Human Immunodeficiency Virus P24 Research Articles

HIV-1 Tat is present in the serum of people living with HIV-1 despite viral suppression

ObjectivesDespite successful human immunodeficiency virus (HIV) control with combination antiretroviral therapy (cART), individuals with HIV still face health risks, including cancers, cardiovascular and neurocognitive diseases. An HIV protein, Tat, is potentially involved in these HIV-related diseases. Previous studies demonstrated circulating Tat in the blood of untreated people with HIV. Here, we measured Tat levels in the serum of cART-treated people with HIV to examine the effect of cART on Tat production. MethodsSerum samples from 63 HIV-positive and 20 HIV-seronegative individuals were analyzed using an ELISA assay that detected Tat concentrations above 2.5 ng/mL. ResultsAmong HIV-positive individuals, the Tat level ranged from 0 to 14 ng/mL. 25.4% (16 out of 63) exceeded the 2.5 ng/mL cut-off, with a median HIV Tat level of 4.518 [3.329-8.120] ng/mL. No correlation was revealed between Tat levels and CD4+ T cell counts, serum HIV RNA, p24 antigen, or anti-Tat levels. ConclusionsDespite cART, circulating HIV Tat persists and may contribute to HIV-related diseases. This emphasizes the need for further research on the mechanisms of Tat action in non-infected cells where it can penetrate upon circulation in the blood.

Sensitive Blood-Based Detection of HIV-1 and Mycobacterium tuberculosis Peptides for Disease Diagnosis by Immuno-Affinity Liquid Chromatography-Tandem Mass Spectrometry: A Method Development and Proof-of-Concept Study.

Novel approaches that allow early diagnosis and treatment monitoring of both human immunodeficiency virus-1 (HIV-1) and tuberculosis disease (TB) are essential to improve patient outcomes. We developed and validated an immuno-affinity liquid chromatography-tandem mass spectrometry (ILM) assay that simultaneously quantifies single peptides derived from HIV-1 p24 and Mycobacterium tuberculosis (Mtb) 10-kDa culture filtrate protein (CFP10) in trypsin-digested serum derived from cryopreserved serum archives of cohorts of adults and children with/without HIV and TB. ILM p24 and CFP10 results demonstrated good intra-laboratory precision and accuracy, with recovery values of 96.7% to 104.6% and 88.2% to 111.0%, total within-laboratory precision (CV) values of 5.68% to 13.25% and 10.36% to 14.92%, and good linearity (r2 > 0.99) from 1.0 to 256.0 pmol/L and 0.016 to 16.000 pmol/L, respectively. In cohorts of adults (n = 34) and children (n = 17) with HIV and/or TB, ILM detected p24 and CFP10 demonstrated 85.7% to 88.9% and 88.9% to 100.0% diagnostic sensitivity for HIV-1 and TB, with 100% specificity for both, and detected HIV-1 infection earlier than 3 commercial p24 antigen/antibody immunoassays. Finally, p24 and CFP10 values measured in longitudinal serum samples from children with HIV-1 and TB distinguished individuals who responded to TB treatment from those who failed to respond or were untreated, and who developed TB immune reconstitution inflammatory syndrome. Simultaneous ILM evaluation of p24 and CFP10 results may allow for early TB and HIV detection and provide valuable information on treatment response to facilitate integration of TB and HIV diagnosis and management.

Electrochemical Biosensor for the Rapid Detection of HIV P24 Protein

Human immunodeficiency virus (HIV) is a deadly virus that propagates rapidly. It leads to acquired immunodeficiency syndrome (AIDS) disease with time. Early detection is one way to decrease the spread of the virus. P24 protein is the capsid protein of HIV available for detection in one’s body two weeks after the virus infection. A nickel-based electrochemical sensor is developed to detect the P24 protein using electrocatalysis. A rotating disk electrode is used to minimize the mass limitations. Also, the sensor is label-free which limits the dependence on immobilization. Cyclic voltammetry (CV), chronoamperometry, and electrochemical impedance spectroscopy (EIS) are used throughout the detection process. The electrochemical method CV is used to produce the catalyst layer on the nickel surface, which paves the way for the other two methods to monitor the current response and resistance of the P24 protein with the nickel surface. The sensor detects the presence of HIV P24 protein in the 1 ms of chronoamperometry. Also, the total assay time is less than other conventional methods. The sensor showed good sensitivity and a lower detection limit of around 70 ng/mL, which makes the sensor more reliable. Also, the sensor showed a linear current response with the increasing concentration of P24 protein. So, this simple biosensor has applications to decrease the infections of HIV.

Controlling the Binding Efficiency of Surface Confined Antibodies through the Design of Mixed Self‐Assembled Monolayers

AbstractA plethora of different electronic and optoelectronic devices have been developed lately, for biosensing applications (e.g., label‐free, fast, and easier to operate) based on a detecting interface accommodating the biorecognition elements, anchored by thiolate self‐assembled monolayers (SAMs) on a gold surface. Here, a surface plasmon resonance (SPR) characterization of anti‐p24 anchored on different SAMs is performed to investigate the effect of the SAM structure on the antibodies’ packing efficiency and the sensors’ analytical figures of merit. Notably, the mixed SAM deposited from a solution 10:1 of 3‐mercaptopropionic acid and 11‐mercaptoundecanoic acid (11MUA) is compared to that resulting from a solution 10:1 of ad hoc synthesized N‐(2‐hydroxyethyl)‐3‐mercaptopropanamide (NMPA)/11MUA. Despite the improvement in the anti‐p24 surface coverage registered using the 11MUA/NMPA SAM, the latter produces a significant decrease in the antibodies’ binding efficiency against human immunodeficiency virus p24 protein. To provide a molecular rationale behind the SPR data, density functional theory calculations are also undertaken. A comprehensive physical view of the main competing phenomena affecting the biorecognition events at a biofunctionalized gold detecting interface is represented here.

Human Immunodeficiency Virus in Adults Undergoing Surgery for Brain Tumors.

IntroductionEarly diagnosis and treatment of human immunodeficiency virus (HIV) can improve outcomes. HIV prevalence in brain tumor patients and the impact of an HIV diagnosis on patient outcomes are poorly understood.Materials and methodsThis was a prospective study of 100 consecutive brain tumor patients admitted to a Greater London Tertiary Neurosurgical center for surgery between January 2021 and October 2021. All brain tumors were included. All patients have appropriately consented. Blood was tested to detect HIV antibodies and p24 antigen. Outcomes were noted at 30-day postoperative follow-up.ResultsIn 100 patients, there was one case of a known HIV-positive, seronegative patient, and no new diagnosis was made, giving a prevalence of 1%. The mean age of patients included was 61.7 ± 13.3, with 57% female. The patient with HIV suffered a postoperative pseudomonas infection, requiring intensive care, additional surgery, and antibiotics. This resulted in an inpatient stay of 55 days - an increase of 274% compared to patients without HIV.ConclusionLiterature regarding the prevalence of HIV in glioma patients is inconclusive, of low quality, and primarily out-of-date. Our literature search found no similar study of rates of HIV in brain tumor patients in the United Kingdom. The incidence of both HIV and brain tumors, particularly glioblastomas, is low.

Visible 405 nm Violet-Blue Light Successfully Inactivates HIV-1 in Human Plasma.

Despite significant advances in ensuring the safety of the blood supply, there is continued risk of transfusion transmitted infections (TTIs) from newly emerging or re-emerging infections. Globally, several pathogen reduction technologies (PRTs) for blood safety have been in development as an alternative to traditional treatment methods. Despite broad spectrum antimicrobial efficacy, some of the approved ultraviolet (UV) light-based PRTs, understandably due to UV light-associated toxicities, fall short in preserving the full functional spectrum of the treated blood components. As a safer alternative to the UV-based microbicidal technologies, investigations into the use of violet-blue light in the region of 405 nm have been on the rise as these wavelengths do not impair the treated product at doses that demonstrate microbicidal activity. Recently, we have demonstrated that a 405 nm violet-blue light dose of 270 J/cm2 was sufficient for reducing bacteria and the parasite in plasma and platelets suspended in plasma while preserving the quality of the treated blood product stored for transfusion. Drawn from the previous experience, here we evaluated the virucidal potential of 405 nm violet-blue light dose of 270 J/cm2 on an important blood-borne enveloped virus, the human immunodeficiency virus 1 (HIV-1), in human plasma. Both test plasma (HIV-1 spiked and treated with various doses of 405 nm light) and control plasma (HIV-1 spiked, but not treated with the light) samples were cultured with HIV-1 permissive H9 cell line for up to 21 days to estimate the viral titers. Quantitative HIV-1 p24 antigen (HIV-1 p24) levels reflective of HIV-1 titers were measured for each light dose to assess virus infectivity. Our results demonstrate that a 405 nm light dose of 270 J/cm2 is also capable of 4–5 log HIV-1 reduction in plasma under the conditions tested. Overall, this study provides the first proof-of-concept that 405 nm violet-blue light successfully inactivates HIV-1 present in human plasma, thereby demonstrating its potential towards being an effective PRT for this blood component safety.

Characterization and distribution of HIV-infected cells in semen

ABSTRACT Semen is a known vector for both human immunodeficiency virus (HIV) infection and transmission. However, the distribution and characteristics of HIV-infected cells in semen remain unclear. Investigating the possibility of transmission through the spermatozoon in semen is of great clinical significance to improve the strategies for exposure prevention and assisted reproduction for HIV-infected partners. Twenty-six HIV-infected patients, including twelve treatment-naïve (TN) patients and fourteen antiretroviral treated (ART) patients, were enrolled in this study. HIV p24 protein in spermatozoa was detected using imaging flow cytometry and immunohistochemistry, and HIV RNA was identified using next-generation RNAscope in situ hybridization. Additionally, we described the rates of HIV-positive spermatozoon and CD4+ T lymphocytes in semen, and found that p24+ spermatozoon were mainly CD4 negative regardless of whether the patients received ART. Of note, p24-positive cells in semen are predominantly spermatozoa, and we confirmed that motile spermatozoa carried HIV into peripheral blood mononuclear cells of healthy men in vitro. Our findings provide evidence regarding the risk of HIV-infected spermatozoa.

Paradigms in HIV vaccine research

Although a large number of preventative human immunodeficiency virus (HIV) vaccine trials have been carried out during the last 30 years, it is remarkable that an effective HIV vaccine has not yet been developed. Research paradigms correspond to theoretical assumptions and particular strategies that scientists use when they try to solve a particular problem. Many paradigms used successfully in vaccinology were ineffective with HIV. For instance: 1) The structure-based reverse vaccinology approach failed because investigators tried to generate a vaccine starting with the antigenic structure of HIV-envelope (Env) epitopes bound to neutralizing monoclonal antibodies (mAbs) derived from HIV-infected individuals. They assumed that this antigenic structure would also possess the immunogenic capacity of inducing in vaccinees a polyclonal antibody (Ab) response with the same neutralizing capacity as the mAb. 2) The structures observed in epitope-paratope crystallographic complexes result from mutually induced fit between the two partners and do not correspond to the structures present in the free molecules before they had interacted. 3) The affinity-matured neutralizing mAbs obtained from chronically infected individuals did not recognize the germline predecessors of these Abs present in vaccinees. 4) The HIV p17 matrix protein that lines the inner surface of the viral membrane is one of the most disordered proteins identified on our planet and this prevents the induced Abs from binding to the glycosylated HIV gp120 protein. 5) Vaccinologists need to solve so-called inverse problems, for instance, guessing what are the multiple causes that produced an earlier wanted beneficial effect such as the absence of deleterious HIV infection in elite controllers. Since the immune system consists of numerous subsystems that have not yet been elucidated, it is impossible to solve the inverse problems posed by each subsystem. 6) Vaccinology is an empirical science that only sometimes succeeds because we do not understand the complex mechanisms that lead to protective immune responses.

Human immunodeficiency virus p24 antigen and antibody, herpes simplex virus-2 IgM and tumor necrosis factor alpha plasma levels in adult female children living in single-parent households: a case-control observational study

Abstract Objective: Human immunodeficiency virus (HIV) p24 antigen and antibody and herpes simplex virus 2 IgM are seromarkers indicating infection with HIV and herpes simplex virus-2 (HSV-2), respectively, whereas tumor necrosis factor α is an inflammatory biomarker that can be triggered by infections. Female children of single parents are faced with many socio-economic challenges that make them vulnerable to sexual influences and prone to sexually transmitted infections. The goal of this work was to determine HIV p24 antigen/antibody, HSV-2 IgM and tumor necrosis factor-α plasma levels in adult female children living in single-parent households. Methods: In this case-control observational study, 100 adult female children living with a single parent (50 living with a single mother and 50 living with a single father; age: 18–22 years) and 100 age-matched women living with both parents were recruited to serve as the test and control groups, respectively. All subjects were negative for acid-fast bacilli, plasmodium, hepatitis C virus, and hepatitis B virus. Human tumor necrosis factor α, HSV-2 IgM, antibody to hepatitis C virus, hepatitis B surface antigen and human immunodeficiency virus p24 antigen and antibody (HIV p24 Ag/Ab) levels were determined by ELISA, while the detection of acid-fast bacilli in sputum and Plasmodium in blood was carried out by optical microscopy. This work was carried out in the Owo/Ose Federal Constituency in Ondo State that shares boundaries with Edo State. The study protocol was approved by the Research and Ethical Committee of the Department of Medical Laboratory Science, Achievers University, Owo, Nigeria (AUO/MLS/2020/127) on August 27, 2020. Results: HIV p24 Ag/Ab was detected in 0 adult female children living with a single mother, 1 (2%) adult female child living with a single father and 1 (1%) adult female child living with both parents. HSV-2 IgM was detected in 9 (18%) adult female children living with a single mother, 13 (26%) adult female children living with a single father, and 5 (10%) adult female children living with both parents. Conclusion: This work shows that adult female children of single parents are vulnerable to sexual influences, and thereby more prone to HSV-2 and possibly HIV, especially adult female children of single fathers.

A triple-layer microchannel array chip with micro through-holes for smartphone based point-of-care testing of biomarker

Colorimetric detection on a microchip frequently suffers the sensitivity problem due to the limited optical path length. Herein, a point-of-care testing (POCT) device based on the colorimetric detection on a microchip with an array of micro through holes for enlarged optical length and an optical imaging system was demonstrated. Each micro through hole has a depth of 2 mm and a volume roughly 0.6 μl, connecting to a microchannel for the efficient reaction. Up to 32 micro through holes were arranged compactly along a round region with a diameter of 13 mm to make them effectively captured by a smartphone camera in a short distance (6.5 cm). For reliable smartphone imaging, a handheld, battery powered optical imaging cassette was made by three-dimensional (3D) printing. A data processing program was also written to realize the detection area selection, RGB (red, green, blue) value reading and detection signal calculation. With the integrated device, a biomarker, human immunodeficiency virus (HIV) p24 antigen in human serum was detected down to 20 pg/ml, much better than that with microchannels. The proposed microchip and the imaging system can be a promising POCT or on-site detection tool based on colorimetric detection.

Donation testing and transfusion transmissible infections

Donation testing and transfusion transmissible infections

HIV p17 enhances T cell proliferation by suppressing autophagy through the p17-OLA1-GSK3β axis under nutrient starvation.

Nutrient starvation is a common phenomenon that occurs during T cell activation. Upon pathogen infection, large amounts of immune cells migrate to infection sites, and antigen-specific T cells are activated; this is followed by rapid proliferation through clonal expansion. The dramatic expansion of cells will commonly lead to nutrient shortage. Cellular autophagy is often upregulated as a way to sustain the body's energy requirements. During infection, human immunodeficiency virus (HIV) co-opts a series of host cell metabolic pathways for replication. Several HIV proteins, such as Env, Nef, and Vpr, have already been reported as being involved in autophagy-related processes. In this report, we identified that the HIV p17 protein acts as a major factor in suppressing the autophagic process in T cells, especially under glucose starvation condition. HIV p17 interacts with Obg-like ATPase 1 (OLA1) and disrupts OLA1-glycogen synthase kinase-3 beta (GSK3β) complex, leading to GSK3β hyperactivation. Consequently, a prior proliferation of HIV-infected T cells under glucose starvation will occur. The inhibition of autophagy also aids HIV replication by antagonizing the antiviral effect of autophagy. Our study shows a new cellular pathway that HIV can hijack for viral spreading by a prior proliferation of HIV-loaded T cells and may provide new therapeutic targets for acquired immunodeficiency syndrome intervention.

Development and optimization of thermal contrast amplification lateral flow immunoassays for ultrasensitive HIV p24 protein detection

Detection of human immunodeficiency virus (HIV) p24 protein at a single pg/ml concentration in point-of-care (POC) settings is important because it can facilitate acute HIV infection diagnosis with a detection sensitivity approaching that of laboratory-based assays. However, the limit of detection (LOD) of lateral flow immunoassays (LFAs), the most prominent POC diagnostic platform, falls short of that of laboratory protein detection methods such as enzyme-linked immunosorbent assay (ELISA). Here, we report the development and optimization of a thermal contrast amplification (TCA) LFA that will allow ultrasensitive detection of 8 pg/ml p24 protein spiked into human serum at POC, approaching the LOD of a laboratory test. To achieve this aim, we pursued several innovations as follows: (a) defining a new quantitative figure of merit for LFA design based on the specific to nonspecific binding ratio (BR); (b) using different sizes and shapes of gold nanoparticles (GNPs) in the systematic optimization of TCA LFA designs; and (c) exploring new laser wavelengths and power regimes for TCA LFA designs. First, we optimized the blocking buffer for the membrane and running buffer by quantitatively measuring the BR using a TCA reader. The TCA reader interprets the thermal signal (i.e., temperature) of GNPs within the membrane when irradiated by a laser at the plasmon resonance wavelength of the particle. This process results in higher detection and quantitation of GNPs than in traditional visual detection (i.e., color intensity). Further, we investigated the effect of laser power (30, 100, 200 mW), GNP size and shape (30 and 100 nm gold spheres, 150 nm gold-silica shells), and laser wavelength (532, 800 nm). Applying these innovations to a new TCA LFA design, we demonstrated that 100 nm spheres with a 100 mW 532 nm laser provided the best performance (i.e., LOD = 8 pg/ml). This LOD is significantly better than that of the current colorimetric LFA and is in the range of the laboratory-based p24 ELISA. In summary, this TCA LFA for p24 protein shows promise for detecting acute HIV infection in POC settings.

Intact Human Immunodeficiency Virus (HIV) Reservoir Estimated by the Intact Proviral DNA Assay Correlates With Levels of Total and Integrated DNA in the Blood During Suppressive Antiretroviral Therapy.

Accurate characterization of the human immunodeficiency virus (HIV) reservoir is imperative to develop an effective cure. HIV was measured in antiretroviral therapy-suppressed individuals using the intact proviral DNA assay (IPDA), along with assays for total or integrated HIV DNA, and inducible HIV RNA or p24. Intact provirus correlated with total and integrated HIV.

Products of adaptive host immune response to viral agents in hyperthermia pediatric patients receiving malaria treatment and the immunoserologic status of their mothers

Study Background: Hyperthermia is a manifestation of disease condition, which may also be as a result of immune response to the presence of infectious agent in the body that can be expressed in the serum/plasma. At times, it is erroneously assumed for Plasmodium infection. Aims and Objectives: This work was designed to express the products of adaptive host immune response to viral agents in hyperthermia pediatric patients receiving malaria treatment and the immunoserologic status of their mothers for effective management and diagnosis. Materials and Methods: One hundred and four hyperthermia children aged 5–9 years receiving treatment in herbal homes were investigated as test, while one hundred age-matched children with normal body temperature were recruited from Owo and Ose local government areas as control subjects. Mothers of the hyperthermia children were also studied. Antihepatitis C virus (HCV), human immunodeficiency virus (HIV) P24 antigen/antibody, anti-HBe, and HBeAg were determined by enzyme-linked immunosorbent assay, Plasmodium by Geimsa thick blood film, and acid-fast bacilli by Ziehl–Neelsen techniques. Results: R Results obtained showed that the frequency of anti-HBe in mothers of hyperthermia patients (10.6% [11]) and in children with normal body temperature (9% [9]) was higher than the frequency of anti-HBe obtained in the hyperthermia herbal home children (4.8% [5]) while the frequency of HBeAg found in hyperthermia herbal home children (14.4% [15]) and their mothers (14.4% [15]) was higher than the results obtained in children with normal body temperature (4% [4]). The frequency of anti-HCV in herbal home hyperthermia children (2.9% [3]) was higher than the results obtained in their mothers (1.9% [2]) and children with normal body temperature (1% [1]). The HIV P24 antigen/antibody expressed by the mothers of herbal home hyperthermia children (3.85% [4]) was higher the results obtained in their children (2.9% [3]), while none of the children with normal body temperature expressed HIV P24 antigen/antibody. The frequency of Plasmodium in mothers of herbal home hyperthermia children (19.2% [20]) was higher than the results obtained in their hyperthermia children (9.6% [10]) and children with normal body temperature (3% [3]). The frequency of HIV in hyperthermia was 2.9% (3), while that of active/replicating hepatitis B virus (HBV) and HCV was 14.4% (HBeAg) and 2.9% (anti-HCV), respectively. Only mothers of hyperthermia herbal home children expressed 1.6% (2) HIV P24Ag/Ab + HBeAg + anti-HBe. None of the mothers of hyperthermia herbal home children expressed HIV P24 antigen/antibody and infected with Plasmodium as against 2.9% (3) and 9.6% (10) obtained in their children. 1.6% (2) of the mothers of hyperthermia herbal home children expressed anti-HCV as against 2.9% (3) of their children who expressed the immunoserologic marker, while the hyperthermia herbal home children and their mothers expressed anti-HBe (4.8%[5] vs. 4.8% [5]) and HBeAg (14.4%[15] vs. 14.4% [15]). Conclusion: Products of adaptive host immune response expressed in hyperthermia pediatric patients receiving malaria treatment and the immunoserologic status of their mothers include HIV P24Ag/Ab, HBeAg, anti-HBe, and anti-HCV, while the frequency of HIV in hyperthermia was 2.9% (3) and that of active/replicating HBV and HCV was 14.4% (HBeAg) and 2.9% (anti-HCV), respectively, with only HBV and HCV traceable to the mothers.

Validation of Spiked Postmortem Blood Samples from Cornea Donors on the Abbott ARCHITECT and m2000 Systems for Viral Infections

Background: Transplantation of human corneal tissue is associated with the potential risk of transmittance of viral infections. In accordance with European directives and federal laws, in Germany each tissue donor has to be tested for infectious diseases such as hepatitis B and C virus (HBV and HCV) and human immunodeficiency virus (HIV) infection. However, most of the currently available CE-marked serologic and nucleic acid screening systems are only validated for antemortem blood. Methods: Twenty related and paired ante- and postmortem blood samples from cornea donors were obtained and subsequently analyzed for hepatitis B surface antigen (HBsAg), hepatitis B antibody (anti-HBc), anti-HCV, HCV RNA, anti-HIV-1/2, and HIV p24 Ag using Abbott test systems. The sera were also spiked with reference materials in concentrations giving low and high positivity for HBV, HCV, and HIV markers. Results: The spiked ante- and postmortem sera from related donors showed similar results for HBsAg, anti-HBc, anti-HCV, HCV RNA, anti-HIV, and HIV p24 Ag, indicating a high stability of viral markers in cadaveric specimens. Three cornea donors had a medical history of HBV infection and revealed anti-HBc at similar levels in the ante- and postmortem sera. In addition, there was a single postmortem sample demonstrating a weak signal of anti-HIV-1 and HIV-1 p24 Ag. False-positive or false-negative results were not detected. The results obtained with the Abbott ARCHITECT analyzer and Abbott RealTime HCV PCR showed no significant differences. Conclusion: The analyzed screening assays are suitable for the detection of infectious markers of HBV, HCV, and HIV at similar levels in spiked ante- and postmortem sera from cornea donors.

Skewed CD39/CD73/adenosine pathway in B cells is associated with innate immune hyperactivation in chronic HIV-1 infection

BackgroundThe CD39/CD73/adenosine suppression pathway correlates with disease progression in patients with chronic human immunodeficiency virus 1 (HIV-1) infection; however, the relationships between this pathway in B cells and innate immune hyperactivation remain largely undefined.MethodsIn this study, we examined CD39 and CD73 expression, and adenosine production by B cells from 136 patients with chronic HIV-1 infection.ResultsThe CD39 and CD73 expression levels on total B cells (and their subsets) decreased continuously in typical progressors (TPs), but were gradually recovered in complete responders. In TPs, the frequencies of CD39+, CD73+, and CD39+CD73+ B cells inversely correlated with HIV-1 viral load and positively correlated with CD4 T cell counts, but also positively associated with CD4/CD8 ratioes and inversely correlated with markers for intestinal epithelial injury and microbial translocation. Furthermore, B cells from TPs showed a markedly reduced capacity to generate CD39/CD73-dependent extracellular adenosine; the loss of this capacity was more serious in patients with acquired immunodeficiency syndrome. In vitro, increased adenosine could inhibit the activation of monocytes and suppress the ability of monocytes and T cells to produce TNF-alpha, regardless of their infection status. Moreover, adenosine could inhibit activation-induced HIV-1 virion production and p24 expression.ConclusionsThese findings provided a new role of B cell pathology in HIV-1 infection, in which the skewed CD39/CD73/adenosine pathway in B cells linked to microbial translocation-induced innate immune hyperactivation, supporting the notion that regulating the adenosine pathway in B cells might be an attractive approach to treat patients with HIV-1-infection.

Benzyl-2-Acetamido-2-Deoxy-α-d-Galactopyranoside Increases Human Immunodeficiency Virus Replication and Viral Outgrowth Efficacy In Vitro.

Glycosylation of host and viral proteins is an important posttranslational modification needed to ensure correct function of glycoproteins. For this reason, we asked whether inhibition of O-glycosylation during human immunodeficiency virus (HIV) in vitro replication could affect HIV infectivity and replication rates. We used benzyl-2-acetamido-2-deoxy-α-d-galactopyranoside (BAGN), a compound that has been widely used to inhibit O-glycosylation in several cell lines. Pretreatment and culture of PHA-blast target cells with BAGN increased the percentage of HIV-infected cells (7.6-fold, p = 0.0115), the per-cell amount of HIV p24 protein (1.3-fold, p = 0.2475), and the viral particles in culture supernatants (7.1-fold, p = 0.0029) compared to BAGN-free cultures. Initiating infection with virus previously grown in the presence of BAGN further increased percentage of infected cells (30-fold, p < 0.0001), intracellular p24 (1.5-fold, p = 0.0433), and secreted viral particles (74-fold, p < 0.0001). BAGN-treated target cells showed less CD25 and CCR5 expression, but increased HLA-DR surface expression, which positively correlated with the number of infected cells. Importantly, BAGN improved viral outgrowth kinetics in 66% of the samples tested, including samples from HIV controllers and subjects in whom no virus could be expanded in the absence of BAGN. Sequencing of the isolated virus indicated no skewing of viral quasi-species populations when compared to BAGN-free culture conditions. BAGN also increased virus production in the ACH2 latency model when used together with latency-reversing agents. Taken together, our results identify BAGN treatment as a simple strategy to improve viral outgrowth in vitro and may provide novel insights into host restriction mechanisms and O-glycosylation-related therapeutic targets for HIV control strategies.

Human immunodeficiency virus-associated cystic lymphoid hyperplasia: An immunohistochemical description.

Cystic lymphoid hyperplasia (CLH) describes benign salivary lymphoepithelial cysts with a strong link to human immunodeficiency virus (HIV) infection and acquired immunodeficiency syndrome (AIDS). The pathogenesis is related to ductal dilatation of entrapped salivary elements due to intranodal HIV-associated atypical lymphoid hyperplasia. Very little is known about the immunophenotypic profile of this entity. This study aims to describe the immunopathological features of a series of CLH cases in HIV-positive patients to clarify the etiopathogenesis. Paraffin-embedded tissue from 25 cases of parotid CLH in HIV seropositive patients was immunohistochemically analyzed with CD3, CD20, CD4, CD8, and p24 using standard procedures. The data are mostly descriptive and were analyzed using EpiInfo (3.5.1) (CDC, Atlanta, USA); significant differences were analyzed using the Student's t-test and the Chi-square test with a statistical significance level of P < 0.05 being used. Immunostaining showed a CD8:CD4 of ~1:1 except in selected cases with decreased CD4 and increased CD8 expression in the interfollicular (IF) areas. p24 staining revealed 100% specificity in HIV-associated CLH. The immunohistochemical description of CD20, CD3, CD4, and CD8 provides an understanding of CLH pathogenesis. CLH of parotid lymph nodes in confirmed HIV-positive patients with 100% specificity with HIV p24 antibody validates the strong association of CLH with HIV and AIDS. The CD4:CD8 ratio was ~1:1; however, increased CD8 expression within IF areas may indicate possible HIV-related CLH as compared to other cystic parotid lesions.

A multi-walled carbon nanotubes based molecularly imprinted polymers electrochemical sensor for the sensitive determination of HIV-p24

To develop a rapid, simple and sensitive method for the determination of human immunodeficiency virus p24 (HIV-p24), a novel molecularly imprinted polymers (MIPs) electrochemical sensor was constructed on the surface of a multi-walled carbon nanotubes (MWCNTs) modified glassy carbon electrode (GCE) by surface polymerization using acrylamide (AAM) as functional monomer, N,N′-methylenebisacrylamide (MBA) as cross-linking agent and ammonium persulphate (APS) as initiator. Each modification step was carefully examined by cyclic voltammetry (CV), differential pulse voltammetry (DPV) and scanning electron microscope (SEM). The proposed MIPs electrochemical biosensor exhibited specific recognition to HIV-p24 and displayed a broad linear detection range from 1.0×10−4 to 2ngcm−3 with a low detection limit of 0.083pgcm−3 (S/N=3). This performance is superior to most HIV-p24 sensors based on other methods. Meanwhile, this sensor possessed of good selectivity, repeatability, reproducibility, stability and was successfully applied for the determination of HIV-p24 in real human serum samples, giving satisfactory results. The accuracy and reliability of the sensor is further confirmed by enzyme-linked immunosorbent assay (ELISA).