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OBJECTIVE: The aim of this study was to examine outcomes of follow-up for persons with discordant fourth-generation HIV screening test results. DESIGN: A retrospective chart review. METHODS: We analyzed the electronic health record at the Medical University of South Carolina for a 10-year period spanning 2012-2022 to identify instances of discordant HIV screening test results, wherein initial antigen/antibody screening was positive, but reflex confirmatory testing for HIV-1 and HIV-2 antibodies was negative. We reviewed individual records to evaluate clinical follow-up and determine if the discordant test represented an acute HIV infection, a false-positive result, or was unresolved. RESULTS: We identified 199 testing instances with discordant results. Most discordant results ( n â=â115) were subsequently determined to reflect a false-positive test, while 56 were unresolved without documented follow-up testing. Twenty-eight cases of acute HIV infection were identified of which 26 were linked to care within a month of initial testing. Two acute HIV cases were not identified in real time leading to delay in diagnosis and care. Testing done in the context of infectious symptoms and testing performed in the emergency department were associated with increased odds of a discordant test ultimately reflecting acute HIV infection. CONCLUSION: These results demonstrate the importance of appropriate and timely follow-up for discordant HIV screening test results.
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Centros Médicos Académicos , Infecciones por VIH , Prueba de VIH , Humanos , Estudios Retrospectivos , Infecciones por VIH/diagnóstico , Masculino , Femenino , Adulto , Prueba de VIH/estadística & datos numéricos , Persona de Mediana Edad , South Carolina , Adulto Joven , Tamizaje Masivo/métodos , Anticuerpos Anti-VIH/sangre , Anciano , Adolescente , VIH-1/aislamiento & purificación , VIH-1/inmunología , VIH-2/inmunologíaRESUMEN
Diagnosing of human immunodeficiency virus (HIV) types 1 and 2 requires a screening with a highly sensitive and specific enzyme immunoassay and a low detection limit for the HIV-1 p24 antigen to minimize the diagnostic window. The objective of the study was to determine the sensitivity, specificity, and p24 limit of detection of the Access HIV combo V2 assay. Retrospective part of sensitivity: 452 HIV-1 positive samples from 403 chronic (9 different HIV-1 group M subtypes, 22 different HIV-1 group M CRFs, and 3 HIV-1 group O), 49 primary HIV-1 infections, 103 HIV-2 positive samples assessed at Pitié-Salpêtrière Hospital, 600 untyped HIV-1, 10 subtype-D, and 159 untyped HIV-2 samples assessed in Bio-Rad Laboratories. Prospective part of clinical specificity: all consecutive samples in two blood donor facilities and Pitié-Salpêtrière (6,570 patients) tested with Access HIV combo V2 and respectively Prism HIV O Plus (Abbott) or Architect HIV Ag/Ab Combo (Abbott) for Ag/Ab screening, and Procleix Ultrio (Gen Probe) for HIV RNA screening. Limit of detection for p24 antigen was assessed on recombinant virus-like particles (10 HIV-1 group M subtypes/CRFs, HIV-1 group O). Sensitivity [95% confidence interval (CI)] of Access HIV combo V2 was 100% (99.63-100) for HIV-1 chronic infection, 100% (98.55-100) for HIV-2 chronic infection, and 100% (93.00-100) for HIV-1 primary infection. Specificity (95% CI) was 99.98 (99.91-100). Limit of detection for p24 antigen was around 0.43 IU/mL [interquartile range (0.38-0.56)], and consistent across the 11 analyzed subtypes/CRFs. Hence, with both high sensitivity and specificity, Access HIV combo V2 is a suitable screening assay for HIV-1/2 infection. IMPORTANCE: Bio-Rad is one of the leading human immunodeficiency virus (HIV) screening test manufacturers. This laboratory released in 2021 their new version of the Access combo HIV test. However, to date, there have been no studies regarding its performance, especially its limit of detection of the diverse p24 antigen. We present the sensitivity (chronic and primary HIV-1 infection and HIV-2 chronic infection), specificity (blood donors and hospitalized patients), and raw data for the p24/seroconversion panels the manufacturer gave to the European agencies.
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Proteína p24 del Núcleo del VIH , Infecciones por VIH , VIH-1 , VIH-2 , Tamizaje Masivo , Sensibilidad y Especificidad , Humanos , Infecciones por VIH/diagnóstico , VIH-1/genética , VIH-1/clasificación , VIH-1/aislamiento & purificación , VIH-1/inmunología , Estudios Retrospectivos , Proteína p24 del Núcleo del VIH/sangre , VIH-2/inmunología , VIH-2/clasificación , VIH-2/genética , VIH-2/aislamiento & purificación , Tamizaje Masivo/métodos , Estudios Prospectivos , Prueba de VIH/métodos , MasculinoRESUMEN
IMPORTANCE: Twenty-five years after the first report that HIV-2 infection can reduce HIV-1-associated pathogenesis in dual-infected patients, the mechanisms are still not well understood. We explored these mechanisms in cell culture and showed first that these viruses can co-infect individual cells. Under specific conditions, HIV-2 inhibits HIV-1 through two distinct mechanisms, a broad-spectrum interferon response and an HIV-1-specific inhibition conferred by the HIV-2 TAR. The former could play a prominent role in dually infected individuals, whereas the latter targets HIV-1 promoter activity through competition for HIV-1 Tat binding when the same target cell is dually infected. That mechanism suppresses HIV-1 transcription by stalling RNA polymerase II complexes at the promoter through a minimal inhibitory region within the HIV-2 TAR. This work delineates the sequence of appearance and the modus operandi of each mechanism.
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Coinfección , Regulación Viral de la Expresión Génica , Duplicado del Terminal Largo de VIH , VIH-1 , VIH-2 , Interferones , ARN Viral , Productos del Gen tat del Virus de la Inmunodeficiencia Humana , Humanos , Coinfección/inmunología , Coinfección/virología , Duplicado del Terminal Largo de VIH/genética , VIH-1/genética , VIH-1/inmunología , VIH-2/genética , VIH-2/inmunología , VIH-2/metabolismo , ARN Viral/genética , Productos del Gen tat del Virus de la Inmunodeficiencia Humana/metabolismo , Interferones/inmunología , Regiones Promotoras Genéticas/genética , Unión Competitiva , ARN Polimerasa II/metabolismo , Transcripción GenéticaRESUMEN
Emerging studies indicate that infusion of HIV-resistant cells could be an effective strategy to achieve a sterilizing or functional cure. We recently reported that glycosylphosphatidylinositol (GPI)-anchored nanobody or a fusion inhibitory peptide can render modified cells resistant to HIV-1 infection. In this study, we comprehensively characterized a panel of newly isolated HIV-1-neutralizing antibodies as GPI-anchored inhibitors. Fusion genes encoding the single-chain variable fragment (scFv) of 3BNC117, N6, PGT126, PGT128, 10E8, or 35O22 were constructed with a self-inactivating lentiviral vector, and they were efficiently expressed in the lipid raft sites of target cell membrane without affecting the expression of HIV-1 receptors (CD4, CCR5 and CXCR4). Significantly, transduced cells exhibited various degrees of resistance to cell-free HIV-1 infection and cell-associated HIV-1 transmission, as well as viral Env-mediated cell-cell fusion, with the cells modified by GPI-10E8 showing the most potent and broad anti-HIV activity. In mechanism, GPI-10E8 also interfered with the processing of viral Env in transduced cells and attenuated the infectivity of progeny viruses. By genetically linking 10E8 with a fusion inhibitor peptide, we subsequently designed a group of eight bifunctional constructs as cell membrane-based inhibitors, designated CMI01â¼CMI08, which rendered cells completely resistant to HIV-1, HIV-2, and simian immunodeficiency virus (SIV). In human CD4+ T cells, GPI-10E8 and its bifunctional derivatives blocked both CCR5- and CXCR4-tropic HIV-1 isolates efficiently, and the modified cells displayed robust survival selection under HIV-1 infection. Therefore, our studies provide new strategies for generating HIV-resistant cells, which can be used alone or with other gene therapy approaches.
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Anticuerpos Anti-VIH/inmunología , Proteína gp41 de Envoltorio del VIH/antagonistas & inhibidores , Infecciones por VIH/terapia , VIH-1/fisiología , Fragmentos de Péptidos/farmacología , Anticuerpos de Cadena Única/inmunología , Fármacos Anti-VIH/farmacología , Anticuerpos ampliamente neutralizantes/genética , Anticuerpos ampliamente neutralizantes/inmunología , Linfocitos T CD4-Positivos/virología , Fusión Celular , Línea Celular , Terapia Genética , Glicosilfosfatidilinositoles , Anticuerpos Anti-VIH/genética , Proteína gp41 de Envoltorio del VIH/inmunología , Inhibidores de Fusión de VIH/farmacología , Infecciones por VIH/virología , VIH-1/inmunología , VIH-2/inmunología , VIH-2/fisiología , Humanos , Microdominios de Membrana , Fragmentos de Péptidos/genética , Receptores del VIH/metabolismo , Virus de la Inmunodeficiencia de los Simios/inmunología , Virus de la Inmunodeficiencia de los Simios/fisiología , Anticuerpos de Cadena Única/genética , Transgenes , Tropismo ViralRESUMEN
HIV-2 is less pathogenic compared to HIV-1. Still, disease progression may develop in aviremic HIV-2 infection, but the driving forces and mechanisms behind such development are unclear. Here, we aimed to reveal the immunophenotypic pattern associated with CD8 T-cell pathology in HIV-2 infection, in relation to viremia and markers of disease progression. The relationships between pathological differences of the CD8 T-cell memory population and viremia were analyzed in blood samples obtained from an occupational cohort in Guinea-Bissau, including HIV-2 viremic and aviremic individuals. For comparison, samples from HIV-1- or dually HIV-1/2-infected and seronegative individuals were obtained from the same cohort. CD8 T-cell exhaustion was evaluated by the combined expression patterns of activation, stimulatory and inhibitory immune checkpoint markers analyzed using multicolor flow cytometry and advanced bioinformatics. Unsupervised multidimensional clustering analysis identified a cluster of late differentiated CD8 T-cells expressing activation (CD38+, HLA-DRint/high), co-stimulatory (CD226+/-), and immune inhibitory (2B4+, PD-1high, TIGIThigh) markers that distinguished aviremic from viremic HIV-2, and treated from untreated HIV-1-infected individuals. This CD8 T-cell population displayed close correlations to CD4%, viremia, and plasma levels of IP-10, sCD14 and beta-2 microglobulin in HIV-2 infection. Detailed analysis revealed that aviremic HIV-2-infected individuals had higher frequencies of exhausted TIGIT+ CD8 T-cell populations lacking CD226, while reduced percentage of stimulation-receptive TIGIT-CD226+ CD8 T-cells, compared to seronegative individuals. Our results suggest that HIV-2 infection, independent of viremia, skews CD8 T-cells towards exhaustion and reduced co-stimulation readiness. Further knowledge on CD8 T-cell phenotypes might provide help in therapy monitoring and identification of immunotherapy targets.
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Linfocitos T CD8-positivos/inmunología , Infecciones por VIH/inmunología , Infecciones por VIH/virología , VIH-2/inmunología , Inmunosenescencia/inmunología , Adulto , Femenino , Seronegatividad para VIH/inmunología , Humanos , Masculino , Persona de Mediana Edad , Viremia/inmunologíaAsunto(s)
Infecciones por VIH/diagnóstico , VIH-1 , Biomarcadores , Encéfalo/diagnóstico por imagen , Recuento de Linfocito CD4 , Confusión/etiología , Diagnóstico Diferencial , Femenino , Fiebre/etiología , Anticuerpos Anti-VIH/sangre , Infecciones por VIH/complicaciones , Infecciones por VIH/virología , VIH-1/inmunología , VIH-1/aislamiento & purificación , VIH-2/inmunología , Humanos , Linfopenia/etiología , Imagen por Resonancia Magnética , Persona de Mediana Edad , Faringitis/etiología , Carga ViralRESUMEN
BACKGROUND: NEW LAV BLOT I and II (LAV I and LAV II), they were only option for human immunodeficiency virus (HIV) confirmatory test, following HIV screening test using HIV Ag/Ab combination test in Japan. We evaluated the performance of Geenius HIV-1/2 Confirmatory Assay (Geenius), both as a confirmatory test and for differentiation between HIV-1 and HIV-2, in comparison with LAV I and LAV II. METHODS: Eighty-nine HIV-1-positive plasma specimens, one anti-HIV-1 low-titer performance panel, 10 seroconversion panels, and two anti-HIV-1/2 combo performance panels were tested. The results were read with the Geenius Reader and by visual reading. RESULTS: All 89 HIV-1-positive plasma specimens were identified as HIV-1-positive using Geenius; this 100% success rate was superior to that with LAV I (95.5% using WHO criteria, 98.9% using CDC criteria). The HIV-1-positive specimens showed low cross-reactivity with HIV-2 lines in Geenius. The sensitivity of Geenius for HIV-1 detection was the same as or greater than that of LAV I, but less than that of Genscreen HIV Ag-Ab ULT, in our analysis of the commercial performance and seroconversion panels. In contrast, five of the 13 HIV-2-positive specimens that had been identified as HIV-positive untypable by visual reading because of their cross-reactivity to HIV-1 lines were successfully identified by the Geenius Reader as HIV-2-positive with cross-reactivity. CONCLUSIONS: Geenius provides strong performance for HIV confirmatory tests and HIV-1 differentiation tests. However, when visual reading is used, its performance in HIV-2 differentiation is less reliable. Because HIV-2 infection has been sporadically reported in Japan, the use of the Geenius Reader is preferable to ensure more reliable HIV-1/HIV-2 differentiation.
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Serodiagnóstico del SIDA/métodos , Infecciones por VIH/diagnóstico , VIH-1/aislamiento & purificación , VIH-2/aislamiento & purificación , Serodiagnóstico del SIDA/instrumentación , Reacciones Cruzadas , Diagnóstico Diferencial , Anticuerpos Anti-VIH/sangre , VIH-1/inmunología , VIH-2/inmunología , Humanos , Japón , Tamizaje Masivo , Reproducibilidad de los Resultados , Sensibilidad y EspecificidadRESUMEN
CD4+ T cells are critical players in the host adaptive immune response. Emerging evidence suggests that certain CD4+ T cell subsets contribute significantly to the production of neutralizing antibodies and help in the control of virus replication. Circulating T follicular helper cells (Tfh) constitute a key T cell subset that triggers the adaptive immune response and stimulates the production of neutralizing antibodies (NAbs). T cells having stem cell-like property, called stem-like memory T cells (Tscm), constitute another important subset of T cells that play a critical role in slowing the rate of disease progression through the differentiation and expansion of different types of memory cell subsets. However, the role of these immune cell subsets in T cell homeostasis, CD4+ T cell proliferation, and progression of disease, particularly in HIV-2 infection, has not yet been elucidated. The present study involved a detailed evaluation of the different CD4+ T cell subsets in HIV-2 infected persons with a view to understanding the role of these immune cell subsets in the better control of virus replication and delayed disease progression that is characteristic of HIV-2 infection. We observed elevated levels of CD4+ Tfh and CD4+ Tscm cells along with memory and effector T cell abundance in HIV-2 infected individuals. We also found increased frequencies of CXCR5+ CD8+ T cells and CD8+ Tscm cells, as well as memory B cells that are responsible for NAb development in HIV-2 infected persons. Interestingly, we found that the frequency of memory CD4+ T cells as well as memory B cells correlated significantly with neutralizing antibody titers in HIV-2 infected persons. These observations point to a more robust CD4+ T cell response that supports B cell differentiation, antibody production, and CD8+ T cell development in HIV-2 infected persons and contributes to better control of the virus and slower rate of disease progression in these individuals.
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Linfocitos T CD8-positivos/inmunología , Infecciones por VIH/inmunología , Infecciones por VIH/virología , VIH-2/inmunología , Células T Auxiliares Foliculares/inmunología , Adolescente , Adulto , Formación de Anticuerpos , Linfocitos B/inmunología , Linfocitos B/metabolismo , Linfocitos T CD8-positivos/metabolismo , Progresión de la Enfermedad , Femenino , Infecciones por VIH/metabolismo , Humanos , Memoria Inmunológica/inmunología , Activación de Linfocitos , Masculino , Persona de Mediana Edad , Células T Auxiliares Foliculares/metabolismo , Adulto JovenRESUMEN
SERINC5 is a potent lentiviral restriction factor that gets incorporated into nascent virions and inhibits viral fusion and infectivity. The envelope glycoprotein (Env) is a key determinant for SERINC restriction, but many aspects of this relationship remain incompletely understood, and the mechanism of SERINC5 restriction remains unresolved. Here, we have used mutants of HIV-1 and HIV-2 to show that truncation of the Env cytoplasmic tail (ΔCT) confers complete resistance of both viruses to SERINC5 and SERINC3 restriction. Critically, fusion of HIV-1 ΔCT virus was not inhibited by SERINC5 incorporation into virions, providing a mechanism to explain how EnvCT truncation allows escape from restriction. Neutralization and inhibitor assays showed ΔCT viruses have an altered Env conformation and fusion kinetics, suggesting that EnvCT truncation dysregulates the processivity of entry, in turn allowing Env to escape targeting by SERINC5. Furthermore, HIV-1 and HIV-2 ΔCT viruses were also resistant to IFITMs, another entry-targeting family of restriction factors. Notably, while the EnvCT is essential for Env incorporation into HIV-1 virions and spreading infection in T cells, HIV-2 does not require the EnvCT. Here, we reveal a mechanism by which human lentiviruses can evade two potent Env-targeting restriction factors but show key differences in the capacity of HIV-1 and HIV-2 to exploit this. Taken together, this study provides insights into the interplay between HIV and entry-targeting restriction factors, revealing viral plasticity toward mechanisms of escape and a key role for the long lentiviral EnvCT in regulating these processes.
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VIH-1/genética , VIH-2/genética , Glicoproteínas de Membrana/genética , Proteínas de la Membrana/genética , Virión/genética , Productos del Gen env del Virus de la Inmunodeficiencia Humana/genética , Antígenos de Diferenciación/genética , Antígenos de Diferenciación/inmunología , Línea Celular , Regulación de la Expresión Génica , Células HEK293 , Infecciones por VIH/genética , Infecciones por VIH/inmunología , Infecciones por VIH/virología , VIH-1/inmunología , VIH-1/patogenicidad , VIH-2/inmunología , VIH-2/patogenicidad , Células HeLa , Humanos , Evasión Inmune , Glicoproteínas de Membrana/inmunología , Proteínas de la Membrana/inmunología , Dominios Proteicos , Isoformas de Proteínas/genética , Isoformas de Proteínas/inmunología , Transducción de Señal , Células Madre , Virión/inmunología , Virión/patogenicidad , Internalización del Virus , Productos del Gen env del Virus de la Inmunodeficiencia Humana/inmunologíaRESUMEN
RATIONALE/STUDY DESIGN: A major challenge in the development of HIV vaccines is finding immunogens that elicit protection against a broad range of viral strains. Immunity to a narrow range of viral strains may protect infants of HIV-infected women or partners discordant for HIV. We hypothesized that immunization to the relevant viral variants could be achieved by exposure to infectious virus during prophylaxis with antiretroviral drugs. To explore this approach in an animal model, macaques were exposed to live virus (SIVmne or HIV-2287) during prophylaxis with parenteral tenofovir and humoral and cellular immune responses were quantified. Subsequently, experimental animals were challenged with homologous virus to evaluate protection from infection, and if infection occurred, the course of disease was compared to control animals. Experimental animals uninfected with SIVmne were challenged with heterologous HIV-2287 to assess resistance to retroviral infection. METHODOLOGY/PRINCIPAL FINDINGS: Juvenile female Macaca nemestrina (N = 8) were given ten weekly intravaginal exposures with either moderately (SIVmne) or highly (HIV-2287) pathogenic virus during tenofovir prophylaxis. Tenofovir protected all 8 experimental animals from infection, while all untreated control animals became infected. Specific non-neutralizing antibodies were elicited in blood and vaginal secretions of experimental animals, but no ELISPOT responses were detected. Six weeks following the cessation of tenofovir, intravaginal challenge with homologous virus infected 2/4 (50%) of the SIVmne-immunized animals and 4/4 (100%) of the HIV-2287-immunized animals. The two SIVmne-infected and 3 (75%) HIV-2287-infected had attenuated disease, suggesting partial protection. CONCLUSIONS/SIGNIFICANCE: Repeated exposure to SIVmne or HIV-2287, during antiretroviral prophylaxis that blocked infection, induced binding antibodies in the blood and mucosa, but not neutralizing antibodies or specific cellular immune responses. Studies to determine whether antibodies are similarly induced in breastfeeding infants and sexual partners discordant for HIV infection and receiving pre-exposure antiretroviral prophylaxis are warranted, including whether these antibodies appear to confer partial or complete protection from infection.
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Antirretrovirales/uso terapéutico , Infecciones por VIH/prevención & control , VIH-2/efectos de los fármacos , Síndrome de Inmunodeficiencia Adquirida del Simio/prevención & control , Virus de la Inmunodeficiencia de los Simios/efectos de los fármacos , Tenofovir/uso terapéutico , Animales , Modelos Animales de Enfermedad , Femenino , Infecciones por VIH/inmunología , VIH-2/inmunología , Humanos , Inmunización/métodos , Macaca nemestrina , Profilaxis Pre-Exposición/métodos , Síndrome de Inmunodeficiencia Adquirida del Simio/inmunología , Virus de la Inmunodeficiencia de los Simios/inmunologíaRESUMEN
Shortly after the first detection of human immundeficiency virus (HIV) infection in USA in 1981, the number of cases have increased gradually from all around the world. Turkey's high capacity for tourism and the unique geographic location extending between Europe and Asia, provides convenience for the passage of individuals across the countries and sexually transmitted infections including HIV, as well. According to the official data of the Ministry of Health; there are 25809 HIV positive and 1958 AIDS cases as of November 30, 2020, after the epidemic started in 1985 in Turkey. Despite the decrease in the number of newly detected HIV cases as a result of serious measures taken for the transmission of infection worldwide, the increase in the number of cases still continues in our country. Shortening the reporting period and starting treatment as soon as possible in the diagnosis of infection is critical for the control of the epidemic. For this purpose, Centers for Disease Control and Prevention (CDC) published a new test algorithm in 2010, which suggested the use of the Geenius™ HIV ½ supplemental assay test instead of western blot tests, which have been used for many years to verify HIV screening test positivity. In this study, we aimed to report the experience of the National HIV-Acquiner Immundeficiency Syndrome(AIDS) and Viral Hepatitis Reference Laboratories of Turkey in the first year of transition to the new HIV algorithm and to evaluate the diagnostic performance of Geenius™ HIV ó and line immunassay (LIA) s. A total of 2090 anti-HIV positive patient sera sent to National HIV-AIDS and Viral Hepatitis Reference Laboratories of Turkey, Ankara for HIV confirmation were included in the study. All samples were retested with a fourth-generation enzyme linked immunosorbent assay (ELISA) test (VIDAS® HIV-1/2 Duo Ultra assay, BioMerieux, France) followed by the confirmatory tests; Geenius™ HIV 1/2 confirmatory assay (BioRad, Redmond, WA) and Line-immunoassay (INNO-LIA HIV ½ Score, Fujirebio, reverse transcriptase polymerase chain reaction (RT-PCR) (artus HI Virus-1 RT-PCR, Qiagen, Germany) test and in-house HIV-2 RNA and proviral DNA PCR. The sensitivity, specificity, and the agreement of the each assay were compared. Cohen's Kappa analysis was used for the evaluation of the agreement between the tests. According to the new algorithm which recommended Geenius™ test besides HIV-1 RNA test, 1707 (81.7%) HIV-1 positive samples were identified. Of these samples; 95.9% and 95.02% were identified as HIV-1 positive by GeeniusTM and INNO-LIA, respectively. However, 2.5% of the positive samples were negative with Geenius™ and 3.5% with INNO-LIA. One and a half percentage (1.5%) of these samples were detected with Geenius™ and 1.4% with INNO-LIA as indeterminant. When all the positive samples determined with ELISA were evaluated; it was detected that,1.3% were indeterminate by Geenius™ test and 2.4% by the INNO-LIA test. When the INNO-LIA test was regarded as the gold standard method; sensitivity, specificity, positive predictive and negative predictive values of the Geenius™ test were as follows; 99.7%, 96.1%, 98.9%, and 99.1%. The agreement between INNO-LIA and Geenius™ tests was found to be 98.95% (κ= 0.969; very good). When the Geenius™ and HIV-1 PCR tests were evaluated together for the confirmation; the sensitivities of Geenius™ and INNO-LIA tests were 99.8% and 98.3%, specificities were 89.8% and 85.3%, respectively. Slight positive bands were detected in the gp36 or gp140 bands, the HIV-2 specific envelope proteins, were detected in seven samples, However, the positivity disappeared after the dilution of the samples and it was accepted as false positivite reaction due to the absence of HIV-2 RNA and proviral DNA in these samples. In conclusion; we concluded that Geenius ™ and INNO-LIA tests have a perfect agreement in HIV diagnosis and due to the rapid and reliable results provided for the HIV test protocol, Geenius™ test can be used safely as an alternative to the immunoblot tests. HIV-1 RNA testing must be performed in all HIV confirmation centers in order to detect acute HIV cases in the fast and early period which are the main reason for the updates in HIV diagnosis.
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Algoritmos , Infecciones por VIH , Inmunoensayo , Anticuerpos Anti-VIH/sangre , Infecciones por VIH/diagnóstico , Infecciones por VIH/epidemiología , Infecciones por VIH/virología , VIH-1/inmunología , VIH-2/inmunología , Humanos , Inmunoensayo/normas , Sensibilidad y Especificidad , Turquía/epidemiologíaRESUMEN
Human macrophages are protected by intrinsic antiviral defenses that provide moderate protection against HIV-1 infection. Macrophages that do become infected can serve as long-lived reservoirs, to disseminate HIV-1 to CD4+ T cells. Infection of macrophages with HIV-1 and HIV-2 is inhibited by constitutive mobilization of antioxidant response master transcription regulator Nrf2. The downstream mediator of this restriction was not identified. Among the tens of genes controlled directly by Nrf2 in macrophages, we found that xCT/SLC7A11, a 12-transmembrane, cystine-glutamate antiporter promotes antiretroviral activity. We show here that depletion of xCT mRNA increases HIV-1 infection. Reconstitution of xCT knock out cells with wild-type xCT but not a transport-deficient mutant restores anti-HIV-1 activity. Pharmacological inhibitors of xCT amino acid transport also increase infection. The block is independent of known restriction factors and acts against HIV-1 and HIV-2. Like the block triggered through Nrf2, xCT function impedes infection immediately before 2-LTR circle formation.
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Sistema de Transporte de Aminoácidos y+/inmunología , Infecciones por VIH , VIH-1 , VIH-2 , Células HEK293 , Infecciones por VIH/inmunología , Infecciones por VIH/virología , VIH-1/inmunología , VIH-1/fisiología , VIH-2/inmunología , VIH-2/fisiología , Células HeLa , Humanos , Leucocitos , Células THP-1RESUMEN
Recently, immunoblots (IBs) have tended to substitute Western blots (WBs) for HIV infection diagnosis. Several studies have confirmed IBs' high sensitivity to confirm HIV infection for every stage. Since the nature and pattern of the antigens of IBs are different from those of WB, the abilities of IBs and WBs to distinguish the stages of recent seroconversion and open-ended chronic infection might differ. We aimed to evaluate the performance of two IBs (INNO-LIA™ HIVI/II, Fujirebio, and Geenius™ HIV1/2 Confirmatory assay, Bio-Rad) to define the stage of infection. We studied 53 patients from the French ANRS CO6 PRIMO cohort. IBs have higher positive rates than WB. However, Geenius was less sensitive than WB and INNO-LIA to detect antibodies to p31 (0% vs 22.6 % and 15.1 %, respectively), so it could wrongly label late Fiebig stage and open-ended chronic infections as recent infections (n = 5/53). For the first time, we provide evidence that centralized WBs associated with an enzyme immunoassay for the identification of recent HIV-1 infection support the establishment of a more accurate diagnosis of primary HIV infection to improve the accuracy of enrollments in cohorts of recent HIV infections useful for epidemiological studies, pathogenesis studies or therapeutic trials.
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Western Blotting , Infecciones por VIH , VIH-1 , Anticuerpos Anti-VIH , Infecciones por VIH/diagnóstico , VIH-1/inmunología , VIH-2/inmunología , Humanos , Inmunoensayo , Sensibilidad y EspecificidadRESUMEN
BACKGROUND: Blood donor care and blood safety require a quick and accurate decision on the presence or absence of Human Immunodeficiency Virus (HIV) infection, based on the proper selection of blood donors, serological and molecular HIV testing as well as western blot test. The aim was investigating the possibility of inclusion of Geenius HIV 1/2 Confirmatory Assay in blood donor testing algorithm in order to shorten test time and decrease the number of indeterminate results. METHODS: A total of 75 archived serum/plasma samples were tested. Their previous serological and molecular HIV results were: 3 negative samples, 7 positive samples, 65 serological indeterminate or positive but confirmatory testing and NAT negative samples. RESULTS: Geenius assay confirmed the presence of antibodies in all blood donors with HIV positive serology and Nucleic Acid Testing (NAT). HIV-1 gp160 and gp41 antibodies were detected in these donors, while p31 and p24 antibodies were not detected in two and three donors, respectively. HIV-2 antibodies gp36 and gp140 were not found. Blood donor with HIV indeterminate or positive serology but negative confirmatory testing and NAT, were negative in Geenius assay. Conclusion The results obtained are consistent with western blot results. The assay proved simple and quick to perform. Studies have confirmed the possibility of introducing Bio-Rad Geenius into a routine blood donor testing protocol.
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Bioensayo/métodos , Donantes de Sangre/estadística & datos numéricos , VIH-1/inmunología , VIH-2/inmunología , Humanos , Estudios ProspectivosRESUMEN
BACKGROUND: Relying on reference laboratories for HIV confirmation testing may lead to delays in treatment and can cause stress for patients who have positive HIV screening results. OBJECTIVE: To internalize HIV-1/HIV-2 antibody differentiation testing within the hospital laboratory. METHODS: We analytically verified an HIV antibody differentiation immunoassay and subsequently compared result turnaround times (TATs) for HIV antibody differentiation and HIV-1 qualitative RNA in the months before and after the test internalization. RESULTS: HIV antibody differentiation was successfully verified. TATs for HIV antibody differentiation and HIV-1 RNA significantly improved, from medians of 40.4 hours and 156.5 hours to medians of 17.7 hours and 56.5 hours, respectively, after the internalization. The 90th-percentile turnaround times declined by 72% and 44%, respectively. CONCLUSIONS: It is feasible for a hospital laboratory to verify HIV antibody-differentiation testing. Its implementation may considerably improve result TATs for the HIV diagnostic algorithm.
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Infecciones por VIH , VIH-1 , Algoritmos , Anticuerpos Anti-VIH , Infecciones por VIH/diagnóstico , VIH-1/inmunología , VIH-2/genética , VIH-2/inmunología , Humanos , ARN ViralRESUMEN
BACKGROUND: HIV, HBV and HCV infections continue to represent major health concerns, especially among key at-risk populations such as men who have sex with men (MSM), people who inject drugs (PWIDs), transgender women (TGW) and sex workers (SW). The objective of the ANRS-CUBE study was to evaluate the acceptability of a healthcare, community-based strategy offering a triple rapid HIV-HBV-HCV testing, and HBV vaccination, targeted at three priority groups (MSM, PWIDs and TGW/SWs), in three community centers, in the Paris area. METHODS: This longitudinal multicentric non-randomized study included all adult volunteers attending one of the three specialized community centers in Paris, between July 2014 and December 2015. HIV, HBV and HCV status and acceptability of HBV vaccination were evaluated. RESULTS: A total of 3662, MSM, 80 PWIDs and 72 TGW/SW were recruited in the three centers respectively. Acceptability of rapid tests was 98.5% in MSM and 14.9% in TGW/SWs, but could not be estimated in PWIDs since the number of users attending and the number of proposals were not recorded. User acceptability of HBV vaccination was weak, only 17.9% of the eligible MSM (neither vaccinated, nor infected) agreed to receive the first dose, 12.2% two doses, 5.9% had a complete vaccination. User acceptability of HBV vaccination was greater in PWIDs and TGW/SWs, but decreased for the last doses (66.7 and 53.3% respectively received a first dose, 24.4 and 26.7% a second dose and 6.7 and 0% a third dose). Fifty-three participants (49 MSM and 4 PWIDs) were discovered HIV positive, more than half with a recent infection. All but two HIV positive participants were linked to appropriate care in less than one month. CONCLUSIONS: Rapid HIV-HCV-HBV screening showed a very high level of acceptability among MSM. Efforts need to be made to improve immediate acceptability for HBV vaccination, especially among MSM, and follow-up doses compliance. Our results show the important role of community centers in reaching targets, often fragile, populations, while also suggesting the need to reinforce on-site human support in terms of testing and vaccination, especially when addressing PWIDs.
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Infecciones por VIH/diagnóstico , VIH-1/inmunología , VIH-2/inmunología , Hepacivirus/inmunología , Virus de la Hepatitis B/inmunología , Hepatitis B Crónica/diagnóstico , Hepatitis C Crónica/diagnóstico , Vacunación , Vacunas Virales/inmunología , Adolescente , Adulto , Servicios de Salud Comunitaria , Femenino , Infecciones por VIH/epidemiología , Infecciones por VIH/virología , Hepatitis B Crónica/epidemiología , Hepatitis B Crónica/virología , Hepatitis C Crónica/epidemiología , Hepatitis C Crónica/virología , Homosexualidad Masculina , Humanos , Estudios Longitudinales , Masculino , Tamizaje Masivo/métodos , Paris/epidemiología , Proyectos Piloto , Estudios Prospectivos , Factores de Riesgo , Pruebas Serológicas , Trabajadores Sexuales , Minorías Sexuales y de Género , Abuso de Sustancias por Vía Intravenosa , Personas Transgénero , Adulto JovenRESUMEN
The main purpose in the diagnosis of human immunodeficiency virus (HIV) infection is to rapidly and accurately identify people with HIV infection. It is recommended that samples that are repeatedly reactive should be verified/supported according to the classical algorithms of international and national guidelines. The recombinant "line immunoassay test (LIA)", which has been used for many years, is studied with the accumulated samples to be cost and labor-effective. In this study, the supplemental recombinant HIV 1/2 LIA (INNO-LIA®, Fujirebio, Ghent, Belgium) used to confirm and differentiate the diagnosis of HIV-1 and HIV-2 infections, and an immunochromatographic supplemental test (Geenius™ (Bio-Rad Laboratories, Marnes-la-Coquette, France) which can provide faster results were compared. One hundred fifty serum samples sent to Ege University Faculty of Medicine Hospital Medical Virology Laboratory with anti-HIV 1/2 and p24 antigen positive and indeterminant results and three HIV-1 positive external quality control samples were included in the study. Samples were tested both with the Geenius™ HIV 1/2 (Bio-Rad Laboratories, Marnes-la-Coquette, France) and recombinant HIV 1/2 LIA (INNO-LIA®, Fujirebio, Ghent, Belgium). HIV 1 viral load was evaluated by using Abbott real-time HIV-1 test in Abbott m200sp system (Abbott Molecular, Wiesbaden, Germany) in plasma samples. In both assays, the results were consistent in 147 samples (96.08%). Six samples that have discordant results were as follows: one sample was LIA HIV-1 positive and Geenius indeterminate, two samples were LIA indeterminant and Geenius HIV-1 positive, and in three samples, LIA was indeterminate and Geenius negative. In two EIA reactive samples (2/97, 2.06%) and three EIA negative samples (3/53, 5.66%) LIA results were indeterminant. Geenius test, on the other hand, correctly identified HIV positive and negative samples. The immunochromatographic test could be used in the diagnostic algorithm of HIV infection, due to its short application time, not being labor intensive, its ability to distinguish HIV-1/2, its high sensitivity/specificity compared to LIA, and the compliance with LIA. However, it should be noted that in acute HIV infection, all analytical antibody tests, become reactive later than the fourth generation enzyme immunoassays.
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Infecciones por VIH , VIH-1 , VIH-2 , Alemania , Anticuerpos Anti-VIH , Infecciones por VIH/diagnóstico , VIH-1/genética , VIH-2/genética , VIH-2/inmunología , Humanos , InmunoensayoRESUMEN
The Mozambique Indicators of Immunization, Malaria and HIV/AIDS (IMASIDA) survey was conducted in 2015 and used a two Enzyme Immunoassay (EIA) (Vironostika HIV-1/2 and Murex HIV-1/2) based algorithm to determine the HIV status of the consented participants. The Mozambique Ministry of Health, with support from the US Centers for Disease Control and Prevention (US CDC), added Bio-Rad Geenius™ HIV-1/2 Supplemental Assay to the IMASIDA HIV testing algorithm to confirm all specimens that were found to be reactive on one or both EIAs. In total 11690 specimens were collected to estimate the proportion of HIV positive samples. Results indicate that the proportion of HIV positive samples based on the concordant positive results of two EIA assays was 21.5% (2518/11690). The addition of the Geenius assay to the IMASIDA HIV testing algorithm demonstrated that 792 (31.5%) of 2518 specimens were false-positive and reduced the proportion of HIV positive samples to 14.7% (1722/11690), demonstrating the importance of including a highly specific HIV test to confirm HIV diagnosis. HIV surveys exclusively based on EIA testing algorithm may result in misleading high prevalence results. Our results demonstrate that more specific confirmatory testing should be added to the EIA-based algorithms to ensure accurate HIV diagnosis and correct HIV prevalence estimate in cross-sectional surveys.
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Infecciones por VIH/diagnóstico , VIH-1/inmunología , VIH-2/inmunología , Técnicas para Inmunoenzimas , Adolescente , Adulto , Algoritmos , Estudios Transversales , Reacciones Falso Positivas , Femenino , Anticuerpos Anti-VIH , Infecciones por VIH/epidemiología , Humanos , Lactante , Masculino , Tamizaje Masivo , Persona de Mediana Edad , Mozambique , Sensibilidad y Especificidad , Adulto JovenRESUMEN
BACKGROUND: The surge of methamphetamine use has been a complicating factor compounding the steeply increasing number of drug overdose deaths in the U.S. Infection from blood-borne viruses including hepatitis B virus (HBV), hepatitis C virus (HCV) and HIV, related to methamphetamine use continue to grow. This study aims to examine the risk factors associated with HBV, HCV and HIV among people who used methamphetamine. METHODS: People who ever used methamphetamine were identified from five National Health and Nutrition Examination Survey (NHANES) cohorts, 2007 to 2016. The outcome was either positive or negative for blood-borne viruses as identified from laboratory tests. Weighted statistics for the combined ten years of data were calculated by multiplying the weighted variable for laboratory measurements by 0.2. We examined the association of sexual activities (sexual partners, sexual identity), drug use behaviors (poly-drug use, injection drug use, frequency of drug use, age started using methamphetamine), demographics, and socio-economic status with blood-borne viruses using bivariate and multivariable logistic regression models. RESULTS: There were 1132 participants representing approximately 11,996,319 persons who ever used methamphetamine in the U.S. Blood-borne viruses' positive rate was 13.0 per 100,000. Multivariable logistic regression analyses showed significant associations of blood-borne infections with age 40-49 years (vs. age 20-29 years, adjusted odds ratio 4.77, 95% CI 1.11-20.55), age 50-59 years (vs. age 20-29 years, 10.25, 2.40-43.82), living within poverty index 1-1.9 (vs. poverty index > = 2, 2.55; 1.19-5.49), living below the poverty threshold (vs. poverty index > = 2, 2.55; 1.11-5.86), having lower than high school education (vs. equal or higher than high school education, 3.13; 1.51-6.46), sexual identity as other than heterosexual (vs. heterosexual, 5.60; 1.72-18.28), using methamphetamine and heroin and cocaine (vs. using methamphetamine alone, 4.24; 1.06-16.92), injection drug use (vs. no injection drug use, 3.15; 1.61-6.16), and started using methamphetamine at age above 25 (vs. started using methamphetamine at age between 10 and 17, 2.09; 1.01-4.35). CONCLUSIONS: Among people who use methamphetamine, those who use polysubstance, or who inject substances, are in urgent need for vaccination and interventions to avoid further harm from blood borne infections.