CD4+CD69+ T cells and CD4+CD25+FoxP3+ Treg cells imbalance in peripheral blood, spleen and peritoneal lavage from pristane-induced systemic lupus erythematosus (SLE) mice

Abstract Background: Adaptive immune cells, including CD4+CD69+ and CD4+CD25+FoxP3+ regulatory T (Treg) cells, are important for maintaining immunological tolerance. In human systemic lupus erythematosus (SLE), CD4+CD25+FoxP3+ Treg cells are reduced, whereas CD69 expression is increased, resulting in a homeostatic immune imbalance that may intensify autoreactive T cell activity. To analyze the mechanisms implicated in autotolerance failure, we evaluated CD4+CD69+ and CD4+CD25+FoxP3+ T cells and interleukin profiles in a pristane-induced SLE experimental model. Methods: For lupus induction, 26 female Balb/c mice received a single intraperitoneal 0.5 ml dose of pristane, and 16 mice received the same dose of saline. Blood and spleen samples were collected from euthanized mice 90 and 120 days after pristane or saline inoculation. Mononuclear cells from peripheral blood (PBMC), peritoneal lavage (PL) and splenocytes were obtained by erythrocyte lysis and cryopreserved for further evaluation by flow cytometry using the GuavaEasyCyte TM HT. After thawing, cells were washed and stained with monoclonal antibodies against CD3, CD4, CD8, CD25, CD28, CD69, FoxP3, CD14 and Ly6C (BD Pharmingen TM). Interleukins were quantified using Multiplex® MAP. The Mann-Whitney test and the Pearson coefficient were used for statistical analysis, and p < 0.05 considered significant. Results: Compared with the controls, SLE-induced animals presented increased numbers of CD4+CD69+ T cells in the blood on T90 and T120 (p = 0.022 and p = 0.008) and in the spleen on T120 (p = 0.049), but there were decreased numbers in the PL (p = 0.049) on T120. The percentage of Treg was lower in blood (p < 0.005 and p < 0.012) on T90 and T120, in spleen (p = 0.043) on T120 and in PL (p = 0.001) on T90. Increased numbers of CD4+ CD69+ T cells in the PL were positively associated with high IL-2 (p = 0.486) and IFN-γ (p = 0.017) levels, whereas reduced Treg cells in the blood were negatively correlated with TNFα levels (p = 0.043) and positively correlated with TGFβ1 (p = 0.038). Conclusion: Increased numbers of CD4+CD69+ T cells and reduced numbers of CD4+CD25+FoxP3+ Treg cells with an altered interleukin profile suggests loss of autotolerance in pristane-induced lupus mice, which is similar to human lupus. Therefore, this model is useful in evaluating mechanisms of cellular activation, peripheral tolerance and homeostatic immune imbalance involved in human SLE.

Serum adipocytokine profile and metabolic syndrome in young adult female dermatomyositis patients

OBJECTIVES: To analyse the frequency of metabolic syndrome in young adult female dermatomyositis patients and its possible association with clinical and laboratory dermatomyositis-related features and serum adipocytokines. METHOD: This cross-sectional study included 35 dermatomyositis patients and 48 healthy controls. Metabolic syndrome was defined according to the 2009 Joint Interim Statement. RESULTS: Patient age was comparable in the dermatomyositis and control groups, and the median disease duration was 1.0 year. An increased prevalence of metabolic syndrome was detected in the dermatomyositis group (34.3% vs. 6.3%; p=0.001). In addition, increased serum adiponectin and resistin levels were noted in contrast to lower leptin levels. In dermatomyositis patients, adipocytokine levels were correlated with the levels of total cholesterol, low-density cholesterol, triglycerides and muscle enzymes. A comparison of dermatomyositis patients with (n=12) and without (n=23) syndrome metabolic revealed that adipocytokine levels were also correlated with age, and that dermatomyositis patients with metabolic syndrome tended to have more disease activity despite similar adipocytokine levels. CONCLUSIONS: Metabolic syndrome is highly prevalent in young adult female dermatomyositis patients and is related to age and disease activity. Moreover, increased serum adiponectin and resistin levels were detected in dermatomyositis patients, but lower serum leptin levels were observed.

Serum adipocytokine profile and metabolic syndrome in young adult female dermatomyositis patients.

OBJECTIVES:: To analyse the frequency of metabolic syndrome in young adult female dermatomyositis patients and its possible association with clinical and laboratory dermatomyositis-related features and serum adipocytokines. METHOD:: This cross-sectional study included 35 dermatomyositis patients and 48 healthy controls. Metabolic syndrome was defined according to the 2009 Joint Interim Statement. RESULTS:: Patient age was comparable in the dermatomyositis and control groups, and the median disease duration was 1.0 year. An increased prevalence of metabolic syndrome was detected in the dermatomyositis group (34.3% vs. 6.3%; p=0.001). In addition, increased serum adiponectin and resistin levels were noted in contrast to lower leptin levels. In dermatomyositis patients, adipocytokine levels were correlated with the levels of total cholesterol, low-density cholesterol, triglycerides and muscle enzymes. A comparison of dermatomyositis patients with (n=12) and without (n=23) syndrome metabolic revealed that adipocytokine levels were also correlated with age, and that dermatomyositis patients with metabolic syndrome tended to have more disease activity despite similar adipocytokine levels. CONCLUSIONS:: Metabolic syndrome is highly prevalent in young adult female dermatomyositis patients and is related to age and disease activity. Moreover, increased serum adiponectin and resistin levels were detected in dermatomyositis patients, but lower serum leptin levels were observed.

No deleterious effect of low dose methotrexate on titanium implant osseointegration in a rabbit model.

OBJECTIVE: To evaluate the effect of low dose methotrexate alone or in combination with glucocorticoid treatment on titanium implant osseointegration. METHODS: Groups of 6-8 adult New Zealand White rabbits were treated for 18 weeks with saline (control), methotrexate, glucocorticoid, or methotrexate plus glucocorticoid. The animals received a titanium implant in the tibia at week 6. Lumbar spine and tibia bone mineral densities were analyzed before and after treatment. Histomorphometric analysis of bone cortical thickness, total bone area around the implant, and % of bone to implant contact was performed. RESULTS: After 18 weeks, the change in the bone mineral density in the lumbar spines and tibias in the methotrexate group was comparable to the control group (0.035 vs. 0.055 g/cm² and 0.021 vs. 0.041 g/cm², respectively). In contrast, both the glucocorticoid group and glucocorticoid plus methotrexate group had significant reductions at both sites. Histomorphometric analysis of the tibia in the control and methotrexate groups revealed no significant changes in cortical thickness (133 vs. 126 µm), total bone area around the implant (33 vs. 30%), or bone to implant contact (40 vs. 38%). In contrast, glucocorticoid group had significant reductions compared to controls in tibia cortical thickness (99 vs. 133 µm), total bone area around the implant (24 vs. 33%), and bone to implant contact (27 vs. 40%). Similar reductions were observed in the glucocorticoid plus methotrexate group. CONCLUSIONS: Our results demonstrate that low dose methotrexate treatment does not affect titanium implant osseointegration, suggesting that this therapy is safe for surgical procedures requiring a titanium implant.

Participação do óxido nítrico no processo inflamatório articular / Nitric oxide participation in the articular inflammatory process

O presente trabalho avalia a participação do óxido nítrico (NO) na fisiopatologia da resposta inflamatória articular. O papel pró-inflamatório deste medidor, tanto no componente vascular quanto no celular foi evidenciado em diferentes modelos experimentais, utilizando animais tratados cronicamente com o inibidor inespecífico de síntese de NO, NG-nitro-L-arginine metil ester (L-NAME, 20mg/Kg/dia/14 dias). Investigamos os mecanismos envolvidos na alteração de parâmetros inflamatórios como: interação leucócito endotélio, expressão de moléculas de adesão, reatividade da microcirculação, parâmetros hemodinâmicos, alterações na permeabilidade vascular, além da interação entre produção de NO e outros mediadores da resposta inflamatória, como eicosanóides, sistema de cininas, citocinas e histamina, bem como influência do tratamento na. hematopoiese e hemograma. Em conjunto, os estudos contribuem para esclarecer o controverso papel do NO no processo inflamatório. /The present study describes the participation of nitric oxide (NO) in the patophisiology of experimental arthritis induced by different antigens through chronicle administration of the unspecific inhibitor of NO synthases NG-nitro-L-arginine metyl ester (L-NAME, 20mg/Kg/day/14 days). The pro-inflammatory effect of NO was observed such as on cellular as on vascular events of the process. The mechanisms involved were assessed by investigations on haematopoeisis and haemogram, leukocyte endothelial interaction, adhesion molecules expression, haemodynamic properties, microvascular reactivity, protein leakage, as well as interaction between NO and eicosanoids, bradykinin, cytokines and histamine production. Together these studies contribute to clarify the controversial role of NO on the inflammatory process...

Atividade das enzimas ciclooxigenases e drogas anti-reumáticas modificadoras de doença / Cyclooxigenase enzyme activity and disease-modifying antirheumatic drugs

Os autores apresentam uma revisão que abrange a ação de drogas anti-reumáticas modificadoras de doença (DMARDS) como metotrexato, cloroquina, sulfasalazina e clorambucil, sobre atividades das enzimas ciclooxigenases 1 e 2. São apresentados resultados comprovando a ação inibitória de metotrexato preferencialmente sobre a atividade de ciclooxigenase-2, o que acrescenta novos conhecimentos sobre o mecanismo antiinflamatório de baixas doses de MTX. Os estudos até aqui realizados abrem novas perspectivas no conhecimento do mecanismo de ação das DMARDS

Participacao dos radicais livres na inflamacao articular / Role of free radicals on the articular inflammatory process

Os autores revisam as principais evidencias sugerindo a participacao dos radicais livres no processo articular inflamatorio. Particularmente, o anion superoxido e seus derivados (EAO - especies ativas derivadas do oxigenio) e o oxido nitrico sao analisados em relacao aos seus mecanismos de producao pelas celulas e tecidos das articulacoes, a sua capacidade de causar destruicao destes tecidos especializados e aos resultados decorrentes da inibicao de sua producao ou atividade

Niveis intra-articulares de oxido nitrico em pacientes com doenca reumatoide / Intraarticular nitric oxide levels in patients with rheumatoid arthritis

O objetivo deste trabalho foi determinar os niveis intra-articulares de nitrito e nitrato, produtos do metabolismo de oxido nitrico, em pacientes com doenca reumatoide (DRe). Liquido sinovial (LS) de pacientes com osteoartrite (OA) foram avaliados para fins comparativos. A casuistica constou de seis pacientes com DRe e quatro pacientes com OA. Foram avaliadas as caracteristicas demograficas como idade e sexo, e clinico-laboratoriais como duracao da doenca, classe funcional e velocidade de hemossedimentacao (VHS). No LS obtido foram realizadas a contagem total e diferencial de leucocitos, e a determinacao dos niveis de nitrito e nitrato pela tecnica de reacao de Griess. Os resultados foram analisados pelos testes de correlacao e "t" de Student...

Participacao dos radicais livres de oxigenio na fisiopatologia da artrite reumatoide / Oxygen free radicals participation in physiopathology of rheumatoid arthritis

Os autores realizam uma revisao sobre os mecanismos capazes de produzir radicais livres derivados do oxigenio e analisam as evidencias provenientes de estudos realizados in vivo e in vitro sugerindo o envolvimento destes radicais na fisiopatologia da artrite reumatoide. Como conclusao e possivel afirmar que, provavelmente, ha uma maior producao de oxidantes nos pacientes portadores desta doenca, com participacao efetiva destes agentes na lesao articular. Entretanto, maiores estudos sao necessarios para avaliar devidamente sua contribuicao para o desenvolvimento da doenca e sobretudo a potencialidade de agentes terapeuticos capazes de intervir com a producao e/ou acao destes oxidantes.