RESUMO
Dendritic cells (DCs) play an essential role in regulation of immune responses. In the periphery, Ag presentation by DCs is critical for adaptive responses; for this reason, DCs are often targets of adjuvants that enhance vaccine responses. Activated mature DCs enhance B cell activation and differentiation by providing cytokines like BAFF and a proliferation-inducing ligand. However, the role of immature DCs in B cell tolerance is not well studied. Recently, mouse immature bone marrow-derived DCs (iBMDCs) have been shown to suppress anti-IgM-induced B cell activation. In this study, we tested the ability of mouse DCs to modulate B cell functions during TLR activation. We found that iBMDCs potently suppressed proliferation and differentiation of various B cell subsets on TLR stimulation. However, iBMDCs did not affect CD40-mediated B cell activation. Optimal suppression of B cell activation by iBMDCs required cell contact via the CD22 receptor on B cells. The B cell suppression was a property of iBMDCs or DCs resident in the bone marrow (BM), but not mature BM-derived DCs or DCs resident in the spleen. Presence of iBMDCs also enhanced the Ag-induced apoptotic response of BM B cells, suggesting that the suppressive effects of iBMDCs may have a role in B cell tolerance.
Assuntos
Subpopulações de Linfócitos B/imunologia , Células da Medula Óssea/imunologia , Células Dendríticas/imunologia , Receptores de Antígenos de Linfócitos T/fisiologia , Transdução de Sinais/imunologia , Animais , Subpopulações de Linfócitos B/citologia , Subpopulações de Linfócitos B/metabolismo , Células da Medula Óssea/citologia , Células da Medula Óssea/metabolismo , Diferenciação Celular/imunologia , Células Cultivadas , Células Dendríticas/citologia , Células Dendríticas/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Camundongos Transgênicos , Receptores de Antígenos de Linfócitos B/antagonistas & inibidores , Receptores de Antígenos de Linfócitos B/fisiologia , Receptores de Antígenos de Linfócitos T/antagonistas & inibidores , Baço/citologia , Baço/imunologia , Baço/metabolismoRESUMO
BACKGROUND: Thiazolidinediones (TZDs) activate peroxisome proliferator-activated receptor gamma (PPARgamma) and are used clinically to help restore peripheral insulin sensitivity in Type 2 diabetes (T2DM). Interestingly, long-term treatment of mouse models of Alzheimer's disease (AD) with TZDs also has been shown to reduce several well-established brain biomarkers of AD including inflammation, oxidative stress and Abeta accumulation. While TZD's actions in AD models help to elucidate the mechanisms underlying their potentially beneficial effects in AD patients, little is known about the functional consequences of TZDs in animal models of normal aging. Because aging is a common risk factor for both AD and T2DM, we investigated whether the TZD, pioglitazone could alter brain aging under non-pathological conditions. METHODS AND FINDINGS: We used the F344 rat model of aging, and monitored behavioral, electrophysiological, and molecular variables to assess the effects of pioglitazone (PIO-Actos(R) a TZD) on several peripheral (blood and liver) and central (hippocampal) biomarkers of aging. Starting at 3 months or 17 months of age, male rats were treated for 4-5 months with either a control or a PIO-containing diet (final dose approximately 2.3 mg/kg body weight/day). A significant reduction in the Ca(2+)-dependent afterhyperpolarization was seen in the aged animals, with no significant change in long-term potentiation maintenance or learning and memory performance. Blood insulin levels were unchanged with age, but significantly reduced by PIO. Finally, a combination of microarray analyses on hippocampal tissue and serum-based multiplex cytokine assays revealed that age-dependent inflammatory increases were not reversed by PIO. CONCLUSIONS: While current research efforts continue to identify the underlying processes responsible for the progressive decline in cognitive function seen during normal aging, available medical treatments are still very limited. Because TZDs have been shown to have benefits in age-related conditions such as T2DM and AD, our study was aimed at elucidating PIO's potentially beneficial actions in normal aging. Using a clinically-relevant dose and delivery method, long-term PIO treatment was able to blunt several indices of aging but apparently affected neither age-related cognitive decline nor peripheral/central age-related increases in inflammatory signaling.
Assuntos
Envelhecimento/efeitos dos fármacos , Tiazolidinedionas/farmacologia , Doença de Alzheimer , Animais , Biomarcadores/análise , Biomarcadores/sangue , Encéfalo/fisiologia , Hipocampo , Hipoglicemiantes/farmacologia , Inflamação , Aprendizagem/efeitos dos fármacos , Fígado , Potenciação de Longa Duração/efeitos dos fármacos , Masculino , Memória/efeitos dos fármacos , Pioglitazona , RatosRESUMO
The homeostatic microenvironment in lung is immunosuppressive and interleukin-10 (IL-10) helps maintain this microenvironment. Despite constitutive production of IL-10 in normal lung, macrophages (MØs) and dentritic cells (DCs) remain capable of responding to microorganisms, suggesting that these innate immune cells have a mechanism to override the immunosuppressive effects of IL-10. Prior studies by the authors revealed that Toll-like receptor (TLR) ligands inhibit IL-10 receptor signaling in alveolar macrophages (AMØs), thereby obviating the immunosuppressive activity of IL-10. This report compares the immunologic phenotypes of AMØs and lung DCs and their ability to respond to IL-10 following exposure to microbial stimuli. IL-10 was constitutively produced by normal lung epithelium and exposure to lipopolysaccharide (LPS) in vivo increased the expression of IL-10 during the first 24 hours. AMØs constitutively produced IL-10 mRNA, whereas both AMØs and LDCs constitutively expressed IL-12 mRNA. AMØs and LDCs, as well as bone marrow-derived MØs and DCs, had reduced capacity to activate STAT3 in response to IL-10 if pretreated with LPS. Inhibition was not associated with decreased expression of IL-10 receptor (IL-10R) and was dependent on the MyD88 signaling pathway. These results demonstrate a common underlying regulatory mechanism in both DCs and MØs by which microbial stimuli can override the immunosuppressive effect of constitutive IL-10 production in the lung.
Assuntos
Células Dendríticas/efeitos dos fármacos , Interleucina-10/genética , Lipopolissacarídeos/farmacologia , Macrófagos Alveolares/efeitos dos fármacos , Mucosa Respiratória/efeitos dos fármacos , Transdução de Sinais/efeitos dos fármacos , Receptor 4 Toll-Like/antagonistas & inibidores , Animais , Células da Medula Óssea/citologia , Células Cultivadas , Células Dendríticas/metabolismo , Feminino , Citometria de Fluxo , Expressão Gênica/efeitos dos fármacos , Imuno-Histoquímica , Ligantes , Pulmão/citologia , Macrófagos Alveolares/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Fator 88 de Diferenciação Mieloide/genética , Fator 88 de Diferenciação Mieloide/metabolismo , Fosforilação , RNA Mensageiro/metabolismo , Mucosa Respiratória/metabolismo , Fator de Transcrição STAT3/genética , Fator de Transcrição STAT3/metabolismo , Receptor 4 Toll-Like/deficiênciaRESUMO
Despite an immunosuppressive lung environment, alveolar macrophages (AM) retain the capacity to respond to microorganisms. This report demonstrates that IL-10, constitutively produced by normal alveolar epithelium, stimulates signal transduction through the IL-10R on AM and that IL-10R function can be inhibited by stimulation of Toll-like receptor (TLR) on AM. IL-10 mRNA and protein were constitutively expressed in normal alveolar epithelium of mice, and IL-10R were constitutively expressed on normal murine AM. Stimulation of AM through TLR2, TLR4, or TLR9 was sufficient to inhibit IL-10R signal transduction, including phosphorylation and nuclear translocation of STAT3 transcription factor. Inhibition of IL-10R function by TLRs was not associated with a decrease in IL-10R expression, but did require expression of the myeloid differentiation factor 88 adaptor protein. Continuous exposure of macrophages to IL-10 caused sustained expression of the chemokine receptors CCR1 and CCR5. However, the addition of TLR ligands inhibited IL-10-induced expression of CCR1 and CCR5. Finally, exposure of macrophages to TLR ligands blocked the ability of IL-10 to inhibit the induction of TNF-alpha by C2-ceramide. These findings demonstrate a novel regulatory mechanism that may allow AM to overcome inhibitory effects of constitutive IL-10 in the lungs that may permit a more effective response to pulmonary infections.