RESUMEN
Inducing humoral, cellular and mucosal immunity is likely to improve the effectiveness of HIV-1 vaccine strategies. Here, we tested a vaccine regimen in pigtail macaques using an intranasal (i.n.) recombinant Fowl Pox Virus (FPV)-gag pol env-IL-4R antagonist prime, intramuscular (i.m.) recombinant Modified Vaccinia Ankara Virus (MVA)-gag pol-IL-4R antagonist boost followed by an i.m SOSIP-gp140 boost. The viral vector-expressed IL-4R antagonist transiently inhibited IL-4/IL-13 signalling at the vaccination site. The SOSIP booster not only induced gp140-specific IgG, ADCC (antibody-dependent cellular cytotoxicity) and some neutralisation activity, but also bolstered the HIV-specific cellular and humoral responses. Specifically, superior sustained systemic and mucosal HIV Gag-specific poly-functional/cytotoxic CD4+ and CD8+ T cells were detected with the IL-4R antagonist adjuvanted strategy compared to the unadjuvanted control. In the systemic compartment elevated Granzyme K expression was linked to CD4+ T cells, whilst Granzyme B/TIA-1 to CD8+ T cells. In contrast, the cytotoxic marker expression by mucosal CD4+ and CD8+ T cells differed according to the mucosal compartment. This vector-based mucosal IL-4R antagonist/SOSIP booster strategy, which promotes cytotoxic mucosal CD4+ T cells at the first line of defence, and cytotoxic CD4+ and CD8+ T cells plus functional antibodies in the blood, may prove valuable in combating mucosal infection with HIV-1 and warrants further investigation.
Asunto(s)
Vacunas contra el SIDA/farmacología , Linfocitos T CD4-Positivos/inmunología , Linfocitos T CD8-positivos/inmunología , Inmunidad Celular/efectos de los fármacos , Inmunidad Humoral/efectos de los fármacos , Inmunización Secundaria , Subunidad alfa del Receptor de Interleucina-4/antagonistas & inhibidores , Productos del Gen env del Virus de la Inmunodeficiencia Humana/farmacología , Animales , Subunidad alfa del Receptor de Interleucina-4/inmunología , Macaca nemestrinaRESUMEN
We have previously identified differences in the presentation and treatment of cancer between patients who live in rural compared with urban areas, but have not yet seen differences in those treated by orthognathic surgery. We hypothesised that patients from areas further away from the hospital face higher costs to attend and may not present with minor problems as often as those who live nearby. We therefore retrospectively reviewed all those (n=216) who had presented for orthognathic surgery over a six-year period (May 2011 to May 2017). The severity of malocclusion and facial asymmetry was established by combining measurements of intraoperative movements. Rurality was measured as the distance from home to the hospital at the time of operation. Those with smaller intraoperative movements (less than 7mm combined movement) lived significantly closer to the hospital as the crow flies (mean difference 15.13 miles, 95% CI 0.20 to 30.48, p=0.05) and could travel there more quickly (mean difference 65minutes 95% CI 9.8 to 121.7, p=0.02) than those with larger movements. Our results suggest that patients with small malocclusions and slight facial asymmetry who live further away from the hospital, may be less likely to present for operation than those who live closer. We explain why socioeconomic class is unlikely to confound our results, and suggest potential ways to minimise the effect observed.
Asunto(s)
Procedimientos Quirúrgicos Ortognáticos/estadística & datos numéricos , Población Rural/estadística & datos numéricos , Población Urbana/estadística & datos numéricos , Adolescente , Adulto , Asimetría Facial/epidemiología , Asimetría Facial/patología , Asimetría Facial/cirugía , Femenino , Accesibilidad a los Servicios de Salud/estadística & datos numéricos , Humanos , Masculino , Maloclusión/epidemiología , Maloclusión/patología , Maloclusión/cirugía , Persona de Mediana Edad , Aceptación de la Atención de Salud/estadística & datos numéricos , Estudios Retrospectivos , Escocia/epidemiología , Índice de Severidad de la Enfermedad , Factores Socioeconómicos , Adulto JovenRESUMEN
Tissue-resident memory (TRM) CD8 T cells survey a range of non-lymphoid mucosal tissues where they rapidly mediate clearance of viral infections at the entry portals. Vaccines that establish CD8 TRM cells in the cervicovaginal mucosa hold promise for effective immunity against sexually transmitted HIV. We demonstrate that HIV-specific CD8 TRM cells can be established in the murine vaginal mucosa using a combined intranasal and intravaginal mucosal immunization with recombinant influenza-HIV vectors. Using in situ tetramer immunofluorescence microscopy, we found that this mucosally administered prime-boost immunization also resulted in the durable seeding of CD8 T cells in the frontline vaginal epithelial compartment as opposed to the vaginal submucosa. Upon cognate antigen recognition within the vaginal mucosa, these HIV-specific CD8 TRM cells rapidly initiated a tissue-wide state of immunity. The activation of HIV-specific CD8 TRM cells resulted in the upregulation of endothelial vessel addressin expression and substantial recruitment of both adaptive and innate immune cells in the vaginal mucosa. These findings suggest that the epithelial localization of HIV-specific CD8 TRM cell populations and their capacity to rapidly activate both arms of the immune system could significantly augment frontline defenses against vaginal HIV infection.
Asunto(s)
Vacunas contra el SIDA/inmunología , Linfocitos T CD8-positivos/inmunología , Endotelio Vascular/fisiología , Infecciones por VIH/inmunología , VIH-1/inmunología , Membrana Mucosa/inmunología , Vagina/inmunología , Inmunidad Adaptativa , Animales , Movimiento Celular , Femenino , Vectores Genéticos , Humanos , Inmunización Secundaria , Memoria Inmunológica , Gripe Humana/genética , Activación de Linfocitos , Ratones , Ratones Endogámicos BALB C , Membrana Mucosa/virología , Especificidad de ÓrganosRESUMEN
PURPOSE: To compare morbidity and in-hospital mortality in patients who underwent surgery for femoral neck fracture on weekends versus on weekdays. METHODS: Records of 90 men and 225 women (mean age, 80.5 years) who underwent surgery for femoral neck fractures on weekends or public holidays (n=110) or on weekdays (n=205) were retrospectively reviewed. The morbidity and in-hospital mortality of the 2 groups were compared. RESULTS: The 2 groups were comparable in terms of age, sex, and time to surgery, but more hemiarthroplasties were performed on weekdays (35.0% vs. 25.0%, p=0.036). Compared with surgery on weekdays, surgery on weekends was associated with increased in-hospital mortality (3.4% vs. 9.1%, p=0.04). None of the potential confounders (age, type of surgery, presenting hospital, and time to surgery) had a significant effect on in-hospital mortality. CONCLUSION: In patients with femoral neck fractures, surgery on weekends was associated with increased in-hospital mortality but not with increased morbidity after adjusting for confounders, compared with surgery on weekdays.
Asunto(s)
Fracturas del Cuello Femoral/complicaciones , Fracturas del Cuello Femoral/mortalidad , Factores de Edad , Anciano , Anciano de 80 o más Años , Australia , Femenino , Fracturas del Cuello Femoral/cirugía , Mortalidad Hospitalaria , Humanos , Masculino , Persona de Mediana Edad , Estudios Retrospectivos , Factores de TiempoRESUMEN
Evidence from the RV144 HIV-1 vaccine trial implicates anti-HIV-1 antibody-dependent cellular cytotoxicity (ADCC) in vaccine-conferred protection from infection. Among effector cells that mediate ADCC are natural killer (NK) cells. The ability of NK cells to be activated in an antibody-dependent manner is reliant upon several factors. In general, NK cell-mediated antibody-dependent activation is most robust in terminally differentiated CD57(+) NK cells, as well as NK cells educated through ontological interactions between inhibitory killer immunoglobulin-like receptors (KIR) and their major histocompatibility complex class I [MHC-I or human leucocyte antigen (HLA-I)] ligands. With regard to anti-HIV-1 antibody-dependent NK cell activation, previous research has demonstrated that the epidemiologically relevant KIR3DL1/HLA-Bw4 receptor/ligand combination confers enhanced activation potential. In the present study we assessed the ability of the KIR2DL1/HLA-C2 receptor/ligand combination to confer enhanced activation upon direct stimulation with HLA-I-devoid target cells or antibody-dependent stimulation with HIV-1 gp140-pulsed CEM.NKr-CCR5 target cells in the presence of an anti-HIV-1 antibody source. Among donors carrying the HLA-C2 ligand for KIR2DL1, higher interferon (IFN)-γ production was observed within KIR2DL1(+) NK cells than in KIR2DL1(-) NK cells upon both direct and antibody-dependent stimulation. No differences in KIR2DL1(+) and KIR2DL1(-) NK cell activation were observed in HLA-C1 homozygous donors. Additionally, higher activation in KIR2DL1(+) than KIR2DL1(-) NK cells from HLA-C2 carrying donors was observed within less differentiated CD57(-) NK cells, demonstrating that the observed differences were due to education and not an overabundance of KIR2DL1(+) NK cells within differentiated CD57(+) NK cells. These observations are relevant for understanding the regulation of anti-HIV-1 antibody-dependent NK cell responses.
Asunto(s)
Anticuerpos Anti-VIH/biosíntesis , VIH-1/inmunología , Antígenos HLA-C/inmunología , Inmunidad Humoral , Células Asesinas Naturales/efectos de los fármacos , Receptores KIR2DL1/inmunología , Alelos , Linfocitos T CD4-Positivos/efectos de los fármacos , Linfocitos T CD4-Positivos/inmunología , Linfocitos T CD4-Positivos/virología , Antígenos CD57/genética , Antígenos CD57/inmunología , Expresión Génica , Anticuerpos Anti-VIH/farmacología , Infecciones por VIH/inmunología , Infecciones por VIH/virología , Antígenos HLA-C/clasificación , Antígenos HLA-C/genética , Prueba de Histocompatibilidad , Humanos , Memoria Inmunológica/efectos de los fármacos , Interferón gamma/biosíntesis , Interferón gamma/inmunología , Células Asesinas Naturales/inmunología , Células Asesinas Naturales/virología , Activación de Linfocitos/efectos de los fármacos , Cultivo Primario de Células , Receptores KIR2DL1/deficiencia , Receptores KIR2DL1/genética , Proteínas Recombinantes/genética , Proteínas Recombinantes/inmunología , Proteínas Recombinantes/farmacología , Productos del Gen env del Virus de la Inmunodeficiencia Humana/genética , Productos del Gen env del Virus de la Inmunodeficiencia Humana/inmunología , Productos del Gen env del Virus de la Inmunodeficiencia Humana/farmacologíaRESUMEN
Natural killer (NK) cell-mediated antibody-dependent cellular cytotoxicity (ADCC) has been linked to protection from HIV infection and slower progression towards AIDS. However, antibody-dependent activation of NK cells results in phenotypical alterations similar to those observed on NK cells from individuals with progressive HIV infection. Activation of NK cells induces matrix metalloproteinase (MMP)-mediated cleavage of cell surface CD16. In the present study we assessed the phenotype and functional profile of NK cells exhibiting post-activation MMP-mediated CD16 cleavage. We found that NK cells achieving the highest levels of activation during stimulation exhibit the most profound decreases in CD16 expression. Further, we observed that educated KIR3DL1(+) NK cells from human leucocyte antigen (HLA)-Bw4-carrying donors exhibit larger decreases in CD16 expression post-activation than the KIR3DL1(-) NK cell subset containing cells educated via other inhibitory receptor/ligand combinations and non-educated NK cells. Lastly, we assessed the ex-vivo expression of CD16 on educated KIR3DL1(+) NK cells and the KIR3DL1(-) NK cell subset from HLA-Bw4-carrying HIV-uninfected and HIV-infected donors. Suggestive of in-vivo activation of KIR3DL1(+) NK cells during HIV infection, CD16 expression was higher on KIR3DL1(+) than KIR3DL1(-) NK cells in uninfected donors but similar on both subsets in HIV-infected donors. These results are discussed in the context of how they may assist with understanding HIV disease progression and the design of immunotherapies that utilize antibody-dependent NK cell responses.
Asunto(s)
Infecciones por VIH/inmunología , Células Asesinas Naturales/inmunología , Metaloproteinasas de la Matriz/inmunología , ARN Viral/sangre , Receptores de IgG/inmunología , Anticuerpos/farmacología , Citotoxicidad Celular Dependiente de Anticuerpos , Progresión de la Enfermedad , Proteínas Ligadas a GPI/genética , Proteínas Ligadas a GPI/inmunología , Expresión Génica , Infecciones por VIH/genética , Infecciones por VIH/patología , Infecciones por VIH/virología , VIH-1/inmunología , Antígenos HLA-B/genética , Antígenos HLA-B/inmunología , Prueba de Histocompatibilidad , Humanos , Inmunofenotipificación , Células Asesinas Naturales/efectos de los fármacos , Células Asesinas Naturales/patología , Células Asesinas Naturales/virología , Activación de Linfocitos , Metaloproteinasas de la Matriz/genética , Fenotipo , Cultivo Primario de Células , Proteolisis , Receptores de IgG/genética , Receptores KIR3DL1/genética , Receptores KIR3DL1/inmunología , Transducción de Señal , Carga ViralRESUMEN
Human immunodeficiency virus (HIV) management is entering a "universal test and treat" phase, although the benefits from this approach in developed world scenarios are uncertain. We analyzed 79 combination anti-retroviral therapy (cART)-naïve HIV-positive individuals who were intensively prospectively followed from 2004 to 2013. We studied HIV-related illnesses, potential HIV transmissions, impact on sexual behavior, and factors impeding earlier cART initiation. Sixty-eight (86 %) subjects commenced cART at a mean of 6.0 years after diagnosis: 71 % with a CD4 T-cell count <350 cells/µl. A significant minority of subjects (29 %) resisted initiation of cART despite physician recommendation for a mean of 18 months. Only one HIV-related illness occurred in a patient who had not previously recorded a CD4 T-cell count <500 cell/µl, totaling 195 person-years of observation. A 40 % increase in sexually transmitted infections (STIs) occurred after commencing cART. We detected six HIV transmissions in our cohort, all of which were before initiating cART and 5 of them had a prior CD4 T-cell count <500 cells/µl. Illnesses related to cART deferral were rare and most HIV transmissions we detected occurred in people with a prior CD4 T-cell count <500 cells/µl. Our study raises concerns about increasing STI rates after cART initiation. Focusing resources on cART initiation among patients with CD4 T-cell counts <500 cells/µl and enhancing safe sexual practices should remain a priority.
Asunto(s)
Terapia Antirretroviral Altamente Activa/efectos adversos , Infecciones por VIH/epidemiología , Enfermedades de Transmisión Sexual/epidemiología , Adulto , Australia/epidemiología , Recuento de Linfocito CD4 , Linfocitos T CD4-Positivos , Estudios de Cohortes , Quimioterapia Combinada , Femenino , Estudios de Seguimiento , Infecciones por VIH/tratamiento farmacológico , Infecciones por VIH/transmisión , Humanos , Masculino , Persona de Mediana Edad , Enfermedades de Transmisión Sexual/prevención & control , Adulto JovenRESUMEN
UNLABELLED: The mechanisms by which hepatitis B virus (HBV) establishes and maintains chronic hepatitis B infection (CHB) are poorly defined. Innate immune responses play an important role in reducing HBV replication and pathogenesis. HBV has developed numerous mechanisms to escape these responses, including the production of the secreted hepatitis B e antigen (HBeAg), which has been shown to regulate antiviral toll-like receptor (TLR) and interleukin-1 (IL-1) signaling. IL-18 is a related cytokine that inhibits HBV replication in hepatoma cell lines and in the liver through the induction of gamma interferon (IFN-γ) by NK cells and T cells. We hypothesized that HBV or HBV proteins inhibit IFN-γ expression by NK cells as an accessory immunomodulatory function. We show that HBeAg protein inhibits the NF-κB pathway and thereby downregulates NK cell IFN-γ expression. Additionally, IFN-γ expression was significantly inhibited by exposure to serum from individuals with HBeAg-positive but not HBeAg-negative chronic HBV infection. Further, we show that the HBeAg protein suppresses IL-18-mediated NF-κB signaling in NK and hepatoma cells via modulation of the NF-κB pathway. Together, these findings show that the HBeAg inhibits IL-18 signaling and IFN-γ expression, which may play an important role in the establishment and/or maintenance of persistent HBV infection. IMPORTANCE: It is becoming increasingly apparent that NK cells play a role in the establishment and/or maintenance of chronic hepatitis B infection. The secreted HBeAg is an important regulator of innate and adaptive immune responses. We now show that the HBeAg downregulates NK cell-mediated IFN-γ production and IL-18 signaling, which may contribute to the establishment of infection and/or viral persistence. Our findings build on previous studies showing that the HBeAg also suppresses the TLR and IL-1 signaling pathways, suggesting that this viral protein is a key regulator of antiviral innate immune responses.
Asunto(s)
Regulación hacia Abajo , Antígenos e de la Hepatitis B/metabolismo , Virus de la Hepatitis B/metabolismo , Hepatitis B/genética , Interferón gamma/genética , Interleucina-18/metabolismo , Adulto , Células Cultivadas , Femenino , Hepatitis B/inmunología , Hepatitis B/virología , Antígenos e de la Hepatitis B/genética , Virus de la Hepatitis B/genética , Interacciones Huésped-Patógeno , Humanos , Interferón gamma/inmunología , Interleucina-18/genética , Células Asesinas Naturales/inmunología , Masculino , Persona de Mediana Edad , Transducción de Señal , Adulto JovenRESUMEN
BACKGROUND: Road traffic crashes are responsible for a vast amount of death and disability in developing countries. This study uses a bottom up, micro-costing approach to determine the cost of road traffic related crashes in South Africa. METHODS: Using the data from one hundred consecutive RTC related admissions to a regional hospital in South Africa we performed a bottom up costing study. To calculate costs patients were reviewed every 48 h and all interventions were recorded for each individual patient. Prices of interventions were obtained from hospital pricelists. A total cost was calculated on an individual basis. RESULTS: The total cost of in-patient care for these patients was US $6,98,850. Upper limb injuries were the most expensive, and the total cost increased with the number of body regions injured. The biggest expenditure was on ward overheads ($2,81,681). Ninety operations were performed - the total cost of theatre time was $1,48,230 and the cost of orthopaedic implants was $1,26,487. CONCLUSION: The cost of care of a RTC victim is significant. In light of the high numbers of RTC victims admitted over the course of the year this is a significant cost burden for a regional hospital to bear. This cost must be taken into account when allocating hospital budgets.
Asunto(s)
Accidentes de Tránsito/economía , Costos y Análisis de Costo/métodos , Costos de Hospital , Centros Traumatológicos/economía , Heridas y Lesiones/economía , Adolescente , Adulto , Anciano , Niño , Preescolar , Femenino , Servicios de Salud/economía , Servicios de Salud/estadística & datos numéricos , Hospitalización/economía , Humanos , Masculino , Persona de Mediana Edad , Prevalencia , Estudios Retrospectivos , Sudáfrica/epidemiología , Heridas y Lesiones/epidemiología , Adulto JovenRESUMEN
Natural killer T cells are a potent mediator of anti-viral immunity in mice, but little is known about the effects of manipulating NKT cells in non-human primates. We evaluated the delivery of the NKT cell ligand, α-galactosylceramide (α-GalCer), in 27 macaques by studying the effects of different dosing (1-100 µg), and delivery modes [directly intravenously (i.v.) or pulsed onto blood or peripheral blood mononuclear cells]. We found that peripheral NKT cells were depleted transiently from the periphery following α-GalCer administration across all delivery modes, particularly in doses of ≥10 µg. Furthermore, NKT cell numbers frequently remained depressed at i.v. α-GalCer doses of >10 µg. Levels of cytokine expression were also not enhanced after α-GalCer delivery to macaques. To evaluate the effects of α-GalCer administration on anti-viral immunity, we administered α-GalCer either together with live attenuated influenza virus infection or prior to simian immunodeficiency virus (SIV) infection of two macaques. There was no clear enhancement of influenza-specific T or B cell immunity following α-GalCer delivery. Further, there was no modulation of pathogenic SIVmac251 infection following α-GalCer delivery to a further two macaques in a pilot study. Accordingly, although macaque peripheral NKT cells are modulated by α-GalCer in vivo, at least for the dosing regimens tested in this study, this does not appear to have a significant impact on anti-viral immunity in macaque models.
Asunto(s)
Galactosilceramidas/farmacología , Células T Asesinas Naturales/efectos de los fármacos , Células T Asesinas Naturales/inmunología , Adyuvantes Inmunológicos/administración & dosificación , Adyuvantes Inmunológicos/farmacología , Animales , Galactosilceramidas/administración & dosificación , Activación de Linfocitos/efectos de los fármacos , Activación de Linfocitos/inmunología , Recuento de Linfocitos , Macaca nemestrina , Enfermedades de los Monos/inmunología , Células T Asesinas Naturales/metabolismo , Síndrome de Inmunodeficiencia Adquirida del Simio/inmunología , Virus de la Inmunodeficiencia de los Simios/inmunologíaRESUMEN
Antibodies (Abs) that mediate antibody-dependent cellular cytotoxicity (ADCC) activity against HIV-1 are of major interest. A widely used method to measure ADCC Abs is the rapid and fluorometric antibody-dependent cellular cytotoxicity (RFADCC) assay. Antibody-dependent killing of a labelled target cell line by PBMC is assessed by loss of intracellular CFSE but retention of membrane dye PKH26 (CFSE-PKH26+). Cells of this phenotype are assumed to be derived from CFSE+PKH26+ target cells killed by NK cells. We assessed the effector cells that mediate ADCC in this assay. Backgating analysis and phenotyping of CFSE-PKH26+ revealed that the RFADCC assay's readout mainly represents CD3-CD14+ monocytes taking up the PKH26 dye. This was confirmed for 53 HIV+plasma-purified IgG samples when co-cultured with PBMC from three separate healthy donors. Emergence of the CFSE-PKH26+ monocyte population was observed upon co-culture of targets with purified monocytes but not with purified NK cells. Image flow cytometry and microscopy showed a monocyte-specific interaction with target cells without typical morphological changes associated with phagocytosis, suggesting a monocyte-mediated ADCC process. We conclude that the RFADCC assay primarily reflects Ab-mediated monocyte function. Further studies on the immunological importance of HIV-specific monocyte-mediated ADCC are warranted.
Asunto(s)
Citotoxicidad Celular Dependiente de Anticuerpos/inmunología , VIH/inmunología , Células Asesinas Naturales/inmunología , Monocitos/inmunología , Complejo CD3/inmunología , Complejo CD3/metabolismo , Línea Celular , Células Cultivadas , Técnicas de Cocultivo , Citometría de Flujo , Fluoresceínas/química , Colorantes Fluorescentes/química , Humanos , Células Asesinas Naturales/química , Células Asesinas Naturales/metabolismo , Leucocitos Mononucleares/citología , Leucocitos Mononucleares/inmunología , Leucocitos Mononucleares/metabolismo , Receptores de Lipopolisacáridos/inmunología , Receptores de Lipopolisacáridos/metabolismo , Microscopía Fluorescente , Monocitos/citología , Monocitos/metabolismo , Compuestos Orgánicos/química , Fagocitosis/inmunología , Análisis de la Célula Individual/métodos , Succinimidas/químicaRESUMEN
Advances in treating and preventing AIDS depend on understanding how human immunodeficiency virus (HIV) is eliminated in vivo and on the manipulation of effective immune responses to HIV. During the development of assays quantifying the elimination of fluorescent autologous cells coated with overlapping 15-mer simian immunodeficiency virus (SIV) or HIV-1 peptides, we made a remarkable observation: the reinfusion of macaque peripheral blood mononuclear cells, or even whole blood, pulsed with SIV and/or HIV peptides generated sharply enhanced SIV- and HIV-1-specific T-cell immunity. Strong, broad CD4+- and CD8+-T-cell responses could be enhanced simultaneously against peptide pools spanning 87% of all SIV- and HIV-1-expressed proteins-highly desirable characteristics of HIV-specific immunity. De novo hepatitis C virus-specific CD4+- and CD8+-T-cell responses were generated in macaques by the same method. This simple technique holds promise for the immunotherapy of HIV and other chronic viral infections.
Asunto(s)
Infecciones por VIH/terapia , VIH/inmunología , Péptidos/inmunología , Síndrome de Inmunodeficiencia Adquirida del Simio/terapia , Virus de la Inmunodeficiencia de los Simios/inmunología , Linfocitos T/inmunología , Animales , Transfusión de Componentes Sanguíneos , Transfusión Sanguínea , Linfocitos T CD4-Positivos/inmunología , Linfocitos T CD8-positivos/inmunología , Modelos Animales de Enfermedad , Antígenos VIH/administración & dosificación , Antígenos VIH/inmunología , Infecciones por VIH/inmunología , Hepatitis C/inmunología , Antígenos de la Hepatitis C/administración & dosificación , Antígenos de la Hepatitis C/inmunología , Inmunoterapia , Macaca nemestrina , Péptidos/administración & dosificación , Proteínas de los Retroviridae/administración & dosificación , Proteínas de los Retroviridae/inmunología , Síndrome de Inmunodeficiencia Adquirida del Simio/inmunologíaRESUMEN
Further advances are required in understanding protection from AIDS by T-cell immunity. We analyzed a set of multigenic simian/human immunodeficiency virus (SHIV) DNA and fowlpox virus priming and boosting vaccines for immunogenicity and protective efficacy in outbred pigtail macaques. The number of vaccinations required, the effect of DNA vaccination alone, and the effect of cytokine (gamma interferon) coexpression by the fowlpox virus boost was also studied. A coordinated induction of high levels of broadly reactive CD4 and CD8 T-cell immune responses was induced by sequential DNA and fowlpox virus vaccination. The immunogenicity of regimens utilizing fowlpox virus coexpressing gamma interferon, a single DNA priming vaccination, or DNA vaccines alone was inferior. Significant control of a virulent SHIV challenge was observed despite a loss of SHIV-specific proliferating T cells. The outcome of challenge with virulent SHIV(mn229) correlated with vaccine immunogenicity except that DNA vaccination alone primed for protection almost as effectively as the DNA/fowlpox virus regimen despite negligible immunogenicity by standard assays. These studies suggest that priming of immunity with DNA and fowlpox virus vaccines could delay AIDS in humans.
Asunto(s)
Vacunas contra el SIDA/inmunología , Virus de la Viruela de las Aves de Corral/inmunología , Infecciones por VIH/prevención & control , Inmunización , Síndrome de Inmunodeficiencia Adquirida del Simio/prevención & control , Vacunas de ADN/inmunología , Vacunas contra el SIDA/administración & dosificación , Animales , Linfocitos T CD4-Positivos/inmunología , Linfocitos T CD8-positivos/inmunología , VIH-1/genética , VIH-1/inmunología , Humanos , Esquemas de Inmunización , Inmunización Secundaria , Interferón gamma/inmunología , Interferón gamma/metabolismo , Activación de Linfocitos , Macaca nemestrina , Vacunas contra el SIDAS/administración & dosificación , Vacunas contra el SIDAS/inmunología , Virus de la Inmunodeficiencia de los Simios/genética , Virus de la Inmunodeficiencia de los Simios/inmunología , Resultado del Tratamiento , Vacunas de ADN/administración & dosificaciónRESUMEN
Delivering attenuated lentivirus vaccines as proviral DNA would be simple and inexpensive. Inoculation of macaques with wild-type simian immunodeficiency virus strain mac239 (SIV(mac239)) DNA or SIV(mac239) DNA containing a single deletion in the 3' nef-long terminal repeat overlap region (nef/LTR) led to sustained SIV infections and AIDS. Injection of SIV(mac239) DNA containing identical deletions in both the 5' LTR and 3' nef/LTR resulted in attenuated SIV infections and substantial protection against subsequent mucosal SIV(mac251) challenge.
Asunto(s)
Virus de la Inmunodeficiencia de los Simios/inmunología , Vacunas de ADN/administración & dosificación , Vacunas Virales/administración & dosificación , Secuencia de Aminoácidos , Animales , Anticuerpos Antivirales/biosíntesis , Secuencia de Bases , Recuento de Linfocito CD4 , Cartilla de ADN , Interferón gamma/biosíntesis , Macaca nemestrina , Datos de Secuencia Molecular , Mutagénesis , Plásmidos , ARN Viral/sangre , Secuencias Repetitivas de Ácidos Nucleicos , Vacunas de ADN/química , Vacunas de ADN/inmunología , Vacunas Virales/química , Vacunas Virales/inmunologíaRESUMEN
BACKGROUND: The aim of this prospective study was to assess the effect of an intensive in-patient rehabilitation programme upon the mobility of amputees. METHODS: All major lower limb amputations between 1997 and 1999 received a pre-operative mobility assessment and, where appropriate, were referred for a vigorous rehabilitation programme. RESULTS: 92 lower limb amputations were performed in 87 patients (57 below knee, 33 above knee, two hip disarticulations). Overall, 63% of patients were able to ambulate independently following rehabilitation. Univariate analysis revealed that the only predictor of mobility was the level of amputation, below knee gaining better mobility than above knee (p=0.002). CONCLUSION: Lower limb amputees should participate in an aggressive in-patient physiotherapy regimen since reasonable mobility can be achieved in the majority of patients.
Asunto(s)
Amputación Quirúrgica/rehabilitación , Miembros Artificiales , Modalidades de Fisioterapia , Calidad de Vida , Adulto , Anciano , Amputación Quirúrgica/métodos , Femenino , Cadera/cirugía , Humanos , Pierna/cirugía , Masculino , Persona de Mediana Edad , Pronóstico , Estudios Prospectivos , Diseño de Prótesis , Ajuste de Prótesis , Recuperación de la Función , Resultado del TratamientoRESUMEN
Gene expression from HIV-based gene therapy vectors or live-attenuated HIV-1 vaccines requires RNA transcription supported by the HIV-1 promoter, the long terminal repeat (LTR). Delivery of live-attenuated HIV-1 vaccines as plasmid DNA would overcome problems associated with production of attenuated HIV-1 strains. We investigated the expression of reporter plasmids and proviral HIV-1 constructs driven by either the HIV-1 LTR or LTRs with deletions in the U3 enhancer regions. LTR-driven plasmids were inoculated by gene gun into both human epidermis ex vivo and macaques in vivo. The HIV-1 LTR drove reporter gene expression in human and macaque skin, although with 15- to 20-fold less efficiency compared to the immediate-early cytomegalovirus promoter. A deleted LTR derived from a naturally attenuated HIV-1 strain infecting a member of the well-characterized Sydney Blood Bank Cohort of long-term nonprogressors was 5-fold less efficient in expression of the reporter gene compared to wild-type LTR. Delivery of proviral wild-type HIV-1 DNA constructs to human skin resulted in recovery of HIV-1 from cells emigrating from the epidermis, providing an ex vivo model of the infectivity of proviral HIV-1 DNA. However, delivery of proviral HIV-1 DNA containing deletions in either the LTR, Nef, or the secondary viral transcription activator,Vpr, significantly reduced HIV-1 replication in this model. The early coexpression of Tat from a second plasmid did not restore replication. Thus, although attenuated lentiviral vaccines might be deliverable as proviral DNA constructs in primate subjects, significant improvements are needed to enhance the efficiency of this method.
Asunto(s)
Vacunas contra el SIDA/genética , Biolística , Regulación Viral de la Expresión Génica , Infecciones por VIH/genética , VIH-1 , Piel/virología , Secuencias Repetidas Terminales/genética , Animales , Citomegalovirus/genética , Genes Inmediatos-Precoces , Genes Reporteros , Genes nef , Humanos , Inyecciones Subcutáneas , Macaca nemestrina , Plásmidos , Transfección , Vacunas Atenuadas/genética , beta-Galactosidasa/biosíntesis , beta-Galactosidasa/genéticaRESUMEN
A number of monkey species, including African green monkeys and African vervet monkeys (Chlorocebus aethiops), are frequently infected in the wild and in captivity with a Simian immunodeficiency virus strain, SIVagm, a primate lentivirus. Up to 50% of African green monkeys are estimated to be infected with SIVagm. SIV strains are very closely related to HIV-2 strains, which are a cause of AIDS in humans, predominantly in western Africa, although cases in Australia have also been reported. It is generally thought that SIV is non-pathogenic in several natural hosts, including African green monkeys. Nevertheless many SIV strains induce a profound immunodeficiency virtually identical to HIV-1 induced AIDS in humans when administered to Asian macaque species such as rhesus (Macaca mulatta) or pigtailed macaques (M nemestrina). SIV infection of Asian macaque species is frequently employed as an animal model for AIDS vaccine studies. In November 1996 a group of 10 African vervet monkeys were imported from the USA for display at Victoria's Open Range Zoo in Werribee. Two animals in this group of monkeys later developed a fatal gastroenteric illness. These diagnoses led us to initiate SIV testing of the colony.
Asunto(s)
Chlorocebus aethiops , ADN Viral/sangre , Brotes de Enfermedades/veterinaria , Síndrome de Inmunodeficiencia Adquirida del Simio/epidemiología , Virus de la Inmunodeficiencia de los Simios/aislamiento & purificación , Animales , Animales de Zoológico , Femenino , Masculino , Reacción en Cadena de la Polimerasa/veterinaria , Síndrome de Inmunodeficiencia Adquirida del Simio/etiología , Virus de la Inmunodeficiencia de los Simios/genética , Victoria/epidemiologíaRESUMEN
The AIDS pandemic is a global emergency and a preventive vaccine is urgently needed. CD4 and CD8 T-cell responses appear important in controlling human immunodeficiency virus (HIV)-1 in humans and simian immunodeficiency virus (SIV) in macaques. The utility of vaccines that induce high levels of SIV- or HIV-specific T cells has recently become clearer. Since T cells recognize virus-infected cells rather than free virus, T-cell-based vaccines only have the capacity to control infections (non-sterilizing immunity) and to prevent continuing or persisting infection. An HIV/SIV infection of macaques that is partially controlled by vaccine-induced T-cell responses permits a critical window of opportunity for the efficient generation and recruitment of additional T- and B-cell immune responses to the incoming viral inoculum. Although CD8-depletion experiments in macaques have defined the utility of CD8 T responses in control of SIV infections in macaques, direct evidence on the utility of either CD4 or CD8 T-cell responses in protective immunity to SIV following vaccination is lacking. The availability of genetically identical macaques would allow cell transfer studies and help define with more certainty the role of cellular immune responses in protection from AIDS. The review also focuses on the development of syngeneic macaques by twinning and cloning technologies.