Human Immunodeficiency Virus Research Articles

Acceptability of Moringa oleifera seeds flour supplement-added to normal diet by Women Living With Human Immunodeficiency Virus (PLWHIV) in resource-limited setting

This article aims to assess the acceptability of Moringa oleifera seed supplementation among HIV-positive women in resource-limited settings. The management of HIV/AIDS in resource-constrained settings presents numerous challenges, highlighting the need for affordable supplements to complement standard anti-retroviral therapy (ART). Moringa oleifera, known for its nutritional and medicinal properties, has been noted for its immunomodulatory, anti-inflammatory, and antiviral effects. A descriptive study utilising a Likert-scale survey evaluated participants' familiarity with Moringa oleifera, perceived health benefits, potential risks, ease of dietary integration, and taste acceptability. The findings revealed a generally positive acceptance of Moringa oleifera supplementation, with a mean acceptability score of 2.5750. Notably, 58.2 per cent of participants found the taste palatable, and 72.2 per cent reported ease of incorporation into their daily diet. A significant 90 per cent expressed willingness to use Moringa oleifera if proven effective in suppressing HIV viral load. The study also highlighted the influence of healthcare provider recommendations, with 77.7 per cent of participants more likely to use Moringa oleifera based on professional advice. In conclusion, Moringa oleifera seed supplementation is generally acceptable among People Living with HIV, particularly when supported by healthcare providers. To enhance its use, the study recommends increasing awareness, engaging healthcare professionals, and conducting further research to maximise its potential health benefits for HIV-positive individuals.

Bioactives from marine resources as natural health products: A review.

The oceans are a rich source of a myriad of structurally different and unique natural products that are mainly found in invertebrates with potential applications in different disciplines. Microbial infection and cancer are the leading causes of death worldwide. Discovery of new sources of therapy for microbial infections is an urgent requirement due to the emergence of pathogenic microorganisms that are resistant to existing therapies. Marine bioactives have demonstrated to be promising sources for the discovery and development of novel antimicrobial and anticancer compounds. Several marine compounds are confirmed to have antibacterial effects and most marine-based antifungal compounds are cytotoxic. Numerous antitumor marine natural products, derived mainly from sponges or molluscs, and also bryozoans and cyanobacteria, exhibit potent antimitotic activity. In addition, marine biodiversity offers some possible leads or new drugs to treat human immunodeficiency virus (HIV). A majority of marine derived drugs are currently in clinical trials or under preclinical evaluation. Furthermore, marine-based drugs, approved by the US Food and Drug Administration (FDA) are available in the market. This review summarizes the sources, mechanisms of action and potential utilization of marine natural products such as peptides, alkaloids, polyketides, polyphenols, terpenoids and sterols as antifungal, antibacterial, antiviral, and anticancer compounds. Significance Statement Utilization of marine bioactives as natural health products leads to crucial advancement in providing dietary supplements, nutraceuticals, functional foods and pharmaceuticals. Their myriad of application promotes health and plays a role in disease risk reduction. Therapeutic potential of potent compounds from marine organisms and use of their bioactives have promising medicinal value for preventing ailments and advancing pharmaceutical and nutraceutical industries. Their utilization benefits human health globally and contributes to the conservation of marine ecosystem in a transformative / sustainable approach.

SPECIES IDENTIFICATION AND IN VITRO ANTIFUNGAL SUSCEPTIBILITY TESTING OF ASPERGILLUS ISOLATED FROM VARIOUS CLINICAL SAMPLES

Objectives: Aspergillus is ubiquitous mold species present in environment in the form of spores. Infection due to Aspergillus is uncommon in immunocompetent individuals unless they possess any abnormality or have undergone any treatment with corticosteroids. In immunocompromised individuals, infection by this fungus is common among people with prolonged neutropenia, transplants, and human immunodeficiency virus infection. The symptoms are diverse, ranging from allergic reactions to invasive aspergillosis and life-threatening complications. The aim of this study is to isolate and identify Aspergillus species obtained from various clinical specimens and perform antifungal susceptibility testing (AFST). Methods: A total of 200 isolates in which positive direct microscopic findings correlated well with culture growth for Aspergillus were included. Antifungal susceptibility was performed by micro broth dilution technique according to clinical laboratory standard institute M38-A2 guidelines. Results: Aspergillus flavus was found to be predominant species followed by Aspergillus fumigatus. AFST was performed, where all Aspergillus strains exhibited low minimum inhibitory concentration (MIC) and minimum effective concentration (MEC) values for most of antifungal drugs tested except for one strain of A. flavus, which exhibited high MIC for voriconazole and high MEC for caspofungin. Conclusion: Polyenes, azoles, and echinocandin resistance are emerging in many parts of the world. Therefore, continued application of AFST combined with morphological characterization for species identification is critical in detecting the emergence of resistance among various Aspergillus species to reduce mortality, morbidity, and overcome treatment failure.

Pityriasis rubra pilaris: A review

Pityriasis rubra pilaris (PRP, syn.: Devergie's disease) is a rare idiopathic papulo-squamous inflammatory skin disease characterized by follicular papules, palmar-plantar keratoderma, scaling plaques of orange-red color with characteristic foci of unaffected skin. Histologically, the alternating pattern of orthokeratosis and parakeratosis is considered a distinctive feature of PRP (staggered hyperkeratosis). A violation of the regulation of innate immunity is a component of the pathogenesis of PRP. Congenital mutations of CARD 14 or concomitant antigens, which can take the form of iatrogenic lesions, infections or malignant neoplasms, can be the initial triggers of the disease, although the etiology is often idiopathic. There are classically five subtypes of the disease, depending on the age of onset and clinical picture. Type VI PRP is associated with infection caused by the human immunodeficiency virus. CARD 14-associated papulosquamous rash and discoid dermatitis of the face represent new clinical phenotypes of PRP. Devergie's disease has a pronounced negative impact on the quality of life, and those who become ill are at increased risk of depression and suicide. Ixekizumab, methotrexate and secukinumab can be considered as systemic drugs of the first choice. If there is a contraindication to immunosuppressive therapy, high doses of isotretinoin are recommended. Topical agents, including calcipotriene, calcineurin inhibitors, emollients and topical corticosteroids, are used as additional therapy.

Changes in early HIV/AIDS mortality rates in people initiating antiretroviral treatment between 2013 and 2023: A 10-year multicenter survival study in Senegal

BackgroundHIV/AIDS-related early mortality has long been a significant challenge. Subsequent to recent policy changes and treatment advancements, we aimed to assess changes in early mortality rates in 2017–19 and 2020-23 compared to 2013–16. MethodsThis is a 10-year multicenter survival study in people living with human immunodeficiency virus having initiated ART between 2013 and 2023. We used frailty-based competing risk models to estimate adjusted early (6-month and one-year) mortality hazard ratios (HRs) in people living with HIV (PwHIV) having initiated ART in 2013–16 (comparator), 2017–19, and 2020–23. ResultsWe enrolled 4006 persons of whom 2281 (56.9 %) were female; median age was 40 years (IQR: 31–50); 635 (15.9 %) were at WHO clinical stage IV and 934 (23.3 %) had a CD4 count <200 cells/mm3. Median follow-up was 80.4 months (IQR: 48.6–106.7). All in all, 463 participants died (4.37 deaths per 100 person-years), including 296 at one year of follow-up (7.4 % [95 % CI: 6.6–8.2]). ART initiation in 2016–19 and 2020–23 was associated with 27 % (adjusted HR [aHR]: 0.73; 95 % CI: 0.55–0.98) and 63 % (aHR: 0.37; 95 % CI: 0.25–0.56) reductions in one-year mortality rates, respectively, compared to the 2013–16 period. ConclusionEarly mortality risk has significantly decreased over time in Senegal. However, the proportion of PwHIV with AIDS-defining conditions remains concerning. Continued efforts to ensure early diagnosis and prompt linkage to care are needed for more impact.

7660 Navigating Hypercalcemia: MAC Treatment Hurdles and the Unforeseen Impact of Vitamin D Supplementation

Abstract Disclosure: S. Noguchi: None. S. Lai: None. A. Rajpal: None. Introduction: Hypercalcemia is a recognized complication in granulomatous disorders such as sarcoidosis and tuberculosis, resulting from increased 1 alpha-hydroxylase activity in macrophages leading to excessive conversion to 1,25(OH)2 vitamin D (calcitriol). Mycobacterium avium complex (MAC) is an infectious granulomatous disease often suspected in immunocompromised individuals, including those with human immunodeficiency virus (HIV). Additionally, vitamin D deficiency is prevalent in the general population, prompting supplementation. We hereby present a patient with advanced HIV and disseminated MAC who developed severe calcitriol-induced hypercalcemia in the setting of high dose vitamin D supplementation. Case description: A 48-year-old male with advanced HIV (CD4 count &amp;lt;20 cells/mm^3) presented for evaluation of abdominal pain and constipation, and was found to have severe hypercalcemia of 14.8 mg/dL. Five months prior, he had restarted anti-retroviral therapy (ART) after having been out of care for over a year and was diagnosed with disseminated MAC for which he was on azithromycin and ethambutol. Cultures drawn during this admission were negative for MAC. Hypercalcemia was possibly due to paradoxical immune reconstitution inflammatory syndrome (IRIS) and with appropriate management his calcium level rapidly normalized to 9.7 mg/dL and remained stable until the recent admission. He represented one year later with a calcium level of 13.8 mg/dL with suppressed parathyroid hormone levels, which was refractory to bisphosphonate therapy, therefore was empirically initiated on dexamethasone. By day 4, his calcium levels improved significantly and was discharged on a taper regimen. Cultures collected this admission later revealed ongoing MAC infection, which raised concern for treatment failure and calcitriol-induced hypercalcemia. Calcitriol levels were elevated at 105 pg/mL, which further supported this hypothesis. Additionally, prior to admission he had been prescribed ergocalciferol 50,000 units weekly for 8 weeks, after which he was transitioned to 2,000 units daily. This likely exacerbated the calcitriol-induced hypercalcemia by providing sudden availability of substrate, 25-OH vitamin D, for the activated macrophages. Conclusion: ART has reduced the incidence of opportunistic infections such as MAC, however mortality remains high. Vigilant monitoring for complications, particularly life-threatening calcitriol-induced hypercalcemia and timely use of corticosteroids, is crucial. This case also highlights the need for careful oversight in high-dose vitamin D supplementation for individuals with granulomatous disease and vitamin D dysregulation. Presentation: 6/2/2024

6411 Ozempic - Oh no! Acute Renal Failure Associated with Initiation of Semaglutide

Abstract Disclosure: K. Barney: None. D. Tahir: None. A. Pannu: None. R. Saeed: None. Background: Semaglutide is a glucagon-like peptide-1 (GLP-1) receptor agonist which has been proven to reduce weight in those with obesity and improve glycemic control in patients with type II diabetes mellitus. The effect of semaglutide on kidney function has not been well established. The phase 3b SUSTAIN trials revealed that when compared with other GLP-1 agonists, semaglutide exhibited more frequent acute kidney injuries [1-3]. The SUSTAIN 6 trial reported two instances of acute renal failure (ARF) in patients with chronic kidney disease, and one case of ARF in a patient with normal kidney function who was treated with semaglutide [3]. Despite these reports, the SUSTAIN 1-9 trials report no significant decline in kidney function associated with semaglutide [3-6]. Case: We present the case of a 58-year-old male with a past medical history of human immunodeficiency virus (HIV), type II diabetes mellitus, and hypertension who presented to the hospital with ARF after initiation of Ozempic (semaglutide) two months prior to presentation. On admission, vitals were stable and physical examination was benign. His pertinent labs were blood urea nitrogen (BUN) 55 mg/dL, creatinine 5.94 mg/dL (baseline 1.2 mg/dL), and urinalysis with 2+ protein. To determine the cause of his renal failure, urine sodium and creatinine were obtained to calculate his fractional excretion of sodium, which was found to be consistent with intrinsic renal disease. Further workup, including renal ultrasound, urine protein, urine eosinophils, complement and free light chain levels, and creatinine kinase, were unrevealing for cause of intrinsic renal injury. He was managed with intravenous fluids and his creatinine improved to 2.03 mg/dL. Given the acute onset of the patient’s renal failure and recent initiation of Ozempic, it was determined that this medication was the cause of his renal failure, and it was discontinued on discharge. He has since followed up with nephrology and endocrinology. He remains off Ozempic and is doing well. Conclusions: Our case highlights a rare but significant adverse effect of semaglutide on renal function. As the use of semaglutide has increased due to its rising popularity for weight loss and glycemic control in type II diabetes mellitus, the potential for renal injury should be considered and discussed with patients. Furthermore, it is imperative that semaglutide associated renal injury be considered as a cause for ARF in patients taking this medication. This case demonstrates that instances of ARF associated with semaglutide treatment are a cause for concern and further exploration. Presentation: 6/1/2024

The relationship between HIV/AIDS and coronary heart disease: A bibliometric analysis.

Acquired immunodeficiency syndrome is a malignant infectious disease caused by the human immunodeficiency virus (HIV). HIV gradually destroys the body's immune system and weakens the body's ability to resist diseases. People living with HIV may have a higher incidence of coronary heart disease than people without HIV. A literature retrieval from January 1, 1993 to October 1, 2023 based on the Web of Science Core Collection database. CiteSpace6.2.R4, VOSviewer v1.6.19, and Microsoft Excel 2019 were utilized for analyzing the following terms: countries, institutions, authors, journals, references, and keywords. There were 1144 articles. The highest number of articles is in the USA, followed by Italy. University of California System, Harvard University, and Johns Hopkins University were the top 3 most productive institutions with publications in this field of research. Journal of Infectious Diseases ranked first with the highest publications (532 records), followed by Immunology (362 records), and Cardiac Cardiovascular Systems (242 records). Keyword co-occurrence analysis showed antiretroviral therapy, myocardial infarction, and protease inhibitors, etc. Keyword cluster analysis obtained 13 categories, which were roughly divided into 3 themes: (1) cardiovascular disease that has occurred or may occur; (2) HIV acquisitions that have occurred; (3) risk factors for cardiovascular disease. The article obtained the hotspots and trends and provided references for subsequent research. Based on the keyword citation burst detection analysis, we speculated that heart failure, risk, subclinical atherosclerosis, infection, and association were the research hotspots in recent years, which had a certain predictive effect on the future research direction.

Differential T-cell profiles determined by Hepatitis B surface antigen decrease among people with Human Immunodeficiency Virus /Hepatitis B Virus coinfection on treatment

BackgroundSeveral studies have reported that combination antiretroviral therapy (cART) enhances the hepatitis B surface antigen (HBsAg) clearance rate in Human Immunodeficiency Virus-1/Hepatitis B Virus (HIV/HBV) coinfected patients, yet the associated immunological characteristics remain unclear.MethodsGlobal and specific immune phenotypic profiles were examined in 48 patients with HIV/HBV coinfection before cART and at 1-year, and 3-year after cART using flow cytometry. In addition, 61 patients with HBV monoinfection were included for comparison.ResultsHBsAg response (sAg-R) was defined as > 0.5 log decrease within six months of cART initiation, and 16 patients achieved it. Patients with sAg-R (the sAg-R group) exhibited distinct immune phenotypes compared to those of HBsAg-retained patients (the sAg-NR group). Notably, patients with sAg-R had lower CD4+ T cell counts and a higher number of HBcAg-specific T cells. Further, the sAg-R group exhibited upregulation of HLA-DR, Ki67, and PD-1 in CD4+ T cells and heightened HLA-DR and T-bet in CD8+ T cells. However, the sAg-R group had fewer TEMRA cells but more TEM and Th17 cells than those in the sAg-NR group. Expression of various markers, including HLA-DR+CD4+, Ki67+CD4+, PD-1+CD4+, CD38+CD8+, HLA-DR+CD8+, TIM-3+CD8+, HBV-specific CD4+ T cell secreting IFN-γ and IL-2, and specific CD8+ T cell secreting IFN-γ and IL-2, correlated with HBsAg decrease.ConclusionThe decline in HBsAg levels during cART in HIV/HBV coinfection involves significant alterations in CD4+ and CD8+ T cells phenotypes, offering a novel perspective on a functional HBV cure.

Large expansion of plasma commensal viruses is associated with SIV pathogenesis in Macaca leonina.

Human immunodeficiency virus-1 (HIV-1) infection disrupts the homeostatic equilibrium between the host and commensal microbes. However, the dynamic changes of plasma commensal viruses and their role in HIV/simian immunodeficiency virus (SIV) pathogenesis are rarely reported. Here, we investigated the longitudinal changes of plasma virome, inflammation levels, and disease markers using an SIV-infected Macaca leonina model. Large expansions of plasma Anelloviridae, Parvoviridae, Circoviridae and other commensal viruses, and elevated levels of inflammation and D-dimer were observed since the chronic phase of SIV infection. Anelloviridae abundance appears to correlate positively with the CD4+ T cell count but negatively with SIV load especially at the acute phase, whereas other commensal viruses' abundances show opposite correlations with the two disease markers. Antiretroviral therapy slightly reduces but does not substantially reverse the expansion of commensal viruses. Furthermore, 1387 primate anellovirus open reading frame 1 sequences of more than 1500 nucleotides were annotated. The data reveal different roles of commensal viruses in SIV pathogenesis.

Antibiotic resistance and draft genome profiles of 10 Streptococcus pneumoniae and 3 Streptococcus pseudopneumoniae strains isolated from the nasopharynx of people living with human immunodeficiency virus in Ghana.

Genomic data on clinically important bacteria species such as Streptococcus pneumoniae and Streptococcus pseudopneumoniae from low- and middle-income countries, including Ghana, are scarce. In this study, we provide data on antimicrobial resistance (AMR) and whole-genome profiles of a collection of streptococci species to support AMR surveillance efforts in the country.

Immune triggers preceding neuralgic amyotrophy.

Infections and vaccinations have been identified as potential immunological triggers of neuralgic amyotrophy (NA), but the exact type and frequency of the preceding agents is unknown. This was a multicentre, prospective, observational, matched case-control study. NA was diagnosed by neuromuscular experts according to validated clinical criteria and electrodiagnostic studies. Clinical data and biological samples of NA patients were collected within 90 days from disease onset between June 2018 and December 2023. All NA patients were asked about prior infection and vaccination in the month before disease onset. Serological tests for hepatitis E virus, human immunodeficiency virus, severe acute respiratory syndrome coronavirus 2, Epstein-Barr virus, cytomegalovirus, parvovirus B19, varicella-zoster virus, Borrelia burgdorferi, Mycoplasma pneumoniae and Bartonella henselae were performed in a central laboratory. Each case was matched with a healthy control for age, sex, place of residence and time of blood collection. Fifty-seven patients and corresponding controls were included. The mean age was 45 years for both groups. NA onset was preceded by a symptomatic infectious trigger confirmed by microbiological tests in 15/57 (26.3%) patients. Coronavirus disease 2019 vaccination was considered a potential trigger in 7/57 (12.3%) subjects. An acute viral infection was associated with a bilateral involvement of the brachial plexus (p = 0.003, Cramèr's V = 0.43). Confirmed immune triggers (infection or vaccination) preceded disease onset in 22/57 (38.6%) NA cases. We suggest to test NA patients in the acute phase for intracellular antigens, especially in the case of concomitant bilateral involvement and hepatitis.

Plasmablastic lymphoma in an HIV positive patient: colonic disease as an unusual presentation.

Plasmablastic lymphoma (PBL) is an aggressive and rare variant of diffuse large B-cell lymphoma associated with human immunodeficiency virus (HIV) infection. Here, we describe a case of PBL of the ascending colon in an HIV positive patient.

MicroRNA Expression in Exosome Extracellular Vesicles as Targets for Personalized Medicine in Diffuse Large B Cell Lymphoma Patients with HIV Infection.

Extracellular Vesicles (EVs), more specifically exosomes (xEVs), have been associated with Diffuse Large B-cell Lymphoma (DLBCL). These xEVs contain a variety of biomolecules, such as proteins and nucleic acids (e.g., microRNA, LncRNA, and DNA). The expressions of these vesicles in the setting of Human Immunodeficiency Virus (HIV) have been linked to disease progression. Studies have explored the use of EVs in more practical clinical settings. Several studies have found that biomolecules within xEVs can serve to detect disease progression. The biomolecule content within xEVs is useful in prognostication and has even been associated with mechanisms of resistance for some DLBCL treatment modalities. This review article explores the role of xEV biomolecule content in DLBCL progression in the context of HIV infection and its applied use in practical disease management.

Review Based Study of Prevalence and Association of Social Determinants with Tuberculosis in Doda Region of Jammu and Kashmir, India

Tuberculosis (TB) continues to rank among the world’s most serious health problems caused by Mycobacterium tuberculosis. There are several factors which are responsible for the exposure of tuberculosis among the communities. Human immunodeficiency virus (HIV) is one of the major factors for the exposure of tuberculosis disease among the persons. Worldwide, TB is one of the leading causes of death among people living with HIV. Tuberculosis is a serious health threat, especially for the people living with HIV. The socio-economic determinants such as unemployment, malnutrition and crowding play a significant role in increasing the rate of infection. In this article, we will discuss the socio-economic determinants of tuberculosis infection and disease. The rapidly growing consensus indicates that in the low and middle income country, the tuberculosis control programs are required with the investment in tuberculosis diagnostics and treatment in addition to action on the social determinants of tuberculosis. Tuberculosis is most common among the disadvantaged group which includes the poor, ethnic minorities as well as hungry.

Study of the protective efficacy of CombiMab-2 against human immunodeficiency virus type 1 in mice humanised with CD4&lt;sup&gt;+&lt;/sup&gt; T-lymphocytes

INTRODUCTION. Despite existing treatment methods, complete eradication of human immunodeficiency virus (HIV) infection remains an unattainable goal due to the high variability of HIV type 1 (HIV-1). HIV infection necessitates life-long administration of antiretroviral medicinal products, which cause serious adverse drug reactions. The development of gene therapy products based on adeno-associated virus (AAV) vectors encoding broadly neutralising antibodies represents a promising direction for creating long-term therapies capable of countering a wide range of viral variants.AIM. This study aimed to evaluate the protective efficacy of CombiMab-2, a medicinal product consisting of a combination of three AAV vectors (AAV9-VRC07-523, AAV9-10-1074, and AAV9-PGDM1400) encoding broadly neutralising antibodies against HIV-1, in a humanised mouse model.MATERIALS AND METHODS. The study used an HIV infection model based on immunodeficient B-NDG mice humanised with human CD4+ lymphocytes (1.5×107 cells per animal) from a leukoconcentrate of a healthy donor. The experiment used two groups of mice, including a control group (3 animals) receiving saline solution and an experimental group (5 animals) receiving CombiMab-2. The medicinal product was administered into different muscles as three separate components six weeks prior to infection. The CCR5-tropic HIV-1 strain was obtained by transfecting HEK293FT cells with the pNL4-3(AD8) plasmid encoding the full-length virus. The authors monitored viral loads in the plasma of animals by reverse transcription polymerase chain reaction and CD4+ lymphocyte counts in the peripheral blood of animals by flow cytometry for four weeks after infection.RESULTS. Six weeks after CombiMab-2 administration, the levels of broadly neutralising antibodies in the serum of humanised mice ranged from 0.17 μg/mL to 4.0 μg/mL. In the control group, the viral load reached 105 copies/mL one week after HIV-1 infection and continued to rise over the next three weeks. In the treatment group, infection developed only in one mouse, which had the lowest antibody titre before infection. No viral load was detected in the remaining mice of the treatment group, which indicated that the medicinal product was effective if serum concentrations of broadly neutralising antibodies reached 0.5 μg/mL or higher.CONCLUSIONS. The tested medicinal product based on three AAV vectors (AAV9-VRC07-523, AAV9-10-1074, and AAV9-PGDM1400) exhibits protective activity against HIV-1 in humanised mice. The presented data allow the authors to consider CombiMab-2 as a promising antiviral agent that can serve as a basis for further pharmaceutical development.

Determinants of Human Immuno Deficiency Virus (HIV) Prevalence in the Central Region of Ghana

Determinants of Human Immuno Deficiency Virus (HIV) Prevalence in the Central Region of Ghana

Host cell glycosylation selects for infection with CCR5- versus CXCR4-tropic HIV-1.

Human immunodeficiency virus type 1 (HIV-1) infection involves a selection bottleneck that leads to transmission of one or a few variants. C-C motif chemokine receptor 5 (CCR5) or C-X-C motif chemokine receptor 4 (CXCR4) can act as coreceptors for HIV-1 viral entry. However, initial infection mostly occurs via CCR5, despite abundant expression of CXCR4 on target cells. The host factors that influence HIV-1 susceptibility and selection during transmission are unclear. Here we conduct CRISPR-Cas9 screens and identify SLC35A2 (a transporter of UDP-galactose expressed in target cells in blood and mucosa) as a potent and specific CXCR4-tropic restriction factor in primary target CD4+ T cells. SLC35A2 inactivation, which resulted in truncated glycans, not only increased CXCR4-tropic infection levels but also decreased those of CCR5-tropic strains consistently. Single-cycle infections demonstrated that the effect is cell-intrinsic. These data support a role for a host protein that influences glycan structure in regulating HIV-1 infection. Host cell glycosylation may, therefore, affect HIV-1 selection during transmission in vivo.

B-224 Rapid, Extraction-Free, and Portable Molecular Detection of HIV-1 and HCV Directly From Whole Blood

Abstract Background Early diagnosis of human immunodeficiency virus-1 (HIV-1) and hepatitis C virus (HCV) is key to preventing viral transmission and improving linkage to treatment. Isothermal amplification methods, such as reverse transcription looped-mediated amplification (RT-LAMP), have enabled molecular diagnostics to expand beyond centralized laboratories. Despite the advances in molecular point-of-care (POC) testing, many of these tests are costly and still depend on a reliable power-source, specialized equipment, and cold-chain storage, making them impractical for resource-limited settings. To overcome these barriers, there is a need for innovative molecular diagnostics for blood-borne pathogens. Here, we propose a simple, extraction-free, portable workflow for detecting HIV-1 and HCV from whole blood within 40 minutes. Methods We developed singleplex RT-LAMP-based tests using modified published primer sets. Contrived whole blood samples containing HIV-1 or HCV virions were diluted in equal parts water and loaded directly into optimized RT-LAMP master mixes. To mitigate cold-chain storage dependence, RT-LAMP reactions were performed using a lyophilized master mix. The reactions were heated for 30 minutes using a hand-held, battery-powered heating device for simultaneous virion lysis and amplification. For simple visual detection, amplification was coupled with a lateral flow-based dipstick test. The analytical sensitivity of each detection test was evaluated using contrived whole blood samples ranging in HIV-1 and HCV 1a viral loads. Additionally, HCV detection was evaluated on genotypes 1b, 2b, and 3a. For result confirmation, a custom CRISPR-Cas12a-based assay was used to verify the presence or absence of pathogen-specific amplicons following amplification. Results The diagnostic workflow can be completed within 40 minutes, including less than 10 minutes of hands-on time. At optimal conditions, the HIV-1 and HCV 1a singleplex tests had lower limits of detections of 4.89 log10 cp/mL and 3.77 log10 IU/mL, respectively. Evaluation of the published literature showed that these viral loads are within the ranges observed during acute HIV-1 and HCV infection. HCV detection was also observed for genotypes 1b and 3a, albeit at a lower sensitivity than genotype 1a. No detection was observed for genotype 2b, presumably due to the lower viral loads of the samples. No cross-reactivity between the two tests was observed. Conclusions Our proposed workflow allows for rapid detection of blood-borne pathogens without the need for complex equipment or cold-chain storage, showing potential for application in resource-limited settings. Additional validation within a clinical setting using fingerstick blood remains to be tested.

A-248 Assessment of Nanopore Sequencing for HIV Genotyping: A Comparative Study with Sanger Sequencing

Abstract Background The HIV drug resistance test, conducted through genotyping of the Human Immunodeficiency Virus (HIV), is a critical tool for assessing drug resistance in virus-infected patients and is commonly performed using Sanger Sequencing methodology (first-generation sequencing). Nanopore sequencing (third-generation sequencing) is a new technology that started to be commercialized a few years ago and has recently undergone significant updates with the new Q20+ chemistry. Therefore, this study aims to evaluate the potential use of nanopore sequencing (MinION) for HIV genotyping. Methods The genomic region encoding HIV protease and transcriptase was amplified by nested PCR using primer pairs modified with unique barcodes in the second round of amplification. Three libraries were assembled, including samples from 22 HIV-positive patients, one negative control (negative control patient), and one no-template control (NTC) per library, totaling 66 HIV-positive samples. The libraries were constructed using the SQK-LSK114 kit and sequenced on a MinION platform using the new Q20+ chemistry in flowcells or flongle R10.4.1. The sequences were assembled and analyzed using HIVdb-NGS (Stanford). Validation tests, such as accuracy and precision, were conducted following WHO recommendations for HIV genotyping. The same samples were also sequenced using the Sanger method for comparison. Results 62 HIV sequences were generated by the MinION and analyzed in the described pipeline. The average sequence coverage per sample was 25,866 x. 41 sequences were classified as type B (63.08%), 12 as type C (18.46%), 8 as type F (12.31%), three as CRF02_AG (4.62%), and 1 as a recombinant B+C (1.54%). All identified subtypes were consistent with those found by the Sanger method. The average similarity between the sequences of each sample (MinION vs Sanger) was 98.68% (minimum 94.96%, maximum 99.95%). Basecalling similarity at relevant positions (associated with resistance) was above 98% in 59 out of 62 samples (95.16%), meeting the acceptance criteria recommended by the WHO. Conclusions Our results suggest that nanopore sequencing (utilizing Q20+ chemistry) can replace Sanger sequencing for HIV genotyping, which is used to monitor the effectiveness of anti-HIV drugs. This transition will allow for a reduction in test costs in laboratories with high demand for HIV drug resistance testing. Additionally, the use of modified primers proposed here eliminates the need for commercial barcodes, resulting in reduced processing time and improved throughput.