RESUMEN
BACKGROUND: Increased intracellular concentration of cyclic AMP (cAMP) in T cells is associated with various immunodeficiency conditions including human immunodeficiency virus (HIV) infection. Several reports indicate a critical role of activated protein kinase A (PKA) in the susceptibility of cells to HIV infection. We have used a cell permeable, stable peptidomimetic version (P3) of the RI-anchoring disruptor (RIAD), which prevents PKA interaction with A-kinase-anchoring proteins (AKAPs). It is known that RIAD peptide abrogates effects of localized cAMP signalling through anchored type I PKA in lymphocytes and prevents murine AIDS (MAIDS) infection when expressed as a transgene in mice. METHODS AND RESULTS: In vitro HIV-infected human peripheral blood mononuclear cells (PBMCs) show reduced levels of p24 and intracellular cAMP in T cells when treated with RIAD peptidomimetic (RIAD-P3). Humanized NOD/SCID/IL2γnull (NSG) mice infected with HIV-1 JRCSF and treated with RIAD-P3 (3·5 mg) once every 2 weeks showed significantly reduced levels of viral load at +28, +42 and +56 days and increased CD4 numbers at +56 days after the start of treatment. RIAD-P3-treated humanized mice had lower levels of intracellular cAMP in T cells sorted from splenocytes. CONCLUSIONS: Treatment with RIAD-P3 limits HIV-1 viral replication and stabilizes CD4 levels by mechanisms involving cAMP/PKA-I pathway in human PBMCs and humanized NSG mice.
Asunto(s)
Infecciones por VIH/tratamiento farmacológico , VIH-1/efectos de los fármacos , Peptidomiméticos/farmacología , Proteínas de Anclaje a la Quinasa A/metabolismo , Animales , Linfocitos T CD4-Positivos/efectos de los fármacos , Linfocitos T CD4-Positivos/metabolismo , Linfocitos T CD4-Positivos/virología , Células Cultivadas , AMP Cíclico/metabolismo , Proteínas Quinasas Dependientes de AMP Cíclico/metabolismo , Proteína p24 del Núcleo del VIH/metabolismo , Humanos , Péptidos y Proteínas de Señalización Intracelular/farmacología , Leucocitos Mononucleares/virología , Ratones SCID , Ratones Transgénicos , Transducción de Señal/fisiología , Carga Viral/efectos de los fármacos , Replicación Viral/efectos de los fármacosRESUMEN
Chronic HIV infection is characterized by chronic immune activation and dysfunctional T cells with elevated intracellular cyclic AMP (cAMP), which inhibits the T cell activation capability. cAMP may be induced by prostaglandin E(2) following lipopolysaccharide (LPS)-induced upregulation of cyclooxygenase type 2 (COX-2) in monocytes due to the elevated LPS levels in patients with chronic HIV infection. This hypothesis was tested using celecoxib, a COX-2 inhibitor, for 12 weeks in HIV-infected patients without antiretroviral treatment in a prospective, open, randomized exploratory trial. Thirty-one patients were randomized in the trial; 27 completed the study, including 13 patients on celecoxib. Celecoxib reduced chronic immune activation in terms of CD38 density on CD8(+) T cells (-24%; P = 0.04), IgA levels (P = 0.04), and a combined score for inflammatory markers (P < 0.05). Celecoxib further reduced the inhibitory surface receptor programmed death 1 (PD-1) on CD8(+) T cells (P = 0.01), including PD-1 on the HIV Gag-specific subset (P = 0.02), enhanced the number of CD3(+) CD4(+) CD25(+) CD127(lo/-) Treg or activated cells (P = 0.02), and improved humoral memory recall responses to a T cell-dependent vaccine (P = 0.04). HIV RNA (P = 0.06) and D dimers (P = 0.07) tended to increase in the controls, whereas interleukin-6 (IL-6) possibly decreased in the treatment arm (P = 0.10). In conclusion, celecoxib downmodulated the immune activation related to clinical progression of chronic HIV infection and improved T cell-dependent functions in vivo.
Asunto(s)
Vacunas contra el SIDA/inmunología , Inhibidores de la Ciclooxigenasa 2/uso terapéutico , Infecciones por VIH/tratamiento farmacológico , Infecciones por VIH/inmunología , VIH-1/efectos de los fármacos , Pirazoles/uso terapéutico , Sulfonamidas/uso terapéutico , Linfocitos T/inmunología , Vacunas contra el SIDA/administración & dosificación , Adulto , Celecoxib , Enfermedad Crónica , Inhibidores de la Ciclooxigenasa 2/administración & dosificación , Progresión de la Enfermedad , Regulación hacia Abajo , Femenino , VIH-1/inmunología , Humanos , Masculino , Persona de Mediana Edad , Pirazoles/administración & dosificación , Sulfonamidas/administración & dosificación , Resultado del TratamientoRESUMEN
The complexity of immunoregulation has focused attention on the CD4+ T "suppressor" regulatory cell (Treg), which helps maintain balance between immunity and tolerance. An immunoregulatory T-cell population that upon activation amplifies cellular immune responses was described in murine models more than 30 years ago; however, no study has yet identified a naturally occurring T "inducer" cell type. Here, we report that the ectoenzyme CD39/NTPDase1 (ecto-nucleoside triphosphate diphosphohydrolase 1) helps to delineate a novel population of human "inducer" CD4+ T cells (Tind) that significantly increases the proliferation and cytokine production of responder T cells in a dose-dependent manner. Furthermore, this unique Tind subset produces a distinct repertoire of cytokines in comparison to the other CD4+ T-cell subsets. We propose that this novel CD4+ T-cell population counterbalances the suppressive activity of suppressor Treg in peripheral blood and serves as a calibrator of immunoregulation.
Asunto(s)
Antígenos CD/inmunología , Apirasa/inmunología , Linfocitos T CD4-Positivos/inmunología , Subgrupos de Linfocitos T/inmunología , Linfocitos T CD4-Positivos/citología , Proliferación Celular , Citocinas/biosíntesis , Citocinas/inmunología , Humanos , Subgrupos de Linfocitos T/citologíaRESUMEN
Recent studies have indicated that Treg contribute to the HIV type 1 (HIV-1)-related immune pathogenesis. However, it is not clear whether T cells with suppressive properties reside within the HIV-1-specific T-cell population. Here, PBMC from HIV-1-infected individuals were stimulated with a 15-mer Gag peptide pool, and HIV-1-specific T cells were enriched by virtue of their secretion of IL-10 or IFN-gamma using immunomagnetic cell-sorting. Neither the IL-10-secreting cells nor the IFN-gamma-secreting cells expressed the Treg marker FOXP3, yet the IL-10-secreting cells potently suppressed anti-CD3/CD28-induced CD4(+) as well as CD8(+) T-cell proliferative responses. As shown by intracellular cytokine staining, IL-10- and IFN-gamma-producing T cells represent distinct subsets of the HIV-1-specific T cells. Our data collectively suggest that functionally defined HIV-1-specific T-cell subsets harbor potent immunoregulatory properties that may contribute to HIV-1-associated T-cell dysfunction.
Asunto(s)
Infecciones por VIH/inmunología , VIH-1/inmunología , Interleucina-10/inmunología , Subgrupos de Linfocitos T/inmunología , Linfocitos T Reguladores/inmunología , Proliferación Celular , Citometría de Flujo , Factores de Transcripción Forkhead/sangre , Factores de Transcripción Forkhead/inmunología , Infecciones por VIH/sangre , Humanos , Separación Inmunomagnética , Interferón gamma/sangre , Interferón gamma/inmunología , Interleucina-10/sangre , Leucocitos Mononucleares/inmunología , Estadísticas no Paramétricas , Subgrupos de Linfocitos T/citología , Linfocitos T Reguladores/citologíaRESUMEN
We have reported previously the design of a RIAD (RI-anchoring disruptor) peptide that specifically displaces PKA (protein kinase A) type I from the AKAP (A-kinase-anchoring protein) ezrin, which is present in the immunological synapse of T-cells. This increases immune reactivity by reducing the threshold for activation and may prove a feasible approach for improving immune function in patients with cAMP-mediated T-cell dysfunction. However, the use of RIAD in biological systems is restricted by its susceptibility to enzymatic cleavage and, consequently, its short half-life in presence of the ubiquitous serum peptidases. In the present study, carefully selected non-natural amino acids were employed in the design of RIAD analogues with improved stability. The resulting peptidomimetics demonstrated up to 50-fold increased half-lives in serum compared with RIAD, while maintaining similar or improved specificity and potency with respect to disruption of PKA type I-AKAP interactions.
Asunto(s)
Proteínas de Anclaje a la Quinasa A/metabolismo , Péptidos/síntesis química , Péptidos/metabolismo , Transducción de Señal/fisiología , Animales , Proteína Tirosina Quinasa CSK , Bovinos , Células Cultivadas , Cromatografía Líquida de Alta Presión , AMP Cíclico/metabolismo , Humanos , Interleucina-2/metabolismo , Ratones , Péptidos/química , Unión Proteica , Proteínas Tirosina Quinasas/metabolismo , Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción , Familia-src QuinasasRESUMEN
OBJECTIVE: The aim of this systematic review was to investigate the potential effect of pictograms on patient adherence to medication therapies. METHOD: PubMed, MEDLINE, Embase, CINAHL, and CENTRAL were searched for relevant articles. Experimental studies testing the use of pictograms in patient counselling regarding medication therapy, which quantitatively measured adherence, were included. RESULTS: Seventeen studies were identified that fulfilled our inclusion criteria. These were heterogeneous with respect to study setting, population size, and the medication regimen tested. All the studies had methodological quality limitations. The pictogram interventions differed with respect to complexity, intervention length, and the measured adherence outcome. Ten studies (58.8 %) reported a statistically significant effect, of the pictogram intervention in question, on patient adherence to medication therapies. Of these, 80 % involved populations at elevated risk for non-adherence. CONCLUSION AND PRACTICE IMPLICATIONS: Pictograms used in combination with written and/or oral information can have a positive impact on patient populations that are highly at risk for non-adherence when counselled on the proper use of medicines.
Asunto(s)
Comunicación en Salud/métodos , Cumplimiento de la Medicación , Educación del Paciente como Asunto , Comprensión , Humanos , Preparaciones FarmacéuticasRESUMEN
Antitumor immunity is mediated by Th1 CD4+ and CD8+ T lymphocytes, which induce tumor-specific cytolysis, whereas Th17 CD4+ T cells have been described to promote tumor growth. Here, we explored the influence of IL-17 on the ability of therapeutic vaccines to induce the rejection of tumors in mice using several adjuvants known to elicit either Th1 or Th17-type immunity. Immunization of mice with Th1-adjuvanted vaccine induced high levels of IFN-γ-producing T cells, whereas injection with Th17-promoting adjuvants triggered the stimulation of both IL-17 and IFN-γ-producing T cells. However, despite their capacity to induce strong Th1 responses, these Th17-promoting adjuvants failed to induce the eradication of tumors. In addition, the systemic administration of IL-17A strongly decreases the therapeutic effect of Th1-adjuvanted vaccines in two different tumor models. This suppressive effect correlated with the capacity of systemically delivered IL-17A to inhibit the induction of CD8+ T-cell responses. The suppressive effect of IL-17A on the induction of CD8+ T-cell responses was abolished in mice depleted of neutrophils, clearly demonstrating the role played by these cells in the inhibitory effect of IL-17A in the induction of antitumor responses. These results demonstrate that even though strong Th1-type responses favor tumor control, the simultaneous activation of Th17 cells may redirect or curtail tumor-specific immunity through a mechanism involving neutrophils. This study establishes that IL-17 plays a detrimental role in the development of an effective antitumor T cell response and thus could strongly affect the efficiency of immunotherapy through the inhibition of CTL responses.