32a has been proposed as a specific marker of latently HIV-infected CD4⁺ T cells. However, CD32a was recently found to be expressed on CD4⁺ T cells of healthy donors, leading to controversy on the relevance of this marker in HIV persistence. Here, we used mass cytometry to characterize the landscape and variation in the abundance of CD32a⁺ CD4⁺ T cells during HIV infection. To this end, we analyzed CD32a⁺ CD4⁺ T cells in primary HIV infection before and after effective combination antiretroviral therapy (cART) and in healthy donors. We found that CD32a⁺ CD4⁺ T cells include heterogeneous subsets that are differentially affected by HIV infection. Our analysis revealed that naive (_N), central memory (_CM), and effector/memory (_Eff/Mem) CD32a⁺ CD4⁺ T-cell clusters that co-express LILRA2- and CD64-activating receptors were more abundant in primary HIV infection and cART stages. Conversely, LILRA2⁻ CD32a⁺ CD4⁺ T-cell clusters of either the T_N, T_CM, or T_Eff/Mem phenotype were more abundant in healthy individuals. Finally, an activated CD32a⁺ CD4⁺ T_Eff/Mem cell cluster co-expressing LILRA2, CD57, and NKG2C was more abundant in all HIV stages, particularly during primary HIV infection. Overall, our data show that multiple abundance modifications of CD32a⁺ CD4⁺ T-cell subsets occur in the early phase of HIV infection, and some of which are conserved after effective cART. Our study brings a better comprehension of the relationship between CD32a expression and CD4⁺ T cells during HIV infection.</p

image_3_Mass Cytometry Analysis Reveals the Landscape and Dynamics of CD32a+ CD4+ T Cells From Early HIV Infection to Effective cART.PDF

<p>CD32a has been proposed as a specific marker of latently HIV-infected CD4⁺ T cells. However, CD32a was recently found to be expressed on CD4⁺ T cells of healthy donors, leading to controversy on the relevance of this marker in HIV persistence. Here, we used mass cytometry to characterize the landscape and variation in the abundance of CD32a⁺ CD4⁺ T cells during HIV infection. To this end, we analyzed CD32a⁺ CD4⁺ T cells in primary HIV infection before and after effective combination antiretroviral therapy (cART) and in healthy donors. We found that CD32a⁺ CD4⁺ T cells include heterogeneous subsets that are differentially affected by HIV infection. Our analysis revealed that naive (_N), central memory (_CM), and effector/memory (_Eff/Mem) CD32a⁺ CD4⁺ T-cell clusters that co-express LILRA2- and CD64-activating receptors were more abundant in primary HIV infection and cART stages. Conversely, LILRA2⁻ CD32a⁺ CD4⁺ T-cell clusters of either the T_N, T_CM, or T_Eff/Mem phenotype were more abundant in healthy individuals. Finally, an activated CD32a⁺ CD4⁺ T_Eff/Mem cell cluster co-expressing LILRA2, CD57, and NKG2C was more abundant in all HIV stages, particularly during primary HIV infection. Overall, our data show that multiple abundance modifications of CD32a⁺ CD4⁺ T-cell subsets occur in the early phase of HIV infection, and some of which are conserved after effective cART. Our study brings a better comprehension of the relationship between CD32a expression and CD4⁺ T cells during HIV infection.</p