RESUMO
INTRODUCTION: Type 1 diabetes is a major risk factor for the development of severe periodontal disease. As diabetes increases in severity, so does the susceptibility to and severity of periodontitis. People with diabetes who have periodontal disease have a harder time maintaining healthy blood glucose levels. Macrophages play an important role in both diabetes and periodontitis. Previous research comparing bone-marrow-derived macrophages (BM-Mvarphi) from diabetic non-obese diabetic (NOD) mice and control mice illustrates that a dysregulation in cytokine, Toll-like receptor (TLR) expression, and cell signaling occurs in the diabetic state. METHODS: This study examines the effect of chronic hyperglycemia on BM-Mvarphi TLR expression and activation, cell signaling, cytokine production, and phagocytic function in the diabetic state, when challenged with the periodontal stimulus Porphyromonas gingivalis lipopolysaccharide (LPS) to further understand how diabetes and associated hyperglycemia may contribute to the increased susceptibility of people with diabetes to periodontitis. RESULTS: When BM-Mvarphi, obtained from diabetic NOD mice, are stimulated with P. gingivalis LPS under hyperglycemic conditions the following changes occur: reduced messenger RNA expression and cell surface expression of TLR2, reduced messenger RNA expression and protein production of tumor necrosis factor-alpha, reduced signal transduction, and a reduction in phagocytic function. All the activity of BM-Mvarphi from diabetic NOD mice was restored when differentiation and stimulation occurred under normoglycemic conditions. DISCUSSION: Diabetic patients in a hyperglycemic state may be generating macrophages that are inherently immunocompromised, contributing to an environment allowing periodontal infections to flourish. As a consequence, people with diabetes who maintain proper control of blood sugar levels may experience an increased immunological benefit when challenged with a periodontal infection.
Assuntos
Diabetes Mellitus Tipo 1/imunologia , Hiperglicemia/imunologia , Macrófagos/imunologia , Fagocitose , Porphyromonas gingivalis/imunologia , Animais , Suscetibilidade a Doenças , Feminino , Hiperglicemia/fisiopatologia , Lipopolissacarídeos/imunologia , Células Progenitoras Linfoides/imunologia , Camundongos , Camundongos Endogâmicos NOD , NF-kappa B/imunologia , Doenças Periodontais/imunologia , Transdução de Sinais/imunologia , Receptor 2 Toll-Like/biossíntese , Fator de Necrose Tumoral alfa/biossínteseRESUMO
The expression of MHC class II molecules on beta-cells of the pancreatic islet has been proposed to play a role in the genesis of insulin-dependent diabetes mellitus in the NOD mouse. We investigated this by immunofluorescent double labeling of islet cells with anti-MHC and anti-CD45 to identify cells of hematopoietic origin. MHC class I expression increased with age on CD45- islet cells. MHC class II expression was not observed on CD45- islet cells at any age; the only cells in the islet that were MHC class II positive were also CD45+. This indicates that all MHC class II-positive cells in the islet are lymphoid cells that infiltrate the islet, whereas the islet endocrine cells express no MHC class II molecules. However, an increase in MHC class I expression occurred on beta-cells, and this may play a role in immunopathogenesis.
Assuntos
Antígenos de Diferenciação/análise , Diabetes Mellitus Experimental/imunologia , Antígenos de Histocompatibilidade Classe II/análise , Antígenos de Histocompatibilidade/análise , Ilhotas Pancreáticas/imunologia , Animais , Diabetes Mellitus Experimental/genética , Feminino , Citometria de Fluxo , Fluoresceína-5-Isotiocianato , Fluoresceínas , Corantes Fluorescentes , Antígenos Comuns de Leucócito , Glicoproteínas de Membrana/análise , Camundongos , Camundongos Mutantes , TiocianatosRESUMO
In studies of immune cell defects in autoimmune diabetes mellitus, we observed that complete Freund's adjuvant (CFA) prevented the onset of diabetes when injected into 8- to 10-wk-old prediabetic nonobese diabetic (NOD) mice. The prevalence of the onset of diabetes in the CFA-injected versus uninjected NOD mice was 2 of 81 (2.5%) vs. 231 of 379 (61%) among females and 2 of 44 (4.5%) vs. 83 of 336 (25%) among males, respectively. The incidence of histologically identifiable insulitis was significantly reduced in CFA-treated prediabetic female NOD mice (18%) compared with the incidence in female age-matched controls (70%). Splenocytes or Mac-(1+)-enriched splenocytes from CFA-treated NOD mice, when cotransferred with splenocytes from diabetic mice, reduced the incidence of diabetes provoked by diabetic splenocytes in vivo. In the spleen, CFA injection induced sustained increases in cell proliferation and an associated major increase in the numbers of an immature cell type that expressed the Mac-1 surface antigen. In CFA-treated NOD mice, lymphocytes derived from the spleen failed to respond in vitro to stimulation by the mitogen concanavalin A or by anti-CD3. When cocultured, Mac-1+ cells, enriched from the splenocytes of CFA-treated mice, suppressed concanavalin A- or anti-CD3-induced proliferation of T lymphocytes derived from either the spleen or thymus of untreated NOD mice. Therefore, treatment with CFA prevents the development of diabetes, and concomitantly, insulitis while stimulating the generation of splenic suppressor cells that are capable of suppressing diabetogenic T-lymphocyte function in vivo and in vitro.
Assuntos
Diabetes Mellitus Experimental/prevenção & controle , Adjuvante de Freund/uso terapêutico , Pancreatopatias/prevenção & controle , Animais , Células Cultivadas , Concanavalina A , Diabetes Mellitus Experimental/genética , Diabetes Mellitus Experimental/imunologia , Ilhotas Pancreáticas/efeitos dos fármacos , Ilhotas Pancreáticas/patologia , Ativação Linfocitária , Camundongos , Camundongos Mutantes , Pancreatopatias/genética , Fenótipo , Estado Pré-Diabético/terapia , Baço/imunologia , Linfócitos T/efeitos dos fármacos , Linfócitos T/imunologiaRESUMO
Although CD8+ T cells play a major role in beta cell destruction in insulin-dependent diabetes in the non-obese diabetic mouse, the T cell autoantigen(s) recognized by such cells remains to be identified. Therefore, an islet-reactive, CD8+ T cell line was generated from islet-infiltrating cells and hybridized by fusion with a CD8+ alphabeta TCR- BW5147 thymoma. In the presence of islets, none of the 12 CD3+ CD8+ T cell hybridomas isolated secreted IL-2/IL-4 or IFNgamma but three were islet specific, as shown by activation induced cell death. Subclone 4A7.7.15 recognized only islets expressing H-2Kd, demonstrated islet-specific inhibition of proliferation and concomitant partial arrest in the G2/M phase of the cell cycle. Further analysis using a panel of cell lines, expressing H-2Kd, and transfected with the cDNA for various putative autoantigens in type 1 diabetes showed that 4A7.7.15 recognizes insulin as an antigen.
Assuntos
Anticorpos Monoclonais , Autoantígenos/imunologia , Linfócitos T CD8-Positivos/imunologia , Diabetes Mellitus Tipo 1/imunologia , Insulina/imunologia , Ilhotas Pancreáticas/imunologia , Animais , Especificidade de Anticorpos , Divisão Celular , Fusão Celular , Células Cultivadas , Citotoxicidade Imunológica , Diabetes Mellitus Tipo 1/patologia , Feminino , Hibridomas/citologia , Hibridomas/imunologia , Interleucina-2/genética , Interleucina-4/genética , Ativação Linfocitária , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Camundongos Endogâmicos NOD , Timoma/imunologia , Neoplasias do Timo/imunologiaRESUMO
BACKGROUND AND AIMS: Ingestion of food stimulates the secretion of incretin peptides glucose-dependent insulinotropic polypeptide (GIP) and glucagon-like peptide 1 to ensure the proper absorption and storage of nutrients. Menin is the 67 kDa protein product of the MEN1 gene recently reported to have a role in metabolism. In this study, we will determine the regulation of menin in the proximal duodenum by food intake and diet in correlation with GIP levels in the proximal duodenum of mice after an 18 h fast followed by 4 and 7 h refeeding and 3 months of high-fat diet. METHODS: A dual luciferase assay was used to determine GIP promoter activity and ELISA was used to measure the levels of GIP after inhibition of menin through small interfering RNA (siRNA) and exposure to MAPK and AKT inhibitors. Colocalization of menin and GIP were determined by immunofluorescence. RESULTS: Menin and GIP expression are regulated by fasting, refeeding and diet in the proximal duodenum. Overexpression of menin in STC-1 cells significantly inhibited GIP mRNA and promoter activity, whereas menin siRNA upregulated GIP levels. Inhibition of GIP expression by the PI3/AKT inhibitor, LY294002, was abrogated in STC-1 cells with reduced menin levels, whereas the MAPK inhibitor, UO126, inhibited the expression of GIP independent of menin. Exposure of STC-1 cells to GIP reduced menin expression in a dose-dependent manner via PI3K-AKT signaling. CONCLUSION: Feeding and diet regulates the expression of menin, which inversely correlates with GIP levels in the proximal duodenum. In vitro assays indicate that menin is a negative regulator of GIP via inhibition of PI3K-AKT signaling. We show menin colocalizing with GIP in K cells of the proximal gut and hypothesize that downregulation of menin may serve as a mechanism by which GIP is regulated in response to food intake and diet.
RESUMO
Type 2 diabetes is associated with normal-to-higher bone mineral density (BMD) and increased rate of fracture. Hyperinsulinemia and hyperglycemia may affect bone mass and quality in the diabetic skeleton. In order to dissect the effect of hyperinsulinemia from the hyperglycemic impact on bone homeostasis, we have analyzed L-SACC1 mice, a murine model of impaired insulin clearance in liver causing hyperinsulinemia and insulin resistance without fasting hyperglycemia. Adult L-SACC1 mice exhibit significantly higher trabecular and cortical bone mass, attenuated bone formation as measured by dynamic histomorphometry, and reduced number of osteoclasts. Serum levels of bone formation (BALP) and bone resorption markers (TRAP5b and CTX) are decreased by approximately 50%. The L-SACC1 mutation in the liver affects myeloid cell lineage allocation in the bone marrow: the (CD3(-)CD11b(-)CD45R(-)) population of osteoclast progenitors is decreased by 40% and the number of (CD3(-)CD11b(-)CD45R(+)) B-cell progenitors is increased by 60%. L-SACC1 osteoclasts express lower levels of c-fos and RANK and their differentiation is impaired. In vitro analysis corroborated a negative effect of insulin on osteoclast recruitment, maturation and the expression levels of c-fos and RANK transcripts. Although bone formation is decreased in L-SACC1 mice, the differentiation potential and expression of the osteoblast-specific gene markers in L-SACC1-derived mesenchymal stem cells (MSC) remain unchanged as compared to the WT. Interestingly, however, MSC from L-SACC1 mice exhibit increased PPARgamma2 and decreased IGF-1 transcript levels. These data suggest that high bone mass in L-SACC1 animals results, at least in part, from a negative regulatory effect of insulin on bone resorption and formation, which leads to decreased bone turnover. Because low bone turnover contributes to decreased bone quality and an increased incidence of fractures, studies on L-SACC1 mice may advance our understanding of altered bone homeostasis in type 2 diabetes.
Assuntos
Densidade Óssea/fisiologia , Antígeno Carcinoembrionário/metabolismo , Diferenciação Celular/fisiologia , Insulina/metabolismo , Fígado/metabolismo , Osteoclastos/metabolismo , Análise de Variância , Animais , Composição Corporal/fisiologia , Reabsorção Óssea/metabolismo , Antígeno Carcinoembrionário/genética , Moléculas de Adesão Celular/metabolismo , Citometria de Fluxo , Hiperinsulinismo/metabolismo , Fator de Crescimento Insulin-Like I/metabolismo , Camundongos , Camundongos Transgênicos , Obesidade/metabolismo , Osteogênese/fisiologia , Ligante RANK/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase ReversaRESUMO
Our previous work examining the importance of insulin receptor (IR) expression on T cells has demonstrated that when T cells from nonobese diabetic mice were sorted into populations expressing a high (IR(High)) and a low (IR(Low)) density of IR, IR(High) T cells rapidly transferred insulitis and diabetes. We have further characterized IR(High) T cells. Both CD4+ and CD8+ cells were detected in the IR(High) T cell population, but IR(High) expression was detected predominantly on CD4+ cells. IRHigh T cells were polyclonal for TCR Vbeta-chain expression. By 3 color flow cytometric analysis, virtually all IR(High) T cells expressed low or negligible levels of CD62L (CD62L(Low)/-) and high levels of CD44 (CD44(High)). The lack of IL-2 receptor and transferrin receptor expression as seen previously, together with the CD62L(Low)/- CD44(High) phenotype suggests that IR(High) T cells are memory cells. However, since only about one quarter of all of the CD62L(Low)/- or CD44(High) T cells were also IR(High), the IR(High) phenotype defines a subpopulation of memory T cells that are aggressively diabetogenic.
Assuntos
Diabetes Mellitus Tipo 1/imunologia , Diabetes Mellitus Tipo 1/metabolismo , Memória Imunológica , Receptor de Insulina/metabolismo , Subpopulações de Linfócitos T/imunologia , Subpopulações de Linfócitos T/metabolismo , Animais , Linfócitos T CD4-Positivos/imunologia , Linfócitos T CD4-Positivos/metabolismo , Feminino , Expressão Gênica , Receptores de Hialuronatos/metabolismo , Selectina L/metabolismo , Ativação Linfocitária , Camundongos , Camundongos Endogâmicos NOD , Fenótipo , Receptores de Antígenos de Linfócitos T alfa-beta/genética , Receptores de Interleucina-2/metabolismo , Receptores da Transferrina/metabolismoRESUMO
Sucrose dissimilation was studied in five strains of Streptococcus mutans. Glucose-adapted strain SL-1 makes acid more slowly from sucrose than from glucose; glucose-adapted strain SL-1 gives diauxie growth kinetics in broth containing limiting amounts of both glucose and sucrose. Thus, at least part of the sucrose dissimilative system appears inducible. Sucrase activity was identified in the 37,000 x g soluble cell fraction of five strains. Its intracellular location implies the presence of sucrose permease. The specific activity of the sucrase is higher in sucrose-adapted cells than in cells adapted to glucose or other sugars, further suggesting its inducibility. The enzyme from strain SL-1 was partially purified by diethylaminoethyl-cellulose chromatography and shown to be a single molecule with a molecular weight of about 48,000. The partially purified enzyme is specific for sucrose and produces equimolar glucose and fructose. Since it degrades raffinose, but not melezitose or other alpha-glucosides, it is an invertase. The invertase has a relatively high K(m) for its substrate and a pH optimum of 5.5 to 6.2. It is activated by inorganic orthophosphate (P(i)), P(i) functioning as a positive effector. Arsenate can substitute for phosphate. Neither the crude cell-free extract nor the partially purified enzyme preparations has detectable sucrose phosphorylase activity. A possible potent role of the invertase in the regulation of sucrose carbon flow in S. mutans is discussed.
Assuntos
Streptococcus/enzimologia , Sacarase/metabolismo , Ácidos/biossíntese , Arsênio/farmacologia , Radioisótopos de Carbono , Sistema Livre de Células , Cromatografia DEAE-Celulose , Cromatografia em Papel , Colorimetria , Eletroforese em Gel de Poliacrilamida , Frutose/biossíntese , Glucose/biossíntese , Glucose/metabolismo , Glucosiltransferases/metabolismo , Concentração de Íons de Hidrogênio , Proteínas de Membrana Transportadoras/metabolismo , Peso Molecular , Fosfatos/farmacologia , Radioisótopos de Fósforo , Especificidade da Espécie , Streptococcus/crescimento & desenvolvimento , Streptococcus/metabolismo , Sacarase/isolamento & purificação , Sacarose/metabolismoRESUMO
CD4(+) T cell responses to glutamic acid decarboxylase (GAD65) spontaneously arise in nonobese diabetic (NOD) mice before the onset of insulin-dependent diabetes mellitus (IDDM) and may be critical to the pathogenic process. However, since both CD4(+) and CD8(+) T cells are involved in autoimmune diabetes, we sought to determine whether GAD65-specific CD8(+) T cells were also present in prediabetic NOD mice and contribute to IDDM. To refine the analysis, putative K(d)-binding determinants that were proximal to previously described dominant Th determinants (206-220 and 524-543) were examined for their ability to elicit cytolytic activity in young NOD mice. Naive NOD spleen cells stimulated with GAD65 peptides 206-214 (p206) and 546-554 (p546) produced IFN-gamma and showed Ag-specific CTL responses against targets pulsed with homologous peptide. Conversely, several GAD peptides distal to the Th determinants, and control K(d)-binding peptides did not induce similar responses. Spontaneous CTL responses to p206 and p546 were mediated by CD8(+) T cells that are capable of lysing GAD65-expressing target cells, and p546-specific T cells transferred insulitis to NOD.scid mice. Young NOD mice pretreated with p206 and p546 showed reduced CTL responses to homologous peptides and a delay in the onset of IDDM. Thus, MHC class I-restricted responses to GAD65 may provide an inflammatory focus for the generation of islet-specific pathogenesis and beta cell destruction. This report reveals a potential therapeutic role for MHC class I-restricted peptides in treating autoimmune disease and revisits the notion that the CD4- and CD8-inducing determinants on some molecules may benefit from a proximal relationship.
Assuntos
Citotoxicidade Imunológica/imunologia , Epitopos de Linfócito T/imunologia , Glutamato Descarboxilase/imunologia , Antígenos H-2/imunologia , Epitopos Imunodominantes/imunologia , Isoenzimas/imunologia , Linfócitos T Citotóxicos/enzimologia , Linfócitos T Citotóxicos/imunologia , Administração Intranasal , Envelhecimento/imunologia , Animais , Animais Recém-Nascidos , Animais Lactentes , Linfócitos T CD8-Positivos/enzimologia , Linfócitos T CD8-Positivos/imunologia , Linhagem Celular , Ciclofosfamida/administração & dosagem , Testes Imunológicos de Citotoxicidade , Diabetes Mellitus Tipo 1/imunologia , Diabetes Mellitus Tipo 1/prevenção & controle , Feminino , Adjuvante de Freund/administração & dosagem , Glutamato Descarboxilase/administração & dosagem , Glutamato Descarboxilase/genética , Injeções Intraperitoneais , Isoenzimas/administração & dosagem , Isoenzimas/genética , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos NOD , Camundongos SCID , Fragmentos de Peptídeos/administração & dosagem , Fragmentos de Peptídeos/imunologia , Ratos , Baço/citologia , Baço/imunologia , Subpopulações de Linfócitos T/enzimologia , Subpopulações de Linfócitos T/imunologia , TransfecçãoRESUMO
This study was undertaken to examine the levels and function of peripheral blood immunoregulatory T cell subpopulations in systemic lupus erythematosus (SLE). T cell subpopulations can be distinguished by the T cell differentiation antigens CD4 (recognized by the monoclonal antibodies OKT4 or Leu3) and CD8 (recognized by the monoclonal antibodies OKT8 or Leu2). All SLE patients tested had normal percentages of CD8 cells in their peripheral blood. The SLE patients, however, fell into two groups based on their CD4 cell numbers. Fifty-five percent of the SLE patients had normal levels of CD4 cells (Group A) and therefore normal CD4/CD8 cell ratios, whereas 45% of the SLE patient population had markedly depressed CD4 cell levels (Group B) and significantly low CD4/CD8 cell ratios. T cells from normal donors and SLE patients were further examined for their ability to stimulate allogeneic normal B/M phi cells to secrete IgM in the presence of pokeweed mitogen (PWM). Utilizing this assay system two forms of immunosuppression were observed: (1) that mediated by high concentrations of purified CD4 cells and (2) that mediated by CD8 cells. High concentrations of purified CD4 cells, added to a constant number of allogeneic normal B/M phi cells, suppressed PWM-stimulated IgM synthesis. Group B SLE patients, with significantly low CD4 cell numbers, had defective CD4 cell-mediated suppression which was concentration dependent. This result was confirmed in a study using identical twins discordant for SLE. In this case CD4 cells from the SLE twin did not induce immunosuppression at a high concentration of CD4 cells whereas similar concentrations of CD4 cells from the normal twin resulted in suppression. SLE patients (Group A) with normal levels of CD4 cells had normally immunosuppressive CD4 cells. Suppression mediated by CD8 cells was demonstrated by the fact that removal of CD8 cells resulted in enhanced IgM synthesis induced by the remaining CD4 cells. Although all the SLE patients in this study had normal peripheral blood levels of CD8 cells, SLE Group A patients had defective CD8 cell suppression whereas CD8 function appeared to be normal in Group B patients. These results suggest that in SLE patients with depressed CD4 cell numbers (Group B) there is a corresponding defect in CD4 cell function. We demonstrate that in SLE Group B patients, defective suppression is due to a subset of T cells that bear the CD4 antigen. The SLE patient population (Group A) with normal CD4/CD8 ratios and normally functioning CD4 cells, however, appear to have normal CD4 cell-mediated suppression but defective CD8 suppressor cell function.
Assuntos
Linfócitos B/classificação , Lúpus Eritematoso Sistêmico/sangue , Linfócitos T/classificação , Formação de Anticorpos , Linfócitos B/imunologia , Humanos , Ativação Linfocitária , Linfócitos T/imunologia , Linfócitos T ReguladoresRESUMO
Sucrase activity was studied in 13 strains of Streptococcus mutans representing the five Bratthall serotypes. Sucrose-adapted cells have sucrase activity in the 37,000 x g-soluble fraction of all strains. The enzyme was identified as invertase (beta-d-fructofuranoside fructohydrolase; EC 3.2.1.26) because it hydrolyzed the beta-fructofuranoside trisaccharide raffinose, giving fructose and melibiose as its products, and because it hydrolyzed the beta-fructofuranoside dissacharide sucrose, giving equimolar glucose and fructose as its products. Invertases of c and e strains exhibit two activity peaks by molecular exclusion chromatography with molecular weights of 45,000 to 50,000 and about 180,000; those of serotypes a, b, and d strains exhibit only a single component of 45,000 to 50,000 molecular weight. The electrophoretic mobility of invertases is different between the serotypes and the same within them. Inorganic orthophosphate (P(i)) has a weak positive effect on the V(max) of invertases of serotypes c and e cells but a strong positive effect on the invertases of serotype b cells; P(i) has a strong positive effect on the apparent K(m) of the invertases of serotype d cells, but has no effect on the V(max); P(i) has a strong positive effect on both the apparent K(m) and V(max) of the invertases of serotype a cells. Thus, the invertases were different between all of the serotypes but similar within the serotypes. These findings support the taxonomic schemes of Coykendall and of Bratthall. It was additionally noted that 37,000 x g-soluble fractions of only serotypes b and c but not serotypes a, d, and e cells have melibiase activity, and it could be deduced that serotype d cells lack an intact raffinose permease system.
Assuntos
Streptococcus mutans/enzimologia , Streptococcus/enzimologia , Sacarase/metabolismo , Sistema Livre de Células , Cromatografia em Gel , Cárie Dentária/enzimologia , Diálise , Eletroforese em Gel de Poliacrilamida , Cinética , Peso Molecular , Sorologia , Especificidade da Espécie , Sacarose/metabolismoRESUMO
In the nonobese diabetic mouse, insulin-dependent diabetes is an autoimmune disease characterized by T cell-mediated invasion and destruction of pancreatic islet beta cells. The importance of insulin receptor (IR) expression in the pathogenesis of diabetes was examined, since it has been shown that the IR is a chemotactic receptor capable of directing cell movement in response to insulin. Using polyclonal antisera to the IR, phenotypic analysis of purified splenic T cells from diabetic mice showed that about 15% of T cells expressed high density IR (IRhigh). In addition, IRhigh T cells were already a dominant phenotype in the insulitis of young prediabetic mice. To determine the ability of IRhigh T cells to transfer diabetes, cells were sorted by flow cytometry before adoptive transfer into young (6- to 8-wk-old) nondiabetic irradiated nonobese mice. Transfer of as few as 3 x 10(6) purified IRhigh T cells alone resulted in rapid onset of insulitis and diabetes, and IRhigh-depleted T cells were essentially unable to passage either insulitis or diabetes. The adoptive transfer of disease was not due to the transfer of activated cells, since removal of IL-2R+ or transferrin R+ cells did not alter diabetes transfer. Therefore, IRhigh T cells are aggressively diabetogenic, suggesting that increased IR expression may provide a mechanism for delivering potentially autoreactive T cells to the islet, regardless of their activation state.
Assuntos
Diabetes Mellitus Tipo 1/imunologia , Diabetes Mellitus Tipo 1/metabolismo , Receptor de Insulina/metabolismo , Linfócitos T/imunologia , Linfócitos T/metabolismo , Animais , Anticorpos , Diabetes Mellitus Tipo 1/etiologia , Feminino , Imunização Passiva , Ilhotas Pancreáticas/imunologia , Ilhotas Pancreáticas/patologia , Masculino , Camundongos , Camundongos Endogâmicos NOD , Receptor de Insulina/imunologia , Baço/imunologia , Subpopulações de Linfócitos T/imunologiaRESUMO
Interleukin-1 beta (IL-1 beta) significantly inhibits insulin secretion from glucose stimulated islet cells. The mechanism for this inhibition has been hypothesized to be due to stimulation of the inducible form of nitric oxide synthase and a resulting increase in nitric oxide (NO) concentration. Ways to block the effect of IL-1 beta have focused on blocking the binding of IL-1 beta to the IL-1 receptor and the use of antioxidants to neutralize increases in NO. This report focuses on a 33 residue peptide synthesized based on the C-terminal region of the IL-1 beta molecule, a reported binding site of the IL-1 beta molecule, and the redoxcycling antioxidant pyrroloquinoline quinone (PQQ). The 33 residue peptide did not function as an antagonist, but as a weak agonist. High concentrations of PQQ itself inhibited glucose-dependent insulin release while low concentrations did not. PQQ had no effect on the actions of IL-1 beta. Three isosteric and isomeric analogues of PQQ were also investigated. One of the PQQ isomers had an inhibitory effect on insulin secretion at low concentrations where PQQ had no effect. These results reflect the sensitivity of islets to oxidative stress.