Viral Load Research Articles

Trends in viral load suppression among HIV patients on antiretroviral therapy (ART) at Asante Mampong Municipal Hospital, Ghana: 2019–2023

BackgroundThe Joint United Nations Programme on Human Immunodeficiency Virus (HIV)/Acquired Immune Deficiency Syndrome (AIDS) (UNAIDS) aims to eradicate AIDS by 2030 through 95:95:95 targets: identifying 95% of persons living with HIV (PLHIV), initiating 95% of those identified on antiretroviral therapy (ART), and ensuring that 95% of those initiated on ART are virally suppressed. Virally suppressed patients pose minimal risk of HIV transmission. ART aims to suppress the HIV-viral load (VL) and increase immunity, reducing morbidity and mortality. This study aimed to determine the trends in VL suppression among HIV patients on ART from 2019 to 2023 at Asante Mampong Municipal Hospital.MethodsThis study adopted a retrospective Hospital-based design in which secondary data from 842 patients on ART from 2019 to 2023 were used. The study design specifically involved conducting serial cross-sectional studies to measure the prevalence of VL suppression each year from 2019 to 2023. This approach allowed the researchers to analyse the annual prevalence of VL suppression among study participants without following individual participants longitudinally throughout the entire period. The data were analysed via STATA version 17.0. Chi-square and logistic regressions were used to determine the associations between VL suppression and the independent variables at p < 0.05 and 95% confidence intervals (CIs).ResultsIn 2019, VL suppression was 79.6%, decreasing to 40.0% in 2020 and then rising to 82.7% in 2021, dropping to 67.8% in 2022 and 66.7% in the first quarter of 2023. Clients aged 40–49, 50–59, and 60–69 years were more likely to have VL suppression [aOR = 4.4 (1.36–14.25), p = 0.013], [aOR = 5.5 (1.65–18.39), p = 0.006] and [aOR = 5.0 (1.42–17.46), p = 0.012], respectively. Clients who were consistently on the same type of ART for more than a year were more likely to have VL suppression [aOR = 10.6 (4.18–26.76), p < 0.001].ConclusionVL suppression was low among patients. Advanced age and being on the same ART for more than 12 months were significantly associated with VL suppression. Health promotion activities are needed for people who have been suppressed to maintain and achieve a lifetime undetectable VL, targeting the younger age group.

Prostaglandin E2 suppresses KCNH1 gene expression and inhibits the proliferation of CaSki cervical cells through its four prostanoid PTGER subtypes

The main risk factor for cervical cancer is the persistent infection of high-risk HPV subtypes, notably HPV16. Another contributing factor is proinflammatory prostaglandin E2 (PGE2), a lipid abundantly found in seminal fluid. PGE2, along with its receptors (PTGER1-4), contributes to cancer development; however, its specific role in the proliferation of cervical cancer models with high HPV16 copy numbers remains unclear. In this study, we investigated the effects of PGE2 on the proliferation of CaSki cells, a cell line with a high HPV16 viral load. Surprisingly, PGE2 inhibited CaSki cell proliferation, while it increased the proliferation of SiHa, HeLa, and C-33 A cervical cancer cells. The effect of PGE2 on CaSki cell proliferation was specific, as estradiol increased cell growth. Furthermore, PGE2 suppressed expression and promoter activity of the cervical tumoral marker KCNH1. To discern the specific role of each receptor in cell proliferation, we generated stable CaSki cell lines overexpressing each receptor alongside control cells with an empty vector. Notably, PGE2 significantly inhibited cell proliferation in all stable transfected CaSki cells, suppressing oncogenic KCNH1 expression and its promoter activity. In conclusion, our findings indicate that PGE2 inhibits the proliferation of CaSki cervical cancer cells with a high HPV16 load, at least in part, by suppressing the expression of the oncogenic KCNH1 gene.

Minimally Modified HIV-1 Infection of Macaques: Development, Utility, and Limitations of Current Models.

Nonhuman primate (NHP) studies that utilize simian immunodeficiency virus (SIV) to model human immunodeficiency virus (HIV-1) infection have proven to be powerful, highly informative research tools. However, there are substantial differences between SIV and HIV-1. Accordingly, there are numerous research questions for which SIV-based models are not well suited, including studies of certain aspects of basic HIV-1 biology, and pre-clinical evaluations of many proposed HIV-1 treatment, prevention, and vaccination strategies. To overcome these limitations of NHP models of HIV-1 infection, several groups have pursued the derivation of a minimally modified HIV-1 (mmHIV-1) capable of establishing pathogenic infection in macaques that authentically recapitulates key features of HIV-1 in humans. These efforts have focused on three complementary objectives: (1) engineering HIV-1 to circumvent species-specific cellular restriction factors that otherwise potently inhibit HIV-1 in macaques, (2) introduction of a C chemokine receptor type 5 (CCR5)-tropic envelope, ideally that can efficiently engage macaque CD4, and (3) correction of gene expression defects inadvertently introduced during viral genome manipulations. While some progress has been made toward development of mmHIV-1 variants for use in each of the three macaque species (pigtail, cynomolgus, and rhesus), model development progress has been most promising in pigtail macaques (PTMs), which do not express an HIV-1-restricting tripartite motif-containing protein 5 α (TRIM5α). In our work, we have derived a CCR5-tropic mmHIV-1 clone designated stHIV-A19 that comprises 94% HIV-1 genome sequence and replicates to high acute-phase titers in PTMs. In animals treated with a cell-depleting CD8α antibody at the time of infection, stHIV-A19 maintains chronically elevated plasma viral loads with progressive CD4+ T-cell loss and the development of acquired immune-deficiency syndrome (AIDS)-defining clinical endpoints. However, in the absence of CD8α+ cell depletion, no mmHIV-1 model has yet displayed high levels of chronic viremia or AIDS-like pathogenesis. Here, we review mmHIV-1 development approaches, the phenotypes, features, limitations, and potential utility of currently available mmHIV-1s, and propose future directions to further advance these models.

Exploring the link between COVID-19 and gut microbiome at the community level

Abstract Introduction/Objective Monitoring SARS-CoV-2 virus through wastewater has been adopted as an effective surveillance approach for combating the COVID-19 pandemic. To understand the interplay between this virus and the human microbiome, this study examined the longitudinal co-dynamics between the virus and the microbiome across various sewage samples representing diverse demographic populations in Houston. Methods/Case Report We collected wastewater through manholes and the viral RNA levels of SARS-CoV-2 were monitored from December 2020 to 2021. The sampling sites serve nursing homes, rehabs, and shelters across the metro Houston area, respectively. The viral load was analyzed by the gold standard RT-qPCR method, and the microbiome load and composition by qPCR and amplicon sequencing of the 16S and ITS DNA. Results (if a Case Study enter NA) Preliminary results on the dynamics of SARS-CoV-2 viral load indicated diverse trends among sampling sites. These viral trends could be classified into three basic categories: flat, up and down, and up, suggesting disparities in COVID-19 infections among communities. A full analysis of the microbiome is underway, but a preview of the data revealed that bacteria and fungi commonly seen in the human gut predominated the wastewater samples. This paves the way to exploring the link between COVID-19 and the gut microbiome, which could shed light on the observed COVID-19 disparities. Conclusion As a proof-of-concept, our sewage bio-monitoring program was able to capture COVID-19 viral, bacterial, and fungal signals longitudinally from manholes serving a wide range of communities. Because these signals are predominantly human-associated, further analysis of these signals could offer insight on COVID-19 related disparities among these communities.

Optimizing Patient Care through Analysis of HIV-1 Genotyping: Insights into Test Ordering Strategies, Clinical Relevance, and Implications for Managing Drug Resistance

Abstract Background Antiretroviral resistance can lead to treatment failure and disease progression, especially in HIV patients. HIV-1 sequencing/genotyping tests (e.g., GenoSure) can be ordered by clinicians to detect HIV drug resistance (HIVDR). Here, we investigated our institution’s ordering practices, clinical relevance, and rates of HIVDR. Methods A total of 94 patients positive for HIV-1 between 01/2019-08/2021 were included (n=46 pre-COVID-19 prior to 03/11/2020; n=48 post-COVID-19). Patients’ charts were reviewed for demographics, laboratory results, clinical presentation, and antiviral management. Student t-test and Fisher exact test were used for statistical analyses. Results HIV-1 genotyping was ordered most by Medicine (67%), Surgery, (7%), Emergency medicine (6%), Neurology and Obstetrics &amp; Gynecology (5% each). Only 70% of patients with GenoSure ordered and sent out had sufficient viral loads for genotyping. Among the patients with insufficient viral loads, the GenoSure test was most ordered by Medicine (68%), Surgery (9%) and Ob/Gyn (9%) departments. Our HIV patient population was predominantly male (70%) and had a mean age of 49 years (range: 25 to 90 years). Lower viral loads (1.8x105 vs. 3.4x105 copies/mL, p=0.05), CD4 counts (174 vs. 196 cells/mcL, p=0.7), CD8 counts (555 vs. 800 cells/mcL, p=0.04), and higher CD4/CD8 ratio (0.312 vs. 0.266, p=0.57) were seen in our post-COVID-19 patients. The overall prevalence of HIVDR was 20%. There was a 9% increase in HIVDR, predominantly in non-nucleotide reverse transcriptase inhibitors (NNRTI), after the COVID-19 pandemic. For example, higher resistance was documented for antiviral drugs Efavirenz (16%), Nevirapine (16%), and Rilpivirine (13%). Increased prevalence of mutations in G190A, K101, K103, and E138 positions which confer resistance to NNRTIs was seen. The GenoSure test results helped optimize antiviral therapy in 47.8% (42/88) of total cases, with an increase of 12.5% in patients from post-COVID-19 era. Infectious disease consultations led to more actionable changes (16% increase) and ordering of the updated version of the GenoSure test that includes integrase inhibitors (20% increase, p=0.005). Conclusion We have implemented stringent clinical criteria including pathologist approval and changed our electronic health record ordering system to ensure GenoSure is appropriately ordered. Diagnostic stewardship of sequencing tests needs to be implemented to ensure actionable clinical changes in the right patient population and to control testing costs. Additionally, our findings revealed increased HIVDR in our patient population post-COVID-19. Continued surveillance and collaboration with antimicrobial stewardship are critical to managing HIVDR and improving patient outcomes, particularly given the significant strain that the COVID-19 pandemic has placed on our healthcare system.

Efficiently Improving Laboratory Workflow with Continuous Sample Loading by Validating the COBAS 5800 for HIV and HCV Viral Loads

Abstract Introduction/Objective A regional veteran affairs medical center (VAMC) was using the COBAS AmpliPrep/COBAS Taqman (Roche, Basel Switzerland) with a throughput of 72 tests/8 hours without continuous loading capability. However, a newer cobas 5800 System (Roche, Basel Switzerland) was validated to allow for continuous loading with better parameters for workflow including 144 tests/per 8 hours with a capacity for continuous loading of samples without batching. As validation experiences between the two instruments is sparse in the literature, the validation is described. Methods/Case Report The validation process included a linearity study using 24 samples with levels (100, 600, 6,000, 60,000, 600,000, 2,000,000, and 6,000,000 copies/mL for HIV and 100, 300, 3,000, 30,000, 300,000, 3,000,000, and 30,000,000 copies/mL for HCV) across the analytical range, a precision study at a low (1,000 copies/mL) and high (100,000 copies/mL for HIV, 1,000,000 copies/ML for HCV) values of a triplicate run over 5 days, an accuracy study based on the precision study, a limit of detection study using 20 control samples at the limit of detection, control monitoring (high and low positive controls) over 9 runs, and a method correlation between the Cobas 5800 and the Cobas AmpliPrep/COBAS Taqman. Results (if a Case Study enter NA) The validation study resulted as expected. For the method correlation, the results were plotted and evaluated with an R2 of 0.937 (HIV from 91 samples) and 0.9646 (HCV from 74 samples). Linearity across the analytical range had an R2 of 0.9967 (HIV) and 0.9950 (HCV). All samples were detected at the limit of detection study. Both the accuracy and precision yielded expected results. The values from running the controls (high and low positive) over 9 runs remained within the expected range and within two standard deviations for all runs. Conclusion The COBAS 5800, which has improved capabilities compared to the COBAS AmpliPrep/COBAS Taqman, was validated for clinical use successfully.

HUMAN IMMUNODEFICIENCY VIRUS’ EXPOSED UNINFECTED NEWBORNS: A VULNERABLE POPULATION WITH SOME DEFECTIVE IMMUNE SYSTEM ELEMENTS AT BIRTH IN YAOUNDE-CAMEROON, CENTRAL AFRICA

Abstract Introduction/Objective The reduction in vertical transmission of human immunodeficiency virus (HIV) in Cameroon, thanks to the success of Option B+ prevention measures, has favored the increase in the rate of HIV exposed uninfected (HEU) infants. The latter, however, present a threefold higher morbidity and mortality than their HIV unexposed and uninfected (HUU) counterparts. With multiple causes, a dysfunction of the immune system would be incriminated but the evidence to demonstrate this and to advocate for a specific follow-up is lacking. This study objective was to determine the effect of in utero exposure to HIV-1 on certain immune elements of HEU newborns. Methods/Case Report A Case-control study was carried out in three reference hospitals in Yaounde. From June 2022 to July 2023, demographical characteristics of 62 HEU newborns of positive parturients with undetectable viral load who gave their consent (cases), and 65 HUU (controls) were taken. Then samples of umbilical cord blood were collected under ethical authorization no. BTC-JIRB2022-021. Then, blood count and differential leukocytes count were performed using DF55 DEMIND and May-Grünwald Giemsa Kit. Excel 2007 and R v4.1.3 softwares were used to record and analyze the data, with significance level for a p-value &amp;lt; 0.05. Results (if a Case Study enter NA) The average APGAR vitality score (appearance, pulse, grimace, activity, respiration) at 1, 5 and 10 min of life was reduced, but not significantly (p = 0.3) in the HEU (8/10, 9/10 and 10/10), compared to the HUU (9/10, and 10/10); as well as head circumference: (33.16 ± 1.64) cm and (34.8 ± 1.52) cm (p = 0.8) respectively. Unlike leukopenia (p=0.24), umbilical monocytosis (p=0.04) and lymphopenia (p=0.001) were significantly linked to HIV exposure in HEU, signs of inflammation from birth. Conclusion HIV exposed uninfected newborns present a drop in immunity and an inflammatory sign at birth, materialised clinically and biologically with a highly significant lymphopenia.

Development of a droplet digital PCR assay for the detection of BK polyomavirus.

The objective of this study was to establish a more sensitive and specific diagnostic method for detecting plasma BK polyomavirus (BKPyV) DNA load in patients after renal transplantation using droplet digital polymerase chain reaction (ddPCR) and to validate the methodology. The linear range, lower limit of detection, accuracy, precision, and specificity of the detection system were evaluated by using the WHO BKPyV standard (7.2 log10 IU/mL) as a reference, in accordance with the relevant documents of the Clinical and Laboratory Standards Institute. Plasma samples were collected from 74 renal transplantation patients with urinary BKPyV-DNA levels exceeding 7 log10 copies/mL. Quantitative PCR (qPCR) and ddPCR were performed, and their diagnostic efficacy for BKPyV-DNA in the diagnosis of BK polyomavirus-associated nephropathy was evaluated using a receiver operating characteristic (ROC) curve. The coefficients of variation for the repeated detection of BKPyV standard DNA were 2.55 and 4.71 at concentrations of 6.2 and 3.2 log10 IU/mL, respectively. The linear range was 2.2-6.2 log10 IU/mL, and the lowest detection limit was 100 IU/mL. By utilizing histopathological examination of renal biopsy as the gold standard for BKPyV diagnosis, the area under the ROC curve of 74 post-transplantation plasma samples detected by the ddPCR system was found to be 0.875 (95% CI: 0.797-0.953, P < 0.01). The optimal threshold was 512.86 copies/mL, with a sensitivity of 90.0% and a specificity of 67.6%. In comparison, the area under the ROC curve for qPCR was 0.668 (95% CI: 0.583-0.752, P < 0.01), with an optimal threshold of 11,481.54 copies/mL, a sensitivity of 35.0%, and a specificity of 100.0%. Pairwise comparison (Delong test) of the ROC curves of the two systems showed a significant difference in the area under the curve, with a difference of 0.207 and a P-value <0.01. The BKPyV nucleic acid detection system, based on ddPCR, is appropriate for the regular monitoring of the BK polyomavirus, specifically in plasma samples containing low viral DNA loads while it provides the benefits of both absolute quantification and high sensitivity.IMPORTANCEIt was previously believed that droplet digital polymerase chain reaction had limitations, including high cost, limited throughput, and cumbersome operation, which hindered its widespread application in clinical practice. However, the current fully automated digital PCR platform, combined with streamlined operations, can detect 96 samples at once, and the entire process can be completed within an hour, laying a solid foundation for its extensive use.

Impacts of indoor masks wearing on air contamination during 10-minute speaking in patients with SARS-CoV-2 omicron variant infection.

In 10-minute speaking, N95 respirators significantly decreased SARS-CoV-2 emissions compared with no-mask wearing. However, SARS-CoV-2 was detected in the air even when wearing N95 and surgical masks in patients with high viral loads. Therefore, universal masking of infected and uninfected persons is important for preventing COVID-19 transmission via the air.

HIV Outcomes and Intervention Experiences of Enlaces Por La Salud: A Personal Health Navigation Intervention Informed by the Transnational Framework.

Latinos in the United States are disproportionately affected by HIV and experience sub-optimal levels of viral suppression. Enlaces Por La Salud is an individual-level intervention implemented by personal health navigators and guided by the transnational framework to improve HIV care outcomes among newly diagnosed and out-of-care Mexican and Mexican American men and transgender women in North Carolina. The purpose of this study was to assess: (1) changes in HIV care and treatment outcomes among Enlaces participants and (2) intervention engagement and experiences. Ninety-one participants were recruited between October 2014 and August 2017 for a single-arm, mixed-methods design including surveys at baseline, 6 and 12 months and qualitative in-depth interviews immediately following the intervention with an embedded cohort (n = 19). Mean participant age was 36.8 years and most identified as cisgender male (90%). Participants were significantly more likely to have an undetectable viral load at baseline (18%) compared to 6-months (78%) (p < .001). Outcomes were sustained but not significantly increased from 6 to 12 months. Intervention engagement was high with 81% completing all six intervention sessions. In qualitative interviews, participants emphasized the importance of their relationship with the personal health navigators, who provided information, instrumental, and emotional support. A person-centered intervention guided by the lived experience of migration could be an effective way to support Latinos with HIV from diverse countries of origin to achieve viral suppression and improve overall wellbeing.

Direct genome sequencing of respiratory viruses from low viral load clinical specimens using the target capture sequencing technology.

The use of metagenomic next-generation sequencing technology to obtain complete viral genome sequences directly from clinical samples with low viral load remains challenging-especially in the case of respiratory viruses-due to the low copy number of viral versus host genomes. To overcome this limitation, target capture sequencing for the enrichment of specific genomes has been developed and applied for direct genome sequencing of viruses. However, as the efficiency of enrichment varies depending on the probes, the type of clinical sample, etc., validation is essential before target capture sequencing can be applied to clinical diagnostics. In this study, we evaluated the utility of target capture sequencing with a comprehensive viral probe panel for clinical respiratory specimens collected from patients diagnosed with SARS-CoV-2 or influenza type A. We focused on clinical specimens containing low copy numbers of viral genomes. Target capture sequencing yielded approximately 180- and 2,000-fold higher read counts of SARS-CoV-2 and influenza A virus, respectively, than metagenomic sequencing when the RNA extracted from specimens contained 59.3 copies/µL of SARS-CoV-2 or 625.1 copies/µL of influenza A virus. In addition, the target capture sequencing identified sequence reads in all SARS-CoV-2- or influenza type A-positive specimens with <26 RNA copies/µL, some of which also yielded >70% of the full-length genomes of SARS-CoV-2 or influenza A virus. Furthermore, the target capture sequencing using comprehensive probes identified co-infections with viruses other than SARS-CoV-2, suggesting that this approach will not only detect a wide range of viruses but also contribute to epidemiological studies.IMPORTANCETarget capture sequencing has been developed and applied for direct genome sequencing of viruses in clinical specimens to overcome the low detection sensitivity of metagenomic next-generation sequencing. In this study, we evaluated the utility of target capture sequencing with a comprehensive viral probe panel for clinical respiratory specimens collected from patients diagnosed with SARS-CoV-2 or influenza type A, focusing on clinical specimens containing low copy numbers of viral genomes. Our results showed that the target capture sequencing yielded dramatically higher read counts than metagenomic sequencing for both viruses. Furthermore, the target capture sequencing using comprehensive probes identified co-infections with other viruses, suggesting that this approach will not only detect a wide range of viruses but also contribute to epidemiological studies.

Prevalence of high-risk human papillomavirus genotypes and viral load correlated with squamous cell inflammation among women in Gabon

BackgroundHigh-risk genotypes of Human Papillomavirus are responsible for 90% of cases of cervical cancer worldwide. Inflammation of squamous cells is mainly linked to HPV. In Gabon, HPV is endemic and circulates among the female population. The study aimed to determine the prevalence of HR-HPV genotypes and to investigate the correlation between squamous cell inflammation and HPV viral load in infected women in Gabon.MethodsThe cross-sectional study was conducted at Libreville University Hospital Center (UHC) and National Public Health Laboratory from March to May 2024 among 399 women. Two cervical smears were taken. Genotype detection was carried out by multiplex fluorescence real-time PCR in the NPHL virology unit. Cytology was carried out in UHC’s anatomic-pathology laboratory. Data were analyzed by SPSS software. Graphs were plotted using Microsoft Excel 2016.ResultsThe prevalence of Human Papillomavirus was 26.1% (95% CI: 22-30.6). The prevalence of HR-HPV genotypes was 24.8%. The most common HR-HPV genotypes were HPV-16/52/18/35/56/58/53/68. The rate of multiple HPV infections was 29.8% and 95.2% for the HR-HPV infection rate. Viral load was significantly correlated with squamous cell inflammation (r = 0.977 and P = 0.001).ConclusionHR-HPV infection remains a concern in women, however early screening is necessary for optimal monitoring and management. HR-HPV viral load is a predictive marker of squamous cell inflammation.

Engaging Diverse African American/Black and Latine Youth and Emerging Adults Living with HIV into Research: Description of Recruitment Strategies and Lessons Learned.

Improving engagement along the HIV care continuum and reducing racial/ethnic disparities are necessary to end the HIV epidemic. Research on African American/Black and Latine (AABL) younger people living with HIV (LWH) is essential to this goal. However, a number of key subgroups are challenging to locate and engage, and are therefore under-represented in research. Primary among these are persons with non-suppressed HIV viral load, severe socioeconomic disadvantage, transgender/gender expansive identities, and refugee/migrant/immigrant populations. Research in community settings is needed to complement studies conducted in medical institutions. The present study describes the efficiency of recruitment strategies used in the community to enroll AABL young and emerging adults LWH ages 19-28 years. Strategies were designed to be culturally responsive and structurally salient. They were: peer-to-peer, social media, classified advertisements (newspaper, craigslist), subway ads, dating apps (Jack'd, Positive Singles), and direct recruitment in community-based organizations. Data were analyzed using mainly descriptive statistics and interpreted using a consensus building approach. We screened 575 individuals in a first step, 409 were eligible (71%), of these 297 presented to the second screening step (73%), but 112 were lost. Almost all presenting at the second step were eligible (98%, 291/297) and 94% enrolled (274/291). Peer-to-peer, dating app (Jack'd), direct recruitment, and craigslist were the most efficient strategies. Recruitment on dating apps was superior to the peer-to-peer approach in yielding eligible participants (OR = 1.5; 95% CI: 0.98-2.3; p = 0.06). The sample enrolled was diverse with respect to HIV viral suppression, gender identify, sexual orientation, immigration status, and barriers to HIV care engagement. We discuss the advantages and disadvantages of each strategy. Recruitment is a vital aspect of research and warrants attention in the empirical literature.

Analyzing Social Support from Facebook on Viral Suppression among Young Black Men Who Have Sex with Men Living with HIV: A Pilot Study.

Social support has been associated with viral suppression among persons living with HIV (PWH). We examined cross-sectional data from young Black men who have sex with men to see if sociodemographic factors, medical history, and egocentric Facebook social support measures are related to viral suppression. Differences between participants were examined using Chi-square, Fisher's exact, or Wilcoxon Rank Sum Tests, with significance set at p = 0.10. Degree centrality (p = 0.074) and Eigenvector centrality (p = 0.087) were significant, indicating that PWH with unsuppressed viral loads had more social connections. These results contrast prior studies in the literature. Further research on online social support for PWH is needed.

Hepatitis E virus prevalence among HIV-2 infected individuals in central Portugal.

To date, no studies comparing the prevalence of hepatitis E virus (HEV) infection between the general and human immunodeficiency virus 2 (HIV-2) populations are available. With the purpose of filling this gap, this prevalence was assessed in the HIV-2 population from central Portugal. HEV seropositivity was 19.4%, which did not differ significantly from that found in the matched control population, and was not associated with CD4 cell count, HIV-2 viral load, and geographic origin or travel history to regions considered highly endemic for HEV. The results suggest that HIV-2 is not a risk factor for HEV infection, neither for an increased occurrence of chronic HEV infection.

Evaluation of CMV T-cell Mediated Immunity in Children with CMV DNAemia After Allogeneic Hematopoietic Cell Transplantation

Abstract Background Cytomegalovirus (CMV) is among the most common infectious complications following allogeneic hematopoietic cell transplantation (allo-HCT). A commercially available CMV T-cell immune panel (CMV-TCIP, Viracor®), reported as percentages of CMV-specific interferon gamma releasing CD4 and CD8 cells (%CMV-CD4 or %CMV-CD8 IFN-γ), may be used to assess CMV cell-mediated immunity (CMV-CMI). However, the utility of this assay among pediatric allo-HCT recipients remains unknown. Methods A retrospective analysis among pediatric allo-HCT recipients (age ≤18 years) from 1/2020 to 11/2023 who developed CMV DNAemia and had CMV-TCIP testing was performed. Per institutional protocol, weekly quantitative plasma CMV PCR was performed until day +100. Clinically significant CMV infection (csCMVi) was defined as CMV viral load (VL) ≥500IU/ml and pre-emptive therapy (PET) was provided for csCMVi. The most frequent indications for CMV-TCIP testing were to evaluate CMV-CMI to inform the need of CMV virus-specific T-cell therapy (CMV-VST) in the setting of resistant/refractory CMV, to monitor CMV-CMI as a proxy of VST expansion post CMV-VST, or to evaluate the need of ongoing secondary antiviral prophylaxis. A %CMV-CD4 or %CMV-CD8 IFN-γ ≤0.2% was categorized as a poor response, whereas a result &amp;gt;0.2% was classified as an adequate response. Specimens with insufficient quality, high background, or discrepant results from %CMV-CD4 or %CMV-CD8 were classified as a discordant response. Demographics, underlying condition, HCT type, conditioning regimen, antiviral agents and use of VST, VL, and CMV-TCIP results were recorded. Descriptive and nonparametric statistics (median, interquartile range [IQR]) were used for analysis. Results Twenty-seven CMV-TCIP results from 10 unique allo-HCT recipients were analyzed. The median age at transplant was 8.5 years (IQR 2.75-14.3), predominantly for hematologic malignancy (70%). CMV donor/recipient (D/R) serostatus were D+/R+ (60%), D-/R+ (30%) and D+/R- (10%). CMV DNAemia developed post-HCT in 80% (8/10) with median onset of 4 days post-HCT (IQR 3-40) and 2 subjects had known DNAemia prior to HCT. Nine (90%) subjects were started on PET for csCMVi, whereas 1 (10%) received antiviral therapy when VL was &amp;lt;500IU/ml. The first CMV-TCIP was obtained at median of 112 days (IQR 42-130) post-HCT. Despite appropriate antiviral therapy, patients with a peak VL ≥500 IU/mL (obtained within 1 week before and after CMV-TCIP testing) demonstrated significantly lower absolute CD4 (median 12 vs. 198 cells/uL, p=0.02) and CD8 (median 79 vs. 443 cells/uL, p=0.04) counts and %CMV-CD4 IFN-γ (0.04% vs. 0.36%, p=0.02) compared to those with a peak VL &amp;lt;500 IU/mL. Conclusion Our results suggest that CMV-CMI, particularly %CMV-CD4 IFN-γ &amp;gt;0.2%, was observed concurrently with downward trending VL, and may serve as a proxy of VST expansion in patients receiving CMV-VSTs. Figure 1A. The distribution of peak VLs over time based on %CMV-CD4 and %CMV-CD8 IFN-γ are demonstrated in Figure 1B. CMV peak VLs were lower with TCIP testing results reported as adequate combined responses compared to those with poor combined responses. In five subjects (50%) who received CMV-VSTs as part of CMV therapy, increased %CMV-CD4 and %CMV-CD8 IFN-γ were observed post-infusion with an observed downward trend in VL.

Asymptomatic malaria parasitaemia and virological non-suppression among children living with HIV in a low transmission area in Accra, Ghana: a cross-sectional study

BackgroundHuman Immunodeficiency Virus (HIV) and malaria are two major diseases in sub-Saharan Africa. Co-infection can significantly impact the clinical outcomes of both conditions. We assessed the proportion of HIV-infected children at Korle Bu Teaching Hospital (KBTH) and Princess Marie Louise Hospital (PML) with malaria parasites. The association between asymptomatic malaria parasitaemia and virological non-suppression was also determined in these children.MethodsThis cross-sectional study of 277 asymptomatic malaria in children receiving care at paediatric HIV clinics at KBTH and PML was conducted from September to November 2022. Patients who had been on antiretroviral therapy (ART) for at least six months were eligible to participate. Structured questionnaires were used to collect socio-demographic information, malaria prevention behaviors, and ART-related data using in-person interviews. Microscopy and PCR were used to screen for malaria, and GeneXpert was used to determine viral load. To examine the determinants of malaria PCR positivity and virological non-suppression, chi-square tests and logistic regression were performed.ResultsThe median age of the participants was 9 years (range: 6–12 years). Males comprised 158 (57%) of the study population. We detected 10 (3.6%) and 21 (7.6%) malaria cases by microscopy and PCR, respectively. Virological non-suppression (VL > 1000 copies/ml) was observed in 82 (29.6%) of the 277 participants. Among the suppressed individuals, 62 (22.4%) exhibited low-level viraemia (VL level 40-1000 copies/ml) and 133 (48%) had undetectable viral load levels. No factors were associated with the presence of malaria PCR positivity carriage. Poor adherence to ART was associated with a five-fold increase in the risk of viral load non-suppression (AOR = 4.89 [CI = 2.00-11.98], p = 0.001).ConclusionThe proportion of children living with HIV with asymptomatic malaria parasitaemia was low. Approximately one-third of the study population had virological non-suppression. The interaction between malaria parasitemia and viral replication may not be the main cause for virological non-suppression in this low transmission area.

New US DHHS Guidelines for Feeding of HIV Exposed Infants: Uptake, Practice Variability and Concerns

Abstract Background Approximately 5000 infants are born to mothers living with HIV annually in the US. In the past, the Center for Disease Control (CDC), American Academy of Pediatrics (AAP), and US Department of Health and Human Services (DHHS) have recommended that people living with HIV (PWH) not breast/chest feed their infants. Recent changes published in January 2023 in the “Section on Infant Feeding for Individuals Living with HIV” in the US DHHS HIV Guidelines, now include that individuals with HIV who are on antiretroviral therapy (ART) with a sustained undetectable viral load (VL) and who choose to breastfeed should be supported in this decision. We evaluated the uptake of these guidelines, and practice variability across providers who care for such mothers and infants. Methods We surveyed Pediatric Infectious Diseases Society members who practice clinically in the US, via an internal email list serve. The ten-question survey assessed uptake of the updated DHHS guidelines and respondents’ clinical practice in three key areas that are subject to interpretation in the guidelines: VL control, mixed feeding, and ART prophylaxis. We used descriptive statistics to examine guideline uptake and variability in practice. We used inductive qualitative thematic analysis to identify themes in open-ended survey responses. Results Of 915 individuals who opened the survey, 165 responded and 129 were included in the final analysis. Respondents represented 86 US clinical centers. Uptake varied: 14 respondents (10.8%) were unaware of the new guidelines, 103 (79.8%) had implemented or were planning to implement the new guidelines, and 12 (9.3%) were not planning to implement the guidelines. The table summarizes variability in pre-delivery VL suppression and mixed feeding criteria used by 117 respondents who are either planning/already implementing guidelines or who are unaware of guidelines. Areas of concern revealed by thematic analysis included insufficient data to support guidelines and lack of specific criteria for implementation. Conclusion We identified substantial heterogeneity in provider uptake and comfort with new breastfeeding guidelines among US pediatric infectious diseases practitioners, and variability in assessment of safety to breastfeed. Further research is essential to explore patient preferences, define necessary follow-up, and address provider concerns regarding insufficient data to support guidelines and lack of specific criteria for implementation.

Antibody-mediated cellular responses are dysregulated in Multisystem Inflammatory Syndrome in Children (MIS-C)

Abstract Background Multisystem Inflammatory Syndrome in Children (MIS-C) is a rare and severe complication of SARS-CoV-2 infection that is characterized by multi-organ involvement and substantial inflammation. The immunological responses in MIS-C are not fully known. Antibody-mediated viral clearance in which innate immune cells interact with antibodies via their Fc receptors to remove the virus is an important mechanism for reducing SARS-CoV-2 viral load. However, little is known about how antibody-mediated cellular responses in MIS-C compared to acute COVID-19 infection and direct testing of cellular function ex vivo to understand the aberrant immune response in MIS-C is limited. Therefore, we focused on the antibody-mediated cellular responses in MIS-C relative to children and adults that had acute COVID-19 infection with spectrum of symptoms. We hypothesized that cells that bind antibodies to help clear the virus are dysfunctional in MIS-C patients, preventing the virus from being fully cleared. Methods We collected samples from MIS-C children along with pediatric and adult subjects with a clinical spectrum of COVID-19 symptoms and uninfected controls. Through these controls, we can compare Fc-mediated antibody functions between ages and clinical symptoms. We utilized high dimension flow cytometry to assess phenotype and function of innate immune cells that bind to antibodies and sequencing to study Fc receptor genetic. This data was then correlated with matched clinical data. Results Monocytes in MIS-C were hyperfunctional for antibody-dependent cellular phagocytosis and production of proinflammatory cytokines. In contrast, natural killer (NK) cells in MIS-C were hypofunctional for antibody-dependent cellular cytotoxicity and cytokine production. To provide some preliminary insight into mechanism, we show multiple ways leading to decreased cytotoxicity for NK cells, including phenotypic exhaustion of NK cells, variants in CD16 (NK cell Fc receptor) genetics that reduce Fc binding to IgG, and cytokine-induced associations. Using a novel anti-CD16 novel bispecific reagent that we developed, we could rescue the hypofunctional antibody-mediated NK cell function in MIS-C to the same levels as control. Conclusion Together, our results reveal dysregulation in antibody-mediated responses that are unique to MIS-C and may contribute to the immune pathology of this disease. Specifically, we find that monocytes are hyperfunctional with NK cells are hypofunctional. In the case of the MIS-C antibody-mediated response, while it is likely that multiple pathogenetic mechanisms contribute to MIS-C, our data suggest an imbalance of the antibody-mediated cellular response as a mechanistic correlate of this rare, but life-threatening outcome of SARS-COV-2 in children. Our results highlight that targeted therapies to reduce viral burden and/or boost NK cells responses while decreasing cytokine levels may be effective in preventing MIS-C.

Cytomegalovirus infection in transplant patients according to pre-transplant risk in a Hospital in Northeastern Mexico

Abstract Background Cytomegalovirus (CMV) is an important cause of morbidity and mortality in patients who receive a hematopoietic stem cell transplant (HSCT). It is possible to stratify the risk of developing the disease due to this agent according to the serological status of the HSCT donor and recipient. In post-transplant patients, significant T lymphocyte dysfunction occurs, so reactivation of CMV is common, which is why it should be monitored after HSCT. Our objective is to determine the association of CMV infection in post-transplant patients according to pre-transplant risk status. Methods Patients who received HSCT from January 2019 to June 2023 were included, they were classified according to their pre-transplant risk status and CMV infection was diagnosed by performing a viral load on day 30. Results 70 patients with hemato-oncological diseases who received an HSCT were included, a median age of 9 years (0-16) was found, predominance of the male sex (60%), the most frequent type of transplant was haploidentical (82.9 %). Of these, 20 (28.5%) had CMV infection, with pre-transplant risk status mostly high risk (85%), of the low-risk patients (25.7%) 10% had the infection and the very high risk (4.3%) only 5% were reported infected. The period during which the infection occurred most frequently was in the early post-pregnancy stage (66.6%) and the majority received treatment with valganciclovir (95%). Of the patients infected with CMV, 15 (75%) were asymptomatic, and 5 (25%) had associated symptoms, with 2 (10%) presenting with urinary symptoms (dysuria, frequency, suprapubic pain, tenesmus), and 2 (10%) with fever, and 1 hematochezia (5%). Regarding clinical outcomes, 17 patients (85%) presented resolution of the disease, 1 (5%) presented therapeutic failure and 1 (5%) died due to multiple organ failure. Conclusion Due to the high prevalence of CMV infection, it is important to maintain a high index of suspicion of this pathology in this population, with the aim of achieving early diagnosis and timely initiation of treatment.