RESUMO
Traumatic brain injury (TBI) is a complex and progressive brain injury with no approved treatments that needs both short- and long-term therapeutic strategies to cope with the variety of physiopathological mechanisms involved. In particular, neuroinflammation is a key process modulating TBI outcome, and the potentiation of these mechanisms by pro-inflammatory gene therapy vectors could contribute to the injury progression. Here, we evaluate in the controlled cortical impact model of TBI, the safety of integrative-deficient lentiviral vectors (IDLVs) or the non-viral HNRK recombinant modular protein/DNA nanovector. These two promising vectors display different tropisms, transduction efficiencies, short- or long-term transduction or inflammatory activation profile. We show that the brain intraparenchymal injection of these vectors overexpressing green fluorescent protein after a CCI is not neurotoxic, and interestingly, can decrease the short-term sensory neurological deficits, and diminish the brain tissue loss at 90 days post lesion (dpl). Moreover, only IDLVs were able to mitigate the memory deficits elicited by a CCI. These vectors did not alter the microglial or astroglial reactivity at 90 dpl, suggesting that they do not potentiate the on-going neuroinflammation. Taken together, these data suggest that both types of vectors could be interesting tools for the design of gene therapy strategies targeting immediate or long-term neuropathological mechanisms of TBI.
Assuntos
Lesões Encefálicas Traumáticas/terapia , Terapia Genética/métodos , Neuroproteção/genética , Animais , Encéfalo/metabolismo , Lesões Encefálicas Traumáticas/genética , Modelos Animais de Doenças , Infusões Intraventriculares , Lentivirus/genética , Masculino , Microglia/metabolismo , Neuroimunomodulação/genética , Neuroimunomodulação/imunologia , Fármacos Neuroprotetores/uso terapêutico , Tecido Parenquimatoso , Ratos , Ratos Wistar , Proteínas Recombinantes/metabolismo , Proteínas Recombinantes/farmacologiaRESUMO
Neural stem cells reside in two neurogenic regions of the adult brain: the dentate gyrus of the hippocampus (DG) and the subventricular zone (SVZ). Their proliferation, differentiation, migration and survival are modulated by intrinsic and extrinsic signals, forming a neurogenic niche. Brain cytokines have only been recently regarded as possible components of this neurogenic niche. In particular, we have demonstrated that transforming growth factor-beta (TGF-beta) has a pro-neurogenic effect in the DG in a model of increased neurogenesis by adrenalectomy. We wanted to test whether TGF-beta has a similar effect in another neurogenic region, namely the SVZ. To test this possibility, adult rats were injected with adenoviral vectors expressing TGF-beta (Ad-TGF) or beta-galactosidase (Ad-bgal) in the SVZ and neurogenesis was evaluated 3 weeks later. We have observed that chronic TGF-beta expression increased neurogenesis in the ipsilateral hemisphere of Ad-TGF but not in Ad-bgal-treated rats compared to their contralateral side. In addition, an unspecific effect of the adenoviral vector per se could not be totally discarded. We conclude, under our experimental conditions, that TGF-beta could enhance adult neurogenesis in the SVZ. This data increase the growing evidence supporting a pro-neurogenic role of anti-inflammatory cytokines in the adult brain.
Assuntos
Neurogênese/genética , Neuroimunomodulação/genética , Neurônios/metabolismo , Células-Tronco/metabolismo , Telencéfalo/metabolismo , Fator de Crescimento Transformador beta/metabolismo , Fatores Etários , Animais , Proliferação de Células , Citocinas/genética , Citocinas/metabolismo , Vetores Genéticos/genética , Tolerância Imunológica/imunologia , Masculino , Neurônios/citologia , Ratos , Ratos Wistar , Células-Tronco/citologia , Telencéfalo/citologia , Transfecção/métodos , Fator de Crescimento Transformador beta/genética , Regulação para Cima/fisiologiaRESUMO
In the present study, we used the electronic version of the von Frey test to investigate the role of cytokines (TNF-alpha and IL-1beta) and chemokines (KC/CXCL-1) in the genesis of mechanical hypernociception during antigen-induced inflammation in mice. The nociceptive test consisted of evoking a hindpaw flexion reflex with a hand-held force transducer (electronic anesthesiometer) adapted with a 0.5 mm(2) polypropylene tip. The intraplantar administration of methylated bovine serum albumin (mBSA) in previously immunized (IM), but not in sham-immunized (SI) mice, induced mechanical hypernociception in a dose-dependent manner. Hypernociception induced by antigen was reduced in animals pretreated with IL-1ra and reparixin (a non-competitive allosteric inhibitor of CXCR2), and in TNF receptor type 1 deficient (TNFR1-/-) mice. Consistently, antigen challenge induced a time-dependent release of TNF-alpha, IL-1beta and KC/CXCL-1 in IM, but not in SI, mice. The increase in TNF-alpha levels preceded the increase in IL-1beta and KC/CXCL1. Antigen-induced release of IL-1beta and KC/CXCL1 was reduced in TNFR1-/- mice, and TNF-alpha-induced hypernociception was inhibited by IL-1ra and reparixin. Hypernociception induced by IL-1beta in immunized mice was inhibited by indomethacin, whereas KC/CXCL1-induced hypernociception was inhibited by indomethacin and guanethidine. Antigen-induced hypernociception was reduced by indomethacin and guanethidine and abolished by the two drugs combined. Together, these results suggest that inflammation associated with an adaptive immune response induces hypernociception that is mediated by an initial release of TNF-alpha, which triggers the subsequent release of IL-1beta and KC/CXCL1. The latter cytokines in turn stimulate the release of the direct-acting final mediators, prostanoids and sympathetic amines.
Assuntos
Quimiocinas/metabolismo , Citocinas/metabolismo , Hiperalgesia/imunologia , Hipersensibilidade Tardia/imunologia , Neuroimunomodulação/imunologia , Nociceptores/imunologia , Animais , Quimiocina CXCL1/imunologia , Modelos Animais de Doenças , Feminino , Hiperalgesia/genética , Hiperalgesia/fisiopatologia , Hipersensibilidade Tardia/genética , Hipersensibilidade Tardia/fisiopatologia , Mediadores da Inflamação/farmacologia , Interleucina-1beta/imunologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Neuroimunomodulação/genética , Medição da Dor , Limiar da Dor , Estimulação Física , Receptores Tipo I de Fatores de Necrose Tumoral/genética , Fator de Necrose Tumoral alfa/imunologia , Regulação para Cima/genética , Regulação para Cima/imunologiaRESUMO
OBJECTIVE: The aim of this work was to analyze beta-adrenergic receptor (betaAR) regulation of T-lymphocyte proliferation in mice according to different thyroid hormone statuses. METHODS: T cells from eu-, hypo- (by propylthiouracil treatment) and hyperthyroid (by thyroxine, T4 administration) mice were purified and specific radioligand binding assays were performed. The effects of the beta-agonist isoproterenol (ISO) on intracellular levels of cyclic AMP (cAMP) were determined. Mitogen-induced T-cell proliferation was measured by [(3)H]-thymidine incorporation. Finally, protein kinase C (PKC) activity in cytosol and membrane fractions were determined using radiolabelled enzymatic substrates. RESULTS: Adecrease or a non-significant increase in betaAR number was found on T lymphocytes from hypo- and hyperthyroid mice compared to euthyroid controls. ISO stimulation of cAMP levels was lower in hypothyroid and higher in hyperthyroid T lymphocytes compared to controls. T-selective mitogen-induced proliferation was increased in T4-treated animals, but decreased in hypothyroid mice. During the peak of proliferation, downregulation of betaAR was observed in all animals. However, a higher or a lower decrease was observed in hyper- and hypothyroid T cells, respectively. In parallel, a higher translocation of PKC activity was observed in hyperthyroid cells, and a lower one was found in hypothyroid lymphocytes with respect to controls. CONCLUSIONS: These results indicate that intracellular signals triggered by mitogen activation, namely PKC, would be related to differential betaAR downregulation in T lymphocytes depending on the thyroid hormone status, contributing to the distinct proliferative responses found in hypo- or hyperthyroidism compared to the euthyroid state.
Assuntos
Proliferação de Células/efeitos dos fármacos , Mitógenos/farmacologia , Neuroimunomodulação/imunologia , Receptores Adrenérgicos beta/efeitos dos fármacos , Linfócitos T/metabolismo , Glândula Tireoide/metabolismo , Agonistas Adrenérgicos beta/farmacologia , Animais , AMP Cíclico/metabolismo , Regulação para Baixo/efeitos dos fármacos , Regulação para Baixo/imunologia , Feminino , Hipertireoidismo/induzido quimicamente , Hipertireoidismo/imunologia , Hipertireoidismo/metabolismo , Hipotireoidismo/induzido quimicamente , Hipotireoidismo/imunologia , Hipotireoidismo/metabolismo , Isoproterenol/farmacologia , Camundongos , Camundongos Endogâmicos BALB C , Neuroimunomodulação/genética , Propiltiouracila/farmacologia , Proteína Quinase C/metabolismo , Transporte Proteico/efeitos dos fármacos , Transporte Proteico/imunologia , Agregação de Receptores/efeitos dos fármacos , Agregação de Receptores/imunologia , Receptores Adrenérgicos beta/imunologia , Receptores Adrenérgicos beta/metabolismo , Linfócitos T/efeitos dos fármacos , Linfócitos T/imunologia , Timidina/metabolismo , Glândula Tireoide/imunologia , Tiroxina/farmacologiaRESUMO
The autoimmune sialadenitis developed by non-obese diabetic (NOD) mice is considered a suitable model to study the ethiopathogenic mechanisms leading to sicca symptoms in Sjögren's syndrome (SS). Evidence supporting a neural rather than immune origin of the secretory dysfunction has been provided. As both nitric oxide and vasoactive intestinal peptide (VIP) are common messengers to nervous and immune systems mediating secretory and inflammatory responses, we examined nitric oxide synthase (NOS) activity with special focus on VIP-mediated effects in salivary glands of NOD mice. We found a decreased NOS activity and expression in major salivary glands of NOD mice with respect to control mice. In addition, there was a deficient VIP-activated signaling associated with a reduced saliva and amylase secretion in response to VIP. Our results support the hypothesis of an impaired balance of neuroimmune interactions in salivary glands as early events to take place in the progressive loss of secretory function of NOD mice.
Assuntos
Neuroimunomodulação/imunologia , Óxido Nítrico Sintase/imunologia , Óxido Nítrico/imunologia , Glândulas Salivares/enzimologia , Síndrome de Sjogren/enzimologia , Peptídeo Intestinal Vasoativo/imunologia , Fatores Etários , Animais , Feminino , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos NOD , Neuroimunomodulação/genética , Óxido Nítrico/biossíntese , Óxido Nítrico Sintase/metabolismo , Óxido Nítrico Sintase Tipo I , Fibras Parassimpáticas Pós-Ganglionares/imunologia , Fibras Parassimpáticas Pós-Ganglionares/metabolismo , Fibras Parassimpáticas Pós-Ganglionares/fisiopatologia , Isoformas de Proteínas/genética , Isoformas de Proteínas/imunologia , Receptores Muscarínicos/imunologia , Saliva/efeitos dos fármacos , Saliva/metabolismo , Glândulas Salivares/imunologia , Glândulas Salivares/inervação , Transdução de Sinais/genética , Transdução de Sinais/imunologia , Síndrome de Sjogren/genética , Síndrome de Sjogren/imunologia , Peptídeo Intestinal Vasoativo/metabolismo , Peptídeo Intestinal Vasoativo/farmacologiaRESUMO
Immunosenescence is discussed in the context of an integrated immune-neuroendocrine homeostatic network. Under this new perspective, immunologic decline during aging appears as part of a multilevel phenomenon which affects intracellular as well as systemic regulatory mechanisms. In particular, the experimental data suggesting that during puberty, sex and adrenal steroids trigger thymus involution by inducing extensive apoptotic death of thymocytes is briefly reviewed. Next, the evidence indicating that aging brings about a progressive disruption in immune-neuroendocrine integration and the possible role of this disruption in the occurrence of age-associated autoimmune phenomena is considered. Finally, the possible mechanisms by which the genome can determine life span as well as the potential involvement of the major histocompatibility complex in this process are discussed.