RESUMO
Using a holistic, process approach, this article brings attention to cultural differences in the prevalence of fluid synchrony in collaboration, at a microanalytic scale of analysis that is embodied in the processes of everyday life. We build on findings that in a number of Indigenous American communities, fluid and harmonious collaboration is prioritized both in community organization at a scale of years and centuries, and in everyday family interactions and researcher-organized tasks at a scale of days, hours, or minutes. We examined whether this sophisticated fluid collaboration could be seen even at a scale of fractions of seconds. At a microscale of 200-millisecond segments, Guatemalan Mayan triads of mothers and children frequently engaged mutually, in fluid synchrony together, when exploring novel objects. They did so more commonly than did European American mother-child triads, who usually engaged solo or in dyads, with one person left out, or resisted each other. This microanalysis of mutuality in family interactions reveals the role of culture in the foundations of thinking and working together in both Mayan and European American communities, and the fruitfulness of considering developmental processes holistically.
Assuntos
Relações Interpessoais , Feminino , Humanos , Masculino , Estados UnidosRESUMO
This paper contrasts two ways that shared thinking can be conceptualized: as negotiation, where individuals join their separate ideas, or collaboration, as people mutually engage together in a unified process, as an ensemble. We argue that these paradigms are culturally based, with the negotiation model fitting within an assumption system of separate entities-an assumption system we believe to be common in psychology and in middle-class European American society-and the collaboration model fitting within a holistic worldview that appears to be common in Indigenous-heritage communities of the Americas. We discuss cultural differences in children's interactions-as negotiation or collaboration-that suggest how these distinct paradigms develop.
Assuntos
Comportamento Cooperativo , Comparação Transcultural , Negociação , Comportamento Social , Atenção , Criança , Humanos , Americanos Mexicanos , Motivação , Estados UnidosRESUMO
Significant sex specific differences in the progression of HIV/AIDS have been reported. Several studies have implicated steroid hormones in regulating host factor expression and modulating HIV transmission and replication. However, the exact mechanism exerted by steroid hormones estrogen and progesterone in the regulation of HIV-1 replication is still unclear. Results from the current study indicated a dose dependent down regulation of HIV-1 replication in monocyte derived macrophages pre-treated with high concentrations of estrogen or progesterone. To elucidate the molecular mechanisms associated with the down regulation of HIV-1 replication by estrogen and progesterone we used PCR arrays to analyze the expression profile of host genes involved in antiviral responses. Several chemokines, cytokines, transcription factors, interferon stimulated genes and genes involved in type-1 interferon signaling were down regulated in cells infected with HIV-1 pre-treated with high concentrations of estrogen or progesterone compared to untreated HIV-1 infected cells or HIV-1 infected cells treated with low concentrations of estrogen or progesterone. The down regulation of CXCL9, CXCL10 and CXCL11 chemokines and IL-1ß, IL-6 cytokines in response to high concentrations of estrogen and progesterone pre-treatment in HIV-1 infected cells was confirmed at the protein level by quantitating chemokine and cytokine concentrations in the culture supernatant. These results demonstrate that a potent anti-inflammatory response is mediated by pre-treatment with high concentrations of estrogen and progesterone. Thus, our study suggests a strong correlation between the down-modulation of anti-viral and pro-inflammatory responses mediated by estrogen and progesterone pre-treatment and the down regulation of HIV-1 replication. These findings may be relevant to clinical observations of sex specific differences in patient populations and point to the need for further investigation.
Assuntos
Estrogênios/fisiologia , HIV/fisiologia , Macrófagos/virologia , Progesterona/fisiologia , Replicação Viral , Humanos , Reação em Cadeia da PolimeraseRESUMO
Cultural research can help to identify strengths of cultural communities that are often viewed through a deficit model. Strengths-based approaches open researchers, practitioners, and the public to seeing the logic and value of cultural practices that vary from mainstream approaches. Strengths-based approaches include and extend beyond concerns for social equity: They are necessary for scientific characterization of human cognitive and social processes as well as for effective educational and societal practices. An example of a cultural strength is the sophisticated collaboration shown by many Indigenous-heritage children from North and Central America, which contrasts with the common practice in middle-class communities of dividing up activities into separate roles. These distinct approaches to working together fit with broader cultural paradigms that offer insights into human development as well as inspiration for alternative approaches. As an anonymous reviewer noted, the strengths of each group can be leveraged to mesh with the strengths of others.
Assuntos
Desenvolvimento Infantil , Educação Infantil/etnologia , Cultura , Família/etnologia , Relações Interpessoais , Aprendizagem , Pré-Escolar , Comunicação , Comportamento Cooperativo , Diversidade Cultural , Humanos , Idioma , PesquisaRESUMO
BACKGROUND: The HIV epidemic is expanding worldwide with an increasing number of distinct viral subtypes and circulating recombinant forms (CRFs). Out of 34 million adults living with HIV and AIDS, women account for one half of all HIV-1 infections worldwide. These gender differences in HIV pathogenesis may be attributed to sex hormones. Little is known about the role of sex hormone effects on HIV Subtypes pathogenesis. The aim of our study was to determine sex hormone effects on replication and transmissibility of HIV subtypes. METHODS: Peripheral blood mononuclear cells (PBMC) and monocyte derived dendritic cells (MDDC) from male and female donors were infected with HIV subtypes A-D and CRF02_AG, CRF01_AE, MN (lab adapted), Group-O, Group-N and HIV-2 at a concentration of 5ng/ml of p24 or p27. Virus production was evaluated by measuring p24 and p27 levels in culture supernatants. Similar experiments were carried out in the presence of physiological concentrations of sex steroid hormones. R5/X4 expressions measured by flow cytometry and transmissibility was evaluated by transfer of HIV from primary dendritic cells (DC) to autologous donor PBMC. RESULTS: Our results from primary PBMC and MDDC from male and female donors indicate in the absence of physiological concentrations of hormone treatment virus production was observed in three clusters; high replicating virus (subtype B and C), moderate replicative virus (subtype A, D, CRF01_AE, Group_N) and least replicative virus (strain MN). However, dose of sex steroid hormone treatment influenced HIV replication and transmission kinetics in PBMC, DCs and cell lines. Such effects were inconsistent between donors and HIV subtypes. Sex hormone effects on HIV entry receptors (CCR5/CXCR4) did not correlate with virus production. CONCLUSIONS: Subtypes B and C showed higher replication in PBMC from males and females and were transmitted more efficiently through DC to male and female PBMC compared with other HIV-1 subtypes, HIV-1 Group O and HIV-2. These findings are consistent with increased worldwide prevalence of subtype B and C compared to other subtypes. Sex steroid hormones had variable effect on replication or transmission of different subtypes. These findings suggest that subtype, gender and sex hormones may play a crucial role in the replication and transmission of HIV.
Assuntos
Hormônios Esteroides Gonadais/farmacologia , Infecções por HIV/transmissão , HIV-1/patogenicidade , Células Cultivadas , Células Dendríticas/virologia , Estrogênios/farmacologia , Feminino , Infecções por HIV/metabolismo , HIV-1/efeitos dos fármacos , Humanos , Células Jurkat/virologia , Leucócitos Mononucleares/virologia , Masculino , Progesterona/farmacologia , Receptores CCR5/metabolismo , Testosterona/farmacologia , Replicação Viral/efeitos dos fármacosRESUMO
IL-12 exerts several regulatory effects on natural killer (NK) cells by activating IL-12 signaling. IL-12 signaling is tightly auto-regulated to control its onset and termination, with prolonged IL-12 treatment resulting in IL-12 hyporesponsiveness. However, the mechanisms underlying IL-12 auto-regulation are still unclear. In this study we report that prolonged IL-12 treatment significantly up-regulates microRNAs (miRNAs), including miR-132, -212, and -200a in primary human NK cells. This up-regulation correlates temporally with gradually decreasing STAT4 levels and decreasing IFN-γ expression, after an initial increase within the first 16 hours of IL-12 treatment. The IL-12 hyporesponsiveness is dependent on IL-12 concentration, and associated up-regulation of miR-132, -212, and -200a. Furthermore, IL-12-hyporesponsive cells regain responsiveness of IFN-γ production 24 hours after IL-12 removal, which correlates with decreases in miR-132, -212, and -200a levels. Overexpression of miR-132, -212, and -200a by transfection into NK cells mimics IL-12 priming, inducing IL-12 hyporesponsiveness, whereas transfection of miR-132, -212, and -200a inhibitors largely abolishes IL-12 induction of IL-12 hyporesponsiveness. These data suggest that miR-132, -212, and -200a up-regulation during prolonged IL-12 treatment, negatively regulates the IL-12 signaling pathway by reducing STAT4 expression in primary human NK cells.
Assuntos
MicroRNAs/genética , Biossíntese de Proteínas/genética , Fator de Transcrição STAT4/genética , Regiões 3' não Traduzidas/genética , Sítios de Ligação/genética , Western Blotting , Células Cultivadas , Relação Dose-Resposta a Droga , Citometria de Fluxo , Regulação da Expressão Gênica/efeitos dos fármacos , Humanos , Interferon gama/genética , Interferon gama/metabolismo , Interleucina-12/farmacologia , Células Matadoras Naturais/efeitos dos fármacos , Células Matadoras Naturais/metabolismo , MicroRNAs/metabolismo , Receptores de Interleucina-12/genética , Receptores de Interleucina-12/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Fator de Transcrição STAT4/metabolismo , Regulação para Cima/efeitos dos fármacosRESUMO
The nuclear matrix protein, MATR3, is a newly-described Rev cofactor whose mechanism of action is only starting to be revealed.
Assuntos
Regulação Viral da Expressão Gênica , Produtos do Gene rev/metabolismo , Infecções por HIV/metabolismo , HIV-1/metabolismo , Proteínas Associadas à Matriz Nuclear/metabolismo , RNA Mensageiro/metabolismo , Proteínas de Ligação a RNA/metabolismo , Produtos do Gene rev/genética , Infecções por HIV/genética , Infecções por HIV/virologia , HIV-1/genética , Humanos , Proteínas Associadas à Matriz Nuclear/genética , RNA Mensageiro/genética , Proteínas de Ligação a RNA/genéticaRESUMO
NK cells are prominent mediators of the immunomodulating and antiangiogenic activity of IL-12. However, the effect of prolonged IL-12 treatment on NK cells is unclear. In this study, we observed that IL-12 initially activates NK cells, but prolonged IL-12 treatment specifically down-regulates IL-12 signaling and induces NK cell apoptosis associated with a significant reduction in cytolytic activity and IFN-γ production in response to further IL-12 stimulation. Further results demonstrate that prolonged IL-12 stimulation of NK cells specifically decreases the level of activated STAT4 protein, a critical IL-12 signaling component, through decreasing STAT4 mRNA and protein levels rather than inducing STAT4 protein degradation. IL-12 treatment induces NK cell activation as well as levels of ROS, but prolonged IL-12 treatment causes ROS accumulation, which in turn, results in the loss of Δψ(m), the release of cytochrome c, and the activation of caspase-3, resulting in NK cell apoptosis. These findings provide new insights into IL-12 regulation in human NK cells, where IL-12 initially promotes NK cell activation but subsequently limits this response through a negative-feedback mechanism.
Assuntos
Apoptose/imunologia , Interleucina-12/imunologia , Células Matadoras Naturais/imunologia , Ativação Linfocitária/imunologia , Espécies Reativas de Oxigênio/imunologia , Fator de Transcrição STAT4/imunologia , Western Blotting , Separação Celular , Regulação para Baixo , Retroalimentação Fisiológica/fisiologia , Citometria de Fluxo , Humanos , Interleucina-12/metabolismo , Células Matadoras Naturais/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Fator de Transcrição STAT4/metabolismo , Transdução de Sinais/imunologiaRESUMO
Host factors interacting with the dengue virus (DENV) 3' UTR are involved in virus replication, but their roles remain poorly understood. We used RNA affinity capture and mass spectrometry to identify p100 as a host cellular protein associated with the DENV 3' UTR. By using RNA immunoprecipitation and confocal immunofluorescence analysis we demonstrated an interaction between p100 and the 3' UTR in DENV-infected cells. We identified the A4 region (the extensive stem-loop structure at the 3' end) as the binding site of p100 by studying deletion mutants. p100 knockdown specifically reduced the levels of viral RNA and viral protein in DENV-infected cells. Furthermore, downregulation of p100 reduced the expression of a heterologously expressed luciferase-3' UTR(DENV) mRNA in an A4-dependent manner, confirming the binding data and the effects of p100 knockdown on viral replication. These results provide evidence that p100 interacts with the 3' UTR of DENV and is required for normal DENV replication.
Assuntos
Regiões 3' não Traduzidas , Vírus da Dengue/patogenicidade , Interações Hospedeiro-Patógeno , Proteínas Nucleares/metabolismo , RNA Viral/metabolismo , Replicação Viral , Sítios de Ligação , Linhagem Celular , Imunoprecipitação da Cromatina , Endonucleases , Perfilação da Expressão Gênica , Genes Reporter , Humanos , Espectrometria de Massas , Microscopia Confocal , Microscopia de Fluorescência , Proteínas Nucleares/isolamento & purificação , Ligação ProteicaRESUMO
Human natural killer (NK) cells constitute an important cellular component of innate immunity, capable of killing infected and transformed cells. The proliferation and activation of NK cells are regulated by various cytokines. Interleukin-18 (IL-18) promotes NK cell activation; however, whether the effects of IL-18 on NK cell are associated with other cytokines is still unknown. In this study, we observed that IL-18 induced NK cell apoptosis and inhibited NK cell expansion in the presence of low concentrations of interleukin-2 (IL-2), while high concentrations of IL-2 overcame these effects of IL-18, and high concentrations of IL-2 promoted the stimulatory activity of IL-18 on NK cells. At a low concentration of IL-2, IL-18 induced NK cell apoptosis in part through activation of the FasL/Fas- and TNFα/TNFR-mediated death receptor signaling by enhancing FasL expression and inhibiting c-FLIP(long) expression. However, high concentrations of IL-2 strongly blocked IL-18-induced NK cell apoptosis through alleviating IL-18-induced FasL expression and activation of Fas-mediated death signaling and increasing anti-apoptosis molecule (Bcl-X(L)). These results reveal that the effects of IL-18 on human NK cell are associated with IL-2 concentration and suggest the importance of IL-2 level in cytokine immunotherapy.
Assuntos
Interleucina-18/imunologia , Interleucina-2/imunologia , Células Matadoras Naturais/imunologia , Apoptose , Proteína Reguladora de Apoptosis Semelhante a CASP8 e FADD/imunologia , Células Cultivadas , Proteína Ligante Fas/imunologia , Humanos , Células Matadoras Naturais/citologiaRESUMO
BACKGROUND: Natural Killer (NK) cells are a crucial component of the host innate immune system with anti-viral and anti-cancer properties. However, the role of NK cells in West Nile virus (WNV) infection is controversial, with reported effects ranging from active suppression of virus to no effect at all. It was previously shown that K562-mb15-41BBL (K562D2) cells, which express IL-15 and 4-1BBL on the K562 cell surface, were able to expand and activate human primary NK cells of normal peripheral blood mononuclear cells (PBMC). The expanded NK cells were tested for their ability to inhibit WNV infection in vitro. RESULTS: Co-culture of PBMC with irradiated K562D2 cells expanded the NK cell number by 2-3 logs in 2-3 weeks, with more than 90% purity; upregulated NK cell surface activation receptors; downregulated inhibitory receptors; and boosted interferon gamma (IFN-gamma) production by approximately 33 fold. The expanded NK (D2NK) cell has strong natural killing activity against both K562 and Vero cells, and killed the WNV infected Vero cells through antibody-dependent cellular cytotoxicity (ADCC). The D2NK cell culture supernatants inhibited both WNV replication and WNV induced cytopathic effect (CPE) in Vero cells when added before or after infection. Anti-IFN-gamma neutralizing antibody blocked the NK supernatant-mediated anti-WNV effect, demonstrating a noncytolytic activity mediated through IFN-gamma. CONCLUSIONS: Co-culture of PBMC with K562D2 stimulatory cells is an efficient technique to prepare large quantities of pure and active NK cells, and these expanded NK cells inhibited WNV infection of Vero cells through both cytolytic and noncytolytic activities, which may imply a potential role of NK cells in combating WNV infection.
Assuntos
Interferon gama/imunologia , Células Matadoras Naturais/metabolismo , Ativação Linfocitária , Febre do Nilo Ocidental/imunologia , Vírus do Nilo Ocidental/fisiologia , Anticorpos Bloqueadores/farmacologia , Técnicas de Cultura de Células , Proliferação de Células , Técnicas de Cocultura , Efeito Citopatogênico Viral/efeitos dos fármacos , Efeito Citopatogênico Viral/imunologia , Citotoxicidade Imunológica/efeitos dos fármacos , Humanos , Interferon gama/biossíntese , Interferon gama/genética , Interferon gama/farmacologia , Células K562 , Células Matadoras Naturais/efeitos dos fármacos , Células Matadoras Naturais/imunologia , Células Matadoras Naturais/patologia , Receptores de Células Matadoras Naturais/metabolismo , Replicação Viral/efeitos dos fármacos , Replicação Viral/imunologia , Febre do Nilo Ocidental/virologia , Vírus do Nilo Ocidental/patogenicidadeRESUMO
BACKGROUND: Implementation of sensitive screening methods for human immunodeficiency virus (HIV) and hepatitis viruses prompts the question of what quantitative risks may result from altered deferral strategies for donation of blood by men who have had sex with men (MSM). STUDY DESIGN AND METHODS: Quantitative probabilistic models were developed to assess changes in the residual risk of transfusion-transmitted HIV and hepatitis B virus (HBV) associated with blood testing and quarantine release errors (QREs) in the initial year of two hypothetical policy scenarios that would allow donations from donors who have abstained from MSM behavior for at least 5 years (MSM5) or at least 1 year (MSM1). RESULTS: The MSM5 and MSM1 models, respectively, predicted annual increases in units of HIV-infected blood of 0.5% (0.03 mean additional units; 95% confidence interval [CI], 0-1) and 3.0% (0.18 mean additional units; 95% CI, 0-1) over current estimated HIV residual risk using recent, nationwide biologic product deviation reports to estimate QRE rates. These estimates are approximately 10-fold lower than estimates based on New York State QRE data from the previous decade. The models predicted smaller increases in infectious HBV donations. CONCLUSIONS: QREs remain the most significant preventable source of risk. More accurate inputs, including the percentage of MSM in the population, the percentage of MSM who have abstained from MSM activity for 1 or 5 years, the prevalence of HIV and HBV in MSM who have abstained from MSM activity for 1 or 5 years, the rate of self-deferral, and QRE rates, are required before making more precise predictions.
Assuntos
Doadores de Sangue , Infecções por HIV/prevenção & controle , Homossexualidade Masculina , Comportamento Sexual , Adolescente , Adulto , Idoso , Infecções por HIV/transmissão , Hepatite B/prevenção & controle , Hepatite B/transmissão , Humanos , Masculino , Pessoa de Meia-Idade , QuarentenaRESUMO
BACKGROUND: Female hormones are known to play an important role in predisposition for many infectious diseases. Recent work suggests there are gender effects in HIV/AIDS progression. Here we ask whether the sex steroid hormone beta-estradiol affects the replication of HIV-1 or the efficacy of a common anti-retroviral drug, Stavudine (D4T). RESULTS: Human PBL were infected with HIV-1 in the presence or absence of combinations of sex steroid hormones and the anti-retroviral drug, D4T. After seven days in culture, viral supernatants were assayed for HIV-1 p24 protein. beta-estradiol resulted in a modest inhibition of HIV-1 replication of approximately 26%. However, 2 nM beta-estradiol increased the amount of HIV-1 replication in the presence of 50 nM D4T from a baseline of 33% (+/- SE = 5.4) to 74% (+/- SE = 5.4) of control virus levels in the absence of drug. Both results were statistically highly significant (p < 0.001). beta-estradiol did not increase the replication of a D4T-resistant strain of HIV in the presence of D4T. The effects were unlikely to be due to general cell inhibition or toxicity because these concentrations of drug and hormone cause no cytotoxicity in PBL as measured by trypan blue exclusion. CONCLUSION: beta-estradiol inhibited both HIV-1 replication in primary human PBL and the antiretroviral efficacy of D4T in PBL cultures. To optimize antiretroviral drug therapy, it may be necessary to monitor patient hormonal status.
Assuntos
Fármacos Anti-HIV/farmacologia , Estradiol/farmacologia , Hormônios Esteroides Gonadais/farmacologia , HIV-1/efeitos dos fármacos , Estavudina/farmacologia , Células Cultivadas , Meios de Cultura/química , Interações Medicamentosas , Feminino , Proteína do Núcleo p24 do HIV/biossíntese , HIV-1/crescimento & desenvolvimento , Humanos , Linfócitos/virologia , Replicação Viral/efeitos dos fármacosRESUMO
The recent finding that inhibitors of PI3/Akt can sensitize HIV infected macrophages to oxidative stress-induced cell death suggest a potential new therapeutic approach to targeting HIV reservoirs.
Assuntos
Apoptose , Inibidores Enzimáticos/farmacologia , HIV-1/efeitos dos fármacos , Macrófagos/efeitos dos fármacos , Macrófagos/virologia , Estresse Oxidativo , Inibidores de Fosfoinositídeo-3 Quinase , Proteínas Proto-Oncogênicas c-akt/antagonistas & inibidores , Ativação Enzimática , Produtos do Gene tat/metabolismo , Produtos do Gene tat/farmacologia , Humanos , Peróxido de Hidrogênio/farmacologia , Macrófagos/enzimologia , Macrófagos/fisiologia , Fosfatidilinositol 3-Quinases/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismoRESUMO
The internet is expanding the realm of scientific publishing to include free and open public debate of published papers. Journals are beginning to support web posting of comments on their published articles and independent organizations are providing centralized web sites for posting comments about any published article. The trend promises to give one and all access to read and contribute to cutting edge scientific criticism and debate.
Assuntos
Internet , Revisão por Pares/métodos , Publicações Periódicas como Assunto/tendências , Editoração/tendências , Publicações Periódicas como Assunto/normasRESUMO
BACKGROUND: West Nile virus (WNV) transmission by transfusion was documented in 2002. Approximately 80 percent of WNV infections are asymptomatic and 1 percent develop severe neurological illness. In animals, Langerhans-dendritic cells support initial viral replication, followed by replication in lymphoid tissues and dissemination to organs and possibly to the CNS. The cellular tropism of WNV infection after transfusion and the particular human blood cells that sustain viral replication remain largely unknown. Whether primary monocyte-derived macrophages (MDMs) support WNV infection-replication and produce infectious virions, with an in vitro system, was investigated. STUDY DESIGN AND METHODS: Elutriated monocytes (CD33+/CD14+) from suitable blood donors were cultured in the presence of macrophage-colony-stimulating factor, infected with WNV-NY99 at different time points, washed, and cultivated for up to 47 days. Supernatants were tested for WNV replication by TaqMan reverse transcription-polymerase chain reaction (RT-PCR), with primers for the envelope and/or 3'NC regions, and by cDNA-PCR to detect WNV minus-strand RNA and for the presence of functional virions by infectivity assays in Vero cells. RESULTS: RT-PCR TaqMan of supernatants demonstrated productive infection of MDMs. Viral load reached 2 to 5 log above baseline in 3 to 6 days and then declined, with detectable viral replication persisting for up to 47 days. WNV minus-strand RNA was detected in Day 4 cultures, indicating active viral replication. Infected MDM cultures showed no cytopathic changes. Supernatants that were TaqMan-positive for the presence of WNV-infected Vero cells and produced cytopathic effects within 3 to 5 days of culture. CONCLUSION: The susceptibility of monocytes-macrophages to productive infection in vitro is compatible with a potential role in initial WNV replication and propagation after transmission by transfusion.
Assuntos
Macrófagos/virologia , Monócitos/virologia , Reação Transfusional , Febre do Nilo Ocidental/prevenção & controle , Febre do Nilo Ocidental/transmissão , Vírus do Nilo Ocidental/isolamento & purificação , Células Cultivadas , Primers do DNA , Humanos , Macrófagos/citologia , Monócitos/citologia , RNA Viral/genética , RNA Viral/isolamento & purificação , Reação em Cadeia da Polimerase Via Transcriptase ReversaRESUMO
We report a novel mechanism, involving up-regulation of the interleukin (IL)-7 cytokine receptor, by which human immunodeficiency virus (HIV) enhances its own production in monocyte-derived macrophages (MDM) in vitro. HIV-1 infection or treatment of MDM cultures with exogenous HIV-1 Tat(86) protein up-regulates the IL-7 receptor (IL-7R) alpha-chain at the levels of steady-state RNA, protein, and functional IL-7R on the cell surface (as measured by ligand-induced receptor signaling). This IL-7R up-regulation is associated with increased amounts of HIV-1 virions in the supernatants of infected MDM cultures treated with exogenous IL-7 cytokine. The overall effect of IL-7 stimulation on HIV replication in MDM culture supernatants is typically in the range of one log and greater. The results are consistent with a model in which HIV infection produces the Tat protein, which in turn up-regulates IL-7R in a paracrine manner. This results in increased IL-7R signaling in response to the IL-7 cytokine, which ultimately promotes early events in HIV replication, including binding/entry and possibly other steps prior to reverse transcription. The results suggest that the effects of IL-7 on HIV replication in MDM should be considered when analyzing and designing clinical trials involving treatment of patients with IL-7 or Tat vaccines.
Assuntos
Produtos do Gene tat/fisiologia , HIV-1/fisiologia , Interleucina-7/fisiologia , Macrófagos/virologia , Modelos Biológicos , Replicação Viral/fisiologia , Células Cultivadas/efeitos dos fármacos , Células Cultivadas/metabolismo , Células Cultivadas/virologia , Genes tat , Transcriptase Reversa do HIV/metabolismo , Humanos , Interleucina-7/efeitos adversos , Interleucina-7/farmacologia , Macrófagos/efeitos dos fármacos , Macrófagos/metabolismo , Comunicação Parácrina , Fator de Transcrição STAT3/metabolismo , Vírion , Replicação Viral/efeitos dos fármacos , Produtos do Gene tat do Vírus da Imunodeficiência HumanaRESUMO
Human immunodeficiency virus type 1 viral protein R (Vpr) is required for viral pathogenesis and has been implicated in T-cell apoptosis through its activation of caspase 3 and caspase 9 and perturbation of mitochondrial membrane potential. To understand better Vpr-mitochondria interaction, we report here the identification of antiapoptotic mitochondrial protein HAX-1 as a novel Vpr target. We show that Vpr and HAX-1 physically associate with each other. Overexpression of Vpr in cells dislocates HAX-1 from its normal residence in mitochondria and creates mitochondrion instability and cell death. Conversely, overexpression of HAX-1 suppressed the proapoptotic activity of Vpr.
Assuntos
Produtos do Gene vpr/metabolismo , Infecções por HIV/virologia , HIV-1/crescimento & desenvolvimento , Mitocôndrias/metabolismo , Proteínas/metabolismo , Proteínas Adaptadoras de Transdução de Sinal , Apoptose , Regulação para Baixo , Células HeLa , Humanos , Ligação Proteica , Produtos do Gene vpr do Vírus da Imunodeficiência HumanaRESUMO
Nucleo-cytoplasmic transport of RNA is one of many cellular pathways whose illumination has progressed hand in hand with understanding of retroviral mechanisms. A recent paper in Cell reports the involvement of an RNA helicase in the pathway by which HIV exports partially spliced and unspliced RNA out of the nucleus. This suggests the ubiquity of RNA helicases in RNA export from the nucleus, and has novel mechanistic implications.
Assuntos
Produtos do Gene rev/metabolismo , HIV-1/genética , RNA Viral/metabolismo , Fármacos Anti-HIV , HIV-1/enzimologia , RNA Helicases/metabolismo , Splicing de RNA , Produtos do Gene rev do Vírus da Imunodeficiência HumanaRESUMO
Correct endoproteolytic maturation of gp160 is essential for the infectivity of human immunodeficiency virus type 1. This processing of human immunodeficiency virus-1 envelope protein, gp160, into gp120 and gp41 has been attributed to the activity of the cellular subtilisin-like proprotein convertase furin. The prototypic furin recognition cleavage site is Arg-X-Arg/Lys-Arg. Arg-Arg-Arg-Arg-Arg-Arg or longer iterations of polyarginine have been shown to be competitive inhibitors of substrate cleavage by furin. Here, we tested polyarginine for inhibition of productive human immunodeficiency virus-1-infection in T-cell lines, primary peripheral blood mononuclear cells, and macrophages. We found that polyarginine inhibited significantly human immunodeficiency virus-1 replication at concentrations that were benign to cell cultures ex vivo and mice in vivo. Using a fluorogenic assay, we demonstrated that polyarginine potently inhibited substrate-specific proteolytic cleavage by furin. Moreover, we verified that authentic processing of human immunodeficiency virus-1 gp160 synthesized in human cells from an infectious human immunodeficiency virus-1 (HIV-1) molecular clone was effectively blocked by polyarginine. Taken together, our data support that inhibitors of proteolytic processing of gp160 may be useful for combating human immunodeficiency virus-1 and that polyarginine represents a lead example of such inhibitors.