OBJECTIVE

The pleiotropic cytokine interleukin (IL)-6 seems to play a pivotal role in sepsis, but contradictory findings in animal models impede a rationale for therapies directed against IL-6. IL-6 signals by two mechanisms via the ubiquitous transmembrane glycoprotein 130 (gp130): "classic" signaling using membrane-bound IL-6 receptor (IL-6R) and trans-signaling using soluble IL-6R (sIL-6R). Trans-signaling is selectively inhibited by soluble gp130 (sgp130). The aim of this study was to systematically compare complete blockade of IL-6 signaling (using a neutralizing anti-IL-6 antibody) and selective blockade of IL-6 trans-signaling (using a fusion protein of sgp130 and the crystallizable fragment of immunoglobulin G1, sgp130Fc) in a standardized cecal ligation and puncture (CLP) sepsis model.

METHODS

Animal study.

METHODS

Animal laboratory.

METHODS

C57BL/6J mice.

METHODS

We performed a 96-hr dose-response study and a 24-hr study to investigate short-term mechanisms. In the 96-hr study, CLP was performed in 120 randomized mice (20 mice received vehicle, 10 mice per dose group). Mice were treated with equimolar doses of sgp130Fc (0.01/0.1/1/10 mg/kg) or anti-IL-6 (0.008/0.08/0.8/8 mg/kg) 24 hrs before CLP. Two additional groups received 0.5 mg/kg sgp130Fc 24 hrs before or 1 mg/kg sgp130Fc 24 hrs after CLP. Survival and activity scores were obtained daily until 96 hrs after CLP. In the 24-hr study, mice were randomized into four groups with 10 animals each (sham/vehicle, CLP/vehicle, CLP/anti-IL-6 [0.8 mg/kg], and CLP/sgp130Fc [1 mg/kg]) and killed after 24 hrs.

RESULTS

In contrast to anti-IL-6, pretreatment with sgp130Fc significantly and dose-dependently increased survival from 45% to 100%. In addition, 1 mg/kg sgp130Fc administered 24 hrs after CLP increased survival from 45% to 80%. Mechanistically, sgp130Fc efficacy was reflected by complete prevention of epithelial cell apoptosis in the jejunum after CLP, which was not achieved with anti-IL-6.

CONCLUSIONS

Selective inhibition of IL-6 trans-signaling by sgp130Fc has considerable potential for the treatment of sepsis and related disorders.

BACKGROUND

Reduction in cardiovascular events with statins has been in part attributed to their anti-inflammatory properties.

OBJECTIVE

Evaluate the effects of atorvastatin on levels of inflammatory markers, such as tumor necrosis factor-alpha (TNF), interleukins (IL-1 and IL-6), soluble intercellular adhesion molecule-1 (sICAM-1) and C-reactive protein (CRP) in hypercholesterolemic patients (LDL-cholesterol >160 mg/dL).

RESULTS

Two lipid-lowering regimens were taken for 8 weeks. One set of patients (n=45, 26 men, average 50 +/- 2 years of age) was subjected to atorvastatin treatment (20-40 mg/day), plus diet recommendation. Another set of patients (n=23, 12 men, average 53 +/- 3 years of age) went through diet recommendation alone. Both groups were recommended to perform standard physical activity. Plasma samples were collected after overnight fasting at baseline and after 8 weeks for ELISA. The use of atorvastatin when compared to diet alone, resulted in significant (P <0.0001) reductions for: LDL-cholesterol (39.9% versus 4.4%), TNF (21.4% versus 2.9%), IL-6 (22.1% versus 2.0%), IL-1 (16.4% versus 2.7%) and sICAM-1 (9.6% versus 0.1%), respectively. The percentage of patients with CRP levels >3 mg/dL in the atorvastatin group fell from 25.0 to 6.7% (P <0.0001) while in the diet group the reduction was not significant.

CONCLUSIONS

In hypercholesterolemic patients, atorvastatin, compared to diet alone resulted in significant reductions in levels of proinflammatory cytokines (TNF, IL-1 and IL-6) as well as in sICAM-1 and CRP. Thus, statin-induced inhibition of inflammatory markers may play an important role in the pharmacological and clinical effects of statins seen in cardiovascular diseases.

NF-kappaB is well established as a key component of the inflammatory response. However, the precise mechanisms through which NF-kappaB activation contributes to inflammatory disease states remain poorly defined. To test the role of NF-kappaB in inflammation, we created a knock-in mouse that expresses a constitutively active form of NF-kappaB p65 dimers. These mice are born at normal Mendelian ratios, but display a progressive, systemic hyperinflammatory condition that results in severe runting and, typically, death 8-<em>20</em> d after birth. Examination of homozygous knock-in mice demonstrates significant increases in proinflammatory cytokines and chemokines. Remarkably, crossing this strain with mice lacking TNF receptor 1 (TNFR1) leads to a complete rescue of the hyperinflammatory phenotype. However, upon aging, these rescued mice begin to display chronic keratitis accompanied by increased corneal expression of TNFalpha, <em>IL</em>-1beta, and MMP-9, similar to that seen in human keratoconjunctivitis sicca (KCS) or "dry eyes." Therefore, our results show that, while constitutively active NF-kappaB can trigger systemic inflammation, it does so indirectly, through increased TNF production. However, certain inflammatory disease states, such as keratitis or KCS, a condition that is seen in Sjogren's syndrome, are dependent on NF-kappaB, but are independent of TNFR1 signaling.

The acute-phase proteins, C-reactive protein and serum amyloid A (SAA), are biomarkers of infection and inflammation. However, their precise role in immunity and inflammation remains undefined. We report in this study a novel property of SAA in the differential induction of Th1-type immunomodulatory cytokines <em>IL</em>-12 and <em>IL</em>-23. In peripheral blood monocytes and the THP-1 monocytic cell line, SAA induces the expression of <em>IL</em>-12p40, a subunit shared by <em>IL</em>-12 and <em>IL</em>-23. SAA-stimulated expression of <em>IL</em>-12p40 was rapid (< or = 4 h), sustainable >> or = <em>20</em> h), potent (up to 3380 pg/ml/10(6) cells in 24 h), and insensitive to polymyxin B treatment. The SAA-stimulated <em>IL</em>-12p40 secretion required de novo protein synthesis and was accompanied by activation of the transcription factors NF-kappaB and C/EBP. Expression of <em>IL</em>-12p40 required activation of the p38 MAPK and PI3K. Interestingly, the SAA-induced <em>IL</em>-12p40 production was accompanied by a sustained expression of <em>IL</em>-23p19, but not <em>IL</em>-12p35, resulting in preferential secretion of <em>IL</em>-23, but not <em>IL</em>-12. These results identify SAA as an endogenous ligand that potentially activates the <em>IL</em>-23/<em>IL</em>-17 pathway and present a novel mechanism for regulation of inflammation and immunity by an acute-phase protein.

Interleukin-6 (<em>IL</em>-6) is a multifunctional cytokine mediating inflammatory or immune reactions. Here we investigated the possible role of <em>IL</em>-6 in the intact or lesioned peripheral nervous system using adult <em>IL</em>-6 gene knockout (<em>IL</em>-6(-/-)) mice. Various sensory functions were tested by applying electrophysiological, morphological, biochemical, and behavioral methods. There was a 60% reduction of the compound action potential of the sensory branch of <em>IL</em>-6(-/-) mice as compared with the motor branch in the intact sciatic nerve. Cross sections of L5 DRG of <em>IL</em>-6(-/-) mice showed a shift in the relative size distribution of the neurons. The temperature sensitivity of <em>IL</em>-6(-/-) mice was also significantly reduced. After crush lesion of the sciatic nerve, its functional recovery was delayed in <em>IL</em>-6(-/-) mice as analyzed from a behavioral footprint assay. Measurements of compound action potentials <em>20</em> d after crush lesion showed that there was a very low level of recovery of the sensory but not of the motor branch of <em>IL</em>-6(-/-) mice. Similar results of sensory impairments were obtained with mice showing slow Wallerian degeneration (Wlds) and a delayed lesion-induced recruitment of macrophages. However, in contrast to WldS mice, in <em>IL</em>-6(-/-) mice we observed the characteristic lesion-induced invasion of macrophages and the upregulation of low-affinity neurotrophin receptor p75 (p75LNTR) mRNA levels identical to those of <em>IL</em>-6(+/+) mice. Thus, the mechanisms leading to the common sensory deficiencies were different between <em>IL</em>-6(-/-) and WldS mice. Altogether, the results suggest that interleukin-6 is essential to modulate sensory functions in vivo.

The anorexia associated with acute and chronic inflammatory or infectious conditions is poorly understood. Our objectives were to explore the anorexigenic effects of interleukin-1 (<em>IL</em>-1) in the rat. Recombinant human (rh) <em>IL</em>-1 beta, murine (rm) <em>IL</em>-1 alpha and to a lesser extent rh<em>IL</em>-1 alpha significantly reduced food intake at greater than or equal to 4.0 micrograms/kg i.p. but not at lower doses, in young (<em>20</em>0-250 g) meal-fed rats on chow diets. The anorexic effect appears to be mediated by prostaglandins since pretreatment with ibuprofen completely blocked it, and a fish oil based diet abolished it, in comparison to corn oil or chow diets. Fish oil feeding also decreased basal and <em>IL</em>-1 stimulated prostaglandin E2 production by tissues in vitro (liver, brain, peritoneal macrophages) and in the whole body. Constant intravenous infusions of lower doses of <em>IL</em>-1 also diminished food intake, though intravenous boluses did not (reflecting rapid renal clearance). Chronic daily administration of <em>IL</em>-1 caused persistent inhibition of food intake for 7-17 d in chow and corn oil fed rats, but had no effect in fish oil fed rats. There was an attenuation of the effect (tachyphylaxis) after 7 d in corn oil and chow fed rats, but slowed weight gain and lower final weights were observed after 17-32 d of daily <em>IL</em>-1. Old (18-<em>20</em> mo Fisher 344) rats showed less sensitivity to <em>IL</em>-1 induced anorexia. In conclusion, <em>IL</em>-1 is anorexigenic in the rat, but this is influenced by the structural form of <em>IL</em>-1, the route and chronicity of administration, the source of dietary fat, and the age of the animal. The ability of prior fat intake to influence the anorexic response to <em>IL</em>-1 represents a novel nutrient-nutrient interaction with potential therapeutic implications.

OBJECTIVE

To determine if plasma concentrations of defined cytokines are increased in women with preeclampsia, and to correlate any increases with the elevated concentrations of the vascular cell adhesion molecule (VCAM)-1.

METHODS

Twenty primigravidas with preeclampsia were compared to <em>20</em> healthy primigravidas. Plasma levels of cytokines, tumor necrosis factor-alpha (TNF alpha), interleukin (<em>IL</em>)-6, <em>IL</em>-8, <em>IL</em>-1 beta, <em>IL</em>-1 receptor antagonist (<em>IL</em>-1ra), granulocyte macrophage-colony-stimulating factor (GM-CSF), and VCAM-1, were measured by enzyme-linked immunosorbent assay.

RESULTS

Concentrations of IL-6 and IL-1ra were significantly higher (P < .01) in preeclamptic women (2.56 and 251.85 pg/mL, respectively) compared to normal pregnant patients (2.06 and 142.00 pg/mL, respectively). There were no significant changes in concentrations of TNF alpha, IL-8, GM-CSF, and IL-1 beta in preeclamptic patients (14.09, 50.52, 125.8, and 2.08 pg/mL, respectively) compared to normal patients (11.96 44.46, 121.3, and 2.01 pg/mL, respectively). Serum concentrations of VCAM-1 were increased in women with preeclampsia (preeclamptic group 841.9 +/- 49.7 ng/mL, control group 560.2 +/- 47.9 ng/mL; t = 3.673, P < .001). Interleukin-6 and IL-1ra concentrations correlated with VCAM-1 concentrations (IL-6: r = 0.539, z = 2.9, P < .005; IL-1ra: r = 0.451, z = 2.428, P < .02).

CONCLUSIONS

Increased cytokine concentrations may contribute to the endothelial damage that occurs with preeclampsia and may explain the mechanism underlying leukocyte activation in this disorder. The increased cytokine concentration may also be responsible for the endothelial adhesion that accompanies preeclampsia.

OBJECTIVE

To investigate the effects of probiotic on intestinal mucosae of patients with ulcerative colitis (UC), and to evaluate the role of probiotic in preventing the relapse of UC.

METHODS

Thirty patients received treatment with sulphasalazine (SASP) and glucocorticoid and then were randomly administered bifid triple viable capsule (BIFICO) (1.26 g/d), or an identical placebo (starch) for 8 wk. Fecal samples were collected for stool culture 2 wk before and after the randomized treatments. The patients were evaluated clinically, endoscopically and histologically after 2 mo of treatment or in case of relapse of UC. p65 and IkappaB expressions were determined by Western blot analysis. DNA-binding activity of NF-kappaB in colonic nuclear extracts was detected by electrophoretic mobility shift assay (EMSA). mRNA expressions of cytokines were identified by semi-quantitative assay, reverse transcriptase- polymerase chain reaction (RT-PCR).

RESULTS

Three patients (<em>20</em>%) in the BIFICO group had relapses during 2-mo follow-up period, compared with 14 (93.3%) in placebo group (P<0.01). The concentration of fecal lactobacilli, bifidobacteria was significantly increased in BIFICO-treated group only (P<0.01). The expressions of NF-kappaB p65 and DNA binding activity of NF-kappaB were significantly attenuated in the treatment group than that in control (P<0.05). The mRNA expression of anti-inflammatory cytokines was elevated in comparison with the control group.

CONCLUSIONS

The probiotic could impede the activation of NF-kappaB, decrease the expressions of TNF-alpha and IL-1beta and elevate the expression of IL-10. These results suggest that oral administration of this new probiotic preparation is effective in preventing flare-ups of chronic UC. It may become a prophylactic drug to decrease the relapse of UC.

Mesenchymal stromal cells (MSCs) are potent regulators of immune responses largely through paracrine signaling. MSC secreted extracellular vesicles (MSC-EVs) are increasingly recognized as the key paracrine factors responsible for the biological and therapeutic function of MSCs. We report the first comprehensive study demonstrating the immunomodulatory effect of MSC-EVs on dendritic cell (DC) maturation and function. MSC-EVs were isolated from MSC conditioned media using differential ultracentrifugation. Human monocyte-derived DCs were generated in the absence or presence of MSC-EVs (<em>20</em> ug/ml) then subjected to phenotypic and functional analysis in vitro. MSC-EV treatment impaired antigen uptake by immature DCs and halted DC maturation resulting in reduced expression of the maturation and activation markers CD83, CD38, and CD80, decreased secretion of pro-inflammatory cytokines <em>IL</em>-6 and <em>IL</em>-12p70 and increased production of anti-inflammatory cytokine TGF-β. MSC-EV treated DCs also demonstrated a diminished CCR 7 expression after LPS stimulation, coupled with a significantly reduced ability to migrate toward the CCR7-ligand CCL21, although they were still able to stimulate allogeneic T cell proliferation in vitro. Through microRNA profiling we have identified 49 microRNAs, which were significantly enriched in MSC-EVs compared to their parent MSCs. MicroRNAs with known effect on DC maturation and functions, including miR-21-5p, miR-142-3p, miR-223-3p, and miR-126-3p, were detected within the top 10 most enriched miRNAs in MSC-EVs, with MiR-21-5p as the third highest expressed miRNA in MSC-EVs. In silico analysis revealed that miR-21-5p targets the CCR7 gene for degradation. To verify these observations, DCs were transfected with miR-21-5p mimics and analyzed for their ability to migrate toward the CCR7-ligand CCL21 in vitro. MiR-21-5p mimic transfected DCs showed a clear trend of reduced CCR7 expression and a significantly decreased migratory ability toward the CCL21. Our findings suggest that MSC-EVs are able to recapitulate MSC mediated DC modulation and MSC-EV enclosed microRNAs may represent a novel mechanism through which MSCs modulate DC functions. As MSCs are currently used in clinical trials to treat numerous diseases associated with immune dysregulation, such as graft-versus-host disease and inflammatory bowel disease, our data provide novel evidence to inform potential future application of MSC-EVs as a cell-free therapeutic agent.

In vivo blockade of the human <em>IL</em>-2R by mAb has been used for immunosuppression in transplantation, therapy for leukemia, and autoimmune diseases. In this study, we report that administration of a humanized <em>IL</em>-2R blocking Ab induced a 4- to <em>20</em>-fold expansion of CD56(bright) regulatory NK cells in uveitis patients over time. The induced CD56(bright) regulatory NK cells from patients exhibited similar phenotype as those naturally occurring CD56(bright) cells. Patients with active uveitis had a significantly lower level of CD56(bright) NK cells compared with normal donors (p < 0.01). In addition, the induced CD56(bright) cells could secrete large amounts of <em>IL</em>-10 whereas CD56(dim) NK cells could not, suggesting that the induction of the CD56(bright) cells may have a beneficial effect on the remission of active uveitis. Our observation may have implications to <em>IL</em>-2R blockade therapy and for the potential role of CD56(bright) regulatory NK cells in autoimmune diseases.

We have examined the ability of different neurotrophic and growth factors to prevent axotomy-induced motoneuron cell death in the developing mouse spinal cord. After postnatal unilateral section of the mouse sciatic nerve, most motoneuron (MN) loss occurs in the lateral motor column of the fourth lumbar segment (L4). Significant axotomy-induced cell death occurred after surgery performed on or before postnatal day (PN) 5. In contrast, no significant cell loss was found when axotomy was performed after PN10. Axotomy on PN2 or PN5 resulted in a 44% loss of L4 motoneurons by 7 days, and a 66% loss of motoneurons by 10 days postsurgery. Implantation of gelfoam presoaked in various neurotrophic factors at the lesion site rescued axotomized motoneurons. Nerve growth factor (NGF), neurotrophin-4/5 (NT-4/5) and ciliary neurotrophic factor (CNTF) rescued <em>20</em>%-30% of motoneurons, whereas brain-derived neurotrophic factor (BDNF), neurotrophin-3 (NT-3), and insulin-like growth factor 1 (IGF-1) rescued virtually all motoneurons from axotomy-induced death. By contrast, platelet-derived growth factor (PDGF)-AA, PDGF-AB, basic fibroblast growth factor (bFGF), and interleukin (<em>IL</em>-6) were ineffective on motoneuron survival following axotomy. NGF, BDNF, NT-3, IGF-1, and CNTF also prevented axotomy-induced atrophy of surviving motoneurons. These data show that mouse lumbar motoneurons continue to be vulnerable to axotomy up to about 1 week after birth and that a number of trophic agents, including the neurotrophins, CNTF, and IGF-1, can prevent the death of these neurons following axotomy.(ABSTRACT TRUNCATED AT 250 WORDS)

Circulating soluble urokinase plasminogen activator receptor (suPAR) reflects the immune and pro-inflammatory status of the HIV-infected patient. Highly active antiretroviral therapy (HAART) suppresses suPAR. Independent of the immune response to HAART, suPAR remains elevated in some HIV-infected patients, reflecting possibly a low-grade pro-inflammatory state. Low-grade inflammation has been implicated in insulin resistance and other features of dysmetabolism. Accordingly it is hypothesized that circulating suPAR is associated with the metabolic status of HIV-infected patients on HAART. Fasting plasma suPAR was determined in 36 normoglycaemic HIV-infected patients on HAART (n = 18 lipodystrophic, and n = 18 non-lipodystrophic) who had estimated insulin sensitivity (Rd) and non-oxidative glucose disposal (NOGM) by euglycaemic hyperinsulinaemic clamps, indirect calorimetry, and glucose tracer infusion. Five patients had circadian suPAR concentrations measured (24 hr, <em>20</em> min-intervals). suPAR and non-HDL-cholesterol were higher and Rd, NOGM, and limb fat were lower in lipodystrophic patients than in non-lipodystrophic patients (P < 0.05). suPAR correlated positively with non-HDL-cholesterol and inversely with Rd, NOGM and limb fat (P < 0.005, n = 36). suPAR also correlated positively with leukocyte count and TNF-alpha (P < 0.01, n = 36) but not with <em>IL</em>-6. In multiple regression analyses suPAR was a stronger predictor of dysmetabolism than TNF-alpha and <em>IL</em>-6. Circadian suPAR did not systematically fluctuate. In conclusion, suPAR may reflect the metabolic status of the HIV-infected patient on HAART, thus linking low-grade inflammation, immune constitution, lipid and glucose metabolism, and fat redistribution. Circadian suPAR concentration appeared stable, suggesting that sampling schedule does not affect measurement. Further studies addressing whether suPAR predicts lipodystrophy and dysmetabolism in HIV-infected patients are warranted.

Using the virus vector derived from a baculovirus of Bombyx mori (Bm), we constructed an infectious recombinant virus carrying the mouse interleukin-3 (<em>IL</em>-3) cDNA placed downstream from the polyhedrin promoter. Silkworms infected in vivo with recombinant virus or the silkworm-derived BmN cell line infected in vitro secreted large amounts of <em>IL</em>-3 into hemolymph or culture medium, respectively. On a per volume basis, about <em>20</em>-fold more activity was found in the culture supernatants of the infected BmN cells and 10000-fold more activity was detected in the hemolymph as compared to supernatants obtained from COS7 monkey cells transfected with plasmid pcD-<em>IL</em>3 using the SV40 early promoter [Yokota et al., Proc. Natl. Acad. Sci. USA 81 (1984) 1070-1074]. Three distinct species of <em>Il</em>-3 of molecular masses, 18, <em>20</em> and 22 kDa were produced and all were converted to a 15-kDa protein by N-glycanase digestion, indicating that silkworm cells glycosylated <em>IL</em>-3. The N-terminal amino acid sequences of the <em>IL</em>-3 purified from tissue culture medium and hemolymph were identical to that of mammalian-derived <em>IL</em>-3, showing that silkworm cells recognized the mammalian signal sequence and cleaved it at the correct position. The purified silkworm-produced <em>IL</em>-3 had biological activities indistinguishable from <em>IL</em>-3 produced by mammalian cells as assessed by mast-cell proliferation assays, colony-formation assays using mouse bone marrow cells, and by receptor-binding assays using [125I]<em>IL</em>-3.

We have studied the effects of human r<em>IL</em>-12 on the proliferation and generation of cytotoxic activity in human CTL precursors. Purified human blood CD8+ T lymphocytes were stimulated overnight with immobilized alpha-CD3 and cultured 3 to 4 additional days under various conditions. The addition of <em>IL</em>-12 resulted in a marked (10- to <em>20</em>-fold), dose-dependent, augmentation of cytotoxicity per cell with a smaller (2-fold) increase in cell number. <em>IL</em>-12 augmentation of proliferation and cytotoxicity of CD8+ T cells was not inhibited by a mAb to the p55 subunit of the <em>IL</em>-2 receptor (alpha-Tac) at a concentration sufficient to block the activity of exogenously added <em>IL</em>-2, indicating that the activity of <em>IL</em>-12 did not require <em>IL</em>-2. Addition of <em>IL</em>-12 at the time of alpha-CD3 activation or 1 day later was highly effective at augmenting cytotoxicity, whereas delayed addition of <em>IL</em>-12 (day 2 or 3) resulted in a smaller increase in CTL activity with no increase in cell number. <em>IL</em>-12 at all doses tested synergized with low dose <em>IL</em>-2 in inducing the proliferation and differentiation of CD8+ T cells. The synergistic effect was not blocked by adding neutralizing serum to IFN-gamma. In contrast to this synergistic effect, <em>IL</em>-12 significantly inhibited the proliferation observed in the presence of higher concentrations of <em>IL</em>-2 (4,500 and 13,500 pg/ml). An inhibitory effect of <em>IL</em>-12 was also observed when <em>IL</em>-12 was added to CD8+ T lymphocytes 3 days subsequent to activation with alpha-CD3 and <em>IL</em>-2. This broad set of potent effects of <em>IL</em>-12 on CD8+ T cell responses suggests that <em>IL</em>-12 may play an important immunoregulatory role on CTL development in vivo and may be a useful tool for manipulating this process in vivo for investigational and immunotherapeutic purposes.

OBJECTIVE

To investigate the relation between neighbourhood socioeconomic and ethnic characteristics with depressive symptoms in a population based sample.

METHODS

Cross sectional data from the CARDIA study, including the Center for Epidemiological Studies depression scale score (CES-D). Neighbourhoods were 1990 US census blocks of 1000 people; six census variables reflecting wealth/income, education, and occupation investigated separately and as a summary score; neighbourhood racial composition (percentage white and black) and individual level income and education were also examined.

METHODS

Participants recruited in 1985/86 from community lists in Birmingham, AL; Chicago, IL; Minneapolis MN; from a health plan in Oakland, CA.

METHODS

3437 adults aged 28-40 years in 1995/96: 24% white men, 27% white women, 20% black men, 29% black women.

RESULTS

For each race-sex group, CES-D was inversely related to neighbourhood score and individual income and education. Associations of neighbourhood score with CES-D became weak and inconsistent after adjusting for individual level factors; personal income remained strongly and inversely associated with CES-D. Age adjusted mean differences (standard errors) in CES-D between the lowest and highest income categories were 3.41 (0.62) for white men, 4.57 (0.64) for white women, 5.80 (0.87) for black men, and 5.74 (0.83) for black women. For both black and white participants, CES-D was associated negatively with percentage of white people and positively with percentage of black people in their census block, before, but not after, adjustment for individual and neighbourhood socioeconomic variables.

CONCLUSIONS

Neither neighbourhood socioeconomic characteristics nor ethnic density were consistently related to depressive symptoms once individual socioeconomic characteristics were taken into account.

OBJECTIVE

To compare the effects of an intravenous infusion of propofol and the alpha-2 adrenoceptor, dexmedetomidine, on inflammatory responses and intraabdominal pressure (IAP) in severe sepsis after abdominal surgery, specifically, serum cytokine levels (interleukin [IL]-1, IL-6, and tumor necrosis factor [TNF]-alpha) and IAP.

METHODS

Prospective, single-center study.

METHODS

University hospital.

METHODS

40 adult ICU patients who had undergone ileus surgery and who were expected to require postoperative sedation and ventilation.

METHODS

Patients received either a loading dose infusion of propofol (Group P; n = 20) one mg/kg over 15 minutes followed by a maintenance dose of one to three mg/kg/hr (n = 20, Group P) or a loading dose of dexmedetomidine of one microg/kg over 10 minutes followed by a maintenance dose of 0.2-2.5 microg/kg/h (n = 20, Group D) at the 24th hour.

METHODS

Biochemical and hemodynamic parameters, cytokine levels, and IAP were recorded before the start of the study and at the 24th and 48th hours.

RESULTS

TNF-alpha levels were significantly lower at the 24th hour (14.66 +/- 4.40 pg/mL vs. 21.21 +/- 11.37 pg/mL, respectively) and at the 48th hour (21.25 +/- 15.85 pg/mL vs. 46.55 +/- 35.99 pg/mL, respectively) in Group D. IL-1 levels were significantly lower at the 24th hour (5.03 +/- 0.15 pg/mL vs. 6.23 +/- 2.09 pg/mL, respectively) and the 48th hour (5.01 +/- 0.37 pg/mL vs. 6.42 +/- 2.76 pg/mL, respectively) in Group D. IL-6 levels were significantly lower at the 24th hour (253.1 +/- 303.6 pg/mL and 511.3 +/- 374.8 pg/mL, respectively) and at the 48th hour (343.5 +/- 393.4 pg/mL and 503.7 +/- 306.4 pg/mL, respectively) in Group D. Intraabdominal pressure also was significantly lower at the 24th hour (12.35 +/- 5.84 mmHg vs. 18.1 +/- 2.84 mmHg, respectively) and the 48th hour (13.9 +/- 6.15 mmHg vs. 18.7 +/- 3.46 mmHg, respectively) in Group D.

CONCLUSIONS

Dexmedetomidine infusion decreases TNF-a, IL-1, and IL-6 levels and IAP more than a propofol infusion.

OBJECTIVE

We provide a current review of the management of advanced renal cell carcinoma.

METHODS

A comprehensive literature review of peer reviewed articles which address the current management of metastatic renal cell carcinoma was performed.

RESULTS

Renal cell carcinoma is the seventh leading cause of cancer, accounting for 3% of malignancies in men. The incidence of renal cell carcinoma has increased significantly by 38% from 1974 through 1990 at least in part related to earlier diagnosis with the common use of new radiological techniques. Cytotoxic chemotherapy remains poor as a treatment alternative. Interferon-alpha produces responses in 15 to <em>20</em>% of patients but clinical usefulness as monotherapy has been surpassed by interleukin-2 (<em>IL</em>-2). <em>IL</em>-2 is the first immunotherapy to produce durable remissions resulting in approval by the Food and Drug Administration. Although high dose bolus <em>IL</em>-2 schedules have the longest followup, <em>IL</em>-2 administered on other schedules may have enhanced efficacy. Randomized trials are attempting to delineate the appropriate role for various doses and schedules.

CONCLUSIONS

Advanced renal cell carcinoma, once a disease relegated to the incurable, during the last decade has evolved into a malignancy that may be associated with cure. The first evidence of this potential is the clear and unequivocal demonstration that IL-2 produces durable complete remissions. Building upon this immunotherapeutic approach the future treatment of renal cell carcinoma will incorporate new immunological technology, including gene, dendritic cell, vaccine and antibody therapy.

TH1 cytokine secretion was examined in response to synthetic peptides of the 85A component of the major secreted, fibronectin-binding antigen 85 complex from Mycobacterium tuberculosis in seven different mouse strains infected with live M. bovis BCG. Twenty-eight overlapping <em>20</em>-mer peptides covering the complete mature 295-amino-acid (AA) protein were synthesized. Significant interleukin-2 (<em>IL</em>-2) and gamma interferon (IFN-gamma) secretion could be measured following in vitro stimulation of spleen cells with these peptides. H-2d haplotype mice reacted preferentially against the amino-terminal half of the protein, i.e., against peptide 5 (AA 41 to 60) and especially against peptide 11 (AA 101 to 1<em>20</em>), which contained an I-Ed binding motif. H-2b haplotype mice, on the other hand, reacted against peptides from both amino- and carboxy-terminal halves of the protein, peptide 25 (AA 241 to 260) being the most potent stimulator of <em>IL</em>-2 and IFN-gamma production. (BALB/c x C57BL/6)F1 animals with the H-2d/b haplotype weakly recognized peptides specific for both parental lines. Finally, CBA/J (H-2k) and major histocompatibility complex class II mutant B6.C.bm12 mice, carrying a mutant I-A beta bm12 allele on an H-2b background, reacted only very weakly to the 85A peptides. Reactive T cells isolated from lungs of BCG-infected H-2b haplotype mice recognized the same epitopes as spleen cells, especially peptide 25. These data confirm previous findings regarding the powerful <em>IL</em>-2 and IFN-gamma-inducing properties of antigen 85 during infection with live M. bovis BCG.

The <em>IL</em>-10 cytokine family has nine members, four of which are located in the <em>IL</em>10 cluster on chromosome 1q32. These cytokines are the immune regulatory cytokine <em>IL</em>-10 itself, and the <em>IL</em>-<em>20</em> subfamily members <em>IL</em>-19, <em>IL</em>-<em>20</em>, and <em>IL</em>-24. <em>IL</em>-10 instructs innate and adaptive immune responses and limits pro-inflammatory responses in order to prevent tissue damage. The <em>IL</em>-<em>20</em> subfamily members are involved in host defense mechanisms, particularly from epithelial cells and seem essential for tissue integrity. Dysregulation of <em>IL</em>-10 family cytokines results in inflammation and autoimmune disease. Here, we discuss cellular source, gene regulation, and receptor complexes of cytokines in the <em>IL</em>10 cluster and their contribution to autoimmune disease and tissue damage.

BACKGROUND

Rheumatoid arthritis (RA) is a genetically complex disease where the response to different treatments varies greatly between different patients. This is the case with the tumour necrosis factor (TNF) blocking agents, where <em>20</em>-40% of patients have been described as non-responders. No predictive markers exist as yet for the prognosis of response.

OBJECTIVE

To analyse whether polymorphisms of several cytokine genes are associated with the responsiveness to TNF blockade with etanercept.

METHODS

123 patients with active RA were treated with etanercept and response rates were determined after three months using American College of Rheumatology (ACR)<em>20</em> and disease activity score (DAS)28 response criteria. Genotyping was done for TNF (-308 TNFA), interleukin (IL)10 (-1087 IL10), transforming growth factor (TGF)beta1 (codon 25 TGFB1), and IL1 receptor antagonist (intron 2 IL1RN).

RESULTS

24 patients (<em>20</em>%) were defined as non-responders owing to their failure to fulfil any of the ACR<em>20</em> or DAS28 response criteria. None of the recorded alleles was alone significantly associated with responsiveness to treatment. However, a certain combination of alleles (-308 TNF1/TNF1 and -1087 G/G) was associated with good responsiveness to etanercept (p<0.05). In addition, a combination of alleles influencing interleukin 1 receptor antagonist (IL1Ra) and TGFbeta1 production (A2 allele for IL1RN and rare C allele in codon 25 of TGFB1 gene) was associated with non-responsiveness (p<0.05).

CONCLUSIONS

Genetic polymorphisms, which may influence the balance of pro- and anti-inflammatory cytokines of relevance for the course of RA, are associated with clinical responsiveness to etanercept treatment.

BACKGROUND

T regulatory cells have a key role in the pathogenesis of autoimmune diseases in different animal models. However, less information is available regarding these cells in human autoimmune thyroid diseases (AITD).

OBJECTIVE

The objective of the study was to analyze different regulatory T cell subsets in patients with AITD.

METHODS

We studied by flow cytometry and immunohistochemistry different T regulatory cell subsets in peripheral blood mononuclear cells (PBMCs) and thyroid cell infiltrates from <em>20</em> patients with AITD. In addition, the function of T(REG) lymphocytes was assessed by cell proliferation assays. Finally, TGF-beta mRNA in thyroid tissue and its in vitro synthesis by thyroid mononuclear cells (TMCs) was determined by RNase protection assay and quantitative PCR.

RESULTS

PBMCs from AITD patients showed an increased percent of CD4+ lymphocytes expressing glucocorticoid-induced TNF receptor (GITR), Foxp3, IL-10, TGF-beta, and CD69 as well as CD69+CD25(bright), CD69+TGF-beta, and CD69+IL-10+ cells, compared with controls. TMCs from these patients showed an increased proportion of CD4+GITR+, CD4+CD69+, and CD69+ cells expressing CD25(bright), GITR, and Foxp3, compared with autologous PBMCs. Furthermore, a prominent infiltration of thyroid tissue by CD69+, CD25+, and GITR+ cells, with moderate levels of Foxp3+ lymphocytes, was observed. The suppressive function of peripheral blood T(REG) cells was defective in AITD patients. Finally, increased levels of TGF-beta mRNA were found in thyroid tissue, and thyroid cell infiltrates synthesized in vitro significant levels of TGF-beta upon stimulation through CD69.

CONCLUSIONS

Although T regulatory cells are abundant in inflamed thyroid tissue, they are apparently unable, in most cases, to downmodulate the autoimmune response and the tissue damage seen in AITD.

Peripheral monocytosis may affect the development of heart failure (HF) after acute myocardial infarction (AMI). Activated toll-like receptor (TLR) 4 in monocytes plays an important role in the synthesis of proinflammatory cytokines. We examined TLR4 expression in monocytes, which may be a possible source of proinflammatory cytokines in AMI. Sixty-five patients with AMI and <em>20</em> healthy subjects (HS) were studied. Monocytes were isolated from peripheral blood on days 1 and 14 after the onset of AMI. TLR4 levels in monocytes were measured using real-time RT-PCR and flow cytometry. Generation capacity was evaluated by TLR4 levels and cytokine concentrations in the culture medium with lipopolysaccharide (LPS) stimulation. On day 1 after onset, baseline levels of TLR4 and plasma proinflammatory cytokines, notably <em>IL</em>-6 and TNF-alpha, were higher in AMI patients than in HS. These levels remained elevated in AMI patients 14 days after onset. Generation capacities of TLR4 and proinflammatory cytokines (<em>IL</em>-2, <em>IL</em>-6, <em>IL</em>-8, <em>IL</em>-10, GM-CSF and TNF-alpha) were increased in AMI patients compared to HS. LPS-stimulated TLR4 levels were positively correlated with <em>IL</em>-6 and TNF-alpha levels in AMI patients. Baseline TLR4 levels and plasma proinflammatory cytokine (<em>IL</em>-6, GM-CSF and TNF-alpha) levels were higher in AMI patients with HF (n = 22) than in those without HF. Generation capacities of TLR4 and proinflammatory cytokines (<em>IL</em>-6, GM-CSF and TNF-alpha) were greater in AMI patients with HF than in those without HF. Activation of TLR4 through a myocytic inflammatory reaction is associated with HF after AMI. These observations suggest that TLR4 signaling in monocytes may play a role in the development of HF after AMI.

Adenovirus (AdV)-mediated gene expression of immune stimulators represents a valuable in vivo approach for gene therapy of human cancer. The expression level of the therapeutic gene is of crucial importance for the efficacy of this type of treatment. Entry of AdV is dependent on the primary adenovirus receptor CAR and the secondary AdV receptor identified earlier to be a member of the integrin family of surface molecules. We have analyzed 14 different human melanoma cell cultures from different stages together with one melanoma cell line for their AdV-mediated transduction and expression efficiency. Recombinant viruses at various concentrations were used for expression of the B7-1 costimulatory molecule under the control of different promoters and the expression levels of B7-1 were analyzed by flow cytometry. AdV-mediated <em>IL</em>-12 expression was measured using a commercial ELISA. Levels of transgene expression were compared with the expression levels of HCAR, the alpha(v)beta3 and alpha(v)beta5 integrins, and HLA class I. In 4 of 14 cell cultures tested, the presence of the primary virus receptor CAR was associated with the high transduction efficiency phenotype when using the B7-1- and <em>IL</em>-12-expressing viruses at a relatively low multiplicity of infection (MOI) of 50. Immunohistochemistry on cryosections from the original biopsies yielded a strong signal specific for CAR. In contrast, cell cultures expressing low or undetectable levels of CAR needed a <em>20</em>- to 40-fold higher viral input to show comparable expression level of B7-1 or <em>IL</em>-12. Expression levels of the transgenes hardly varied when using different promoters and no association was observed with the presence or absence of HLA class I molecules or with the expression levels of integrins.

Francisella tularensis is a gram-negative intracellular bacterium and the causative agent of the zoonotic disease tularemia. F. tularensis is a category A select agent and thus a potential agent of bioterrorism. Whereas an F. tularensis live, attenuated vaccine strain (LVS) is the basis of an investigational vaccine, this vaccine is not licensed for human use because of efficacy and safety concerns. In the present study, we immunized mice with isolated native outer membrane proteins (OMPs), ethanol-inactivated LVS (iLVS), or purified LVS lipopolysaccharide (LPS) and assessed the ability of each vaccine preparation to protect mice against pulmonary challenge with the virulent type A F. tularensis strain SchuS4. Antibody isotyping indicated that both Th1 and Th2 antibody responses were generated in mice after immunization with OMPs or iLVS, whereas LPS immunization resulted in only immunoglobulin A production. In survival studies, OMP immunization provided the greatest level of protection (50% survival at <em>20</em> days after infection with SchuS4), and there were associated 3-log reductions in the spleen and liver bacterial burdens (compared to nonvaccinated mice). Cytokine quantitation for the sera of SchuS4-challenged mice indicated that OMP and iLVS immunizations induced high levels of tumor necrosis factor alpha and interleukin-2 (<em>IL</em>-2) production, whereas only OMP immunization induced high levels of <em>IL</em>-10 production. By comparison, high levels of proinflammatory cytokines, including RANTES, granulocyte colony-stimulating factor, <em>IL</em>-6, <em>IL</em>-1alpha, <em>IL</em>-12p40, and KC, in nonvaccinated mice indicated that these cytokines may facilitate disease progression. Taken together, the results of this study demonstrate the potential utility of an OMP subunit (acellular) vaccine for protecting mammals against type A F. tularensis.