RESUMO
AIM: Experimental autoimmune encephalomyelitis (EAE) is a CD4(+)-mediated autoimmune pathology of the central nervous system (CNS) that is used as a model for the study of the human neuroinflammatory disease, multiple sclerosis. During the development of EAE, auto-reactive Th1 and Th17 CD4(+) T cells infiltrate the CNS promoting inflammatory cells recruitment, focal inflammation and tissue destruction. In this sense, statins, agents used to lower lipid levels, have recently shown to exert interesting immunomodulatory function. In fact, statins promote a bias towards a Th2 response, which ameliorates the clinical outcome of EAE. Additionally, simvastatin can inhibit Th17 differentiation. However, many other effects exerted on the immune system by statins have yet to be clarified, in particular during neuroinflammation. Thus, the aim of this study was to investigate the effects of simvastatin on the development of experimental autoimmune encephalomyelitis. METHODS: Mice were immunized with MOG(35-55) and EAE severity was assessed daily and scored using a clinical scale. Cytokine secretion by mononuclear cells infiltrating the CNS was evaluated by flow cytometry. RESULTS: Simvastatin (5 mg/kg/day) improved clinical outcome, induced an increase in TGF-ß mRNA expression and inhibited IL-6, IL-12p40, IL-12p70, RANTES and MIP-1ß secretion (p < 0.05). This was accompanied by a significant decrease in CNS inflammatory mononuclear cell infiltration, with reduced frequencies of both Th1 and Th17 cells. Simvastatin inhibited the proliferation of T lymphocytes co-cultured with primary microglial cells. CONCLUSIONS: Simvastatin treatment promotes EAE clinical amelioration by inhibiting T cell proliferation and CNS infiltration by pathogenic Th1 and Th17 cells.
Assuntos
Encefalomielite Autoimune Experimental/tratamento farmacológico , Sinvastatina/farmacologia , Células Th1/efeitos dos fármacos , Células Th17/efeitos dos fármacos , Animais , Diferenciação Celular/efeitos dos fármacos , Diferenciação Celular/imunologia , Proliferação de Células/efeitos dos fármacos , Sistema Nervoso Central/efeitos dos fármacos , Sistema Nervoso Central/imunologia , Quimiocina CCL5/imunologia , Encefalomielite Autoimune Experimental/imunologia , Feminino , Inflamação/tratamento farmacológico , Inflamação/imunologia , Subunidade p40 da Interleucina-12/imunologia , Interleucina-6/imunologia , Ativação Linfocitária/efeitos dos fármacos , Ativação Linfocitária/imunologia , Camundongos , Camundongos Endogâmicos C57BL , Esclerose Múltipla/tratamento farmacológico , Esclerose Múltipla/imunologia , Sinvastatina/imunologia , Células Th1/imunologia , Células Th17/imunologia , Fator de Crescimento Transformador beta/imunologiaRESUMO
The aim of this study was to investigate if the aerobic training (AT) reverses airway remodeling (AR) in an asthma model. BALB/c were divided into four groups: control (unsensitized and untrained); ovalbumin (OVA: sensitized and untrained); AT (unsensitized and trained) and OVA + AT. Allergic inflammation was induced with intraperitoneal and OVA inhalation. AT (low intensity; 5×/week; 60 min/session) was performed at 7, 15, and 30 days. Leukocyte counting in the bronchoalveolar lavage fluid; the expression of IL-5, eotaxin, RANTES, intercellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1); AR features (airway smooth muscle, epithelium thickness, collagen and elastic fibers, mucus production); and AR inducers (transforming growing factor-beta, osteopontin, vascular endothelial growth factor). OVA induced an increase in leukocyte airway migration and increased AR features (P < 0.05). After 7 days, AT reversed the OVA-induced eosinophil and macrophage airway migration, the expression of IL-5, eotaxin, RANTES, ICAM-1, VCAM-1, and all AR inducers. However, total reversion of the AR features and inducers and airway inflammation occurred only after 15 days of AT compared with the OVA groups (P < 0.05) and the effects were maintained until the 30th day. AT reverses AR after 15 days and this effect is preceded by the inhibition of leukocyte migration and occurs simultaneously with the reduction in the expression of inflammatory mediators and AR inducers.
Assuntos
Remodelação das Vias Aéreas/imunologia , Asma/imunologia , Brônquios/imunologia , Condicionamento Físico Animal , Remodelação das Vias Aéreas/fisiologia , Animais , Asma/induzido quimicamente , Asma/metabolismo , Asma/patologia , Brônquios/patologia , Líquido da Lavagem Broncoalveolar/química , Líquido da Lavagem Broncoalveolar/citologia , Movimento Celular , Quimiocina CCL5/imunologia , Quimiocinas CC/imunologia , Doença Crônica , Colágeno/metabolismo , Modelos Animais de Doenças , Tecido Elástico/patologia , Eosinófilos/imunologia , Molécula 1 de Adesão Intercelular/metabolismo , Interleucina-5/imunologia , Leucócitos , Macrófagos Alveolares/imunologia , Camundongos , Camundongos Endogâmicos BALB C , Muco/metabolismo , Músculo Liso/patologia , Osteopontina/metabolismo , Ovalbumina/toxicidade , Mucosa Respiratória/patologia , Fator de Crescimento Transformador beta/imunologia , Fator de Crescimento Transformador beta/metabolismo , Molécula 1 de Adesão de Célula Vascular/metabolismo , Fator A de Crescimento do Endotélio Vascular/metabolismoRESUMO
BACKGROUND: The purpose of this study was to evaluate the relationship between chemokines and dendritic cells (DCs) in human chronic periodontitis (CP). METHODS: Gingival samples were obtained from 23 individuals with CP, and six samples of normal mucosa (NM) overlapping the third molar were used to control for the chemokine levels. Periodontal examination was conducted. Immunohistochemistry was performed for Factor XIIIa(+) and cluster of differentiation (CD)1a(+) immature DCs and CD83(+) mature DCs. Levels of the CC chemokine ligand (CCL)2, CCL3, CCL5, CCL19, CCL20, and CXC chemokine ligand (CXCL)8 were measured in gingival tissues using enzyme-linked immunosorbent assay. Inflammatory infiltrate, DCs, chemokines, classification of human CP, and clinical parameters were correlated and compared. RESULTS: The expression of CCL2 and CCL20 was positively correlated with increased densities of CD1a(+) DCs. CCL3 and CXCL8 were positively related to the clinical attachment level. CCL3, CCL5, CCL19, and CXCL8 levels increased in the gingival samples of patients with CP compared with NM, whereas CCL20 levels increased in advanced CP compared with mild-moderate CP. CONCLUSIONS: More CD1a(+) immature DCs are related to CCL2 and CCL20. CCL3 and CXCL8 chemokines are related to a greater severity of human CP.
Assuntos
Quimiocinas CC/imunologia , Periodontite Crônica/imunologia , Células Dendríticas/imunologia , Adulto , Idoso , Antígenos CD/imunologia , Antígenos CD1/imunologia , Contagem de Células , Quimiocina CCL19/imunologia , Quimiocina CCL2/imunologia , Quimiocina CCL20/imunologia , Quimiocina CCL3/imunologia , Quimiocina CCL5/imunologia , Periodontite Crônica/classificação , Periodontite Crônica/patologia , Fator XIIIa/análise , Feminino , Gengiva/imunologia , Hemorragia Gengival/imunologia , Humanos , Imunoglobulinas/imunologia , Interleucina-8/imunologia , Masculino , Glicoproteínas de Membrana/imunologia , Pessoa de Meia-Idade , Mucosa Bucal/imunologia , Perda da Inserção Periodontal/imunologia , Bolsa Periodontal/imunologia , Adulto Jovem , Antígeno CD83RESUMO
The complex interplay between cytokines and chemokines regulates innate and adaptive immune responses against pathogens; specifically, cytokine and chemokine expression drives activation of immune effector cells and their recruitment to tissue infection sites. Herein, we inoculated dogs with Leishmania braziliensis antigens plus saponin (the LBSap vaccine), as well as with the vaccine components, and then used real-time PCR to evaluate the kinetics of dermal expression of mRNAs of cytokines (IL-12, IFN-γ, TNF-α, IL-4, IL-13, TGF-ß and IL-10) and chemokines (CCL2, CCL4, CCL5, CCL21 and CXCL8) 1, 12, 24 and 48 h after inoculation. We also evaluated the correlation between cytokine and chemokine expression and dermal cellularity. The LBSap vaccine induced high levels of IL-12 and IL-10 expression at 12 and 24 h, respectively. Furthermore, we observed positive correlations between IL-12 and IL-13 expression, IFN-γ and IL-13 expression, and IL-13 and TGF-ß expression, suggesting that a mixed cytokine microenvironment developed after immunization with the vaccine. Inoculation with the saponin adjuvant alone induced a chemokine and cytokine expression profile similar to that observed in the LBSap group. CCL4 and CXCL8 chemokine expression was up regulated by the LBSap vaccine. CCL5 expression was initially highest in the LBSap group, but at 48 h, expression was highest in the LB group. Information about the kinetics of the immune response to this vaccine gained using this dog model will help to elucidate the mechanisms of and factors involved in a protective response against Leishmania infection and will aid in establishing rational approaches for the development of vaccines against canine visceral leishmaniasis.
Assuntos
Quimiocinas/imunologia , Citocinas/imunologia , Derme/imunologia , Vacinas contra Leishmaniose/imunologia , Animais , Quimiocina CCL4/genética , Quimiocina CCL4/imunologia , Quimiocina CCL5/genética , Quimiocina CCL5/imunologia , Quimiocinas/genética , Citocinas/genética , Derme/metabolismo , Derme/parasitologia , Doenças do Cão/imunologia , Doenças do Cão/parasitologia , Doenças do Cão/prevenção & controle , Cães , Feminino , Expressão Gênica/imunologia , Interações Hospedeiro-Patógeno/imunologia , Imunização , Interleucina-10/genética , Interleucina-10/imunologia , Interleucina-12/genética , Interleucina-12/imunologia , Interleucina-8/genética , Interleucina-8/imunologia , Leishmania braziliensis/imunologia , Leishmania braziliensis/fisiologia , Vacinas contra Leishmaniose/administração & dosagem , Leishmaniose Visceral/genética , Leishmaniose Visceral/imunologia , Leishmaniose Visceral/veterinária , Masculino , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Saponinas/administração & dosagem , Saponinas/imunologia , Fatores de TempoRESUMO
Chagas' disease is caused by Trypanosoma cruzi infection and is characterized by chronic fibrogenic inflammation and heart dysfunction. Chemokines are produced during infection and drive tissue inflammation. In rats, acute infection is characterized by intense myocarditis and regression of inflammation after control of parasitism. We investigated the role of CCL3 and CCL5 during infection by using DNA vaccination encoding for each chemokine separately or simultaneously. MetRANTES treatment was used to evaluate the role of CCR1 and CCR5, the receptors for CCL3 and CCL5. Vaccination with CCL3 or CCL5 increased heart parasitism and decreased local IFN-gamma production, but did not influence intensity of inflammation. Simultaneous treatment with both plasmids or treatment with MetRANTES enhanced cardiac inflammation, fibrosis and parasitism. In conclusion, chemokines CCL3 and CCL5 are relevant, but not essential, for control of T. cruzi infection in rats. On the other hand, combined blockade of these chemokines or their receptors enhanced tissue inflammation and fibrosis, clearly contrasting with available data in murine models of T. cruzi infection. These data reinforce the important role of chemokines during T. cruzi infection but suggest that caution must be taken when expanding the therapeutic modulation of the chemokine system in mice to the human infection.
Assuntos
Cardiomiopatia Chagásica/imunologia , Quimiocina CCL3/imunologia , Quimiocina CCL5/imunologia , Trypanosoma cruzi/imunologia , Animais , Cardiomiopatia Chagásica/patologia , Coração/parasitologia , Miocárdio/patologia , Ratos , Ratos Sprague-Dawley , Trypanosoma cruzi/patogenicidade , Vacinas de DNA/administração & dosagemRESUMO
The Herpes simplex virus-1 (HSV-1) is responsible for several clinical manifestations in humans, including encephalitis. To induce encephalitis, C57BL/6 mice were inoculated with 10(4) plaque-forming cells of HSV-1 by the intracranial route. Met-RANTES (regulated upon activation, normal T cell expressed and presumably secreted) (10 microg/mouse), a CC chemokine family receptor (CCR)1 and CCR5 antagonist, was given subcutaneously the day before, immediately after, and at days 1, 2, and 3 after infection. Treatment with Met-RANTES had no effect on the viral titers. In contrast, intravital microscopy revealed that treatment with Met-RANTES decreased the number of leukocytes adherent to the pial microvasculature at days 1 and 3 after infection. The levels of the chemokines CCL3, CCL5, CXCL1, and CXCL9 increased after infection and were enhanced further by the treatment with Met-RANTES. Treatment with a polyclonal anti-CCL5 antibody 2 h before the intravital microscopy decreased leukocyte adhesion in the microcirculation of infected mice. In conclusion, CCL5, a chemokine that binds to CCR1 and CCR5, is essential for leukocyte adhesion during HSV-1 encephalitis. However, blocking of CCR1 and CCR5 did not affect HSV-1 replication, suggesting that other immune mechanisms are involved in the process of infection control.
Assuntos
Movimento Celular , Quimiocina CCL5/imunologia , Encefalite por Herpes Simples/imunologia , Encefalite por Herpes Simples/virologia , Leucócitos/patologia , Animais , Anticorpos/farmacologia , Encéfalo/efeitos dos fármacos , Encéfalo/imunologia , Encéfalo/patologia , Encéfalo/virologia , Adesão Celular/efeitos dos fármacos , Movimento Celular/efeitos dos fármacos , Quimiocina CCL5/farmacologia , Chlorocebus aethiops , Modelos Animais de Doenças , Encefalite por Herpes Simples/patologia , Endotélio/efeitos dos fármacos , Endotélio/virologia , Herpesvirus Humano 1/fisiologia , Migração e Rolagem de Leucócitos/efeitos dos fármacos , Leucócitos/efeitos dos fármacos , Leucócitos/virologia , Meninges/irrigação sanguínea , Meninges/efeitos dos fármacos , Meninges/patologia , Meninges/virologia , Camundongos , Camundongos Endogâmicos C57BL , Células Vero , Carga ViralRESUMO
BACKGROUND: Kinins are important mediators of inflammation and act through stimulation of two receptor subtypes, B1 and B2. Leukocyte infiltration contributes to the pathogenesis of autoimmune inflammation in the central nervous system (CNS), occurring not only in multiple sclerosis (MS) but also in experimental autoimmune encephalomyelitis (EAE). We have previously shown that the chemokines CCL2 and CCL5 play an important role in the adhesion of leukocytes to the brain microcirculation in EAE. The aim of the present study was to evaluate the relevance of B2 receptors to leukocyte-endothelium interactions in the cerebral microcirculation, and its participation in CNS inflammation in the experimental model of myelin-oligodendrocyte-glycoprotein (MOG)(35-55)-induced EAE in mice. METHODS: In order to evaluate the role of B2 receptor in the cerebral microvasculature we used wild-type (WT) and kinin B2 receptor knockout (B2-/-) mice subjected to MOG(35-55)-induced EAE. Intravital microscopy was used to investigate leukocyte recruitment on pial matter vessels in B2-/- and WT EAE mice. Histological documentation of inflammatory infiltrates in brain and spinal cords was correlated with intravital findings. The expression of CCL5 and CCL2 in cerebral tissue was assessed by ELISA. RESULTS: Clinical parameters of disease were reduced in B2-/- mice in comparison to wild type EAE mice. At day 14 after EAE induction, there was a significant decrease in the number of adherent leukocytes, a reduction of cerebral CCL5 and CCL2 expressions, and smaller inflammatory and degenerative changes in B2-/- mice when compared to WT. CONCLUSION: Our results suggest that B2 receptors have two major effects in the control of EAE severity: (i) B2 regulates the expression of chemokines, including CCL2 and CCL5, and (ii) B2 modulates leukocyte recruitment and inflammatory lesions in the CNS.
Assuntos
Quimiocina CCL2/imunologia , Quimiocina CCL5/imunologia , Encefalomielite Autoimune Experimental/imunologia , Leucócitos/imunologia , Receptor B2 da Bradicinina/imunologia , Animais , Encéfalo/irrigação sanguínea , Encéfalo/metabolismo , Encéfalo/patologia , Quimiocinas/metabolismo , Citocinas/metabolismo , Encefalomielite Autoimune Experimental/patologia , Feminino , Glicoproteínas/imunologia , Migração e Rolagem de Leucócitos , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Microvasos/metabolismo , Microvasos/patologia , Glicoproteína Mielina-Oligodendrócito , Fragmentos de Peptídeos/imunologia , Receptor B2 da Bradicinina/genéticaRESUMO
PROBLEM: Several studies indicate that RANTES (regulated on activation, normal T cell expressed and secreted) is able to downregulate T-cell responses which suggest it might be relevant for fetal tolerance induction. However, the role of RANTES in pregnancy had not been established. Here we investigate RANTES regulation during early pregnancy and potential failures leading to losses of pregnancies. METHOD OF STUDY: RANTES and progesterone levels were determined in sera and feto-placental units from high resorption rate CBA/JxDBA/2 pregnant females and compared with CBA/JxBALB/c normal pregnant mice. RANTES in vitro modulation was also studied in nulliparous, primiparous and multiparous CBA/J and BALB/c cells in response to paternal alloantigen and progesterone stimulation. RESULTS: Nulliparous CBA/J females were quantitatively deficient in RANTES sera levels, whereas pregnancies with male BALB/c or DBA/2 increased its production. However, feto-placental units from CBA/J females are high producers of progesterone and RANTES. CONCLUSION: These data suggest that the beneficial effect of RANTES on feto-maternal interface requires an optimal concentration range and might be modulated by progesterone, hence exacerbated placental expression could be associated with high resorption rate.
Assuntos
Aborto Espontâneo/imunologia , Quimiocina CCL5/imunologia , Progesterona/imunologia , Aborto Espontâneo/sangue , Aborto Espontâneo/metabolismo , Animais , Quimiocina CCL5/sangue , Quimiocina CCL5/metabolismo , Feminino , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos CBA , Camundongos Endogâmicos DBA , Placenta/imunologia , Placenta/metabolismo , Gravidez , Progesterona/biossíntese , Progesterona/sangueRESUMO
BACKGROUND: We have recently isolated two distinct components from Ascaris suum adult worms with different effects on the immune system: the allergenic protein of A. suum (APAS-3), which induces IgE antibody production, and suppressive protein of A. suum (PAS-1), which inhibits humoral and cellular immune responses induced by unrelated antigens. In this study, we investigated the immunomodulatory effect of PAS-1 on a murine model of asthma induced by APAS-3. METHODS: BALB/c mice were immunized twice with APAS-3 or APAS-3 plus PAS-1 by the intraperitoneal and subcutaneous route (on days 0 and 7) and challenged twice with the same antigens intranasally (days 14 and 21). Two days after the last challenge, the allergic airway inflammation was evaluated by cellular migration, eosinophil peroxidase (EPO) activity, cytokine and chemokine production and pulmonary mechanical parameters. RESULTS: The allergenic properties of APAS-3 were confirmed by the stimulation of anaphylactic IgE and IgG1 antibody production and eosinophilic airway inflammation and hyper-responsiveness. On the other hand, PAS-1-treated mice showed a marked suppression of cellular migration and EPO activity that correlated well with a significant reduction in the levels of IL-4, IL-5, eotaxin and RANTES in the bronchoalveolar lavage (BAL) fluid. In contrast, considerable amounts of IL-10 were observed in the BAL fluid of PAS-1-treated mice. Airway hyper-responsiveness was obtained in APAS-3-immunized mice, but the conductance of the respiratory system was restored to normal values in the presence of PAS-1. CONCLUSION: These results indicate that A. suum allergenic protein APAS-3 induces a T helper 2-type immune response and, consequently, eosinophilic airway inflammation and hyper-responsiveness. Moreover, the modulatory protein PAS-1 has a marked suppressive effect on this response, and the inhibition of cytokine (IL-4, IL-5) and chemokine (eotaxin and RANTES) release, probably because of the presence of IL-10, may contribute to this effect.
Assuntos
Ascaris suum/imunologia , Asma/imunologia , Proteínas de Helminto/imunologia , Alérgenos/imunologia , Anafilaxia/imunologia , Animais , Líquido da Lavagem Broncoalveolar/imunologia , Inibição de Migração Celular , Quimiocina CCL11 , Quimiocina CCL5/imunologia , Quimiocinas CC/imunologia , Fatores Quimiotáticos de Eosinófilos/imunologia , Modelos Animais de Doenças , Peroxidase de Eosinófilo/imunologia , Eosinófilos/imunologia , Feminino , Imunoglobulina E/imunologia , Imunoglobulina G/imunologia , Interleucinas/imunologia , Pulmão/imunologia , Camundongos , Camundongos Endogâmicos BALB C , Células Th2/imunologiaRESUMO
In the present study, we investigated the involvement of macrophage-inflammatory protein-1alpha (MIP-1alpha)[CC chemokine ligand 3 (CCL3)], MIP-1beta[CCL4], regulated on activation, normal T expressed and secreted (RANTES)[CCL5], and CC chemokine receptors (CCRs) on neutrophil migration in murine immune inflammation. Previously, we showed that ovalbumin (OVA)-triggered neutrophil migration in immunized mice depends on the sequential release of tumor necrosis factor alpha (TNF-alpha) and leukotriene B(4)(LTB(4)). Herein, we show increased mRNA expression for MIP-1alpha[CCL3], MIP-1beta[CCL4], RANTES[CCL5], and CCR1 in peritoneal cells harvested from OVA-challenged, immunized mice, as well as MIP-1alpha[CCL3] and RANTES[CCL5] but not MIP-1beta[CCL4] proteins in the peritoneal exudates. OVA-induced neutrophil migration response was muted in immunized MIP-1alpha[CCL3](-/-) mice, but it was not inhibited by treatment with antibodies against RANTES[CCL5] or MIP-1beta[CCL4]. MIP-1alpha[CCL3] mediated neutrophil migration in immunized mice through induction of TNF-alpha and LTB(4) synthesis, as these mediators were detected in the exudates harvested from OVA-challenged immunized wild-type but not MIP-1alpha[CCL3](-/-) mice; administration of MIP-1alpha[CCL3] induced a dose-dependent neutrophil migration, which was inhibited by treatment with an anti-TNF-alpha antibody in TNF receptor 1 (p55(-/-))-deficient mice or by MK 886 (a 5-lipoxygenase inhibitor); and MIP-1alpha[CCL3] failed to induce LTB(4) production in p55(-/-) mice. MIP-1alpha[CCL3] used CCR1 to promote neutrophil recruitment, as OVA or MIP-1alpha[CCL3] failed to induce neutrophil migration in CCR1(-/-) mice, in contrast to CCR5(-/-) mice. In summary, we have demonstrated that neutrophil migration observed in this model of immune inflammation is mediated by MIP-1alpha[CCL3], which via CCR1, induces the sequential release of TNF-alpha and LTB(4). Therefore, whether a similar pathway mediates neutrophil migration in human immune-inflammatory diseases, the development of specific CCR1 antagonists might have a therapeutic potential.