Prolonged Highly Active Antiretroviral Therapy Research Articles

Particular Aspects on Applying Nanocarbon Quantum Dots for HIV Inhibition and Theranostics: a Review

Viral infection is a globally leading health issue, causing significantly unfavourable mortality with an adversely decreasing socio-economic growth. To solve those infections of global pandemic HIV specifically, the current utilization of highly active antiretroviral therapy (HAART) in human deficiency virus (HIV) theranostics has remarkably improved the life’s duration of patients infected by human immunodeficiency virus (HIV); however, the unfortunate drawbacks in combination with prolonged HAART therapy need to be used continuously along patient’s lifetime. Additionally, RNA virus of COVID-19 is also associated with viral pneumonia and acute respiratory distress syndrome causing significant morbidity and mortality. Meanwhile, many scientific researchers have explored the successive novelty of carbon quantum dots (CQDs) as alternative to HIV or other related viruses theranostics in the field of antiviral drugs research, but the attempt has been still challenging to introduce perfect antiviral CQDs with excellent biocompatibility, drug resistance, and safety at several areas in the virus’s life cycle. On the contrary, CQDs-based nano-therapy is currently promising because those carbon quantum dots had multiple favourable properties, including significant antiviral response effects, water-soluble activity, color-tunable fluorescence, high yield, low cytotoxic behaviour, and promising biocompatibility. In this review, some effectively recent progress of promising CQDs forviral inhibition and theranostics explored by many studies are systematically summarized.

Particular Aspects on Applying Nanocarbon Quantum Dots for HIV Inhibition and Theranostics: a Review

Viral infection is a globally leading health issue, causing significantly unfavourable mortality with an adversely decreasing socio-economic growth. To solve those infections of global pandemic HIV specifically, the current utilization of highly active antiretroviral therapy (HAART) in human deficiency virus (HIV) theranostics has remarkably improved the life’s duration of patients infected by human immunodeficiency virus (HIV); however, the unfortunate drawbacks in combination with prolonged HAART therapy need to be used continuously along patient’s lifetime. Additionally, RNA virus of COVID-19 is also associated with viral pneumonia and acute respiratory distress syndrome causing significant morbidity and mortality. Meanwhile, many scientific researchers have explored the successive novelty of carbon quantum dots (CQDs) as alternative to HIV or other related viruses theranostics in the field of antiviral drugs research, but the attempt has been still challenging to introduce perfect antiviral CQDs with excellent biocompatibility, drug resistance, and safety at several areas in the virus’s life cycle. On the contrary, CQDs-based nano-therapy is currently promising because those carbon quantum dots had multiple favourable properties, including significant antiviral response effects, water-soluble activity, color-tunable fluorescence, high yield, low cytotoxic behaviour, and promising biocompatibility. In this review, some effectively recent progress of promising CQDs forviral inhibition and theranostics explored by many studies are systematically summarized.

Diffusion tensor magnetic resonance imaging of white matter integrity in patients with HIV-associated neurocognitive disorders.

BackgroundThis study investigated the efficacy and neurotoxicity of highly active antiretroviral therapy (HAART) by evaluating white matter (WM) injury using diffusion tensor magnetic resonance imaging (DTI) in patients with human immunodeficiency virus (HIV)-associated neurocognitive disorders (HAND).MethodsForty-six patients with HAND underwent DTI before and every six months during HAART treatment. DTI data, including fractional anisotropy (FA) and mean diffusivity (MD) values of structural WM before and after HAART, were compared. The relationship between DTI values and plasma viral loads was tested. MD was more sensitive than FA for evaluating WM injury in HAND-positive patients.ResultsFollowing 12 months of HAART, increased MD values (compared to 6 months of HAART) were observed in the right temporal lobe, right parietal lobe, right occipital lobe, right anterior limb of the internal capsule, right lenticular nucleus, the right cerebral peduncle, left caudate nucleus, left dorsal thalamus, and left posterior limb of the internal capsule. MD values in the left genu of the internal capsule (r=0.350, P=0.017) and left corona radiata (r=0.338, P=0.021) were positively correlated with plasma viral loads.ConclusionsDTI may be useful for assessing the efficacy and neurotoxicity of HAART in HAND-positive patients. Starting HAART may halt WM injury; however, prolonged HAART could worsen WM injury, highlighting the importance of optimal HAART duration.

Two types of poor immunological responder showing distinct responses to long-term HAART

ObjectivesMost previous studies on poor immunological responders (PIRs) have been performed on one cohort at one time-point following highly active antiretroviral therapy (HAART). The aim of this study was to investigate whether there are different subtypes of PIR and whether a certain population might achieve better immune reconstitution following longer HAART. MethodsThis study was designed as an ambispective cohort study, including a 4–5-year retrospective study and a 2-year prospective follow-up investigation. Thymic output, activated T cell and regulatory T cell (Treg) subset frequencies, expression levels of interferon-stimulated genes, and plasma concentrations of neopterin were determined at 4–5 years and 6–7 years following HAART initiation. ResultsPIRs were subdivided into two populations after 4–5 years of HAART, according to the kinetics of T cell recovery. Type II PIRs exhibited a significantly lower percentage of naïve CD4+ T cells and CD31+ naïve CD4+ T cells compared with type I PIRs. After an additional 2 years of HAART treatment, type I PIRs showed a better outcome than type II PIRs. Furthermore, it was found that 2 years of additional HAART could persistently improve thymic output. ConclusionsThe two PIR subgroups are different in terms of immune characteristics and the response to prolonged HAART.

Diffusion Tensor MR Imaging of White Matter Integrity in Patients with HIV-Associated Neurocognitive Disorders

Background: Effective highly active antiretroviral therapy (HAART) could prevent progression of white matter (WM) injury. However, prolonged use of HAART may induce neurotoxicity. This study investigated the efficacy and neurotoxicity of HAART by evaluating WM injury using diffusion tensor magnetic resonance imaging (DTI) in patients with HIV-associated neurocognitive disorders (HAND). Methods: Forty-six patients with HAND underwent DTI before and every six months during HAART treatment. DTI data, including fractional anisotropy (FA) and mean diffusivity (MD) values of structural WM before and after HAART, were compared. The relationship between DTI values and plasma viral loads was also evaluated. Findings: MD was more sensitive than FA for evaluating WM injury in HAND-positive patients. Following 12 months of HAART, increased MD values (compared to 6 months of HAART) were observed in the right temporal lobe, right parietal lobe, right occipital lobe, right anterior limb of the internal capsule, right lenticular nucleus, right cerebral pedunculus, left caudate nucleus, left dorsal thalamus, and left posterior limb of the internal capsule. MD values in the left genu of the internal capsule (r = 0.350, P < 0.05) and left corona radiata (r = 0.338, P < 0.05) were positively correlated with plasma viral loads. Interpretation: DTI may be useful for assessing efficacy and neurotoxicity of HAART in HAND-positive patients. Initiating HAART may halt WM injury; however, prolonged HAART could worsen WM injury, highlighting the importance of optimal HAART duration. Funding: Guangxi Science and Technology Department Key Research and Development Programs, Youth Science Foundation of Guangxi Medical University and International Communication of Guangxi Medical University Graduate Education. Declaration of Interest: None. Ethical Approval: All experiments performed are endorsed by the Ethics Committee of Guangxi Medical University and complied with the Declaration of Helsinki.

Serum Biomarkers For Risk Of Cardiovascular Disease In Patients On Highly Active Antiretroviral Therapy In Homa-Bay County Referral Hospital, Kenya

Abstract Background: Human Immunodeficiency Virus (HIV) continues to be a major public health problem in both developing and developed nations. With the introduction of highly active antiretroviral therapy (HAART), a decline in morbidity and mortality from HIV has been observed. However, there is some evidence that HAART increases the risk of cardiovascular diseases (CVD) in people living with HIV. To assess this evidence, the levels of serum biomarkers for risk of CVD were measured across different HAART durations among HIV-positive individuals. Methods: This was a descriptive cross-sectional study conducted at the Homa-Bay County Referral Hospital, Kenya. The study population consisted of male and female HIV-positive individuals on HAART between 18 and 45 years of age. Participants were enrolled into the study after consenting and meeting the recruitment criteria. 120 individuals participated in the study and provided blood samples, which were used in the biochemical analysis. Data was analyzed using descriptive statistics. Results: The majority of the participants (67.5%) had been on HAART for more than sixty months. Most of the study participants had total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein (HDL-C), glycated hemoglobin A1c (HbA1c), lipoprotein-associated phospholipase A2 (Lp-PLA2) and myeloperoxidase (MPO) levels within the reference intervals ( 1.0 mmol/L, 21.2 ng/mL – 167, and 21.4 – 229 ng/mL respectively). The proportions that had deranged levels were as follows; 14.2% (TC), 5.8% (LDL-C), 2.5% (HDL-C), 4.2% (HbA1c), 24.2% (Lp-PLA2) and 44.9% (MPO). Most of the participants with deranged levels of serum biomarkers for risk of CVD had been on HAART for more than 60 years. Conclusion: Our findings demonstrate a high proportion of deranged levels of some serum biomarkers are associated with increased risk of CVD in prolonged HAART. We recommend conducting prospective association studies in this population to assess the linkage between high proportions of the levels of these biomarkers to durations time of HAART.

Detection of Simian Immunodeficiency Virus in Semen, Urethra, and Male Reproductive Organs during Efficient Highly Active Antiretroviral Therapy.

A number of men receiving prolonged suppressive highly active antiretroviral therapy (HAART) still shed human immunodeficiency virus (HIV) in semen. To investigate whether this seminal shedding may be due to poor drug penetration and/or viral production by long-lived cells within male genital tissues, we analyzed semen and reproductive tissues from macaques chronically infected with simian immunodeficiency virus mac251 (SIVmac251) who were treated for 4 months with HAART, which was intensified over the last 7 weeks with an integrase inhibitor. We showed that a subset of treated animals continued shedding SIV in semen despite efficient HAART. This shedding was not associated with low antiretroviral drug concentrations in semen or in testis, epididymis, seminal vesicles, and prostate. HAART had no significant impact on SIV RNA in the urethra, whereas it drastically reduced SIV RNA levels in the prostate and vas deferens and to a lesser extent in the epididymis and seminal vesicle. The only detectable SIV RNA-positive cells within the male genital tract after HAART were urethral macrophages. SIV DNA levels in genital tissues were not decreased by HAART, suggesting the presence throughout the male genital tract of nonproductively infected cells. In conclusion, our results demonstrate that 4 months of HAART induced variable and limited control of viral infection in the male reproductive organs, particularly in the urethra, and suggest that infected long-lived cells in the male genital tract may be involved in persistent seminal shedding during HAART. These results pave the way for further investigations of male genital organ infection in long-term-treated infected individuals. A substantial subset of men receiving prolonged HAART suppressing viral loads in the blood still harbor HIV in semen, and cases of sexual transmission have been reported. To understand the origin of this persistence, we analyzed the semen and male reproductive tissues from SIV-infected macaques treated with HAART. We demonstrated that persistent seminal shedding was not linked to poor drug penetration in semen or semen-producing prostate, seminal vesicle, epididymis, and testis. We revealed that HAART decreased SIV RNA to various extents in all male genital organs, with the exception of the urethra, in which SIV RNA(+) macrophages were observed despite HAART. Importantly, HAART did not impact SIV DNA levels in the male genital organs. These results suggest that infection of male genital organs, and particularly the urethra, could be involved in the release of virus in semen during HAART.

Benefícios do exercício físico para pacientes com HIV/AIDS

The Acquired Immunodeficiency Syndrome (AIDS), characterized by a compromised immune system, is caused by the Human Immunodeficiency Virus (HIV). The advent of the Highly Active Antiretroviral Therapy (HAART) has significantly increased the survival of HIV-infected patients, but the infection and collateral effects of prolonged HAART may provoke adverse consequences tothe patients’ health. The objective of this study was to review the literature with regard to the effects of physical exercise as a complementary therapeutic strategy to the adverse consequences of prolonged HAART and HIV infection.The following issues have been discussed: cardiovascular responses, lipodystrophy, dyslipidaemia, psychological markers, wasting syndrome and muscle function, immune system and osteopenia / osteoporosis. In conclusion, the available evidence indicates that the regular practice of supervised exercise can be considered as an effective alternative to prevent and reduce the adverse effects of HIV‑infection and prolonged HAART, increasing the overall fitness without compromising the immune function.

Cholangiocarcinoma in a Young Woman With Perinatally Acquired HIV

Cholangiocarcinoma in a Young Woman With Perinatally Acquired HIV

Effect of prolonged HAART on oral colonization with Candidaand candidiasis

BackgroundProgressive cell-mediated immunodeficiency with decrease of CD4+ lymphocyte count to less than or equal to 200 cells/mm3 is a major risk factor for colonization with Candida species and development of candidiasis. Oropharyngeal candidiasis may occur in up to 90% of human immunodeficiency virus (HIV)-infected patients during the course of the disease. This study is to determine the effect of prolonged highly active antiretroviral therapy (HAART) on oropharyngeal colonization with Candida species and oral candidiasis.MethodsA prospective, longitudinal follow-up study in HIV-infected patients receiving HAART.ResultsThe mean CD4+ count increased from 232.5 to 316 cells/mm3 and the proportion of patients whose CD4+ count less than 200 cells/mm3 decreased from 50.0% to 28.9% (p = 0.0003) in patients receiving HAART for at least 2 years. The prevalence of oral candidiasis decreased from 10.6% to 2.1% (p = 0.004). The decrease in Candida colonization was less impressive, falling from 57.8% to 46.5 % (p = 0.06). Of the 142 patients enrolled in at least two surveys, 48 (33.8%) remained colonized with Candida and 42 (29.6%) remained negative. In the remaining 52 patients, 34 switched from culture positive to negative, and an increase in CD4+ lymphocytes was noted in 91.2% of them. Among the 18 patients who switched from culture negative to positive, 61.1% also demonstrated an increase in CD4+ lymphocyte count (p = 0.01).ConclusionThese findings indicate that HAART is highly effective in decreasing oral candidiasis in association with a rise in CD4+ lymphocyte counts, but only marginally effective in eliminating Candida from the oropharynx.

Persistence of distinct HIV-1 populations in blood monocytes and naive and memory CD4 T cells during prolonged suppressive HAART

Reservoirs of HIV-1 are a major obstacle to virus eradication. There is therefore a need to clearly understand the molecular nature of the virus populations that persist in patients with sustained suppression of plasma viraemia on highly active antiretroviral therapy (HAART). We performed a detailed analysis of the genotypes of HIV-1 quasispecies isolated from highly purified blood cell types taken from three selected patients with sustained undetectable viral loads on HAART for 7 years. We used polychromatic flow cytometry to sort naive and memory CD4 T cells, CD14 monocytes, and CD56+CD3- natural killer (NK) cells from the total peripheral blood mononuclear cells after 7 years of HAART. Clonal analysis was used to determine coreceptor use and drug-resistance genotypes of HIV-1 quasispecies in the sorted blood cell types. We detected HIV-1 DNA in memory and naive CD4 T cells and in CD14 monocytes, but not in the CD56+CD3- NK cells. Phylogenetic analysis demonstrated that the various blood cells types of two of the three patients harboured genetically distinct HIV-1 quasispecies. Drug-resistance mutations were also distributed differently from one cell type to another. This compartmentalization suggests a minimal virus trafficking between blood cell types during suppressive HAART. We observed a cell-specific compartmentalization of the residual virus populations during prolonged suppressive HAART. The coexistence of numerous HIV-1 quasispecies with different resistance genotypes and coreceptor use in cellular reservoirs may be relevant for future antiretroviral treatment strategies.

Association between highly active antiretroviral therapy and hypertension in a large cohort of men followed from 1984 to 2003

UO1-AI-35041: To examine the impact of HIV infection and highly active antiretroviral therapy on systolic and diastolic hypertension. Cohort study with semi-annual assessment of the outcome. We studied 5578 participants of the Multicenter AIDS Cohort Study with blood pressure measurements obtained between 1984 and 2003. The primary outcomes were systolic hypertension (SH; systolic blood pressure > 140 mmHg) and diastolic hypertension (DH; diastolic blood pressure > 90 mmHg). Statistical analyses were performed using multiple logistic regression with robust variance estimation. Of the 84 813 person-visits available for analysis, 7.3 and 8.0% showed SH and DH, respectively. Controlling for age, race, body mass index, and smoking, HIV positive men not taking antiretroviral therapy were significantly less likely than HIV negative men to have SH [odds ratio (OR), 0.79; 95% confidence interval (CI), 0.70-0.89], as were men taking mono/combination therapy (OR, 0.69; 95% CI, 0.59-0.80). The prevalence of SH among men taking highly active antiretroviral therapy (HAART) for less than 2 years was similar to that among HIV negative men (OR, 1.06; 95% CI, 0.87-1.30), but was significantly higher thereafter; for 2 to 5 years of HAART (OR, 1.51; 95% CI, 1.25-1.82) and for more than 5 years of HAART (OR, 1.70; 95% CI, 1.34-2.16). In contrast, DH was not significantly higher among men with prolonged HAART use compared to that among HIV negative controls. Prolonged HAART use was significantly associated with a higher prevalence of SH. This finding suggests that individuals taking HAART may be at increased risk of developing hypertension-related conditions and underscores the importance of blood pressure monitoring among these individuals.

Cytopenias in HIV infection: mechanisms and alleviation of hematopoietic inhibition.

Hematopoietic abnormalities including anemia, cytopenias, and alterations of the stem cell plasticity in the bone marrow microenvironment commonly occur in HIV infected patients. These observations suggest that HIV-1 infection may affect processes important during early stages of hematopoiesis or stem cell differentiation. Hematopoietic abnormalities may be caused by altered stem cell differentiation possibly due to abnormal lineage specific expression of certain cellular genes such as cytokines relevant to hematopoiesis. These cytokines could affect regulatory signals important in hematopoiesis. However, in HIV infected individuals, it is not only the virus but also the highly active antiretroviral therapy (HAART) that both contribute to persistent hematopoietic suppression and ensuing cytopenias. Even if a lowering of HIV replication by HAART were to occur in infected individuals, prolonged HAART by itself and/or appearance of drug resistant mutants can contribute to hematopoietic suppression and resulting cytopenias. However, confounding factors such as opportunistic infections, immune mediated effects, or the consequences of prolonged physiological stress, which could contribute to decreased hematopoiesis in patients or other individuals, make the causative role of HIV in vivo, uncertain. The severe combined immunodeficient mouse transplanted with human fetal thymus and liver tissues (SCID-hu) is a small animal model which mimics HIV infection in humans, and is useful to determine the mechanisms of HIV-1 induced hematopoietic inhibition and development of drug therapies for interventions of stem cell differentiation. Further, SCID mouse serves as a useful small animal recipient of human progenitor cells and also allows us to study the differentiation of these cells in vivo. Results from our studies are expected to provide relief for HIV infected individuals from hematopoietic inhibition and ensuing cytopenias.

Hydroxyurea plus Didanosine as Maintenance Therapy for HIV-Infected Patients on Long-Term Successful Highly Active Antiretroviral Therapy

Background: Toxicity and quality of life issues have moved to delay the initiation of highly active antiretroviral therapy (HAART) and to explore novel treatment strategies for HIV infection. The switch to simpler regimens or treatment discontinuation has been attempted with limited success. The combination of hydroxyurea (HU) plus didanosine (ddI) is a simple regimen that might be able to restrain virus replication for long periods of time and could be an acceptable option as maintenance therapy in patients on prolonged successful HAART. Method: The combination of HU (500 mg bid) plus ddI (400 mg qd) was offered to participants with viral load (VL) <50 HIV RNA copies/mL and CD4 counts >350 cells/μL for more than 6 months under HAART. The prior HAART regimen was resumed if VL rose to >5,000 copies/mL and/or the CD4 count fell to <200 cells/μL after being on HU + ddI maintenance therapy. Results: A total of 187 participants replaced HAART with HU + ddI. In an intent-to-treat analysis at 48 weeks, 109 (58%) and 77 (41%) patients had VL below 5,000 and 500 HIV RNA copies/mL, respectively. The mean CD4 count dropped from 809 ± 283 to 573 ± 270 cells/μL, while 77% of patients remained above 350 cells/μL. The proportion of participants with hypercholesterolemia declined from 70% to 46% (p < .001), while those with hypertriglyceridemia fell from 36% to 21% (p < .05). Significant improvements in lipohypertrophy and lipoatrophy were observed in 52% and 64% of participants, respectively. Grade 3-4 toxicities appeared in 20 patients (11%), including 3 cases of pancreatitis and 1 of peripheral neuropathy. Prior history of VL >5 log, CD4 counts <200 cells/μL, and ddI experience were independently associated with lower response to HU + ddI maintenance therapy. Conclusion: The combination of HU + ddI may be a satisfactory maintenance therapy for more than half of patients on successful HAART who want to alleviate drug-related toxicities and/or pill burden. Patients with metabolic and/or body-shape abnormalities might particularly benefit from switching to this simple regimen.

Analysis of human immunodeficiency virus type 1 gene expression in latently infected resting CD4+ T lymphocytes in vivo.

In individuals with human immunodeficiency virus type 1 (HIV-1) infection, a small reservoir of resting memory CD4(+) T lymphocytes carrying latent, integrated provirus persists even in patients treated for prolonged periods with highly active antiretroviral therapy (HAART). This reservoir greatly complicates the prospects for eradicating HIV-1 infection with antiretroviral drugs. Therefore, it is critical to understand how HIV-1 latency is established and maintained. In particular, it is important to determine whether transcriptional or posttranscriptional mechanisms are involved. Therefore, HIV-1 DNA and mRNAs were measured in highly purified populations of resting CD4(+) T lymphocytes from the peripheral blood of patients on long-term HAART. In such patients, the predominant form of persistent HIV-1 is latent integrated provirus. Typically, 100 HIV-1 DNA molecules were detected per 10(6) resting CD4(+) T cells. Only very low levels of unspliced HIV-1 RNA ( approximately 50 copies/10(6) resting CD4(+) T cells) were detected using a reverse transcriptase PCR assay capable of detecting a single molecule of RNA standard. Levels of multiply spliced HIV-1 RNA were below the limit of detection (<50 copies/10(6) cells). Only 1% of the HIV-1 DNA-positive lymphocytes in this compartment could be induced to up-regulate HIV-1 mRNAs after cellular activation, indicating that most of the proviral DNA in resting CD4(+) T cells either carries intrinsic defects precluding transcription or is subjected to transcriptional control mechanisms that preclude high-level production of multiply spliced mRNAs. Nevertheless, by inducing T-cell activation, it is possible to isolate replication-competent virus from resting CD4(+) T lymphocytes of all infected individuals, including those on prolonged HAART. Thus, a subset of integrated proviruses (1%) remains competent for high-level mRNA production after cellular activation, and a subset of these can produce infectious virus. Measurements of steady-state levels of multiply spliced and unspliced HIV-1 RNA prior to cellular activation suggest that infected resting CD4(+) T lymphocytes in blood synthesize very little viral RNA and are unlikely to be capable of producing virus. In these cells, latency appears to reflect regulation at the level of mRNA production rather than at the level of splicing or nuclear export of viral mRNAs.

Impact of highly active antiretroviral therapy and immunologic status on hepatitis C virus quasispecies diversity in human immunodeficiency virus/hepatitis C virus-coinfected patients.

This study analyzes the effect of highly active antiretroviral therapy (HAART), and thus immunologic status, on hepatitis C virus (HCV) load and quasispecies diversity in patients coinfected with the human immunodeficiency virus (HIV) and HCV. Three cohorts of coinfected patients were analyzed retrospectively over a period of 7 to 10 months: group A was antiretroviral drug naïve at baseline and then on HAART for the remainder of the study, group B did not receive antiretroviral therapy at any point, and group C was on HAART for the entire study. HCV quasispecies diversity was analyzed by sequencing hypervariable region 1. In a longitudinal analysis, there was no significant change from baseline in any immunologic, virologic, or quasispecies parameter in any of the three groups. However, in comparison to groups A and B, group C had significantly higher CD4+- and CD8+-cell counts, a trend toward a higher HCV load, and significantly increased number of HCV clones, entropy, genetic distance, and ratio of nonsynonymous substitutions per nonsynonymous site to synonymous substitutions per synonymous site (Ka/Ks). In addition, CD4+-cell count was positively correlated with HCV load, genetic distance, and Ka. Interestingly, patients infected with HCV genotype 2 or 3 had a significantly higher CD4+-cell count, HCV load, genetic distance, and Ka/Ks than those infected with genotype 1. These results suggest that there is no immediate effect of HAART on HCV but that, with prolonged HAART, immune restoration results in an increase in HCV load and quasispecies diversity.

Efecto del tratamiento antirretroviral de gran actividad en la carga viral del virus de la hepatitis C en los primeros tres años de seguimiento

Highly active antiretroviral therapy (HAART) has been considered to be ineffective in controlling hepatitis C viremia in the first year of treatment. The present study evaluates the effect of prolonged HAART upon HCV-VL. We selected those cases with positive serology for HCV, never been treated with protease inhibitors (PIs) at baseline, which after the introduction of HAART maintained undetectable HIV viral loads (HIV-VL) uninterruptedly for at least three years. Determinations were made of HCV-VL at baseline and after two and three years of HIV-VL suppression. Spearman coefficient and Friedman test were used. Of these patients, 21 satisfied the inclusion criteria. HCV-VL remained without significant changes and with relatively stable levels throughout the three-year follow-up period. However, two patients showed HCV-VL negative conversion after two years, that persisted after three years. Patients who respond to HAART maintain stable HCV-VL. Some subjects may exhibit HCV RNA clearance in the first two years of treatment, maintaining this situation after three years.

Persistence of replication-competent HIV in both memory and naive CD4 T cell subsets in patients on prolonged and effective HAART

To investigate the phenotypic features of infected lymphocytes in patients on prolonged and effective highly active antiretroviral therapy (HAART). We examined highly purified subsets of memory and naive CD4 T lymphocytes for the presence of replication-competent virus. In 11 highly selected HAART-treated patients, we isolated highly purified CD45RO CD45RA CD4 T cells using a magnetic bead-based procedure. In some patients, a subsequent cell separation according to CD62L expression was performed. We quantified total viral DNA in freshly isolated T-cell subsets. To verify whether the virus was replication-competent, HIV RNA was measured in supernatants following cell activation. HIV DNA was detectable in the CD45RO and CD45RA CD4 T-cell subsets in 100% and 90% of the patients tested, respectively. In central memory CD45ROCD62L, effector memory CD45RO+CD62L-, truly naive CD45RACD62L, and CD45RA+CD62L- CD4 T cells, HIV DNA was found in 100%, 55%, 88%, and 50% of the patients tested respectively. HIV DNA was significantly higher in the CD45RO fraction than in the CD45RA subset and in the CD45ROCD62L fraction than in the three other CD45RA/ROCD62L+/- subsets. Detectable HIV RNA was found in the culture supernatants of CD45RO and CD45RA CD4 T-cell subsets in 80% and 66% of the patients tested respectively, and in CD45ROCD62L, CD45RO+CD62L-, CD45RACD62L, and CD45RA+CD62L- CD4 T cells in 100%, 100%, 100% and 50% of the patients tested respectively. In patients on prolonged and effective HAART, the pool of infected CD4 T lymphocytes consists predominantly of memory cells but also contains naive cells.

In patients on prolonged HAART, a significant pool of HIV infected CD4 T cells are HIV-specific.

To examine the antigen specificities of HIV reservoir CD4 T cells in patients on prolonged and effective highly active antiretroviral therapy (HAART). Five HIV-infected patients, who were highly adherent to antiretroviral treatment, were selected on the basis of long-term undetectable plasma viral RNA on unmodified HAART. To investigate the antigen specificities of infected memory CD4 T cells, we examined the capacity of recall antigens, including HIV antigens, to induce virus production by peripheral blood mononuclear cells (PBMC). To quantify CD4 T cells infected by replication-competent virus, and to determine their antigen specificities, we used a limiting dilution-based culture assay. CD8 T cell-depleted PBMC at several cell densities were activated by using Tuberculin purified protein derivative, cytomegalovirus, or HIV-1 p24 with and without HIV-1 Nef. We found that the pool of infected CD4 T cells includes HIV-specific cells with apparent frequencies between 5- and 100-fold higher than those of the common specificities for cytomegalovirus or Tuberculin. Our findings suggest that a significant proportion of replication-competent HIV-infected CD4 T cells in these patients are memory cells directed against HIV determinants. This may provide a rationale for the therapeutic use of recombinant HIV antigens to reduce the pool of HIV-reservoir cells.

Evidence for human immunodeficiency virus type 1 replication in vivo in CD14(+) monocytes and its potential role as a source of virus in patients on highly active antiretroviral therapy.

In vitro studies show that human immunodeficiency virus type 1 (HIV-1) does not replicate in freshly isolated monocytes unless monocytes differentiate to monocyte-derived macrophages. Similarly, HIV-1 may replicate in macrophages in vivo, whereas it is unclear whether blood monocytes are permissive to productive infection with HIV-1. We investigated HIV-1 replication in CD14(+) monocytes and resting and activated CD4(+) T cells by measuring the levels of cell-associated viral DNA and mRNA and the genetic evolution of HIV-1 in seven acutely infected patients whose plasma viremia had been <100 copies/ml for 803 to 1,544 days during highly active antiretroviral therapy (HAART). HIV-1 DNA was detected in CD14(+) monocytes as well as in activated and resting CD4(+) T cells throughout the course of study. While significant variation in the decay slopes of HIV-1 DNA was seen among individual patients, viral decay in CD14(+) monocytes was on average slower than that in activated and resting CD4(+) T cells. Measurements of HIV-1 sequence evolution and the concentrations of unspliced and multiply spliced mRNA provided evidence of ongoing HIV-1 replication, more pronounced in CD14(+) monocytes than in resting CD4(+) T cells. Phylogenetic analyses of HIV-1 sequences indicated that after prolonged HAART, viral populations related or identical to those found only in CD14(+) monocytes were seen in plasma from three of the seven patients. In the other four patients, HIV-1 sequences in plasma and the three cell populations were identical. CD14(+) monocytes appear to be one of the potential in vivo sources of HIV-1 in patients receiving HAART.