Current HIV Research - Volume 19, Issue 2, 2021

In 2014, The Joint United Nations Program on HIV and AIDS (UNAIDS) has set an ambitious target code-named 90-90-90, which aims to ensure that 90% of all people living with HIV will know their state, 90% of all people diagnosed will receive sustained antiretroviral therapy, and 90% of all people receiving ART will have viral suppression by 2020. Since 2014, many tests and treatment programs have been developed to achieve the above goals worldwide. In 2019, it was reported that many developed countries can reach the target with the right strategies, as well as regions that are still far from the targets. It has been reported that the fourth 90 should be one of the targets related to HIV infection in recent years. This view, beyond virological suppression, was towards developing programs that would enable people living with HIV to live not only longer but also healthy. The socio-cultural and economic obstacles to reach the targets may vary according to geographical regions, but it is clear that COVID-19 disease, which has taken the whole world under the influence since 2019, is a major obstacle to the 90-90-90 targets worldwide. Difficulties in the diagnosis and access to ART and treatment nonadherence which may be encountered more frequently due to many factors may threaten both the health of people living with HIV and public health. The COVID-19 pandemic has disrupted many programs developed in the fight against the HIV epidemic. Considering COVID-19 disease and future epidemics that may create a chaotic environment, analyzing the difficulties experienced in the pandemic retrospectively, and determining new strategies that will bring appropriate solutions to the problems will play an important role in the proper management of future issues.

Background: Human immunodeficiency virus (HIV) infection continues to expand worldwide, and a significant proportion of infection is still undiagnosed. Recent studies have addressed the impact and feasibility of ‘opt-out’ HIV screening in Emergency Departments (EDs) in urban settings at high HIV prevalence, whereas little is known about the yield of implementing ‘targeted’ HIV testing, especially in low-prevalence areas. Objective: The present study undertakes a scoping review of research carried out on the implementation of targeted HIV screening of adult in EDs to determine the impact, feasibility and acceptability of HIV testing in different HIV prevalence settings. Design: Online databases (EMBASE, MEDLINE) were used to identify papers published between 2000 to 2020. A three-concept search was employed with HIV (HIV, Human immunodeficiency virus infection, HIV infections), targeted testing (Target, screening or testing) and emergency medicine (Emergency Service, emergency ward, A, accident and emergency or Emergency Department) (28^th February 2020). Only full-text articles written in English, French, Spanish or Italian and using impact and/or feasibility and/or acceptability of the program as primary or secondary outcomes were analysed. Results: The search provided 416 articles. Of these, 12 met inclusion criteria and were included in the final review. Most of the included studies were carried out in the United States (n=8; 67%) and in areas of high HIV prevalence (n=11; 92%). Three (20%) were randomized control studies. While the rate of newly diagnosed HIV cases varied widely (0.03-2.2%), likely due to methodological heterogeneity between studies, the linkage of new HIV diagnosis was often high (80-100%) and median CD4+ cell count was always greater than 200 cells per microliter. Targeted HIV screening was found to be cost-effective (out of 2 studies) and well accepted by participants (out 2 studies). Conclusions: Targeted HIV screening at the ED can be impactful, feasible and well accepted, but often requires extra funding and staff. Most previous work has focused on areas of high disease prevalence.

Background: Chronic infection by HIV evolves with a vascular inflammatory action causing endothelial dysfunction. The action of the virus, as well as the side effects of antiretroviral drugs, contribute to the progression of cardiovascular diseases. The present study aimed to evaluate the percentage of collagen fibers and the density of mast cells, chymase and tryptase, in aortas of patients with and without HIV, and also patients with and without atherosclerosis. Methods: Aortic fragments were obtained from autopsied patients aged 22-69 years and selected regardless of the cause of death or underlying disease. The samples were divided into four groups, (1) Group with HIV and with atherosclerosis; (2) Group with HIV and without atherosclerosis; (3) Group without HIV and with atherosclerosis; (4) Group without HIV and without atherosclerosis (Control). The percentage of collagen fibers was analyzed in the intima-media layer and the density of mast cells was analyzed in all aortic layers. Graphpad Prism 5.0^® software was used for statistical analysis. Results: There were more collagen fibers in HIV patients, with or without atherosclerosis. The group with HIV and atherosclerosis presented a higher density of chymase and tryptase mast cells. The correlation between collagen fibers and age was negative in the non-HIV group and with atherosclerosis. Conclusion: The inflammatory process resulting from HIV infection may be relevant in the alteration of aortic collagen fibers and in triggering or accelerating atherosclerosis. The study is important because HIV patients have increased risks for the development of cardiovascular diseases, and follow-up is necessary to prevent such diseases.

Background: Ethanol has been shown to increase oxidative stress, drug efflux transporter expression, and promote HIV progression. Macrophages, which express drug efflux transporters, serve as an essential sanctuary site for HIV. The antiretroviral drug lopinavir, a protease inhibitor, is a substrate of the drug efflux transporters P-glycoprotein and multidrug resistance-associated protein 1. The NF-ΚB signaling pathway is associated with inflammation and drug efflux transporter expression. Objective: To examine the effects of ethanol on drug efflux transporters and HIV replication of macrophages and develop strategies to increase the efficacy of the protease inhibitor. Methods: The expression of PGP and MRP1 was examined with western blot. The NF- ΚB inhibition was assessed with nuclear western blot. LC-MS/MS and p24 ELISA were used to assess intracellular LPV and viral replication. Results: Ethanol at 40mM slightly increased drug efflux transporter PGP and MRP1 expression in activated macrophages. IKK-16, an NF- ΚB inhibitor, counteracted the increased transporter expression caused by ethanol exposure. MK571, an MRP1 inhibitor, and IKK-16 significantly increased intracellular LPV concentration with or without ethanol treatment. MK571 significantly increased LPV efficacy in suppressing viral replication with or without ethanol treatment. A decreasing trend and a significant decrease were observed with IKK-16+LPV treatment compared with LPV alone in the no ethanol treatment and ethanol treatment groups, respectively. Conclusion: In activated macrophages, inhibiting drug efflux transporter MRP1 activity and reducing its expression may represent a promising approach to suppress viral replication by increasing intracellular antiretroviral concentrations. However, different strategies may be required for ethanolrelated vs. untreated groups.

Background: HIV-1 TAT protein is essential for the regulation of viral genome transcription. The first exon of TAT protein has a fundamental role in the stimulation of the extrinsic and intrinsic apoptosis pathways, but its anti-HIV activity is not clear yet. Methods: In the current study, we firstly cloned the first exon of the TAT coding sequence in the pET-24a expression vector and then protein expression was done in the Rosetta expression host. Next, the expressed TAT protein was purified by Ni-NTA column under native conditions. After that, the protein yield was determined by Bradford kit and NanoDrop spectrophotometry. Finally, the cytotoxicity effect and anti-Scr-HIV-1 activity of the recombinant TAT protein alone and along with Tenofovir drug were assessed by MTT and ELISA, respectively. Results: The recombinant TAT protein was successfully generated in E. coli, as confirmed by 13.5% SDS-PAGE and western blotting. The protein yield was ~150-200 μg/ml. In addition, the recombinant TAT protein at a certain dose with low toxicity could suppress Scr-HIV replication in the infected HeLa cells (∼30%) that was comparable with a low toxic dose of Tenofovir drug (∼40%). It was interesting that the recombinant TAT protein could enhance anti-HIV potency of Tenofovir drug up to 66%. Conclusion: Generally, a combination of TAT protein and Tenofovir drug could significantly inhibit HIV-1 replication. It will be required to determine their mechanism of action in the next studies.

Background: Cognitive impairment in patients with human immunodeficiency virus (HIV) is associated with higher morbidity. The prevalence of the metabolite changes in the brain associated with cognitive impairment in anti-retroviral therapy naïve patients with HIV is unknown. Objective: To estimate the prevalence of the neurometabolites associated with cognitive impairment in antiretroviral therapy (ART) naïve patients with HIV. Methods: We conducted a cross-sectional study among ART naïve patients with HIV aged 18-50 years in a tertiary care center in India. Cognition was tested using the Post Graduate Institute battery of brain dysfunction across five domains; memory, attention-information processing, abstraction executive, complex perceptual, and simple motor skills. We assessed the total N-acetyl aspartyl (tNAA), creatine (tCr) and glutamate + glutamine (Glx) using 3T magnetic resonance spectroscopy. Cognitive impairment was defined as an impairment in ≥2 domains. Results: Among 43 patients eligible for this study, the median age was 32 years (IQR 29, 40) and 30% were women. Median CD4 count and viral load were 317 cells/μL (IQR 157, 456) and 9.3 copies/ μL (IQR 1.4, 38), respectively. Impairment in at least one cognitive domain was present in 32 patients (74.4%). Impairment in simple motor skills and memory was present in 46.5% and 44% of patients, respectively. Cognitive impairment, defined by impairment in ≥2 domains, was found in 22 (51.2%) patients. There was a trend towards higher concentration of tNAA (7.3 vs. 7.0 mmol/kg), tGlx (9.1 vs. 8.2 mmol/kg), and tCr (5.5 vs. 5.2 mmol/kg) in the frontal lobe of patients with cognitive impairment vs. without cognitive impairment but it did not reach statistical significance (p>0.05 for all). There was no difference in the concentration of these metabolites in the two groups in the basal ganglia. Conclusion: There is a high prevalence of cognitive impairment in ART naïve patients with HIV. There is no difference in metabolites in patients with or without cognitive impairment. Further studies, with longitudinal follow-up are required to understand the underlying pathophysiological mechanisms.

Background: Inhibition of HIV-I protease enzyme is a strategic step for providing better treatment in retrovirus infections, which avoids resistance and possesses less toxicity. Objectives: In the course of our research to discover new and potent protease inhibitors, 3D-QSAR (CoMFA and CoMSIA) models were generated using 3 different alignment techniques, including multifit alignment, docking based and Distill based alignment for 63 compounds. Novel molecules were designed from the output of this study. Methods: A total of 3 alignment methods were used to generate CoMFA and CoMSIA models. A Distill based alignment method was considered a better method according to different validation parameters. A 3D-QSAR model was generated and contour maps were discussed. The biological activity of designed molecules was predicted using the generated QSAR model to validate QSAR. The newly designed molecules were docked to predict binding affinity. Results: In CoMFA, leave one out cross-validated coefficient (q²), conventional coefficient (r²) and predicted correlation coefficient (r²Predicted) values were found to be 0.721, 0.991 and 0.780, respectively. The best obtained CoMSIA model also showed significant cross-validated coefficient (q²), conventional coefficient (r²) and predicted correlation coefficient (r²Predicted) values of 0.714, 0.987 and 0.721, respectively. Steric and electrostatic contour maps generated from CoMFA and hydrophobic and hydrogen bond donor and hydrogen bond acceptor contour maps from CoMSIA models were used to design new and bioactive protease inhibitors by incorporating bioisosterism and knowledge-based structure-activity relationship. Conclusion: The results from both these approaches, ligand-based drug design and structure-based drug design, are adequate and promising to discover protease inhibitors.

Background: HIV-1 Nef is an important accessory protein with multiple effector functions. Genetic studies of the HIV-1 Nef gene show extensive genetic diversity and the functional studies have been carried out mostly with Nef derived from regions dominated by subtype B (North America & Europe). Objective: This study was carried out to characterize genetic variations of the Nef gene from HIV-1 infected individuals from North India and to find out their functional implications. Methods: The unique representative variants were sub-cloned in a eukaryotic expression vector and further characterized with respect to their ability to downregulate cell surface expression of CD4 and MHC-1 molecules. Results: The phylogenetic analysis of Nef variants revealed sequence similarity with either consensus subtype B or B/C recombinants. Boot scan analysis of some of our variants showed homology to B/C recombinant and some to wild type Nef B. Extensive variations were observed in most of the variants. The dN/dS ratio revealed 80% purifying selection and 20% diversifying selection implying the importance of mutations in Nef variants. Intracellular stability of Nef variants differed greatly when compared with wild type Nef B and C. There were some variants that possessed mutations in the functional domains of Nef and responsible for its differential CD4 and MHC-1 downregulation activity. Conclusion: We observed enhanced biological activities in some of the variants, perhaps arising from amino acid substitutions in their functional domains. The CD4 and MHC-1 down-regulation activity of Nef is likely to confer immense survival advantage allowing the most rare genotype in a population to become the most abundant after a single selection event.

Background: CRF01_AE and CRF07_BC are the two major HIV-1 virus strains circulating in China. The proportion of dominant subtypes (CRF01_AE and CRF07_BC) among MSM in Jiangsu province was over 80%. A large number of URFs have been found in China in recent years. Objective: This study aimed to report on novel HIV-1 recombinants. Methods: We constructed Phylogenetic trees using the maximum likelihood (ML) method with 1000 bootstrap replicates in IQ-TREE 1.6.8 software and determined recombination breakpoints using SimPlot 3.5.1. Results: We identified a novel, second-generation HIV-1 recombinant (JS020202) between CRF01_ AE and CRF07_BC. The analysis of near full-length genome (NFLG) showed there were at least 8 breakpoints in the virus, which differed from any previously identified CRF and URF around the world. Conclusion: Novel diverse CRF01_AE/07_BC suggested the complexity trends of HIV-1 genetics. The emergency situation of diverse recombinant strains should be monitored continuously.

Background: Concurrence of talaromycosis, an infection caused by the opportunistic fungal pathogen Talaromyces marneffei and Kaposi sarcoma, a common vascular tumor, is a rare but severe medical condition in patients infected with the human immunodeficiency virus (HIV). Despite poor outcomes, the clinical characteristics and management strategies for HIV-infected patients with comorbid Kaposi sarcoma and talaromycosis have not been well documented. Case presentation: A 33-year-old HIV-positive male patient presented to the Department of Infectious Diseases at Wenzhou Central Hospital with cough, sputum expectoration, hemoptysis, rashes on the feet and violaceous plaques in the oral cavity. Chest computed tomography (CT) showed bilateral nodules, patchy shadows and lymphadenectasis. Skin biopsy and histopathological examination indicated Kaposi sarcoma. T. marneffei was isolated from blood cultures and suggested talaromycosis. The patient’s overall conditions significantly improved following initiation of combination antiretroviral therapy (cART) and chemotherapy for Kaposi sarcoma and antifungal treatment for talaromycosis. Conclusion: Severe medical conditions such as Kaposi sarcoma and talaromycosis may coexist in HIV-infected patients and pose an increased risk of mortality. Etiological diagnosis and treatment are the keys to the successful management of HIV-infected patients with these concurrent conditions.