Genetic recombination is believed to assist HIV-1 diversification and escape from host immunity antiviral therapies, yet this process remains largely unexamined within the natural target-cell populations. We developed a method for measuring directly that employs reporter viruses bearing functional enhanced yellow fluorescent protein (YFP) cyan (CFP) genes in which produces modified GFP gene fluorescence infected cells. These allow simultaneous quantification of dynamics infection,...

No single universal definition of emergency department (ED) overcrowding exists. The authors hypothesize that a previously developed site-sampling form for academic ED is valid model to quantify in institutions and can be used develop validated short correlates with overcrowding.A 23-question was designed based on input from physicians at eight medical schools representative EDs nationwide. A total 336 site-samplings centers were conducted 42 computer-generated random times over three-week...

Immunogenic, broadly reactive epitopes of the HIV-1 envelope glycoprotein could serve as important targets adaptive humoral immune response in natural infection and, potentially, components an acquired deficiency syndrome vaccine. However, variability exposed and a combination highly effective envelope-cloaking strategies have made identification such problematic. Here, we show that chemokine coreceptor binding site from clade A, B, C, D, F, G, H circulating recombinant form (CRF)01, CRF02,...

In human immunodeficiency virus (HIV)-positive individuals, the vast majority of infected peripheral blood cells and lymph node may be latently or nonproductively infected. The vpr open reading frame HIV-1 encodes a 15-kDa virion-associated protein, Vpr. gene has been shown to increase replication in T monocyte/macrophages vitro. We have previously reported that expression various tumor lines leads growth inhibition differentiation, indicating Vpr function as regulator cellular...

During cell-to-cell transmission of human immunodeficiency virus type 1 (HIV-1), many viral particles can be simultaneously transferred from infected to uninfected CD4 T cells through structures called virological synapses (VS). Here we directly examine how cell-free and infections differ initiated with in the number genetic copies that are transmitted one generation next, i.e., inheritance. Following exposure HIV-1-expressing cells, show target high uptake much more likely become infected....

The vpr gene product of human immunodeficiency virus (HIV) and simian is a virion-associated regulatory protein that has been shown using mutant viruses to increase replication, particularly in monocytes/macrophages. We have previously can directly inhibit cell proliferation induce differentiation, events linked the control HIV also replication but not wild-type type 1 (HIV-1) was compatible with cellular (D. N. Levy, L. S. Fernandes, W. V. Williams, D. B. Weiner, Cell 72:541-550, 1993)....

ABSTRACT The ability of human immunodeficiency virus type 1 (HIV-1) to establish latent infections in cells has received renewed attention owing the failure highly active antiretroviral therapy eradicate HIV-1 vivo. Despite much study, molecular bases latency and reactivation are incompletely understood. Research on would benefit from a model system that is amenable rapid efficient analysis through which compounds capable regulating may be conveniently screened. We describe novel reporter...

Abstract Background The integration of HIV-1 DNA into cellular chromatin is required for high levels viral gene expression and the production new virions. However, majority remains unintegrated generally considered a replicative dead-end. A limited amount early from has been reported, but replication does not proceed further in cells which contain only DNA. Multiple infection common, that are productively infected with an integrated provirus frequently also Here we examine influence on...

Abstract Human and simian immunodeficiency viruses (HIV SIV) exploit follicular lymphoid regions by establishing high levels of viral replication dysregulating humoral immunity. Follicular regulatory T cells (T FR ) are a recently characterized subset lymphocytes that influence the germinal centre response through interactions with helper FH ). Here, utilizing both human rhesus macaque models, we show impact HIV SIV infection on number function. We find proportionately numerically expand...

ABSTRACT Percentages of activated T cells correlate with HIV-1 disease progression, but the underlying mechanisms are not fully understood. We hypothesized that HLA-DR + CD38 (DR 38 ) CD4 produce majority due to elevated expression CCR5 and CXCR4. In phytohemagglutinin (PHA)-stimulated CD8-depleted peripheral blood mononuclear (PBMC) infected green fluorescent protein (GFP) reporter viruses, DR − constituted (R5)-tropic (median, 62%) CXCR4 (X4)-tropic HIV-1-producing 61%), although cell...

Abstract HIV-1 replication is concentrated within CD4+ T cells in B cell follicles of secondary lymphoid tissues during asymptomatic disease. Limited data suggest that a subset follicular helper (TFH) germinal centers (GC) highly permissive to HIV-1. Whether GC TFH are the major virus–producing vivo has not been established. In this study, we investigated permissivity ex by spinoculating and culturing tonsil with GFP reporter viruses. Using flow cytometry, higher percentages...

HIV-1 integration is prone to a high rate of failure, resulting in the accumulation unintegrated viral genomes (uDNA) vivo and vitro. uDNA can be transcriptionally active, circularized are biochemically stable non-proliferating cells. Resting, CD4 T cells prime targets infection latently infected resting major barrier HIV cure. Our prior studies demonstrated that generates infectious virions when cell activation follows rather than precedes infection.Here, we characterize primary dynamics...

P-selectin glycoprotein ligand-1 (PSGL-1) is a dimeric, mucin-like, 120-kDa that binds to P-, E-, and L-selectins. PSGL-1 expressed primarily on the surface of lymphoid myeloid cells up-regulated during inflammation mediate leukocyte tethering rolling endothelium for migration into inflamed tissues. Although it has been reported expression inhibits HIV-1 replication, mechanism PSGL-1-mediated anti-HIV activity remains be elucidated. Here we report in virions blocks infectivity particles by...

Retroviruses pack multiple genes into relatively small genomes by encoding several in the same genomic region with overlapping reading frames. Both sense and antisense HIV-1 transcripts contain open frames for known functional proteins as well numerous alternative (ARFs). At least some ARFs have potential to encode of unknown function, their antigenic properties can be considered cryptic epitopes (CEs). To examine extent active immune response virally encoded CEs, we analyzed human leukocyte...

A major goal in HIV eradication research is characterizing the reservoir cells that harbor presence of antiretroviral therapy (ART), which reseed viremia after treatment stopped. In general, it assumed consists CD4(+) T express no viral proteins. However, recent findings suggest this may be an overly simplistic view and contribute to a diverse population includes both CD4(-) cells. study, we directly infected resting used fluorescence-activated cell sorting (FACS) fiber-optic array scanning...

Human immunodeficiency virus can spread through target cells by transmission of cell-free or directly from cell-to-cell via formation virological synapses. Although has been described as much more efficient than infection, the relative contribution two pathways to growth during multiple rounds replication remains poorly defined. Here, we fit a mathematical model previously published and newly generated in vitro data, determine that free-virus synaptic contribute approximately equally population.

HIV-1 infection of resting CD4 T cells plays a crucial and numerically dominant role during virus transmission at mucosal sites subsequent acute replication cell depletion. Resveratrol pterostilbene are plant stilbenoids associated with several health-promoting benefits. has been shown to inhibit the viruses, including herpes simplex viruses 1 2, papillomaviruses, severe respiratory syndrome virus, influenza virus. Alone, resveratrol does not activated cells, but it synergize nucleoside...

During chronic HIV infection, viral replication is concentrated in secondary lymphoid follicles. Cytotoxic CD8 T cells control extrafollicular regions, but not the follicle. Here, we show CXCR5hiCD44hiCD8 are a regulatory subset differing from conventional cells, and constitute majority of This subset, follicular (CD8 TFR), expand SIV exhibit enhanced expression Tim-3 IL-10, express less perforin compared to cells. TFR modestly limit helper (TFH), impair TFH IL-21 production via Tim-3,...

Three genomic libraries were constructed using a mixture of DNA from Solanum phureja Juz. & Buk., and S. chacoense Bitt. Two the enriched for ATT GT repeats (a 27-fold enrichment was achieved). In total, 3500 clones conventional library, 1000 library ATT, 12 000 one screened with five different repeat motifs, total 18 primer pairs obtained. Another group obtained SSR-containing sequences in public databases (18 utilized). From among 30 newly developed pairs, previously published ones,...

Parathyroid hormone-related protein (PTHrP) is a secreted that acts as an autocrine and paracrine mediator of cell proliferation differentiation. In addition to its biological activity mediated through signal transduction cascades, there evidence for intracellular role PTHrP in cycle progression apoptosis. These effects are mid-region nuclear targeting sequence (NTS) localizes the region nucleolus where ribonucleoprotein complexes form vivo. this work, we show endogenous, transfected, andin...

ABSTRACT Integration is a central event in the replication of retroviruses, yet ≥90% HIV-1 reverse transcripts fail to integrate, resulting accumulation unintegrated viral DNA cells. However, understanding what role, if any, plays natural history has remained elusive. Unintegrated reported possess limited capacity for gene expression restricted early products and considered replicative dead end. Although majority peripheral blood CD4 + T cells are refractory infection, nonactivated present...

HIV-1 hijacks and disrupts many processes in the cells it infects order to suppress antiviral immunity facilitate its replication. Resting CD4 T are important early targets of infection which must overcome intrinsic barriers viral Although resting refractory vitro, local environmental factors within lymphoid mucosal tissues such as cytokines replication while maintaining state. These can be utilized vitro study cells. In vivo, migration naïve central memory into is dependent upon expression...

Mutation rates of human immunodeficiency virus type 1 (HIV-1) genomes have been estimated using purified reverse transcriptase or single-round infection system. Since small sequences were used as templates, the overall mutation could only be extrapolated and biological significance mutations is unknown. For direct estimation HIV-1 understanding potential influences mutations, we obtained 19 complete nearly full-length proviral from single-round-infected adherent cells lymphocytes by a lambda...