RESUMO
Bazyler, CD, Mizuguchi, S, Zourdos, MC, Sato, K, Kavanaugh, AA, DeWeese, BH, Breuel, KF, and Stone, MH. Characteristics of a national level female weightlifter peaking for competition: A case study. J Strength Cond Res 32(11): 3029-3038, 2018-This study investigated physiological and performance changes of a national-level 69 kg female weightlifter after 3 competition phases over a 28-week training period. The athlete first trained for a regional championship (weeks 1-12), followed by a local competition (weeks 13-23) and the national championship (weeks 24-28). Body mass, vastus lateralis cross-sectional area (CSA), and unloaded and loaded squat jump performance were assessed weekly during each 4-week competition phase. Serum biomarkers and dynamic midthigh pulls were assessed before and after each competition phase. Weightlifting performance goals were met for the regional championship (total = 200 kg) and the local competition (total = 193 kg), but not the national championship (total = 196 kg). She lost more body mass in preparation for Nationals (-6.0 kg) compared with regionals (-2.5 kg) and the local competition (+2.2 kg). Vastus lateralis CSA very likely decreased after Nationals (precision = 99%, effect size = 2.08). Her testosterone:cortisol ratio likely increased (88%, 2.64), whereas interleukin-6 (79%, 2.47) and tumor necrosis factor-alpha (81%, 3.59) likely decreased after Nationals. Serum myostatin (99%, 1.95) and decorin (99%, 1.96) very likely decreased after the local competition. Unloaded squat jump height likely increased the week of regionals (89%, 0.95) and the local competition (99%, 1.83), whereas unloaded and loaded squat jump height possibly (69%, 0.99) and likely (82%, 1.52) decreased the week of Nationals. Dynamic midthigh pull vertical displacement likely increased after regionals (93%, 0.84) and likely decreased after Nationals (94%, 0.87). These findings indicate that biomarkers of stress, inflammation, and hypertrophy are related to changes in training volume-load; however, performance measures are needed to assess competition preparedness. Considering the reductions in muscle CSA corresponding with the large reductions in body mass and underperformance at the national championship, sport scientists, and coaches should instruct weightlifters to not attempt large losses in body mass (e.g., >3 kg) close to competition (e.g., <1week).
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Adaptação Fisiológica , Condicionamento Físico Humano , Músculo Quadríceps/anatomia & histologia , Levantamento de Peso/fisiologia , Atletas , Desempenho Atlético/fisiologia , Tamanho Corporal , Decorina/sangue , Feminino , Humanos , Hidrocortisona/sangue , Interleucina-6/sangue , Força Muscular , Miostatina/sangue , Músculo Quadríceps/diagnóstico por imagem , Treinamento Resistido , Testosterona/sangue , Fator de Necrose Tumoral alfa/sangue , Adulto JovemRESUMO
Sepsis is a frequent complication in critical illness. The mechanisms that are involved in initiation and propagation of the disease are not well understood. Scavenger receptor A (SRA) is a membrane receptor that binds multiple polyanions such as oxidized LDL and endotoxin. Recent studies suggest that SRA acts as a pattern recognition receptor in the innate immune response. The goal of the present study was to determine the role of SRA in polymicrobial sepsis. SRA deficient (SRA(-/-)) and C57BL/6JB/6J (WT) male mice were subjected to cecal ligation and puncture (CLP) to induce polymicrobial sepsis. NFκB activity, myeloperoxidase activity, and co-association of SRA with toll like receptor (TLR) 4 and TLR2 was analyzed in the lungs. Spleens were analyzed for apoptosis. Serum cytokines and chemokines were assayed. Blood and peritoneal fluid were cultured for aerobic and anaerobic bacterial burdens. Long-term survival was significantly increased in SRA(-/-) septic mice (53.6% vs. 3.6%, p < 0.05) when compared to WT mice. NFκB activity was 45.5% lower in the lungs of SRA(-/-) septic mice versus WT septic mice (p < 0.05). Serum levels of interleukin (IL)-5, IL-6, IL-10 and monocyte chemoattractant protein -1 were significantly lower in septic SRA(-/-) mice when compared to septic WT mice (p < 0.05). We found that SRA immuno-precipitated with TLR4, but not TLR2, in the lungs of WT septic mice. We also found that septic SRA(-/-) mice had lower bacterial burdens than WT septic mice. SRA deficiency had no effect on pulmonary neutrophil infiltration or splenocyte apoptosis during sepsis. We conclude that SRA plays a pivotal, and previously unknown, role in mediating the pathophysiology of sepsis/septic shock in a murine model of polymicrobial sepsis. Mechanistically, SRA interacts with TLR4 to enhance the development of the pro-inflammatory phenotype and mediate the morbidity and mortality of sepsis/septic shock.
Assuntos
Coinfecção/imunologia , Receptores Depuradores Classe A/metabolismo , Sepse/imunologia , Receptor 4 Toll-Like/metabolismo , Animais , Apoptose , Líquido Ascítico/microbiologia , Carga Bacteriana , Sangue/microbiologia , Ceco/cirurgia , Quimiocinas/sangue , Quimiocinas/imunologia , Coinfecção/microbiologia , Coinfecção/mortalidade , Citocinas/sangue , Citocinas/imunologia , Regulação da Expressão Gênica , Pulmão/imunologia , Pulmão/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , NF-kappa B/metabolismo , Infiltração de Neutrófilos , Peroxidase , Receptores Depuradores Classe A/deficiência , Receptores Depuradores Classe A/genética , Sepse/microbiologia , Sepse/mortalidade , Choque Séptico/microbiologia , Baço/imunologia , Baço/metabolismo , Receptor 2 Toll-Like/metabolismoRESUMO
The purpose of this study was to determine whether changes in collegiate weightlifters' external training load, biochemical markers, and jumping performance correlate to changes in items of the Short Recovery and Stress Scale (SRSS) throughout four microcycles. Twelve well-trained weightlifters (8 males, 4 females; age 24.30 ± 4.36 yr; height 170.28 ± 7.09 cm; body mass 81.73 ± 17.00 kg) with at least one year of competition experience participated in the study. Measurements included hydration, SRSS, biochemical analysis of blood (cortisol [C], creatine kinase [CK]), and unloaded and loaded squat jumps (SJ), and volume-load displacement. Pearson correlation coefficients were calculated between the changes in SRSS items and all other variables. The alpha criterion for all analyses was set at p ≤ 0.05. Negative relationships were observed between changes in SRSS recovery items and C (r = -0.608 to -0.723), and unloaded and loaded SJ height and peak power (r = -0.587 to -0.636). Positive relationships were observed between changes in several SRSS stress items and C (r = 0.609 to 0.723), CK (r = 0.922), and unloaded and loaded SJ height and peak power (r = 0.583 to 0.839). Relationships between changes in some SRSS items and cortisol agree with previous findings highlighting C as an indicator of training stress. Nonetheless, the non-significant relationships between changes in SRSS items, training volume and biochemical markers disagree with previous findings. This may partly be explained by the smaller undulations in training volume in the current study, which were characteristic of typical training. Further, relationships between changes in some SRSS items and jumping performance were opposite of what was expected indicating athletes' perception of their stress and recovery state does not always correspond with their ability to perform.
RESUMO
Background Abnormal renal hemodynamic responses to salt-loading are thought to contribute to salt-sensitive (SS) hypertension. However, this is based largely on studies in anesthetized animals, and little data are available in conscious SS and salt-resistant rats. Methods and Results We assessed arterial blood pressure, renal function, and renal blood flow during administration of a 0.4% NaCl and a high-salt (4.0% NaCl) diet in conscious, chronically instrumented 10- to 14-week-old Dahl SS and consomic SS rats in which chromosome 1 from the salt-resistant Brown-Norway strain was introgressed into the genome of the SS strain (SS.BN1). Three weeks of high salt intake significantly increased blood pressure (20%) and exacerbated renal injury in SS rats. In contrast, the increase in blood pressure (5%) was similarly attenuated in Brown-Norway and SS.BN1 rats, and both strains were completely protected against renal injury. In SS.BN1 rats, 1 week of high salt intake was associated with a significant decrease in renal vascular resistance (-8%) and increase in renal blood flow (15%). In contrast, renal vascular resistance failed to decrease, and renal blood flow remained unchanged in SS rats during high salt intake. Finally, urinary sodium excretion and glomerular filtration rate were similar between SS and SS.BN1 rats during 0.4% NaCl and high salt intake. Conclusions Our data support the concept that renal vasodysfunction contributes to blood pressure salt sensitivity in Dahl SS rats, and that genes on rat chromosome 1 play a major role in modulating renal hemodynamic responses to salt loading and salt-induced hypertension.
Assuntos
Hipertensão Renal , Animais , Pressão Sanguínea , Hipertensão/induzido quimicamente , Hipertensão/genética , Rim/fisiologia , Ratos , Ratos Endogâmicos BN , Ratos Endogâmicos Dahl , Cloreto de Sódio , Cloreto de Sódio na DietaRESUMO
Heterotrimeric Gi proteins play a role in lipopolysaccharide (LPS) and Staphylococcus aureus (SA) activated signaling leading to inflammatory mediator production. We hypothesized that genetic deletion of Gi proteins would alter cytokine and chemokine production induced by LPS and SA. LPS- and heat killed SA-induced cytokine and chemokine production in splenocytes from wild type (WT), Galpha(i2) (-/-) or Galpha(i1/3) (-/-) mice were investigated. LPS- or SA-induced production of TNFalpha, IL-6, IFNgamma, IL-12, IL-17, GM-CSF, MIP-1alpha, MCP-1, MIG and IP-10 were significantly increased (1.2 to 33 fold, p<0.05) in splenocytes harvested from Galpha(i2)(-/-) mice compared with WT mice. The effect of Galpha(i) protein depletion was remarkably isoform specific. In splenocytes from Galpha(i1/3) (-/-) mice relative to WT mice, SA-induced IL-6, IFNgamma, GM-CSF, and IP-10 levels were decreased (59% to 86%, p<0.05), whereas other LPS- or SA-stimulated cytokines and chemokines were not different relative to WT mice. LPS- and SA-induced production of KC were unchanged in both groups of the genetic deficient mice. Splenocytes from both Galpha(i2) (-/-) and Galpha(i1/3) (-/-) mice did not exhibit changes in TLR2 and TLR4 expression. Also analysis of splenic cellular composition by flow cytometry demonstrated an increase in splenic macrophages and reduced CD4 T cells in both Galpha(i2) (-/-) and Galpha(i1/3) (-/-) mice relative to WT mice. The disparate response of splenocytes from the Galpha(i2) (-/-) relative to Galpha(i1/3) (-/-) mice therefore cannot be attributed to major differences in spleen cellular composition. These data demonstrate that G(i2) and G(i1/3) proteins are both involved and differentially regulate splenocyte inflammatory cytokine and chemokine production in a highly Gi isoform specific manner in response to LPS and Gram-positive microbial stimuli.
Assuntos
Quimiocinas/biossíntese , Citocinas/biossíntese , Subunidade alfa Gi2 de Proteína de Ligação ao GTP/fisiologia , Subunidades alfa Gi-Go de Proteínas de Ligação ao GTP/fisiologia , Lipopolissacarídeos/farmacologia , Baço/metabolismo , Staphylococcus aureus/fisiologia , Animais , Células Cultivadas , Subunidade alfa Gi2 de Proteína de Ligação ao GTP/genética , Subunidades alfa Gi-Go de Proteínas de Ligação ao GTP/genética , Regulação da Expressão Gênica , Camundongos , Camundongos Knockout , Isoformas de Proteínas , Sensibilidade e Especificidade , Baço/citologia , Receptor 2 Toll-Like/metabolismo , Receptor 4 Toll-Like/metabolismoRESUMO
Previous studies have demonstrated that bacterial lipopolysaccharide (LPS) and heat killed Staphylococcus aureus (SA) activation of inflammatory cells depended in part upon activation of heterotrimeric Gi proteins. It has also been shown that (1 --> 3) beta-D-glucan can suppress inflammatory cell activation by microbial products although the cellular mechanism of the glucan effect remains to be clearly defined. We hypothesized that Gi proteins function as a common convergent signaling pathway for both LPS and SA leading to monocyte mediator production. Additionally, we hypothesized that soluble glucan suppresses LPS and SA induced cytokine production via Gi protein coupled signaling. Human THP-1 promonocytic cells were pretreated with pertussis toxin (PTx, 100 ng/ml or 1 microgram/ml) 6 hours prior to stimulation with LPS (10 microgram/ml) and SA (10 microgram/ml) and/or soluble glucan (10 microgram/ml). Both LPS and SA significantly (p < 0.05) induced cytokine production IL-6 > TNF alpha > IL-1 beta > GM-CSF > IL-10 > IFN gamma. The induction of these cytokines was significantly (p < 0.05) suppressed by PTx. Glucan treatment alone had no effect on cytokine production but suppressed (P < 0.05) LPS and SA induced cytokines. PTx further augmented (p > 0.05) the inhibitory effect of glucan on the LPS and SA induced cytokine expression. The data support the hypothesis that Gi proteins function as a common signaling protein for both LPS and SA induction of pro-and anti-inflammatory cytokines and that soluble glucan effectively suppresses cytokine production to the microbial stimuli. In contrast, the effect of soluble glucan on inhibiting cellular activation by LPS and SA is Gi protein independent.
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Citocinas/biossíntese , Proteínas de Ligação ao GTP/fisiologia , Lipopolissacarídeos/efeitos adversos , beta-Glucanas/metabolismo , Técnicas de Cultura de Células , Infecções por Bactérias Gram-Negativas/imunologia , Infecções por Bactérias Gram-Positivas/imunologia , Humanos , Inflamação , Monócitos , Proteoglicanas , Choque Séptico/fisiopatologia , Transdução de Sinais , Infecções Estafilocócicas/imunologia , Staphylococcus aureusRESUMO
Mast cells are highly responsive cells that are capable of secreting a variety of inflammatory mediators, including histamine, heparin, serine proteases, leukotrienes, prostaglandins, and thromboxanes. Studies from several laboratories have demonstrated that mast cells have the capacity to produce a variety of cytokines in response to various stimuli. Characterization of the cytokine profiles in mast cells has routinely been determined by the performance of individual enzyme-linked immunosorbent assays. This process is expensive, time-consuming, and requires a great deal of material to characterize multiple cytokines. In this chapter, we describe a multiplex cytokine assay to detect 17 cytokines simultaneously in 50 microL of culture supernatant derived from stimulated human cord blood-derived mast cells.
Assuntos
Bioensaio/métodos , Citocinas/análise , Mastócitos/química , Mastócitos/metabolismo , Sangue Fetal/citologia , Humanos , Ionomicina/metabolismo , Ionóforos/metabolismo , Acetato de Tetradecanoilforbol/metabolismoRESUMO
Two experiments were performed to determine whether administration of ergotamine tartrate altered embryo development (Exp. 1) and uterine competency to establish pregnancy (Exp. 2) in beef cattle. Animals were fed daily either 0 (CON) or 40 mug/kg body weight of ergotamine tartrate (ERGOT). Following a 30-d period on respective diets, animals in Exp. 1 were artificial inseminated at estrus (d = 0) and single embryo recoveries performed on day 7; whereas, animals in Exp. 2 received two frozen-thawed embryos on day 7. As an indicator of ergotamine effects, prolactin was decreased throughout both experiments in ERGOT compared to CON animals (p<0.05). Furthermore, rectal temperature (RT) tended to increase during both experiments in ERGOT compared to CON (p= 0.06). In Exp. 1, embryo recovery (p=0.08) and the percentage of transferable embryos (p=0.09) tended to be greater for CON than for ERGOT. Percentage of embryos that developed to compacted morula or greater was increased for CON compared to ERGOT heifers (p<0.05). In Exp. 2, pregnancy rates of transferred embryos did not differ between treatment groups. Thus, administration of ERGOT to simulate fescue toxicosis altered developmental potential of embryos, but does not appear to affect uterine competency to establish pregnancy.
Assuntos
Bovinos/embriologia , Embrião de Mamíferos/efeitos dos fármacos , Embrião de Mamíferos/fisiologia , Ergotamina/administração & dosagem , Desenvolvimento Fetal/efeitos dos fármacos , Poaceae/microbiologia , Animais , Temperatura Corporal , Dieta , Transferência Embrionária/veterinária , Ergotamina/toxicidade , Feminino , Modelos Biológicos , Poaceae/efeitos adversos , Gravidez , Prolactina/sangue , Útero/efeitos dos fármacos , Útero/fisiologiaRESUMO
Glucans are fungal cell wall glucose polymers that are released into the blood of infected patients. The role of glucans in infection is unknown. We examined serum glucan and cytokine levels in intensive care unit (ICU) patients with infections. There was an inverse correlation (p<0.001) between serum glucan levels and interleukin (IL)-2), IL-4, tumor necrosis factoralpha (TNFalpha) and granulocyte macrophage-colony stimulating factor (GM-CSF) levels in infected ICU patients. The correlation between serum cytokines and serum glucan was only observed at glucan concentrations <40 pg/ml. No change was observed at serum glucan levels of >40 pg/ml. There was no correlation between serum glucan levels and systemic levels of IL-1beta, IL-5, IL-6, IL-8, IL-10 or IFNgamma. Interestingly, blood borne glucans did not suppress systemic cytokine levels in infected ICU patients, instead they were maintained at control levels. We conclude that circulating glucans may prevent cytokine upregulation in response to infection. This may represent an adaptive response to septic injury.
Assuntos
Estado Terminal , Citocinas/sangue , Glucanos/sangue , Infecções/sangue , Hormônio Adrenocorticotrópico/sangue , Adulto , Feminino , Hormônio do Crescimento Humano/sangue , Humanos , Hidrocortisona/sangue , Infecções/fisiopatologia , Unidades de Terapia Intensiva , Masculino , Pessoa de Meia-Idade , Prolactina/sangue , Estudos ProspectivosRESUMO
Phosphoinositide-3-kinase (PI3K)/Akt dependent signaling has been shown to improve outcome in sepsis/septic shock. There is also ample evidence that PI3K/Akt dependent signaling plays a crucial role in maintaining normal cardiac function. We hypothesized that PI3K/Akt signaling may ameliorate septic shock by attenuating sepsis-induced cardiac dysfunction. Cardiac function and survival were evaluated in transgenic mice with cardiac myocyte specific expression of constitutively active PI3K isoform, p110α (caPI3K Tg). caPI3K Tg and wild type (WT) mice were subjected to cecal ligation/puncture (CLP) induced sepsis. Wild type CLP mice showed dramatic cardiac dysfunction at 6 hrs. Septic cardiomyopathy was significantly attenuated in caPI3K CLP mice. The time to 100% mortality was 46 hrs in WT CLP mice. In contrast, 80% of the caPI3K mice survived at 46 hrs after CLP (p<0.01) and 50% survived >30 days (p<0.01). Cardiac caPI3K expression prevented expression of an inflammatory phenotype in CLP sepsis. Organ neutrophil infiltration and lung apoptosis were also effectively inhibited by cardiac PI3k p110α expression. Cardiac high mobility group box-1 (HMGB-1) translocation was also inhibited by caPI3K p110α expression. We conclude that cardiac specific activation of PI3k/Akt dependent signaling can significantly modify the morbidity and mortality associated with sepsis. Our data also indicate that myocardial function/dysfunction plays a prominent role in the pathogenesis of sepsis and that maintenance of cardiac function during sepsis is essential. Finally, these data suggest that modulation of the PI3K/p110α signaling pathway may be beneficial in the prevention and/or management of septic cardiomyopathy and septic shock.
Assuntos
Miocárdio/enzimologia , Fosfatidilinositol 3-Quinase/metabolismo , Sepse/enzimologia , Sepse/terapia , Animais , Apoptose/genética , Apoptose/fisiologia , Western Blotting , Citocinas/sangue , Ecocardiografia , Hemodinâmica/fisiologia , Camundongos , Camundongos Transgênicos , Peroxidase/metabolismo , Fosfatidilinositol 3-Quinase/genética , Sepse/microbiologiaRESUMO
Heterotrimeric G(i) proteins play a role in signalling activated by lipopolysaccharide (LPS), Staphylococcus aureus (SA) and group B streptococci (GBS), leading to production of inflammatory mediators. We hypothesized that genetic deletion of G(i) proteins would alter cytokine and chemokine production induced by LPS, SA and GBS stimulation. LPS-induced, heat-killed SA-induced and heat-killed GBS-induced cytokine and chemokine production in peritoneal macrophages from wild-type (WT), Galpha(i2) (-/-) or Galpha(i1/3) (-/-) mice were investigated. LPS induced production of tumour necrosis factor-alpha (TNF-alpha), interleukin-6 (IL-6), IL-10 and interferon-gamma-inducible protein-10 (IP-10); SA induced TNF-alpha, and IL-1beta production; and GBS induced TNF-alpha, IL-6, IL-1beta, macrophage inflammatory protein-1alpha (MIP-1alpha) and keratinocyte chemoattract (KC) production were all decreased (P < 0.05) in Galpha(i2) (-/-) or Galpha(i1/3) (-/-) mice compared with WT mice. In contrast to the role of G(i) proteins as a positive regulator of mediators, LPS-induced production of MIP-1alpha and granulocyte-macrophage colony-stimulating factor (GM-CSF) were increased in macrophages from Galpha(i1/3) (-/-) mice, and SA-induced MIP-1alpha production was increased in both groups of Galpha(i) protein-depleted mice. LPS-induced production of KC and IL-1beta, SA-induced production of GM-CSF, KC and IP-10, and GBS-induced production of IL-10, GM-CSF and IP-10 were unchanged in macrophages from Galpha(i2) (-/-) or Galpha(i1/3) (-/-) mice compared with WT mice. These data suggest that G(i2) and G(i1/3) proteins are both involved and differentially regulate murine inflammatory cytokine and chemokine production in response to both LPS and Gram-positive microbial stimuli.
Assuntos
Quimiocinas/biossíntese , Lipopolissacarídeos/imunologia , Macrófagos Peritoneais/imunologia , Staphylococcus aureus/imunologia , Streptococcus agalactiae/imunologia , Animais , Células Cultivadas , Citocinas/biossíntese , Subunidades alfa de Proteínas de Ligação ao GTP/imunologia , Genótipo , Camundongos , Camundongos KnockoutRESUMO
Glucans are immunomodulatory carbohydrates found in the cell walls of fungi and certain bacteria. We examined the pharmacokinetics of three water-soluble glucans (glucan phosphate, laminarin, and scleroglucan) after oral administration of 1 mg/kg doses in rats. Maximum plasma concentrations for glucan phosphate occurred at 4 h. In contrast, laminarin and scleroglucan showed two plasma peaks between 0.5 and 12 h. At 24 h, 27 +/- 3% of the glucan phosphate and 20 +/- 7% of the laminarin remained in the serum. Scleroglucan was rapidly absorbed and eliminated. The liver did not significantly contribute to the clearance of plasma glucan. Biological effects were further studied in mice. Following oral administration of 1 mg, glucans were bound and internalized by intestinal epithelial cells and gut-associated lymphoid tissue (GALT) cells. Internalization of glucan by intestinal epithelial cells was not Dectin-dependent. GALT expression of Dectin-1 and toll-like receptor (TLR) 2, but not TLR4, increased following oral administration of glucan. Oral glucan increased systemic levels of interleukin (IL)-12 (151 +/- 15%) in mice. Oral glucan administration also increased survival in mice challenged with Staphylococcus aureus or Candida albicans. These data demonstrate that orally administered water-soluble glucans translocate from the gastrointestinal (GI) tract into the systemic circulation. The glucans are bound by GI epithelial and GALT cells, and they modulate the expression of pattern recognition receptors in the GALT, increase IL-12 expression, and induce protection against infectious challenge.
Assuntos
Glucanos/farmacologia , Imunidade Inata/efeitos dos fármacos , Absorção Intestinal , Administração Oral , Animais , Disponibilidade Biológica , Candidíase/imunologia , Citocinas/biossíntese , Glucanos/administração & dosagem , Glucanos/farmacocinética , Lectinas Tipo C , Masculino , Proteínas de Membrana/análise , Proteínas de Membrana/fisiologia , Camundongos , Camundongos Endogâmicos ICR , Proteínas do Tecido Nervoso/análise , Proteínas do Tecido Nervoso/fisiologia , Nódulos Linfáticos Agregados/metabolismo , Ratos , Ratos Sprague-Dawley , Receptores Imunológicos/análise , Infecções Estafilocócicas/imunologia , Receptor 2 Toll-LikeRESUMO
LPS pretreatment of human pro-monocytic THP-1 cells induces tolerance to secondary LPS stimulation with reduced TNFalpha production. However, secondary stimulation with heat-killed Staphylococcus aureus (HKSa) induces priming as evidenced by augmented TNFalpha production. The pro-inflammatory cytokine, IFNgamma, also abolishes suppression of TNFalpha in LPS tolerance. The effect of LPS tolerance on HKSa and IFNgamma-induced inflammatory mediator production is not well defined. We hypothesized that LPS, HKSa and IFNgamma differentially regulate pro-inflammatory mediators and chemokine production in LPS-induced tolerance. THP-1 cells were pretreated for 24 h with LPS (100 ng/ml) or LPS (100 ng/ml) + IFNgamma (1 microg/ml). Cells were subsequently stimulated with LPS or HKSa (10 microg/ml) for 24 h. The production of the cytokines TNFalpha, IL-6, IL-1beta, and GMCSF and the chemokine IL-8 were measured in supernatants. LPS and HKSa stimulated TNFalpha (3070 +/- 711 pg/ml and 217 +/- 9 pg/ml, respectively) and IL-6 (237 +/- 8.9 pg/ml and 56.2 +/- 2.9 pg/ml, p < 0.05, n = 3, respectively) in control cells compared to basal levels (< 25 pg/ml). LPS induced tolerance to secondary LPS stimulation as evidenced by a 90% (p < 0.05, n = 3) reduction in TNFalpha. However, LPS pretreatment induced priming to HKSa as demonstrated by increased TNFalpha (2.7 fold, from 217 to 580 pg/ml, p < 0.05, n = 3 ). In contrast to suppressed TNFalpha, IL-6 production was augmented to secondary LPS stimulation (9 fold, from 237 to 2076 pg/ml, p < 0.01, n = 3) and also primed to HKSa stimulation (62 fold, from 56 to 3470 pg/ml, p < 0.01, n = 3). LPS induced IL-8 production and to a lesser extent IL-1beta and GMCSF. LPS pretreatment did not affect secondary LPS stimulated IL-8 or IL-1beta, although HKSa stimulation augmented both mediators. In addition, IFNgamma pretreatment reversed LPS tolerance as evidenced by increased TNFalpha levels while IL-6, IL-1beta, and GMCSF levels were further augmented. However, IL-8 production was not affected by IFNgamma. These data support our hypothesis of differential regulation of cytokines and chemokines in gram-negative- and gram-positive-induced inflammatory events. Such changes may have implications in the pathogenesis of polymicrobial sepsis.
Assuntos
Quimiocinas/metabolismo , Tolerância Imunológica/fisiologia , Lipopolissacarídeos/imunologia , Fator de Necrose Tumoral alfa/metabolismo , Quimiocinas/imunologia , Fator Estimulador de Colônias de Granulócitos e Macrófagos/imunologia , Fator Estimulador de Colônias de Granulócitos e Macrófagos/metabolismo , Humanos , Tolerância Imunológica/imunologia , Interleucina-1/imunologia , Interleucina-1/metabolismo , Interleucina-6/imunologia , Interleucina-6/metabolismo , Interleucina-8/imunologia , Interleucina-8/metabolismo , Lipopolissacarídeos/metabolismo , Staphylococcus aureus/imunologia , Fator de Necrose Tumoral alfa/imunologiaRESUMO
OBJECTIVES: Hormones and cytokines are known to act as regulatory messengers between the neuroendocrine and immune systems. The innate immune system identifies infectious agents by means of pattern-recognition receptors. These receptors recognize pathogen-specific macromolecules called pathogen-associated molecular patterns. Fungal cell wall glucans nonspecifically stimulate various aspects of innate immunity via interaction with membrane receptors on immune-competent cells. Glucans are also released into the systemic circulation of patients with fungal infections. Recent evidence confirms the existence of glucan-specific receptors on cells outside the immune system. We hypothesized that glucans may directly interact with pituitary cells as an early signaling event in fungal infections. METHODS: We characterized the receptor-mediated interaction of glucan derived from Candida albicans with pituitary cells using surface plasmon resonance. Prolactin levels were assayed by commercial ELISA. TLR2, TLR4 and CD14 mRNA levels were assessed by RT-PCR. RESULTS: A single glucan-specific binding site was identified on rodent somatomammotroph (K(D) = 3.9 microM) and human folliculostellate cell (K(D) = 3.6 microM) membranes. Coincubation of glucan with somatomammotroph cells for 72 h significantly (p < 0.01) increased prolactin accumulation by 56-62% over that observed in cells treated with media alone. Glucan also increased TLR4 and CD14 gene expression in human folliculostellate cells. CONCLUSIONS: Pituitary cells directly recognize and respond to fungal cell wall glucans resulting in stimulation of pituitary cell TLR4 and CD14 gene expression. In addition, glucan stimulates secretion of prolactin, a hormone that plays an important role in the response to infection.