Cells In HIV-infected Patients Research Articles

Clinical significance of B7-H3 expression in circulating CD4+CD25high T cells, CD14+ monocytes, and plasma for the progression of HIV infection

BackgroundB7-H3 is an important immune checkpoint molecule that plays a negative role in immune regulation. This study was aimed to explore B7-H3 expression in HIV-infected patients and its clinical significance.MethodsTo explore the expression and clinical significance of B7-H3 in HIV-infected patients, we investigated the B7-H3 expression pattern and the correlation of B7-H3 expression with clinical parameters of HIV-infected patients with different levels of CD4+ T cells. To assess the role of B7-H3 in regulating the function of T cells in HIV infection, we performed a proliferation assay and T cell function test in vitro.ResultsB7-H3 expression in HIV-infected patients was significantly higher than that in healthy controls. mB7-H3 expression on CD4+CD25high T cells and CD14+ monocytes increased with disease progression. mB7-H3 expression on CD4+CD25high T cells and monocytes was negatively correlated with lymphocyte count, CD4+T cell count, and positively correlated with HIV viral load in HIV-infected patients. when the number of CD4+ T cells in HIV-infected patients was ≥ 200/µL, sB7-H3 and mB7-H3 expression levels on CD4+CD25high T cells and monocytes were negatively correlated with lymphocyte count, CD4+T cell count. sB7-H3 and mB7-H3 expression on monocytes were positively correlated with HIV viral load. B7-H3 inhibited the proliferation of lymphocytes and the secretion of IFN-γ in vitro, especially the ability of CD8+ T cells to secrete IFN-γ.ConclusionsB7-H3 played an important negative regulatory role in anti-HIV infection immunity. It could be used as a potential biomarker for the progression of HIV infection and a novel target for the treatment of HIV infection.

SMAD2/3 Phosphorylation Is Downregulated in T Cells in HIV-Infected Patients.

Persistent inflammation contributes to the exhaustion of immune system and non-AIDS-defining events in HIV-infected patients. Transforming growth factor β (TGF-β) is generally considered an anti-inflammatory cytokine. It is unclear that why high-level TGF-β coexists with chronic inflammation during HIV infection. In this study, it was found that HIV-infected patients had lower proportion of phosphorylated SMAD2/3-positive cells among total CD3+ T cells and subsets of CD3+CD8+ and CD3+CD8- T cells when compared with health subjects. The findings implied that phosphorylation of SMAD2/3 is inhibited in HIV-infected patients, and that disturbance of TGF-β/SMAD2/3 signaling pathway may be involved in HIV-related chronic inflammation.

HIV Infection Induces Aberrant Myeloid Lineage Differentiation

AbstractIntroduction: Numerous studies on hematopoietic changes caused by HIV infection have focused on immune cell depletion, while research on other hematopoietic lineages development have been sparse and inconclusive. In this study, we used precise markers to detect changes in the distribution of bone marrow hematopoietic stem/progenitor cells in HIV-infected patients and analyzed the dynamic changes of mature cells of each lineages in 782 patients with disease progression. Furthermore, we sought to explore the potential relationship between abnormal development of these lineage cells and survival of HIV patients. Our study provides evidence for a more comprehensive understanding of the effects of HIV on hematopoiesis. Methods: We included 782 HIV-infected patients who underwent bone marrow aspiration from January 2016 to October 2020 in Zhongnan Hospital, and were followed up until January 2022. Examination including flow cytometry and bone marrow aspiration were performed to detect the development of hematopoietic cells. 509 cases were tested for lymphocyte subsets. According to CD3+CD4+ cell count, 325 patients (63.9%) were in the very low group, 61 patients (12.0%) in the low group, 55 patients (10.8%) in the medium group, and 68 patients (13.4%) in the high group. By January 2022, 84 patients had been lost to follow-up and 425 patients had accurate and complete follow-up data. We analyzed the risk factors for HIV death in 425 patients and established a predictive model. Results: Flow cytometry analysis showed an increased proportion of granulocytemonocyte progenitors (GMP) but a decreased proportion of common lymphoid progenitors (CLP) in HIV patients. In myelogram, the percentage of granulocytes and granulocytes/erythroid cells (G/E) increased with the decrease of CD3+CD4+ count (both P<0.001). In addition to the increase in cell proportion, we also observed that the morphologic abnormalities in granulocyte increased with the progression of the disease, including toxic granulation, cytoplasmic vacuolization, Intracellular and extracellular plasma (P<0.001), Döhle bodies (P=0.033) and increased lobulation in megakaryocyte (P=0.048). Subsequent LASSO binary logistic regression also confirmed that the increased granulocyte ratio was an important factor in patients' disease. Conclusion: HIV infection directly or indirectly induces bias on myeloid lineage differentiation. Increased granulocyte ratio is an important risk factor for disease aggravation and death in HIV patients. Our research provide a new perspective for evaluating the status and prognosis of HIV patients by focusing on the changes of myeloid granulocytes. Disclosures No relevant conflicts of interest to declare.

HIV fragments detected in Kaposi sarcoma tumor cells in HIV-infected patients.

Kaposi sarcoma (KS) is a malignant vascular neoplasm caused by KS-associated herpesvirus (KSHV) infection. HIV plays a major role in KS pathogenesis. KS in HIV usually produces more malignant features than classic KS. Despite the close KS-HIV relationship, no study has reported the existence of HIV in KS tissue. We used ddPCR to detect HIV and KSHV in HIV+ KS samples and classic KS control. We verified KS cell types through immunohistochemistry and applied hypersensitive in situ hybridization (ISH) to detect HIV and KSHV in tumor cells. Furthermore, we co-stained samples with ISH and immunohistochemistry to identify HIV and KSHV in specific cell types. Regarding pathological stages, the KS were nodular (58.3%), plaque (33.3%), and patch (8.3%) tumors. Moreover, ddPCR revealed HIV in 58.3% of the KS samples. ISH revealed positive Pol/Gag mRNA signals in CD34 + tumor cells from HIV + patients (95.8%). HIV signals were absent in macrophages and other inflammatory cells. Most HIV + KS cells showed scattered reactive particles of HIV and KSHV. We demonstrated that HIV could infect CD34 + tumor cells and coexist with KSHV in KS, constituting a novel finding. We hypothesized that the direct KSHV-HIV interaction at the cellular level contributes to KS oncogenesis.

Splenectomy in Patients with Visceral Leishmaniasis Resistant to Conventional Therapy and Secondary Prophylaxis: A Retrospective Cohort.

Some patients with visceral leishmaniasis (VL), or kala-azar, suffer relapses and low quality of life despite adequate drug therapy, especially those co-infected with HIV. Occasionally, physicians indicate splenectomy, but the benefit of the procedure needs to be analyzed systematically. Therefore, a retrospective open cohort study was conducted in Teresina, Brazil. Inpatients from a reference hospital with relapsing VL who had a rescue splenectomy between 2012 and 2019 after the nationally recommended drug therapy failed were studied. The procedure's risks and benefits were assessed in a limited-resource setting. The primary outcomes were surgical complications, complete blood count, CD4+ cell count, hospitalizations, survival time, and medical complications preceding death. Thirteen adult patients received medical and surgical indications of splenectomy (12 men and one woman). Eleven had HIV infection. Two had early and two had late complications. Four died, all of whom were infected with HIV. An additional HIV-coinfected patient, apart from the cohort, died just before surgery. The death rate after surgery was 13.3 overall and 22.1 per 100 person-years among HIV-infected patients (31% overall and 36%, respectively). The impressive rise of complete blood counts and reduction of blood transfusions and hospitalizations were observed among all patients. Also, a meaningful increase in CD4+ cells in HIV-infected patients was noted. Splenectomy may benefit patients with relapsing VL. However, before performing splenectomy, available combined drug therapy for VL should be tried.

Minimum redundancy maximal relevance gene selection of apoptosis pathway genes in peripheral blood mononuclear cells of HIV-infected patients with antiretroviral therapy-associated mitochondrial toxicity

BackgroundWe previously identified differentially expressed genes on the basis of false discovery rate adjusted P value using empirical Bayes moderated tests. However, that approach yielded a subset of differentially expressed genes without accounting for redundancy between the selected genes.MethodsThis study is a secondary analysis of a case–control study of the effect of antiretroviral therapy on apoptosis pathway genes comprising of 16 cases (HIV infected with mitochondrial toxicity) and 16 controls (uninfected). We applied the maximum relevance minimum redundancy (mRMR) algorithm on the genes that were differentially expressed between the cases and controls. The mRMR algorithm iteratively selects features (genes) that are maximally relevant for class prediction and minimally redundant. We implemented several machine learning classifiers and tested the prediction accuracy of the two mRMR genes. We next used network analysis to estimate and visualize the association among the differentially expressed genes. We employed Markov Random Field or undirected network models to identify gene networks related to mitochondrial toxicity. The Spinglass model was used to identify clusters of gene communities.ResultsThe mRMR algorithm ranked DFFA and TNFRSF1A, two of the upregulated proapoptotic genes, on the top. The overall prediction accuracy was 86%, the two mRMR genes correctly classified 86% of the participants into their respective groups. The estimated network models showed different patterns of gene networks. In the network of the cases, FASLG was the most central gene. However, instead of FASLG, ABL1 and LTBR had the highest centrality in controls.ConclusionThe mRMR algorithm and network analysis revealed a new correlation of genes associated with mitochondrial toxicity.

Results of immunoprophylaxis of HIV-infected patients with 13-valent conjugated pneumococcal vaccine

To assess changes in the composition of the microflora of the upper respiratory tract and indicators of cellular immunity 1 year after the administration of 13-valent conjugated pneumococcal vaccine (PCV13) in adult HIV-infected patients. Were recruited 100 participants of both sexes (50% male and 50% female). All patients underwent microbiological and immunological (determination of the level of CD3+, CD3+CD4+, CD3+CD8+, CD19+, CD45RO+ peripheral blood lymphocytes) examinations before vaccine administration and after 12 months. Immunization with PCV13 leads to a statistically significant decrease in the carriage of Streptococcus pneumoniae 1 year after vaccination (p=0.012). After 1 year after the administration of PCV13, the patients showed a statistically significant increase in the total number of T-lymphocytes, T-helpers, and cytotoxic T-lymphocytes in comparison with pre-vaccination levels. A statistically significant increase in the level of CD45RO+ lymphocytes was found 1 year after the administration of PCV13 (p0.0001). S. pneumoniae was found on the mucous membrane of the posterior pharyngeal wall in 16% of the participants, indicating its high prevalence in HIV-infected patients. Also, representatives of enterobacteria and Candida spp. were found in smears. (22 and 15% of participants, respectively). One year after the vaccine administration, pneumococcus was isolated from 5 participants, which is statistically significantly lower than before immunization (p=0.012). After the introduction of PCV13, there is a statistically significant increase in the total number of T-lymphocytes, T-helpers and cytotoxic T-lymphocytes 1 year after immunization. However, there is no statistically significant increase in the B-lymphocyte population. In addition, PCV13 leads to the formation of immunological memory cells in HIV-infected patients. Thus, immunoprophylaxis with PCV13 in adult HIV-infected patients leads to a decrease in the carriage of S. pneumoniae, and also promotes the stimulation of the T-cell link of the immune system and stimulates the formation of immunological memory cells.

Glucose uptake by CD4&lt;sup&gt;+&lt;/sup&gt;Т cell subsets in HIV-infected patients receiving highly active antiretroviral therapy (haart)

Metabolism of glucose, a universal biosynthetic substrate in CD4+T lymphocytes, is increased in HIV-infection. However, the issue of heterogeneity in glucose consumption by the CD4+T cell subsets remains unresolved. The aim of this work was to assess glucose uptake by resting and cycling naive and memory CD4+T cells in HIV-infected patients receiving antiretroviral therapy. We examined 47 subjects, 26 of whom were infected with HIV and were administered antiretroviral drugs. The following indices were determined: the number of CD4+T lymphocytes, size of naive (CD4+CD45R0-) and memory (CD4+CD45R0+) T cell subsets, frequencies of resting (CD71-) and cycling (CD71+) cell forms in each subset, the glucose fluorescent analog (2-NBDG) uptake by various CD4+T cell subsets. It was shown that, regardless of HIV status, the CD4+T cells are active glucose consumers. Cycling lymphocytes, compared with resting cells, uptake more biosynthetic substrate. We have revealed a trend for increased glucose uptake in HIV-infected patients when compared with healthy individuals. Memory cells, in comparison with naive lymphocytes, consume the substrate more actively, independent on HIV status. At the same time, naive CD4+T lymphocytes of HIV-infected individuals capture more glucose than the corresponding cell subset in non-infected donors. Cycling naive CD4+T lymphocytes of HIV-positive subjects are more active consumers of glucose than the analogues in healthy subjects. No differences were found between HIV-positive and HIV-negative groups for intensity of substrate consumption by the cycling memory CD4+T cells. Thus, in treated HIV-infected patients, CD4+T cells seem to uptake more glucose than similar cell subpopulations in healthy people, which, apparently, is mediated by the activity of resting naive lymphocytes. The data obtained indicate that metabolic characteristics in resting T-cells are instable and may change depending on the substrate availability.

Изучение Т-клеточного звена иммунитета у ВИЧ-инфицированных пациентов с гельминтозами

Objective. To analyze T cell-mediated immunity (subpopulations of CD3+, CD4+, and CD8+ lymphocytes) in HIV-infected patients with helminthiasis receiving or not receiving antiretroviral therapy (ART). Patients and methods. This study included 159 individuals; 100 of them had subclinical stage 3 HIV infection and helminthiasis (caused by different helminths) and were divided into two experimental groups depending on whether they received ART or not. The control group 1 comprised 29 HIV-positive people without helminthiasis not receiving ART, whereas the control group 2 included 30 HIV-negative people with helminthiasis. Patients in the experimental groups and control group 2 were followed up during the study and received anthelmintic treatment. The assessment of the immune status using monoclonal antibodies against specific antigens of subpopulations of T-lymphocytes (CD3+, CD4+ cells, cytotoxic T-cells-suppressors-CD8+ and CD4+/CD8+ ratio) was performed in the laboratory of the Republican Center for the Prevention and Control of AIDS and Infectious Diseases, Ufa, Russian Federation. Results. In this study, we evaluated the relative count of CD3+, CD4+, and CD8+ T cells in HIV-infected patients with helminthiasis at different time-pints during 6 months. We observed significant differences in the CD3+, CD3+CD4+, and CD3+CD8+ cell count between HIV-infected patients with helminthiasis on ART and without ART. Patients in the experimental group on ART demonstrated a significantly lower CD3+ cell count compared to patients in the experimental group not on ART (2.3 times lower; p &lt; 0.01), as well as lower CD3+CD4+ cell count (1.5 times lower; p &lt; 0.05) and CD4+CD8+ cell count (1.9 times lower; p &lt; 0.05). Our findings suggest that HIV-infected people with helminthiasis on ART are more likely to have their CD3+, CD4+, and CD8+ Т-cell count normalized than those not receiving ART. Conclusion. The assessment of the immune status (T-cell medicated immunity) in the study groups demonstrated that HIVinfected patients with helminthiasis on ART presented with a gradual increase of the relative CD3+ cell count throughout the study (62.5 ± 5.6 %), whereas HIV-infected patients with helminthiasis receiving no ART presented with a gradual decrease of the relative CD3+ cell count (28.0 ± 4%). HIV-infected patients without helminthiasis and receiving no ART (control group 1) also demonstrated a decrease of the relative CD3+ cell count (28.1 ± 3.5%). We also observed a clear trend towards the normalization of relative CD3+CD4+ T-cell count (41.4 ± 8.2%) in HIV-infected patients with helminthiasis on ART, while patients from other groups (including HIV-infected patients with helminthiasis without ART and individuals in both control groups) demonstrated a tendency to a steady decline in the relative CD3+CD4+ cell count at all time-points. We also found that HIV-infected patients with helminthiasis on ART had their CD4+/CD8+ ratio back to almost normal by month 6 (45.7 ± 3.7%), whereas HIV-infected patients with helminthiasis without ART and patients from the control group 1 had their mean CD4+CD8+ ratio gradually decreasing throughout the study (27.5 ± 4.9% and 30.5 ± 7.1% respectively). The parameters of T-cell mediated immunity (subpopulations of CD3+, CD4+, and CD8+ lymphocytes) showed a more pronounced tendency to normalization in HIV-infected patients with helminthiasis who received ART. Our findings suggest that in HIV-infected patients with helminthiasis on ART, compensatory mechanisms of T-lymphocytes predominate, in contrast to HIV-infected patients with helminthiasis receiving no ART and the control group of HIV-infected patients receiving no ART, in whom we observed immunodeficiency of different grades. Key words: HIV infection, helminthiasis, T-cell immunity

Immunocytochemical characteristic of the endometrial mucosa HIV-infected women

Relevance. Human immunodeficiency virus (HIV) infection is an independent factor in reduced fertility and a risk factor for miscarriage. There are some date an endometrial receptivity of HIV-infected patients has changed that plays an important role in embryo invasion, but the true reasons for the decrease in fertility rate in HIV infection remain unknown.&#x0D; Aim. Study of the expression of CD20, CD56 and TLR9 antigens on uterine epithelial cells of HIV-infected patients and the effectiveness of treatment for chronic endometritis by sodium nucleospermate.&#x0D; Materials and methods. This parallel-group study was done at two centres in the Russia. Participants were adults women aged 26 to 49 years (mean age 33.352.9 years), who were HIV-infected (n=12) and HIV-negative (22). An immunocytochemical study of endometrial biopsies taken on the 710th day of the menstrual cycle before and after treatment was done. The course of treatment with sodium nucleospermate was 42 days.&#x0D; Results. The expression level of CD56 and TLR9 in HIV-infected patients was 7.640.92% and 0.330.18%, respectively, and significantly differed from the expression levels in HIV-seronegative patients. There was a decrease in the expression levels of CD20 and CD56 and an increase in the expression levels of TLR9 in all groups of patients after treatment with sodium nucleospermate.&#x0D; Conclusion. A decrease TLR9 expression on uterine epithelial cells in HIV-infected patients showing lack of ability of innate immunity to eliminate pathogens associated with subclinical inflammation and it correlates with an increase in the expression of markers of chronic endometritis.

Prevalence and Risk Factors of Cytopenia in HIV-Infected Patients before and after the Initiation of HAART.

Background Cytopenia is a frequent hematological disorder in patients with human immunodeficiency virus (HIV) infection. However, the distribution and risk factors of cytopenia in patients starting highly active anti-retrovirus treatment (HAART) and the effect of HAART on cytopenia were not fully investigated. Methods From November 2004 to August 2016, a retrospective study was conducted to evaluate the prevalence of cytopenia in 4325 HAART-naïve patients. Risk factors of cytopenia at baseline and on recovery from cytopenia were analyzed using logistic regression analysis after 24 months of HAART in Beijing Ditan Hospital. Results The prevalence of cytopenia was 19.1% in HIV-naïve patients. Risk factors for cytopenia in HAART-naïve patients were a CD4 cell count<200 cells/μL, femaleness, WHO stage IV, coinfection with hepatitis B virus (HBV), BMI <18.5 kg/m2, a viral load ≥100,000 copies/ml, and age ≥40 years. In total, 70.2% and 76.4% of patients with cytopenia recovered after 6 and 24 months of HAART, respectively. The predictors of patients without normal blood cells after 24 months HAART were a CD4 cell count of <200 cells/μL, femaleness, WHO stage IV, coinfection with hepatitis B virus (HBV), BMI <18.5 kg/m2, a viral load ≥100,000 copies/ml, and age ≥40 years. In total, 70.2% and 76.4% of patients with cytopenia recovered after 6 and 24 months of HAART, respectively. The predictors of patients without normal blood cells after 24 months HAART were a CD4 cell count of <200 cells/Conclusion Early detection could decrease the prevalence of HIV-related cytopenia, while starting HAART as early as possible seems to be effective for normalization of the blood cells in HIV-infected patients.

Antiretroviral therapy partially improves the abnormalities of dendritic cells and lymphoid and myeloid regulatory populations in recently infected HIV patients

This study aimed to evaluate the effects of antiretroviral therapy on plasmacytoid (pDC) and myeloid (mDC) dendritic cells as well as regulatory T (Treg) and myeloid-derived suppressor (MDSC) cells in HIV-infected patients. Forty-five HIV-infected patients (20 of them with detectable HIV load −10 recently infected and 10 chronically infected patients-, at baseline and after antiretroviral therapy, and 25 with undetectable viral loads) and 20 healthy controls were studied. The influence of HIV load, bacterial translocation (measured by 16S rDNA and lipopolysaccharide-binding protein) and immune activation markers (interleukin –IL- 6, soluble CD14, activated T cells) was analyzed. The absolute numbers and percentages of pDC and mDC were significantly increased in patients. Patients with detectable viral load exhibited increased intracellular expression of IL-12 by mDCs and interferon -IFN- α by pDCs. Activated population markers were elevated, and the proportion of Tregs was significantly higher in HIV-infected patients. The MDSC percentage was similar in patients and controls, but the intracellular expression of IL-10 was significantly higher in patients. The achievement of undetectable HIV load after therapy did not modify bacterial translocation parameters, but induce an increase in pDCs, mDCs and MDSCs only in recently infected patients. Our data support the importance of early antiretroviral therapy to preserve dendritic and regulatory cell function in HIV-infected individuals.

HIV infection drives immune activation of angiogenic T cells

Abstract In patients with HIV infection, chronic immune activation, systemic inflammation and endothelial dysfunction have been identified as the main drivers of cardiovascular risk/disease. In addition, recent reports have shown that, T cells with angiogenic properties (Tang) contribute to vascular repair by promoting new vessels formation in vivo and in vitro. In the present study, we hypothesized that HIV drives immune activation of Tang cells. We measured the frequency and phenotype of circulating Tang cells defined as CXCR4+CD31+. Patients with HIV infection and well controlled viral replication by cART (n= 25) showed a decreased proportion of both CD4 and CD8 Tangs cells when compared to healthy controls (n= 12), although this difference was more striking in the CD8 Tang subset. CD8 Tang but not CD4 Tang cells were significantly decreased in HIV negative patients whom are undergoing PrEP (Pre-Exposure Prophylaxis) therapy, (n=5). In addition, we found that: angiogenic T cells as defined by the surface markers CXCR4 and CD31 are a heterogenous population including naïve, memory and terminal effector memory phenotypes. Tang cells of HIV infected patients showed an activated phenotype with increased expression of granzymes A (GZA) and B (GZB) when compared to healthy controls. Lastly, CD4 and CD8 Tang cells show increased expression of CD49f (integrin a-6). The frequency of Tang GZA+GZB+ in patients with HIV infection was positively associated with 10yr cardiovascular risk score (ASCVD), CD4 Tang (R= 0.51, p= 0.02) and CD8 Tang (R= 0.6, p&amp;lt; 0.01). These results suggest that in the context of HIV infection, angiogenic T cells possess an immune activated phenotype with a proinflammatory potential that may alter their ability of vascular repair

NK cells in treated HIV-infected children display altered phenotype and function

Chronic HIV infection is known to trigger a population redistribution and alteration in the functional capacity of natural killer (NK) cells. Because of improved antiretroviral treatments, there are rising numbers of adolescents and young adults worldwide who are living with HIV infection since birth. We sought to determine how NK-cell phenotypic and functional subsets are altered in treated pediatric patients. NK cells were contrasted among 29 HIV-unexposed and uninfected controls (5-19years), 23 HIV-exposed but uninfected patients (3-19years), and 25 HIV-infected patients (3-19years) using multiparametric flow cytometry. Although most NK-cell markers did not differ, activating receptors such as NKp46, DNAX accessory molecule-1, and NKG2C and stimulatory receptors such as CD2 and CD11c were expressed by a higher frequency of NK cells in HIV-infected patients than in controls. Interestingly, there were less differences between HIV-infected and HIV-exposed but uninfected children. There was an inverse relationship between CD4/CD8 T-cell ratio (as a marker of disease progression) and CD11c and NKG2C frequency and CD69 upregulation on stimulation among HIV-infected patients. A chronic NK-cell activation phenotype persists in HIV-infected children receiving antiretroviral therapy and is associated with declining CD4/CD8 T-cell ratios. Alower CD4/CD8 T-cell ratio was associated with higher baseline granzyme B (P=.0068; R2=0.29) and degranulation potential (P=.022; R2=0.22) in stimulated NK cells. Thus, NK cells in HIV-infected children receiving treatment have reduced functional potential and an activated phenotype that distinguishes them from uninfected children.

Dual TLR2 and TLR7 agonists as HIV latency-reversing agents.

The presence of a reservoir of latently infected cells in HIV-infected patients is a major barrier towards finding a cure. One active cure strategy is to find latency-reversing agents that induce viral reactivation, thus leading to immune cell recognition and elimination of latently infected cells, known as the shock-and-kill strategy. Therefore, the identification of molecules that reactivate latent HIV and increase immune activation has the potential to further these strategies into the clinic. Here, we characterized synthetic molecules composed of a TLR2 and a TLR7 agonist (dual TLR2/7 agonists) as latency-reversing agents and compared their activity with that of the TLR2 agonist Pam2CSK4 and the TLR7 agonist GS-9620. We found that these dual TLR2/7 agonists reactivate latency by 2 complementary mechanisms. The TLR2 component reactivates HIV by inducing NF-κB activation in memory CD4+ T cells, while the TLR7 component induces the secretion of TNF-α by monocytes and plasmacytoid dendritic cells, promoting viral reactivation in CD4+ T cells. Furthermore, the TLR2 component induces the secretion of IL-22, which promotes an antiviral state and blocks HIV infection in CD4+ T cells. Our study provides insight into the use of these agonists as a multipronged approach targeting eradication of latent HIV.

Reduction in terminally differentiated T cells in virologically controlled HIV-infected aging patients on long-term antiretroviral therapy

Several studies have shown an increased accumulation of terminally differentiated T cells during HIV infection, suggestive of exhaustion/senescence, causing dysregulation of T cell homeostasis and function and rapid HIV disease progression. We have investigated whether long-term antiretroviral therapy (ART), which controls viremia and restores CD4 T cell counts, is correlated with reduction in terminally differentiated T cells, improved ratios of naïve to memory and function of T cells in 100 virologically controlled HIV-infected patients. We show that while the median frequencies of terminally differentiated CD4+ and CD8+ T cells (CD28-, CD27-, CD57+ and CD28-CD57+), were higher in the virologically controlled HIV-infected patients’ cohort compared with uninfected individuals’ cohort, the frequencies of these cells significantly decreased with increasing CD4 T cell counts in HIV-infected patients. Although, the naïve CD4+ and CD8+ T cells were lower in HIV patients’ cohort than uninfected cohort, there was a significant increase in both naïve CD4+ and CD8+ T cells with increasing CD4 T cell counts in HIV-infected patients. The underlying mechanism behind this increased naïve CD4+ and CD8+ T cells in HIV-infected patients was due to an increase in recent thymic emigrants, CD4+CD31+, as compared to CD4+CD31-. The CD4+ T cells of HIV-infected patients produced cytokines, including IL-2, IL-10 and IFN-γ comparable to uninfected individuals. In conclusion, virologically controlled HIV-infected patients on long-term ART show a significant reduction in terminally differentiated T cells, suggestive of decreased exhaustion/senescence, and improvement in the ratios of naïve to memory and function of T cells.

Histone deacetylase inhibitor chidamide promotes reactivation of latent human immunodeficiency virus by introducing histone acetylation

Highly active antiretroviral therapy can reduce the human immunodeficiency virus (HIV) viral load in the plasma to undetectable levels. However, because of the presence of latent HIV reservoirs, it is difficult to completely eradicate HIV in infected patients. Our objective was to assess the potency of chidamide, a novel histone deacetylase inhibitor recently approved for cancer treatment by the China Food and Drug Administration, to reactivate latent HIV-1 via histone acetylation. Viral reactivities of chidamide were accessed in 2 latent HIV pseudotype virus cell reporter systems (J-Lat Tat-green fluorescent protein clone A72 and TZM-bl), a latently infected full-length HIV virus cell system (U1/HIV), and resting CD4+ T cells from 9 HIV-infected patients under highly active antiretroviral therapy with undetectable viral load. Chidamide was able to increase HIV expression in each cell line, as evidenced by green fluorescent protein, luciferase activity, and p24, as well as to reactivate latent HIV-1 in primary CD4+ T cells of HIV-infected patients. Histone acetylation adjacent to the HIV promoter in A72 cells was determined by chromatin immunoprecipitation. Chidamide was able to increase histone H3 and H4 acetylation at the HIV promoter. In brief, chidamide induced the reactivation of latent HIV in pseudotype virus reporter cells, latently infected cells, and primary CD4+ T cells, making this compound an attractive option for future clinical trials.

IL-18 and Stem Cell Factor affect hematopoietic progenitor cells in HIV-infected patients treated during primary HIV infection

The impact of early antiretroviral therapy (ART) during Primary HIV Infection (PHI) on the hematopoietic progenitor cells (HPCs) homeostasis is not available. This study aimed to characterize HPCs and their relationship with cytokines regulating progenitors function in ART-treated patients with PHI.We enrolled HIV infected patients treated with ART during PHI. Circulating HPCs, Lymphoid-HPCs (L-HPCs) frequency and plasmatic concentrations of IL-7, IL-18 and Stem Cell Factor (SCF) were analysed at baseline and after 6 months of therapy.ART introduction during PHI restored the decline of L-HPCs, induced a decrease in the level of pro-inflammatory IL-18 cytokine and a parallel increase of SCF. Moreover, L-HPCs frequency positively correlated with IL-18 at baseline, and with SCF after 6 months of therapy, suggesting that different signals impact L-HPCs expansion and maintenance before and after treatment. Finally, the SCF receptor expression on HPCs decreased after early ART initiation.These insights may open new perspectives for the evaluation of cytokine-driven L-HPCs expansion and their impact on the homeostasis of hematopoietic compartment during HIV infection.

Immunoassay and molecular methods to investigate DNA methylation changes in peripheral blood mononuclear cells in HIV infected patients on cART

ABSTRACTThis study aimed to investigate the influence of antiretroviral therapy on methylation markers, in a group of HIV infected, heavily treated patients. Immune and molecular methods were used to investigate potential changes in methylation profile in DNA isolated from peripheral blood mononuclear cells collected from antiretroviral-experienced HIV infected patients and healthy controls. The percentage of 5-methylcytosine was inversely correlated with proviral DNA and active replication while DNMT1 (p = 0.01) and DNMT3A (p = 0.004) independently correlated with active viral replication. DNMT3A expression increased with total treatment duration (p = 0.03), number of antiretroviral drugs ever used (p = 0.003), and cumulative exposure to protease inhibitors (p = 0.02) even in currently HIV undetectable patients.

In vivo analysis of the effect of panobinostat on cell-associated HIV RNA and DNA levels and latent HIV infection.

BackgroundThe latent reservoir in resting CD4+ T cells presents a major barrier to HIV cure. Latency-reversing agents are therefore being developed with the ultimate goal of disrupting the latent state, resulting in induction of HIV expression and clearance of infected cells. Histone deacetylase inhibitors (HDACi) have received a significant amount of attention for their potential as latency-reversing agents.ResultsHere, we have investigated the in vitro and systemic in vivo effect of panobinostat, a clinically relevant HDACi, on HIV latency. We showed that panobinostat induces histone acetylation in human PBMCs. Further, we showed that panobinostat induced HIV RNA expression and allowed the outgrowth of replication-competent virus ex vivo from resting CD4+ T cells of HIV-infected patients on suppressive antiretroviral therapy (ART). Next, we demonstrated that panobinostat induced systemic histone acetylation in vivo in the tissues of BLT humanized mice. Finally, in HIV-infected, ART-suppressed BLT mice, we evaluated the effect of panobinostat on systemic cell-associated HIV RNA and DNA levels and the total frequency of latently infected resting CD4+ T cells. Our data indicate that panobinostat treatment resulted in systemic increases in cellular levels of histone acetylation, a key biomarker for in vivo activity. However, panobinostat did not affect the levels of cell-associated HIV RNA, HIV DNA, or latently infected resting CD4+ T cells.ConclusionWe have demonstrated robust levels of systemic histone acetylation after panobinostat treatment of BLT humanized mice; and we did not observe a detectable change in the levels of cell-associated HIV RNA, HIV DNA, or latently infected resting CD4+ T cells in HIV-infected, ART-suppressed BLT mice. These results are consistent with the modest effects noted in vitro and suggest that combination therapies may be necessary to reverse latency and enable clearance. Animal models will contribute to the progress towards an HIV cure.