RESUMEN
Studies have shown that vaccine vectors and route of immunization can differentially activate different arms of the immune system. However, the effects of different HIV vaccine immunogens on mucosal inflammation have not yet been studied. Because mucosal sites are the primary route of HIV infection, we evaluated the cervico-vaginal inflammatory cytokine and chemokine levels of Mauritian cynomolgus macaques following immunization and boost using two different SIV vaccine immunogens. The PCS vaccine delivers 12 20-amino acid peptides overlapping the 12 protease cleavage sites, and the Gag/Env vaccine delivers the full Gag and full Env proteins of simian immunodeficiency virus. We showed that the PCS vaccine prime and boosts induced short-lived, lower level increases of a few pro-inflammatory/chemotactic cytokines. In the PCS-vaccine group only the levels of MCP-1 were significantly increased above the baseline (P = 0.0078, Week 6; P = 0.0078, Week 17; P = 0.0234; Week 51) following multiple boosts. In contrast, immunizations with the Gag/Env vaccine persistently increased the levels of multiple cytokines/chemokines. In the Gag/Env group, higher than baseline levels were consistently observed for IL-8 (P = 0.0078, Week 16; P = 0.0078, Week 17; P = 0.0156, Week 52), IL-1ß (P = 0.0234, Week 16; P = 0.0156, Week 17; P = 0.0156, Week 52), and MIP-1α (P = 0.0313, Week 16; P = 0.0156, Week 17; P = 0.0313, Week 52). Over time, repeated boosts altered the relative levels of these cytokines between the Gag/Env and PCS vaccine group. 18 weeks after final boost with a higher dosage, IP-10 levels (P = 0.0313) in the Gag/Env group remained higher than baseline. Thus, the influence of vaccine immunogens on mucosal inflammation needs to be considered when developing and evaluating candidate HIV vaccines.
Asunto(s)
Cuello del Útero/efectos de los fármacos , Citocinas/metabolismo , Productos del Gen env/administración & dosificación , Productos del Gen gag/administración & dosificación , Mediadores de Inflamación/metabolismo , Vacunas contra el SIDAS/administración & dosificación , Síndrome de Inmunodeficiencia Adquirida del Simio/prevención & control , Virus de la Inmunodeficiencia de los Simios/inmunología , Vagina/efectos de los fármacos , Animales , Cuello del Útero/inmunología , Cuello del Útero/metabolismo , Femenino , Productos del Gen env/genética , Productos del Gen env/inmunología , Productos del Gen env/toxicidad , Productos del Gen gag/genética , Productos del Gen gag/inmunología , Productos del Gen gag/toxicidad , Macaca fascicularis , Membrana Mucosa/efectos de los fármacos , Membrana Mucosa/inmunología , Membrana Mucosa/metabolismo , Vacunas contra el SIDAS/genética , Vacunas contra el SIDAS/inmunología , Vacunas contra el SIDAS/toxicidad , Síndrome de Inmunodeficiencia Adquirida del Simio/inmunología , Síndrome de Inmunodeficiencia Adquirida del Simio/virología , Factores de Tiempo , Vacunación , Vacunas Sintéticas/administración & dosificación , Vacunas Sintéticas/genética , Vacunas Sintéticas/inmunología , Vacunas Sintéticas/toxicidad , Vagina/inmunología , Vagina/metabolismoRESUMEN
Mesangial cell (MC) hyperplasia and accumulation of extracellular matrix are the predominant features of HIV-associated nephropathy (HIVAN). Since mice transgenic for HIV-1 genes show renal lesions mimicking HIVAN, we studied the effect of HIV-1 gp160 protein on cultured murine MC (MMC) proliferation and apoptosis. HIV-1 gp160 protein stimulated (p < 0.001) MMC proliferation when compared with control MMCs. This effect of gp160 protein peaked at a concentration of 0.01 microg/ml. MMCs treated with a higher concentration of gp160 protein (0.1 microg/ml) or for a prolonged period of time (72 h) showed apoptosis rather than cell proliferation. These studies were further confirmed by DNA fragmentation and end labeling assays. gp160 also enhanced apoptosis in human MCs. Tumor necrosis factor (TNF)-alpha enhanced (p < 0.001) MMC apoptosis, and anti-TNF-alpha antibodies inhibited gp160-induced MMC apoptosis. In addition, gp160 protein attenuated MMC expression of Bcl-2 mRNA expression. These results suggest that gp160-induced apoptosis may be affected in part by the release of TNF-alpha and associated with attenuated mRNA expression of Bcl-2 by MMCs.
Asunto(s)
Mesangio Glomerular/efectos de los fármacos , Mesangio Glomerular/patología , Proteínas gp160 de Envoltorio del VIH/toxicidad , VIH-1/patogenicidad , Nefropatía Asociada a SIDA/etiología , Nefropatía Asociada a SIDA/patología , Animales , Apoptosis/efectos de los fármacos , Bromodesoxiuridina/metabolismo , División Celular/efectos de los fármacos , Células Cultivadas , ADN/biosíntesis , Fragmentación del ADN/efectos de los fármacos , Expresión Génica/efectos de los fármacos , Productos del Gen gag/toxicidad , Genes bcl-2/efectos de los fármacos , Mesangio Glomerular/metabolismo , Humanos , Cinética , Ratones , Pruebas de Neutralización , ARN Mensajero/genética , ARN Mensajero/metabolismo , Factor de Necrosis Tumoral alfa/antagonistas & inhibidores , Factor de Necrosis Tumoral alfa/farmacologíaRESUMEN
In this study, we tested the hypothesis that human immunodeficiency virus (HIV)-1-derived peptides augment the neurotoxicity of excitatory amino acid agonists in vivo in postnatal day (PND) 7 rats. Stereotaxic intracerebral injections of the excitatory amino acid agonist N-methyl-D-aspartate (NMDA), alone or coinjected with an HIV-derived recombinant fusion peptide envelope gag (env-gag) were performed in PND-7 rats [group I: 5 nmol NMDA, n = 20; group II: 5 nmol NMDA + low-dose (1 or 50 ng) env-gag, n = 27; group III: 5 nmol NMDA + high-dose (100 ng) env-gag, n = 20], and brain injury was evaluated on PND 12. Based on histopathology scoring and measurements of hippocampal cross-sectional areas in the injected and contralateral hemispheres, coinjection of 100 ng of env-gag with 5 nmol of NMDA markedly increased the severity of resulting injury (p < 0.002, comparing histopathology scores; p < 0.003, comparing interhemispheric differences in hippocampal areas). These data suggest that in the developing nervous system HIV neurotoxicity may result, at least in part, from overactivation of excitatory amino acid receptors, and that perinatal rodent models may provide clinically relevant insights about the pathophysiology of HIV-mediated brain injury.
Asunto(s)
Lesiones Encefálicas/etiología , Productos del Gen env/toxicidad , Productos del Gen gag/toxicidad , VIH-1/patogenicidad , N-Metilaspartato/toxicidad , Complejo SIDA Demencia/etiología , Animales , Animales Recién Nacidos , Lesiones Encefálicas/patología , Modelos Animales de Enfermedad , Hipocampo/lesiones , Hipocampo/patología , Ratas , Proteínas Recombinantes de Fusión/toxicidadRESUMEN
HGP-30-KLH vaccine in alum at doses of 10, 25, 50, and 100 micrograms/kg administered intramuscularly at weeks 0, 4, and 10 appear well-tolerated clinically. Local pain at the injection site, appears to be the main clinical toxicity. Laboratory parameters are not affected by administration of the vaccine candidate except for perhaps mild urinalysis abnormalities at the highest dose. This vaccine candidate has no apparent immunotoxicity and does not appear to affect lymphocyte populations or T-cell functional studies. Low levels and transient antibodies develop in a minority of subjects early after immunization with the vaccine candidate. These responses were observed in the lowest dose range. Higher doses, and longer follow-up will be needed to confirm this observation. T-cell proliferative responses to KLH and KLH-HGP-30 are consistent and may not be dose dependent, but the proliferative responses are variable and more data need to be accumulated. Preliminary, there appears to be an HGP-30-induced CTL response of HGP-30-coated EBV-transformed autologous B cell lines. This study was approved under an IND for the California Department of Health Services' Food and Drug Branch. They have provided excellent support and regulatory guidelines for this project. Future work will extend and confirm these initial observations.