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1.
Clin Exp Immunol ; 177(2): 391-403, 2014 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-24673402

RESUMO

The interplay between diet and immune parameters which could affect type 1 diabetes (T1D) pathogenesis is not sufficiently clarified. Intestinal up-regulation of the activating receptor natural killer group 2D (NKG2D) (CD314) and its ligands is a hallmark of coeliac disease. However, the direct effect of gluten on NKG2D expression is not known. We studied, by fluorescence activated cell sorter (lymphoid tissues) and reverse transcription-quantitative polymerase chain reaction (intestine and pancreatic islets), if a gluten-free diet (GF diet) from 4 weeks of age or a gluten-free diet introduced in breeding pairs (SGF diet), induced changes in NKG2D expression on DX5(+) (CD49b) natural killer (NK) cells, CD8(+) T cells and in intestinal and islet levels of NKG2D and ligands in BALB/c and non-obese diabetic (NOD) mice. Gluten-free NOD mice had lower insulitis (P < 0·0001); reduced expression of NKG2D on DX5(+) NK cells in spleen and auricular lymph nodes (P < 0·05); and on CD8(+) T cells in pancreas-associated lymph nodes (P = 0·04). Moreover, the level of CD71 on DX5(+) NK cells and CD8(+) T cells (P < 0·005) was markedly reduced. GF and SGF mice had reduced expression of NKG2D and DX5 mRNA in intestine (P < 0·05). Differences in intestinal mRNA expression were found in mice at 8, 13 and 20 weeks. Intestinal expression of NKG2D ligands was reduced in SGF mice with lower expression of all ligands. In isolated islets, a SGF diet induced a higher expression of specific NKG2D ligands. Our data show that a gluten-free diet reduces the level of NKG2D and the expression of NKG2D ligands. These immunological changes may contribute to the lower T1D incidence associated with a gluten-free diet.


Assuntos
Dieta Livre de Glúten , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos NOD , Subfamília K de Receptores Semelhantes a Lectina de Células NK/metabolismo , Animais , Antígenos CD/metabolismo , Linfócitos T CD8-Positivos/imunologia , Linfócitos T CD8-Positivos/metabolismo , Diabetes Mellitus Tipo 1/genética , Diabetes Mellitus Tipo 1/imunologia , Diabetes Mellitus Tipo 1/metabolismo , Modelos Animais de Doenças , Feminino , Expressão Gênica , Imunofenotipagem , Mucosa Intestinal/metabolismo , Intestinos/imunologia , Intestinos/patologia , Ilhotas Pancreáticas/imunologia , Ilhotas Pancreáticas/metabolismo , Ilhotas Pancreáticas/patologia , Células Matadoras Naturais/imunologia , Células Matadoras Naturais/metabolismo , Ligantes , Tecido Linfoide/metabolismo , Camundongos , Subfamília K de Receptores Semelhantes a Lectina de Células NK/genética , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Receptores da Transferrina/metabolismo
2.
Clin Exp Immunol ; 166(2): 251-7, 2011 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-21985371

RESUMO

Toll-like receptors are pattern-recognition receptors of the innate immune system that are activated during viral, bacterial or other infections, as well as during disease progression of type 1 and type 2 diabetes. Toll-like receptor 5 (TLR-5) specifically recognizes bacterial infection through binding of flagellin from pathogenic bacteria such as Salmonella and Listeria species. We have found that the expression of TLR5 is up-regulated by glucose activation of isolated islets of Langerhans, in contrast to other investigated TLRs (TLR-2, -3, -4, -6 and -9. Stimulation of islets with 10 mm glucose increased the levels of TLR5 mRNA 10-fold (P=0·03) and the TLR-5 protein levels twofold (P=0·04). Furthermore, the protein level of downstream signalling molecule myeloid differentiation primary response gene 88 (MyD88) increased 1·6-fold (P=0·01). Activation of TLR-5 in islets lead to a marked reduction of both stimulated and basal secretion of insulin, as well as an increase in production of nitric oxide, proinflammatory cytokines, anti-inflammatory heat-shock protein and major histocompatibility complex (MHC) class I transporter. We observe no effects of TLR-5 activation on islet survival. We suggest that this regulation by TLR-5 might be beneficial during serious infection such as sepsis by limiting the activity of beta cells during peaks of insulin demand to counteract beta cell damage.


Assuntos
Glucose/metabolismo , Ilhotas Pancreáticas/metabolismo , Receptor 5 Toll-Like/metabolismo , Animais , Linhagem Celular Tumoral , Citocinas/biossíntese , Proteínas de Choque Térmico/biossíntese , Inflamação/imunologia , Inflamação/metabolismo , Insulina/metabolismo , Secreção de Insulina , Complexo Principal de Histocompatibilidade/efeitos dos fármacos , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Fator 88 de Diferenciação Mieloide/biossíntese , Fator 88 de Diferenciação Mieloide/metabolismo , Óxido Nítrico/biossíntese , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Ratos , Ratos Endogâmicos Lew , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Receptor 5 Toll-Like/genética , Receptor 5 Toll-Like/imunologia
3.
Mamm Genome ; 11(4): 288-91, 2000 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-10754104

RESUMO

We have studied the process of tspy gene silencing in murine evolution. We have isolated functional tspy sequences from Apodemus agrarius, A. sylvaticus, A. flavicollis, and Mus platythrix (subgenus Pyromys) and nonfunctional tspy sequences from species of the subgenus Mus. We present two alternative models as to how tspy may have lost its function in the murine lineage.


Assuntos
Evolução Biológica , Proteínas de Ligação a DNA/genética , Inativação Gênica , Proteínas Nucleares , Fatores de Transcrição , Cromossomo Y , Animais , Sequência de Bases , Primers do DNA , Camundongos , Dados de Sequência Molecular , Reação em Cadeia da Polimerase , Proteína da Região Y Determinante do Sexo
4.
Z Orthop Ihre Grenzgeb ; 136(2): 132-7, 1998.
Artigo em Alemão | MEDLINE | ID: mdl-9615975

RESUMO

PURPOSE OF THE STUDY: The aim of this study was to evaluate bone blood flow of the proximal femur during experimentally induced ischemia and to document the ability of epiphyseal and metaphyseal reperfusion. METHODS: 11 pigs (danish landrace) were used to investigate the effect of tamponade by increased joint pressure (Dextran 70) on the perfusion of the femoral head. Additional 8 pigs were used as control. The blood flow in the hip joint was studied by means of the microsphere technique. The flow was determined before, during and after intraarticular pressure increase. With the "radioactive tracer microsphere"-method the blood flow of the epiphysis, metaphysis and proximal femoral corticalis could be measured. RESULTS: In the epiphyseal femoral head the initial blood flow rate, 11.7 ml/min/100 g, was not significant different from that of the control side (11.1 ml/min/100 g). The blood flow decreased in the ischemic phase to 1.8 ml/min/100 g followed by reperfusion to 13.5 ml/min/100 g (p < 0.01). The bone blood flow of reperfusion was not significant different from that of the initial blood flow rate but in 2 cases a "blow out" of the epiphyseal bone blood flow was seen. The proximal femoral metaphysis showed the highest of the measured intraosseous flow rates (17.9 resp. 23.3 ml/min/100 g). During ischemia and reperfusion of the epiphysis bone blood flow of the metaphysis remained the same. The proximal femoral corticalis showed the lowest of the measured intraosseous flow rates. The operated (10.1 ml/min/100 g) and contralateral hip side (11.7 ml/min/100 g) showed no significant differences in the initial blood flow rate. During ischemia and reperfusion the blood flow of the proximal corticalis showed no significant difference to the initial blood flow corresponding to the metaphysis. CONCLUSIONS: Our study demonstrates disturbances of the circulation of different regions of the femoral head during intraarticular pressure increase and following pressure decrease of the growing pig. 2 "blow outs" document a vulnerable proximal epiphysis already after a 6-hour ischemia. Additional minor "bone quality" in cases of certain diseases (kidney transplantation, leukemia) and special administration of drugs (corticosteroids) seem to create an additional vulnerability of the proximal femoral head. The experiment proves to be a reliable model for decreasing the blood flow of the growing epiphysis temporarily and to document the beginning of normal reperfusion. With this model it is possible to examine the vulnerability of the epiphyseal perfusion after different diseases and under the influence of different medication.


Assuntos
Necrose da Cabeça do Fêmur/fisiopatologia , Cabeça do Fêmur/irrigação sanguínea , Isquemia/fisiopatologia , Traumatismo por Reperfusão/fisiopatologia , Animais , Velocidade do Fluxo Sanguíneo/fisiologia , Pressão Sanguínea/fisiologia , Débito Cardíaco/fisiologia , Modelos Animais de Doenças , Epífises/irrigação sanguínea , Fluxo Sanguíneo Regional/fisiologia , Suínos
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