Current HIV Research - Volume 20, Issue 4, 2022

Background: Currently, there is a global contemplation to end the AIDS epidemic by 2030. HIV-2 poses unique challenges to this end. The burden of HIV-2 is higher in resource-limited countries, and it is intrinsically resistant to NNRTI drugs. In addition, there is no FDA-approved plasma viral load assay to monitor disease progression and therapeutic efficacy. To overcome these challenges, we have developed and evaluated an in-house quantitative HIV-2 viral load assay. Methods: Blood samples were collected from 28 HIV-2 treatment-naïve monoinfected individuals and tested using an in-house qPCR HIV-2 viral load assay. The extracted RNA was amplified using Quantifast pathogen + IC kit. Results: The in-house qPCR has a limit of detection of 695 copies/ml. The intra- and inter-assay variation (% CV) of the assay was 0.61 and 0.95, respectively. The in-house assay quantified HIV-2 NIBSC accurately (1000 IU) with a mean of 1952 copies/mL. Among the 28 samples tested by in-house qPCR assay, 11 (39.2%) samples were quantified, whereas 17 (60.7%) samples were not detected. In comparison with Altona RealStar HIV-2 RT PCR and Exavir Load RT assay, the results were 96.4% and 69.6% concordant, respectively. No significant (p = 0.99 and p = 0.13) difference in quantifying viral load between the three assays. Based on clinical and immunological (CD4) staging, the performance characteristics were comparable. Conclusion: To the best of our knowledge, this is the first in-house qPCR developed in India. The performance characteristics of the in-house assay are comparable to the commercial assays, and they can be used assertively to monitor HIV-2 patients.

Background: Lockdown measures for controlling the COVID-19 epidemic were enforced in China between January and May 2020. Previous studies showed a decrease in HIV high- Risk Behaviors (HRBs) and updated testing during the lockdown, but little is known about these behaviors during the post-lockdown period. Objective: We conducted quantitative and qualitative assessments of HIV-related behaviors among MSM during the lockdown and post-lockdown periods in Changsha, south-central China. Methods: Face-to-face structured interviews with open-ended questions were conducted using the TimeLine Follow Back (TLFB) method for collecting retrospective data on frequencies of HRBs and testing. McNemar’s Chi-square test and Wilcoxon signed-rank test were used to comparing frequencies of behaviors between lockdown (January-May 2020) and post-lockdown periods (June- October 2020). Content analysis was used to analyze qualitative data on the reasons for rebounding HRBs and testing. Results: Of 159 MSM participants, 64% had at least one HRB during the post-lockdown period. Men had increased condomless sex (from 24% to 35%), multiple partners (23% to 35%), and substance abuse (16% to 27%) between the two study periods due to the negative emotions and increased use of social networks during the lockdown. HIV testing frequency also increased from 37% to 66% due to resuming routine testing services in the community-based organizations and increased HRBs among MSM during post-lockdown. Conclusion: After lifting the lockdown measures, MSM had rebounding HRBs and uptake of testing. Effective preventive measures and healthcare services should be available to MSM after the lockdown measures are lifted.

Background: Vaccines have had a fundamental impact in containing the ongoing Coronavirus Disease 2019 (COVID-19) pandemic. However, there are few efficacy data relating to frail patients, including the HIV-positive patient. Objective: This study evaluated the Severe Acute Respiratory Syndrome Coronavirus 2 (SARSCoV- 2) serum neutralization in People Living with HIV (PLWH) compared to a cohort of healthy volunteers both vaccinated with BNT162b2. Methods: A serum sample was then withdrawn 14-21 days after the second dose of the vaccine and a serum neutralization assay was performed on Vero E6 cells. The experiments were performed using two strains of SARS-CoV-2 as 20A.EU1 and B.1.617.2. Results: PLWH on Antiretroviral Therapy (ART) showed a vaccine response comparable to the healthy subjects. No correlation between CD4 count or CD4/CD8 and neutralizing antibodies (NTAbs) has been found. No differences in NT-Abs between patients with CD4 nadir above or under 200 cells/μl have been found. In both cohorts, vaccine-elicited serum better neutralized 20A.EU1 than B.1.617.2 strain. Conclusion: PLWH in ART and with good immuno-virological recovery showed a vaccine response comparable to that of healthy subjects and regardless of their immunological status at HIV infection diagnosis. However, larger studies are needed to confirm our results and to evaluate the vaccine response even in patients with low CD4 counts.

Background: Microbial Translocation (MT) and altered gut microbiota are involved in immune activation and inflammation, whereas immune checkpoint proteins play an important role in maintaining immune self-tolerance and preventing excessive immune activation. Objective: This study aims to investigate the relationship between plasma phage load and immune homeostasis in people living with HIV(PLWH). Methods: We recruited 15 antiretroviral therapy (ART)-naive patients, 23 ART-treated (AT) patients, and 34 Healthy Participants (HP) to explore the relationship between the plasma phage load and immune checkpoint proteins. The Deoxyribonucleic Acid (DNA) load of the lambda (λ) phage was detected using fluorescence quantitative Polymerase Chain Reaction (PCR). The Immune Checkpoints (ICPs) were detected using multiplex immunoassay. Results: Our study demonstrated that the plasma phage load was increased in people living with HIV (PLWH) (P<0.05), but not in the ART-naive and AT groups (P>0.05). Plasma ICPs, including cluster of differentiation 27 (CD27), soluble glucocorticoid-induced Tumor Necrosis Factor (TNF) receptor (sGITR), soluble cluster of differentiation 80 (sCD80), sCD86, soluble glucocorticoidinduced TNF receptor-related ligand (sGITRL), soluble induced T-cell Costimulatory (sICOS), sCD40, soluble toll-like receptor 2 (sTLR2), and sCD28, were markedly decreased among the ARTnaive group (P<0.05) but not in the AT and HP groups (P>0.05). The plasma phage load was positively correlated with ICP and C-reactive protein (CRP) levels in PLWH (P<0.05). Conclusion: Our study indicated that the plasma phage load in PLWH was positively related to the expression of ICPs and inflammation, which may be used as a promising marker for the immune level of PLWH.

Background: Many HIV-infected individuals have achieved undetectable viral load and increased CD4 T cell counts due to the success of Antiretroviral Therapy (ART). However, HIV persists in resting T cells, monocytes/macrophages and other quiescent cells. Furthermore, the HIV- 1 vpr accessory gene may play an important role in the persistence of HIV in these infected patients. Objectives: Therefore, we characterized the HIV-1 vpr gene from PBMC DNA of 14 HIV-infected older patients on long-term ART with mostly undetectable viral load and increased CD4 T cell counts. Methods: Peripheral Blood Mononuclear Cells (PBMC) were isolated from 14 HIV-infected individuals, followed by extraction of genomic DNA, amplification of HIV-1 vpr gene by polymerase chain reaction (PCR), cloning of vpr gene in TOPO vector and characterization of correct size recombinant inserts containing vpr genes. An average of 13 clones were sequenced from each patient, followed by sequence analysis by bioinformatic tools. Results: Phylogenetic analysis of 182 vpr sequences demonstrated that the vpr sequences of each patient were well separated and discriminated from other patients’ sequences and formed distinct clusters. The vpr sequences showed a low degree of viral heterogeneity, lower estimates of genetic diversity and about half of the patients’ sequences were under positive selection pressure. While the majority of the vpr deduced amino acid sequences from most patients contained intact open reading frames, several sequences, mostly from two patients, had stop codons. Numerous patient-specific and common amino acid motifs were found in deduced vpr sequences. The functional domains required for vpr activity, including virion incorporation, nuclear import of pre-integration complex and cell cycle arrest, were generally conserved in most vpr sequences. Several of the known Cytotoxic T-lymphocytes (CTL) epitopes in vpr showed variation in our patients’ sequences. Conclusion: In summary, a low degree of genetic variability, conservation of functional domains and variations in CTL epitopes were the features of vpr sequences from the 14 HIV-infected older patients with controlled viremia on long-term ART.

Background: Recently, male hypogonadism was reported to be prevalent in people living with HIV (PLWH), even in cases diagnosed based on the serum free testosterone level (fTST). However, studies on the management of PLWH showing the relationship between male hypogonadism and lifestyle-associated diseases, are sparse. Objective: This study evaluated the relationship between serum fTST levels and lifestyle-related diseases in virologically stable PLWH. Methods: This study was a retrospective cohort single-center study. The study included HIVinfected men on antiretroviral therapy, with available data on serum fTST levels at Teikyo University Hospital between June 2020 and September 2020. Clinical information was collected at the time of fTST measurement. A simple regression analysis was used to identify continuous variables significantly associated with serum fTST levels. Student’s t-test and Mann-Whitney U test were also used to identify non-continuous variables that were significantly correlated with serum fTST levels. Results: Sixty male patients were evaluated. The median age was 47 (40-62) years. Low serum fTST levels were significantly associated with old age, low hemoglobin and total cholesterol levels, and high hemoglobin A1c levels. Non-use of INSTI and comorbid hypertension were also significantly associated with low serum fTST levels. Conclusion: Hypertension and the serum hemoglobin A1c level as a standard parameter for diabetes was significantly associated with low serum fTST levels in Japanese male PLWH. This study suggested that sex-hormone replacement therapy could be a preferred option for PLWH with low serum fTST levels to manage their long-term complications.

Introduction: The analyses of local risks at sub-national levels and tailored interventions are being emphasized for effective HIV management. Andhra Pradesh (AP) is a high HIV prevalence state in India, with the second-highest number of People with HIV (PWH) and a consistent decline in HIV prevalence at the state level. Probing further into the region and district-wise levels and trends of HIV prevalence will provide critical insights into sub-regional epidemic patterns. Hence, this paper analyzes the regional and district-level trends of HIV prevalence among pregnant women attending antenatal clinics (ANC) from 2003 to 2019 in AP, South India. Methods: HIV prevalence data collected from pregnant women in AP during HIV Sentinel Surveillance (HSS) between 2003 and 2019 was used for trend analysis. The consistent sites were grouped into three regions (Coastal Andhra, Rayalaseema and Uttar Andhra), totaling 39 sites, including 21 rural and 18 urban sites. Regional and district-level HIV prevalence was analyzed using the Chisquare trend test, and spatial analysis was done using QGIS software. For the last three HSS rounds, HIV prevalence based on sociodemographic variables was calculated to understand the factors contributing to HIV positivity in each region. Results: In total, 143,211 pregnant women were recruited. HIV prevalence in AP was 0.30% (95% CI: 0.22 - 0.39) in 2019. The prevalence was 0.31%, 0.35% and 0.22% in Coastal Andhra, Rayalaseema and Uttar Andhra, respectively. HIV prevalence had significantly (P < 0.05) declined in all regions. The overall trend indicated that the HIV prevalence was higher among older pregnant women and in urban regions. However, recent trends consistently report HIV positivity among young and new pregnant mothers, illiterates, and rural regions. Conclusion: The overall trend indicated that the HIV prevalence was higher among older pregnant women and in urban regions. However, recent trends consistently report HIV positivity among young and new pregnant mothers, illiterates, and rural regions. Identifying the contextual risk patterns associated with HIV transmission will further improve HIV preventive and management programs among the general population.

Background: Leukopenia, a rare adverse effect of Fingolimod therapy, paves the way for opportunistic infections. In this study, we reported rare fingolimod associated cryptococcal meningitis. Case Presentation: A 39-year-old woman with RRMS was referred to the emergency department. The patient's major complaints were headache, fever, weakness, and progressive loss of consciousness within the last two days prior to the referral. The patient had a history of hospitalization due to RRMS [two times]. In the second hospitalization, interferon Beta-1a was replaced with Fingolimod. Using polymerase chain reaction, Cryptococcus neoformans was detected in CSF. Liposomal amphotericin B and fluconazole [800 mg per day] were started. Six weeks later, the patient was discharged without any major complaints. Conclusion: Albeit fingolimod associated cryptococcal meningitis is a rare event, Fingolimod therapy in patients with MS should be performed cautiously. Regular follow-ups may give rise to a timely diagnosis of probable fingolimod associated cryptococcal meningitis. Fingolimod therapy can lead to lymphocytopenia and various infections. We, therefore, suggest that intermittent blood lymphocyte counts as well as monitoring of clinical manifestations among MS patients treated with Fingolimod to avoid additional neurological and physical disabilities in these patients.