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1.
PLoS Pathog ; 15(12): e1008177, 2019 12.
Artículo en Inglés | MEDLINE | ID: mdl-31821379

RESUMEN

Human immunodeficiency virus (HIV) can adapt to an individual's T cell immune response via genomic mutations that affect antigen recognition and impact disease outcome. These viral adaptations are specific to the host's human leucocyte antigen (HLA) alleles, as these molecules determine which peptides are presented to T cells. As HLA molecules are highly polymorphic at the population level, horizontal transmission events are most commonly between HLA-mismatched donor/recipient pairs, representing new immune selection environments for the transmitted virus. In this study, we utilised a deep sequencing approach to determine the HIV quasispecies in 26 mother-to-child transmission pairs where the potential for founder viruses to be pre-adapted is high due to the pairs being haplo-identical at HLA loci. This scenario allowed the assessment of specific HIV adaptations following transmission in either a non-selective immune environment, due to recipient HLA mismatched to original selecting HLA, or a selective immune environment, mediated by matched donor/recipient HLA. We show that the pattern of reversion or fixation of HIV adaptations following transmission provides insight into the replicative cost, and likely compensatory networks, associated with specific adaptations in vivo. Furthermore, although transmitted viruses were commonly heavily pre-adapted to the child's HLA genotype, we found evidence of de novo post-transmission adaptation, representing new epitopes targeted by the child's T cell response. High-resolution analysis of HIV adaptation is relevant when considering vaccine and cure strategies for individuals exposed to adapted viruses via transmission or reactivated from reservoirs.


Asunto(s)
Adaptación Biológica/genética , Infecciones por VIH/genética , VIH-1/genética , Transmisión Vertical de Enfermedad Infecciosa , Adaptación Biológica/inmunología , Adulto , Niño , Preescolar , Evolución Molecular , Femenino , Infecciones por VIH/inmunología , Infecciones por VIH/transmisión , VIH-1/inmunología , Secuenciación de Nucleótidos de Alto Rendimiento , Humanos , Lactante , Masculino , Persona de Mediana Edad
3.
Analyst ; 142(9): 1569-1580, 2017 May 02.
Artículo en Inglés | MEDLINE | ID: mdl-28386613

RESUMEN

Diagnosis of asymptomatic malaria poses a great challenge to global disease elimination efforts. Healthcare infrastructure in rural settings cannot support existing state-of-the-art tools necessary to diagnose asymptomatic malaria infections. Instead, lateral flow immunoassays (LFAs) are widely used as a diagnostic tool in malaria endemic areas. While LFAs are simple and easy to use, they are unable to detect low levels of parasite infection. We have developed a field deployable Magnetically-enabled Biomarker Extraction And Delivery System (mBEADS) that significantly improves limits of detection for several commercially available LFAs. Integration of mBEADS with leading commercial Plasmodium falciparum malaria LFAs improves detection limits to encompass an estimated 95% of the disease reservoir. This user-centered mBEADS platform makes significant improvements to a previously cumbersome malaria biomarker enrichment strategy by improving reagent stability, decreasing the processing time 10-fold, and reducing the assay cost 10-fold. The resulting mBEADS process adds just three minutes and less than $0.25 to the total cost of a single LFA, thus balancing sensitivity and practicality to align with the World Health Organization's ASSURED criteria for point-of-care (POC) testing.


Asunto(s)
Biomarcadores/análisis , Inmunoensayo , Malaria Falciparum/diagnóstico , Óxido Ferrosoférrico , Humanos , Límite de Detección , Microesferas , Plasmodium falciparum
4.
J Virol ; 86(8): 4213-21, 2012 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-22258246

RESUMEN

After initiation of antiretroviral therapy (ART), HIV loads and frequencies of HIV epitope-specific immune responses decrease. A diverse virus-specific T cell receptor (TCR) repertoire allows the host to respond to viral epitope diversity, but the effect of antigen reduction as a result of ART on the TCR repertoire of epitope-specific CD8(+) T cell populations has not been well defined. We determined the TCR repertoires of 14 HIV-specific CD8(+) T cell responses from 8 HIV-positive individuals before and after initiation of ART. We used multiparameter flow cytometry to measure the distribution of memory T cell subsets and the surface expression of PD-1 on T cell populations and T cell clonotypes within epitope-specific responses from these individuals. Post-ART, we noted decreases in the frequency of circulating epitope-specific T cells (P = 0.02), decreases in the number of T-cell clonotypes found within epitope-specific T cell receptor repertoires (P = 0.024), and an overall reduction in the amino acid diversity within these responses (P < 0.0001). Despite this narrowing of the T cell response to HIV, the overall hierarchy of dominant T cell receptor clonotypes remained stable compared to that pre-ART. CD8(+) T cells underwent redistributions in memory phenotypes and a reduction in CD38 and PD-1 expression post-ART. Despite extensive remodeling at the structural and phenotypic levels, PD-1 was expressed at higher levels on dominant clonotypes within epitope-specific responses before and after initiation of ART. These data suggest that the antigen burden may maintain TCR diversity and that dominant clonotypes are sensitive to antigen even after dramatic reductions after initiation of ART.


Asunto(s)
Infecciones por VIH/inmunología , VIH/inmunología , Receptores de Antígenos de Linfocitos T/genética , Linfocitos T/inmunología , Fármacos Anti-VIH/farmacología , Fármacos Anti-VIH/uso terapéutico , Terapia Antirretroviral Altamente Activa , Epítopos de Linfocito T/inmunología , Variación Genética/efectos de los fármacos , Variación Genética/inmunología , Infecciones por VIH/tratamiento farmacológico , Humanos , Epítopos Inmunodominantes/inmunología , Memoria Inmunológica , Activación de Linfocitos/efectos de los fármacos , Activación de Linfocitos/inmunología , Receptor de Muerte Celular Programada 1/metabolismo , Receptores de Antígenos de Linfocitos T/inmunología
5.
J Immunol ; 186(12): 6871-85, 2011 Jun 15.
Artículo en Inglés | MEDLINE | ID: mdl-21562156

RESUMEN

HIV epitope-specific T cell responses are often comprised of clonotypic expansions with distinct functional properties. In HIV(+) individuals, we measured programmed death-1 (PD-1) and IL-7Rα expression, MHC class I tetramer binding, cytokine production, and proliferation profiles of dominant and subdominant TCR clonotypes to evaluate the relationship between the composition of the HIV-specific T cell repertoire and clonotypic phenotype and function. Dominant clonotypes are characterized by higher PD-1 expression and lower C127 expression compared with subdominant clonotypes, and TCR avidity positively correlates with PD-1 expression. At low peptide concentrations, dominant clonotypes fail to survive in culture. In response to stimulation with peptides representing variant epitopes, subdominant clonotypes produce higher relative levels of cytokines and display greater capacity for cross-recognition compared with dominant clonotypes. These data indicate that dominant clonotypes within HIV-specific T cell responses display a phenotype consistent with ongoing exposure to cognate viral epitopes and suggest that cross-reactive, subdominant clonotypes may retain greater capacity to suppress replication of viral variants as well as to survive in the absence of strong antigenic signaling.


Asunto(s)
Antígenos CD/inmunología , Proteínas Reguladoras de la Apoptosis/inmunología , Reacciones Cruzadas/inmunología , VIH-1/inmunología , Subunidad alfa del Receptor de Interleucina-7/inmunología , Linfocitos T/patología , Linfocitos T/virología , Presentación de Antígeno/inmunología , Antígenos CD/análisis , Proteínas Reguladoras de la Apoptosis/análisis , Células Clonales/patología , Células Clonales/virología , Epítopos de Linfocito T/metabolismo , Infecciones por VIH/inmunología , Humanos , Subunidad alfa del Receptor de Interleucina-7/análisis , Receptor de Muerte Celular Programada 1 , Receptores de Antígenos de Linfocitos T/metabolismo , Linfocitos T/inmunología
6.
Blood ; 116(3): 396-405, 2010 Jul 22.
Artículo en Inglés | MEDLINE | ID: mdl-20424187

RESUMEN

Flexibility of the HIV-specific T-cell receptor repertoire is a hallmark of HIV-1 infection. Altered differentiation of HIV-specific CD45RO(+)/CCR7(-) (TemRO) CD8(+) effector-memory T cells into CD45RA(+)/CCR7(-) (TemRA) CD8(+) effector-memory T cells as well as increased expression of the senescence marker CD57 has been frequently observed HIV-1 infection, but the structural relationship between clonal expansion and T-cell differentiation has not been defined. In this study, we demonstrate that HIV-specific clonotypes have differing degrees of TemRA differentiation but always maintain a significant proportion of TemRO-phenotype cells. These data indicate that structural constraints of the TCR/peptide major histocompatibility complex interaction play a central role in the TemRA differentiation of HIV-specific CD8(+) T cells in chronic HIV-1 infection. Clonotypes with a predominantly TemRA phenotype had a substantial fraction of cells without expression of CD57; and in contrast to the high clonotypic variability of TemRA differentiation, expression of CD57 was highly correlated among T-cell clonotypes within epitope-specific responses, indicating TCR-independent expression of CD57 in vivo. Our data highlight the importance of the structural composition of the TCR repertoire for the effector-memory differentiation of the immune response in chronic viral infections and suggest that TCR-dependent and -independent homeostasis shapes the pathogen-specific effector-memory repertoire in vivo.


Asunto(s)
Linfocitos T CD8-positivos/inmunología , Linfocitos T CD8-positivos/patología , Infecciones por VIH/inmunología , Secuencia de Aminoácidos , Antígenos CD57/metabolismo , Diferenciación Celular/inmunología , Células Clonales/inmunología , Células Clonales/patología , Infecciones por VIH/patología , VIH-1 , Humanos , Memoria Inmunológica , Antígenos Comunes de Leucocito/metabolismo , Datos de Secuencia Molecular , Fenotipo , Receptores de Antígenos de Linfocitos T/genética , Receptores de Antígenos de Linfocitos T/metabolismo , Receptores CCR7/metabolismo , Subgrupos de Linfocitos T/inmunología , Subgrupos de Linfocitos T/patología
7.
J Virol ; 83(17): 8616-27, 2009 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-19515764

RESUMEN

Expression of HLA-B57 is associated with restricted replication of human immunodeficiency virus (HIV), but the mechanism for its protective effect remains unknown. If this advantage depends upon CD8 T-cell recognition of B57-restricted epitopes, mother-to-child transmission of escape mutations within these epitopes could nullify its protective effect. However, if the B57 advantage is largely mediated by selection for fitness-attenuating viral mutations within B57-restricted epitopes, such as T242N in TW10-Gag, then the transmission of such mutations could facilitate viral control in the haploidentical infant. We assessed the consequences of B57-associated mutations on replication capacity, viral control, and clinical outcome after vertical transmission in 13 mother-child pairs. We found that expression of HLA-B57 was associated with exceptional control of HIV during infancy, even when mutations within TW10 and most other B57-restricted epitopes were transmitted, subverting the natural immunodominance of HLA-B57. In contrast, most B57-negative infants born to B57-positive mothers progressed rapidly to AIDS. The presence of T242N led to a reproducible reduction in viral fitness, as demonstrated by in vitro assays using NL4-3 constructs encoding p24 sequences from individual mothers and infants. Associated compensatory mutations within p24-Gag were observed to reverse this impairment and to influence the propensity of T242N to revert after transmission to B57-negative hosts. Moreover, primary failure to control viremia was observed in one infant to whom multiple compensatory mutations were transmitted along with T242N. These parallel in vivo and in vitro data suggest that HLA-B57 confers its advantage primarily by driving and maintaining a fitness-attenuating mutation in p24-Gag.


Asunto(s)
Proteína p24 del Núcleo del VIH/inmunología , Infecciones por VIH/inmunología , VIH/crecimiento & desarrollo , VIH/inmunología , Antígenos HLA-B/inmunología , Transmisión Vertical de Enfermedad Infecciosa , Mutación Missense/inmunología , Secuencia de Aminoácidos , Animales , Preescolar , Progresión de la Enfermedad , Femenino , VIH/genética , Proteína p24 del Núcleo del VIH/genética , Infecciones por VIH/transmisión , Infecciones por VIH/virología , Humanos , Lactante , Masculino , Datos de Secuencia Molecular , Análisis de Secuencia de ADN
8.
Blood ; 107(6): 2373-83, 2006 Mar 15.
Artículo en Inglés | MEDLINE | ID: mdl-16322475

RESUMEN

T-cell receptor (TCR) diversity of virus-specific CD8+ T cells likely helps prevent escape mutations in chronic viral infections. To understand the dynamics of the virus-specific T cells in more detail, we followed the evolution of the TCR repertoire specific for a dominant HLA-B*08-restricted epitope in Nef (FLKEKGGL) in a cohort of subjects infected with HIV. Epitope-specific CD8+ T cells used structurally diverse TCR repertoires, with different TCRbeta variable regions and with high amino acid diversity within antigen recognition sites. In a longitudinal study, distinct Vbeta populations within the HIV-specific TCR repertoire expanded simultaneously with changes in plasma viremia, whereas other Vbeta populations remained stable or even decreased. Despite antigenic variation in some subjects, all subjects had the consensus sequence present during the study period. Functional analysis of distinct Vbeta populations revealed differences in HIV-specific IFN-gamma secretion ex vivo as well as differences in tetramer binding, indicating functional heterogeneity among these populations. This contrasts with findings in a subject on antiretroviral therapy with suppression of viremia to less than 50 copies/mL, where we observed long-term persistence of a single clonotype. Our findings illustrate the flexibility of a heterogeneous HIV-1-specific CD8+ TCR repertoire in subjects with partial control of viremia.


Asunto(s)
Linfocitos T CD8-positivos/inmunología , VIH/inmunología , Receptores de Antígenos de Linfocitos T/inmunología , Especificidad del Receptor de Antígeno de Linfocitos T , Adulto , Anciano , Variación Antigénica , Linfocitos T CD8-positivos/virología , Células Clonales/inmunología , Estudios de Cohortes , Secuencia de Consenso , Infecciones por VIH/inmunología , Humanos , Epítopos Inmunodominantes , Interferón gamma/metabolismo , Estudios Longitudinales , Persona de Mediana Edad , Viremia/prevención & control
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