Patients with H&N tumours treated with surgery, chemo- and radiotherapy also underwent an immunologic therapy with timopentina to evaluate clinic and immunologic efficacy during a 1-year follow-up. Twenty-five patients were recorded in this study divided at random into two groups. In group A the patients were administered timopentina (50 mg/3 times per week/6 weeks) subcutaneously in 4 o 5 cycles during the year. Group B were not administered timopentina. The immunologic state was assessed through investigation of the following: Evaluation of PBL and their T and B cell subpopulations Phagocytosis and blastigenesis Surface receptor and soluble receptor of IL2 NK activity IL1 production. The immunologic values of the two groups were correlated against a control group of twenty non-neoplastic patients. Our study revealed a better immunologic conditions at the end of follow-up in patients treated with timopentina compared to the other patients.

This study examined the combinatory effect on graft survival of neonatal pig pancreatic cell clusters (NPCC) with nordihydroguaiaretic acid (NDGA), a 5-lipoxygenase inhibitor, with systemic CTLA4Ig expression, with local CTLA4Ig and with interleukin-1 (IL-1) receptor antagonist (IL-1ra) expression using a pig to mouse model. About 2000 NPCCs, which were infected with both adenoviruses carrying CTLA4Ig and IL1-1ra genes (each 500 pfu/NPCC), were transplanted beneath the kidney capsule of diabetic BALB/c mice. Two days before transplantation, the recipient mice were either injected with (group C, n = 4; group D, n = 6) or without (group A, n = 7; group B, n = 9) 1 x 10(13) pfu/kg body weight of adenovirus carrying the CTLA4Ig gene. Mice in groups B and D received daily injections of NDGA (20 mg/kg body weight) subcutaneously for 4 weeks. Blood glucose levels less than 200 mg/dL were defined to be normoglycemic and the transplant termed as a functioning graft for the purpose of calculating mean graft function time (MFT). Four weeks posttransplantation, an intraperitoneal glucose tolerance test (IPGTT) was performed to calculate the area under the curve (AUC). Blood glucose levels in groups C and D were significantly lower than groups A and B at 1, 2, and 3 weeks after transplantation. Graft MFT and AUC of IPGTT in group D were significantly different from those in groups A and B. Our data suggested that a high dosage of systemic expression of CTLA4Ig was effective to enhance xenograft survival and that in it was reinforced by a combination with the macrophage inhibitor NDGA.

As soon as a pathogen invades through the physical barriers of its corresponding host, host mounts a series of protective immune response to get rid of the invading pathogen. Host's pattern recognition receptors (PRR), localized at the cellular surface, cytoplasm and also in the nucleus; recognises pathogen associated molecular patterns (PAMPs) and plays crucial role in directing the immune response to be specific. Inflammatory responses are among the earliest strategies to tackle the pathogen by the host and are tightly regulated by multiple molecular pathways. Inflammasomes are multi-subunit protein complex consisting of a receptor molecule viz. NLRP3, an adaptor molecule- Apoptosis-associated speck-like protein containing a CARD (ASC) and an executioner caspase. Upon infection and/or injury; inflammasome components assemble and oligomerizes leading to the auto cleavage of the pro-caspase-1 to its active form. The activated caspase-1 cleaves immature form of the pro-inflammatory cytokines to their mature form e.g. IL1-β and IL-18 which mount inflammatory response. Moreover, C-terminal end of the Gasdermin D molecule is also cleaved by the caspase-1. The activated N-terminal Gasdermin D molecule form pores in the infected cells leading to their pyroptosis. Hence, inflammasomes drive inflammation during infection and controls the establishment of the pathogen by mounting inflammatory response and activation of the pyroptotic cell death.

The intercellular adhesion molecule 1 (ICAM-1) can be induced on many different cell types by a set of various modulators (IL1 beta, TNF, LPS, IFN-gamma), which are released during the inflammatory process. We have investigated the possibility that other factors, related to the stress and biophysical perturbations of the inflammatory response, may also modulate ICAM-1. Here, we report that heavy metals, in particular zinc, can enhance the expression of the ICAM-1 gene on cells actively involved at different levels during inflammation. Kinetic studies of ICAM-1 gene expression shows a maximum level of induction 4 h after treatment with metals, followed by a rapid decrease to basal levels within 12 h. The effect on enhanced gene expression is mostly due to a rapid increase of the transcriptional rate as shown by nuclear run-on experiments. In B lymphoblastoid cells, but not in fibroblasts, the increase in RNA expression seems significantly greater that the subsequent increase in protein expression, suggesting that a further point of post-transcriptional regulation of ICAM-1 occurs and may be linked to the cellular specificity. may be linked to the cellular specificity.

BACKGROUND

Cellular rejection infiltration of the interstitium is the basic histological finding in biopsies of transplanted kidneys, and leukostasis in the muscular arteries and glomeruli is an important sign of exacerbating rejection. For better understanding and more accurate interpretation the authors used immunohistochemistry.

RESULTS

The authors examined 282 tissue specimens from 208 grafts using the two- or three-step immunoenzyme method with 28 mono- or polyclonal antibodies specific for a series of differentiation and activation leukocytic antigens, adhesion molecules and selected cytokines. In the compact component of the rejection infiltrate CD4+ lymphocytes with expression of CD 45 RA antigen predominated while in the disperse component there were mostly macrophages (CD68, 14, 11b); their number correlated significantly with the parenchymatous damage, similarly as intraarterial and glomerular accumulation. The disperse infiltrate and adherent cells expressed CD45 RO (rarely CD25) and integrin molecules of the series CD11 and CD49 CD57+ lymphocytes penetrated into the tubules but did not accumulate in the blood vessels. As to adhesive molecules of the "Ig superfamily", CD106 (VCAM-1) was more important than CD54 (ICAM-1) and its arterial and mesangial expression correlated with the rejection damage. Evidence of cytokines (IL1, IL2, TNF alpha, beta) did provide neither unequivocal results nor correlations.

CONCLUSIONS

Immunohistochemistry improves considerably the accuracy of bioptic evaluation of rejection nephropathy and some antigens (e.g., CD68, CD14, CD45 RO., CD57, CD106) are suitable for diagnostic practice. With their aid it is easier to evaluate the activity of rejection, assess the probability of vascular lesions in specimens without affected vessels and detect more sensitively intravascular stasis and adhesion of leukocytes.

OBJECTIVE

The association between the interleukin IL1 beta gene polymorphisms SNP-511 and SNP+3953 and susceptibility to the development of Hashimoto's thyroiditis among adult Caucasian-Polish population were analyzed.

METHODS

The group studied comprised of 115 unrelated patients with Hashimoto's thyroiditis (112 women and 3 men, mean age 53.3 years). All patients were euthyroid on thyroid replacement therapy, had extremely high serum anti-TPO levels and in 53 patients anti-TG levels were also increased. The control group consisted of 103 healthy blood donors without raised anti-TPO antibodies, in whom a personal and familial history of thyroid, autoimmune and inflammatory diseases was excluded. No goiter or thyroid dysfunction was found.2 polymorphisms of the IL1 beta were studied by PCR-RFLP analysis. To confirm the accuracy of the method used, randomly selected patients were analyzed by direct sequencing.

RESULTS

In both groups allele frequencies were in Hardy-Weinberg equilibrium. The significant statistical differences between the frequency of C and T allele for both SNPs (C-511T and C+3953T) in the group studied and in the controls were found (p=0.0081; OR=1.846; 95% CI: 1.183-2.878 and p=0.0099; OR=1.953; 95% CI: 1.183-3.224).The frequencies of the genotype C-511C compared to C-511T and T-511T as well as C+3953C compared to C+3953T and T+3953T also differed significantly (p=0.0057; OR=2.248; 95% CI: 1.292-3.912 and p=0.0043; OR=2.338; 95% CI: 1.305-4.191) between patients and controls.

CONCLUSIONS

An association between the SNPs of the IL1 beta and susceptibility to Hashimoto's thyroiditis among the group of Caucasian-Polish population studied was found.

<A<em>b</em>stractText>MicroRNA-155 (miR-155) regulates inflammatory cytokines, however its role in Dia<em>b</em>etic neuropathy (DN) remains unexplored.</A<em>b</em>stractText><A<em>b</em>stractText>A strain of mice (d<em>b</em>/d<em>b</em>) having type II dia<em>b</em>etes were studied for expression of miR-155 in plasma and in sciatic nerves. The miR-155 mimic treated mice were studied for effect on motor and sensory nerve conduction velocities along with <em>b</em>lood perfusion in sciatic nerves and response to thermal stimuli test. The mice were evaluated for density of <em>b</em>lood vessels, quantity of intra-epidermal nerve fi<em>b</em>ers (IENF), diameters of axons & thickness of myelin sheath of sciatic nerves. Bioinformatics analysis was done to confirm target genes of miR-155.</A<em>b</em>stractText><p><div>(<em>b</em>)RESULTS</<em>b</em>)</div>The d<em>b</em>/d<em>b</em> mice showed significant suppression of miR-155 in sciatic nerves. The treatment of miR-155 mimic elevated levels of miR-155 in <em>b</em>oth sciatic nerves and plasma; it also enhanced the <em>b</em>lood flow in sciatic nerves and velocities of conduction for <em>b</em>oth sensory and motor nerves. The treatment showed significant decrease in the threshold to thermal stimuli in d<em>b</em>/d<em>b</em> mice. A significant improvement in density of perfused <em>b</em>lood vessels was o<em>b</em>served, along with elevation of IENF and thickness of myelin and axon diameters of sciatic nerves. The treatment attenuated levels of TNF-α, iNOS, <em>IL1</em>-<em>β</em> and Ym1. Microarray analysis showed that the treatment decreased the expression of proinflammatory genes TRAF2 and Notch2, SORT1 and were identified as target <em>b</em>y <i>in silico</i> studies.</p><A<em>b</em>stractText>Treatment of miR-155 mimic in d<em>b</em>/d<em>b</em> mice attenuated DN, suppressed dia<em>b</em>etic associated proinflammatory genes and confirmed miR-155 mimic as therapeutic strategy for treating DN.</A<em>b</em>stractText>

OBJECTIVE

Therapy with antibiotics, dexamethasone, and supportive intensive care has improved the prognosis of pneumococcal meningitis, but mortality remains high. Here, we investigated an adjunctive combination therapy of the non-bacteriolytic antibiotic daptomycin plus several anti-inflammatory agents to identify the currently most promising adjunctive combination therapy for pneumococcal meningitis.

METHODS

C57BL/6 mice were infected by injection of pneumococci into the cisterna magna. Treatment was begun 21 h after infection, and consisted of ceftriaxone plus (a) dexamethasone, (b) dexamethasone plus daptomycin, (c) daptomycin, (d) daptomycin plus an anti-IL1 antibody, (e) daptomycin plus roscovitine, or (f) daptomycin plus an anti-C5 antibody. Animals were followed until 45 h after infection. Furthermore, adjunctive daptomycin plus anti-C5 antibodies were assessed in a long-term follow-up.

RESULTS

Adjunctive treatment with daptomycin and an anti-C5 antibody was superior to adjunctive dexamethasone and reduced disease symptoms (clinical score 1.1 ± 1.1 versus 5.0 ± 2.7, p < 0.0083), improved explorative activity (open field test 17.8 ± 8.2 versus 7.4 ± 4.3 crossed fields/2 minutes, p < 0.0083), and reduced hearing impairment (thresholds for click stimulus 96.1 ± 14.7 versus 114.8 ± 9.3 dB SPL, p < 0.0083) in the acute stage. Furthermore, explorative activity (14.4 ± 7.3 crossed fields/2 minutes versus 6.3 ± 7.2, p < 0.05) and cognitive function (t-maze test, exploration time previously unknown alley 72.4 ± 14.3 versus 48.7 ± 25.6%, p < 0.05) was improved at 2 weeks after infection. Treatment with daptomycin plus an anti-IL-1β antibody or roscovitine was not of significant benefit in comparison to adjunctive therapy with dexamethasone.

CONCLUSIONS

An adjunctive combination of the non-lytic antibiotic daptomycin plus an anti-C5 antibody was superior to standard therapy with adjunctive dexamethasone in the treatment of pneumococcal meningitis.

BACKGROUND

Monascus-fermented products have featured in Chinese cuisine for thousands of years and are widely used as food colourants and dietary materials in many Asian countries. Rice and dioscorea fermented with Monascus purpureus NTU 568 have health-promoting attributes in vitro and in vivo. The aim of this study was to investigate the immunomodulatory and antioxidant effects of polysaccharides from red mould rice (RMRP) and red mould dioscorea (RMDP) in Raw 264.7 cells.

RESULTS

The results showed the antioxidant capabilities (including scavenging, chelating, inhibition of lipid peroxidation, and reducing power) of RMRP and RMDP at a concentration of 10 mg mL(-1). RMRP and RMDP also stimulated cell proliferation, nitric oxide production, phagocytosis and cytokine production (including IL1-β, IL-6 and TNF-α) in Raw 264.7 cells.

CONCLUSIONS

These findings demonstrate that RMRP and RMDP have antioxidant and immunomodulation potential to be developed as novel dietary supplements.

beta 2-Adrenergic agonists are widely prescribed for the symptomatic relief of asthma, but are not thought to alter the underlying pathogenesis. However, it has been suggested that salmeterol, a new beta-agonist with prolonged bronchodilatory action, may have anti-inflammatory properties. A double-blind crossover study of 4 weeks of inhaled salmeterol versus placebo was performed using a chemiluminescence assay to measure peripheral phagocyte function before and after each treatment period. Circulating cytokines [interleukin-1 beta (IL1 beta), IL4, IL6, IL2 receptor (IL2R)] were also measured. Although salmeterol caused a significant improvement in spirometry, there was no apparent modulation of phagocyte or cytokine activity. No evidence was obtained to support a clinically significant anti-inflammatory action of salmeterol.

The incidence of symptomatic radiation induced lung pneumonitis (RILP), a major dose limiting side effect of thoracic radiotherapy, is in the range of 15-40%. Therapeutic options for the prevention and treatment of RILP are limited. Hence there is a need for developing novel radioprotectors to prevent RILP which can be patient compliant. This study sought to evaluate the efficacy of oral 3,3'-diselenodipropionic acid (DSePA), a novel selenocystine derivative to prevent RILP. C3H/HeJ (pneumonitis responding) mice received a single dose of 18 Gy, whole thorax irradiation and a subset were treated with DSePA orally (2.5 mg/kg), three times per week beginning 2 h post irradiation and continued till 6 months. DSePA delayed onset of grade ≥ 2 RILP by 45 days compared to radiation control (~105 versus ~60 days). It also reversed the severity of pneumonitis in 3/10 radiation treated mice leading to significant improvement in asymptomatic survival compared to radiation control (~180 versus ~102 days). DSePA significantly (p < 0.05) reduced the radiation-mediated infiltration of polymorphonuclear neutrophils (PMN) and elevation in levels of cytokines such as IL1-β, ICAM-1, E-selectin, IL-17 and TGF-β in the bronchoalveolar lavage fluid. Moreover DSePA lowered PMN-induced oxidants, maintained glutathione peroxidase activity and suppressed NF-kB/IL-17/G-CSF/neutrophil axis in the lung of irradiated mice. Additionally, this compound did not protect A549 (lung cancer) derived xenograft tumor from radiation exposure in SCID mice. DSePA offers protection to normal lung against RILP without affecting radiation sensitivity of tumors. It has the potential to be developed as an oral agent for preventing RILP.

We cultured isolated islets from human or porcine origin in the presence or absence of IL1 and TNFα and studied cytoprotective effects of two structurally different PBR ligands. Storage of pig or human islets in the presence of cytokines significantly lowered the fraction of vital beta-cells. Compared with cytokine incubations PK11195 alone or in combination with cytokines was effective to prevent cytokine induced cell death. The data indicate that cold storage in the presence of PK11195 may further protect beta-cells from cytokine induced cell death. This ligand may be helpful to preserve beta-cell survival before transplantation.

Considerable evidence supports the existence of a bidirectional communication between the immune system and the hypothalamo-pituitary-adrenal (HPA) axis. In the present study, we examined the interleukin-1 beta (IL1 beta)-mediated regulation of pro-opiomelanocortin (POMC) at a cellular level, from secretion to gene expression, using murine anterior pituitary corticotroph tumor (AtT20) cells as a model system. The regulatory effects of IL1 beta were compared to those of the classical POMC regulator, corticotropin-releasing hormone (CRH). IL1 beta was found to evoke an early, preferential release of beta-lipotropin (beta LPH) which was accompanied by elevations in POMC heteronuclear (hn)RNA and c-fos and c-jun mRNAs. IL1 beta also elicited a late, preferential release of beta LPH which was associated with only an enhanced expression of POMC hnRNA. Additionally, IL1 beta stimulated an intermediate, preferential release of beta-endorphin (beta E) which was not accompanied by any changes in gene expression. In marked contrast to IL1 beta, CRH evoked an early, preferential beta E secretory response which was associated with elevations in POMC hnRNA and c-fos mRNA. CRH also elicited a late, preferential beta E release which was associated with only an enhanced POMC hnRNA expression. These findings show that although both IL1 beta and CRH activate the corticotrophs, they elicit dramatically different patterns in the regulation of the biochemical dynamics of POMC. Such distinct patterns of corticotroph activation in response to IL1 beta or CRH exposure in vivo would allow the pituitary not only to indicate that it has been activated, but also how it has been activated. This characteristic may be critically important in the function of the HPA axis and in the interaction of the HPA axis with the immune system.

IL4 has been shown to differentially modulate HIV1 replication in primary cells of the monocyte/macrophage lineage. Its effects on chronic HIV1 infection, however, are unknown. To address IL4-mediated effects on promonocytic cells chronically infected with HIV1, U1 cells were incubated in the presence or absence of IL4 together with cytokines that are known to induce both HIV1 and IL8 expression. IL4 enhanced IL1 beta-induced HIV1 and IL8 expression in promonocytic U1 cells, whereas it suppressed their expression induced by cytokines IL6, GM-CSF and to a small extent, TNF alpha. IL4 suppressed IFN gamma-induced IL8 production with increasing IL4 concentration, while HIV1 p24 core antigen production was suppressed at low IL4 input (0.1 and 1 U/ml) but was substantially enhanced at a high IL4 input concentration (10 U/ml). Results showed that the immunosuppressive cytokine IL4 can behave variably in modulating HIV1 and IL8 expression, depending on both the inducing cytokine and the input concentration of IL4.

We have previously demonstrated that continuous infusion of low molecular weight (LMW) heparin delays autoantibody production and development of lupus nephritis in (NZBxNZW)F1 (B/W) mice. In this study we investigated the effect of LMW heparin on renal cytokine and chemokine expression and on nucleosome-mediated activation of nucleosome-specific splenocytes. Total mRNA extracted from kidneys of heparin-treated or -untreated B/W mice was analysed by qPCR for the expression of several cytokines, chemokines, and Toll-like receptors. Splenocytes taken from B/W mice were stimulated with nucleosomes with or without the presence of heparin. Splenocyte cell proliferation as thymidine incorporation and the expression of costimulatory molecules and cell activation markers were measured. Heparin treatment of B/W mice reduced the in vivo expression of CCR2, IL1 β , and TLR7 compared to untreated B/W mice. Nucleosome-induced cell proliferation of splenocytes was not influenced by heparin. The expression of CD80, CD86, CD69, CD25, CTLA-4, and TLR 2, 7, 8, and 9 was upregulated upon stimulation by nucleosomes, irrespective of whether heparin was added to the cell culture or not. In conclusion, treatment with heparin lowers the kidney expression of proinflammatory mediators in B/W mice but does not affect nucleosomal activation of splenocytes.

BACKGROUND

Leishmaniasis causes alterations and lesions in the genital system, which leads to azoospermia and testicular atrophy in animals during the chronic phase of the infection. The aim of this study was to reveal the kinetics of Leishmania chagasi infection in the genital system of male golden hamsters (Mesocricetus auratus).

METHODS

Animals were intraperitoneally inoculated with amastigotes from L. chagasi. At different time points animals were euthanized and genital organs processed for histo-pathological, qPCR, cytokines and testosterone detection assays.

RESULTS

Our results showed a high parasite load in testis, followed by an increase of pro-inflammatory cytokines IL1-β, TNF-α and IFN-γ, and testosterone. Subsequently, IL-4 expression was upregulated and basal parasite persistence in testis was observed using the experimental approach.

CONCLUSIONS

Extracellular amastigotes migrated to the epididymis posing as a potential major factor of parasite persistence and venereal transmission of L. chagasi infection in hamsters.

Previously, we have shown that paraformaldehyde-fixed monocytes are able to fully complement, in terms of [3H]dThd incorporation, a primary stimulus delivered to purified T cells by monoclonal antibodies (mAb) reacting with CD3 or CD2 molecules. Here, we show that depending on the stimulus used (CD3 mAb or different pairs of CD2 mAb) HLA class I molecules from monocytes are directly involved in complementary signals provided to T cells. This was evidenced by the following observations: (a) mAb reacting with the heavy or light chain of class I molecules, or their Fab fragments, completely blocked proliferation of peripheral blood lymphocytes (PBL) activated by CD3 mAb; (b) mAb against the heavy chain of HLA class I but not against beta 2-microglobulin partially blocked (approximately equal to 50%) PBL activation by the CD2 "GT2 + T111" mAb pair but did not block activation by CD2 "D66 + T111" mAb; (c) this pattern of inhibition was observed when anti-class I mAb were used in the soluble phase or when they were bound to monocytes subsequently fixed with paraformaldehyde and cultivated with purified autologous T cells; (d) fixed monocytes are able to restore interleukin (IL) 2 receptor expression on purified T cells stimulated by CD3 mAb or CD2 "GT2 + T111", contrary to anti-HLA class I mAb-pretreated monocytes. The inhibitory effects of anti-HLA class I mAb bound to monocytes were not found to be reversed by recombinant IL2 or recombinant IL1. We assume that HLA class I would be involved in two or more signals delivered to T cells by monocytes, the requirement in those signals depending on the initial stimulus applied to T cells.(ABSTRACT TRUNCATED AT 250 WORDS)

The model of liver cirrhosis was induced by CCl4 and alcohol in rats, which were subjected to splenectomy or given Tuftsin. The isolated and purified liver parenchymal, Kupffer and Ito cells were cultured with CCl4 and splenic conditional fluid or Tuftsin. The RNA isolated from the liver tissues of cirrhosis animals were hybridized with five kinds of cDNA probes. In this study we explored the mechanism of spleen's promoting effects on the liver cirrhosis formation at the whole body, cellular and molecular levels. The result showed that in cirrhosis model, the levels of IL1, IL6 and TNF alpha in serum of rats in imitative splenectomy or Tuftsin group were significantly increased compared to those in splenectomy group (P < 0.05). Cell culture showed that if medium contained CCl4 and splenic conditional fluid or Tuftsin, its cells can secrete more fibronectin, laminin and collagen I than those cultured in medium only contained CCl4 (P < 0.05). Slot blot hybridization showed that the RNA isolated from liver of rats in imitative splenectomy or Tuftsin group hybridized with probes of TNF alpha, IL1 beta, TGF beta and Collagen I had a more high density picture of X-ray than that isolated from liver of rats in the splenectomy group. Should be it a complex process for spleen to promote liver cirrhosis formation, in which the TGF beta gene expression enhancement may be the key of splenic effects on liver fibrosis.

CD8+ T cells are the principal cellular mediators of beta cell destruction in the NOD mouse. Molecular mediators include perforin and granzymes from the cytotoxic granule, Fas ligand and pro-inflammatory cytokines. Our studies in NOD mice have shown that beta cell-specific CD8+ T cells use both the perforin and Fas pathway in vitro. Reducing antigen presentation on beta cells, for example by reducing class I MHC expression by overexpression of SOCS1, protects beta cells in vivo. Perforin deficiency effectively reduces diabetes in NOD mice but in NOD8.3 mice other mechanisms compensate. We have been unable to identify a major role for direct toxicity of cytokines in NOD mice. However, in the LCMV glycoprotein model they may be more important. Deficiency of IL1 or TNF or Fas has a protective effect (greatest for TNF deficiency) but this appears to be due to effects of these cytokines on the immune response rather than on the beta cell. Combinations of interventions, for example, beta cell overexpression of SOCS1 combined with IL1 deficiency may be highly protective. It should be possible to define all the molecular mediators of beta cell destruction, and it may be possible to inhibit at least some of these.

The aim of this study was to analyze the association between single nucleotide polymorphism (SNP) in IL1B (rs1143 634) and IL1RN (rs2234677) with chronic low back pain (LBP) in chronic post-traumatic stress disorder (PTSD). A total of 406 war veterans from 1991-1995 war in Croatia participated in this study. They were divided into four groups, according to psychiatric interview, psychometric testing and the presence of LBP, verified by the imaging of lumbar area, into: (i) war veterans suffering from PTSD and LBP (N = 102), (ii) war veterans suffering from PTSD only (N = 108), (iii) war veterans suffering from LBP only (N = 99) and (iv) healthy controls (N =97). Each subject provided a blood sample for IL1B and IL1RN polymorphism testing. We found no association of rs1143634 in IL1-B with LBP Permutation test showed significant association of rs1143634 in IL1-RN with LBP group and presence of wild type allele A was protective in LBP group. The same SNP (rs1143634) in IL1-B was associated with the intensity of pain. No other associations were observed between these two markers and self-reported measures evaluating PTSD and pain symptoms. These results suggest the potential role of cytokine network in the pathogenesis of chronic PTSD and LBP, although the direct causative pathway remains unclear. The alteration of cytokine network on the level of the brain, spinal medulla and the spine may be responsible for modulation of pain and the occurrence of LBP

This study evaluates the consequences of antiretroviral treatment of the acute simian immunodeficiency virus (SIV) primary infection on virus load and cytokine responses. Four cynomolgus macaques were inoculated intravenously with a pathogenic primary isolate (SIVmac251). Animals were pretreated with 10.8 mg/kg/day of dideoxyinosine (ddI) from 4 days before inoculation, and treatment was continued for 28 days. Proinflammatory (IL6, <em>IL1</em> <em>beta</em> and TNF alpha) and antiinflammatory (<em>IL1</em>0) cytokine and lymphokine (IL2, IL4 and IFN gamma) polymerase chain reaction (PCR) ratios were monitored in unmanipulated peripheral blood mononuclear cells (PBMCs) during acute infection by using a semiquantitative reverse transcription (RT)-PCR method. PBMC-associated virus loads were dramatically reduced compared to those of placebo-treated macaques. Nevertheless, a transient rise in IL6, <em>IL1</em> <em>beta</em>, TNF alpha and <em>IL1</em>0 mRNA expression was observed in PBMCs. IL2, IL4 and IFN gamma mRNAs were either undetectable or weakly detectable throughout the study, with no major changes. Despite a dramatic reduction in the acute viral loads in ddI-treated monkeys, early cytokine mRNA profiles were comparable to those of untreated SIVmac251-infected monkeys. Contrary to what was previously evidenced during primary infection with an attenuated SIV clone, no increase in IL2 and IL4 mRNA was detected in PBMCs of the ddI-treated monkeys, although these monkeys exhibited virus loads similar to those evidenced in macaques infected by attenuated SIV. These data indicate that differential lymphokine expression patterns found in pathogenic and Nef-truncated SIV-infected monkeys may not be strictly dependent on virus load levels.

BACKGROUND

Nasal polyposis is characterised by persistent inflammation of the upper airways. Autophagy has been implicated in many chronic inflammatory diseases. Whether autophagy plays a role in nasal polyp (NP) inflammation is completely unknown and deserves investigation.

METHODS

LC3 and COX-2 expression, the common autophagy and inflammation indicators, respectively, was analysed by immunoblotting in fresh tissues of NP and control nasal mucosa (NM). Primary cultures of NP-derived fibroblasts (NPDFs) and NMDFs were established for in vitro studies. Autophagy was induced by amino acid starvation and LC3 ectopic overexpression or inhibited by 3-methyladenine in the fibroblasts. Inflammation was induced by IL1-β and TNF-α. LC3 and COX-2 expression was confirmed in NP specimens by immunohistochemistry.

RESULTS

LC3 expression was decreased while COX-2 expression was significantly increased in fresh NP tissues compared with the NM control. In NMDFs and NPDFs, autophagy induction by starvation and LC3 overexpression downregulated COX-2 expression. Conversely, autophagy inhibition by 3-methyladenine enhanced COX-2 expression. However, IL1-β and TNF-α had no effect on autophagy. Immunohistochemical studies on the NP specimens showed that most displayed low LC3 expression, whereas COX-2 was highly expressed in >50% of the specimens. Examination of two consecutive NP sections from the same tissue blocks revealed a negative correlation between LC3 and COX-2 expression.

CONCLUSIONS

Autophagy is deficient in NP tissues and COX-2 is negatively regulated by autophagy in NP-derived fibroblasts. Since COX-2 is essential for the production of pro-inflammatory mediators, this study might help interpret persistent mucosal inflammation in NP. Attenuation of inflammation by restoring autophagy might be a therapeutic strategy for treating NP.

Using the subcutaneous chamber model of infection, we showed previously that a mixture of four endodontic pathogens (EP: P. intermedia, F. nucleatum, S. intermedius and P. micra) are able to persist without clearance for up to seven days, while a non-pathogenic oral species, S. mitis, was substantially cleared in this time. Here we have compared the cytokine response inside the chambers against these microorganisms. A majority of cytokines tested (17/24) showed different patterns of expression. Several cytokines had a peak of expression at 2 h after infection in response to the EP, while none showed this pattern in S. mitis infections. Chemokines were uniformly present at similar or higher levels in response to S. mitis, with redundant expression of CXCR2 ligands, while several growth/survival factors were present at higher levels in EP infections. Protease activity expressed by EP may be responsible for the lower levels of some chemokines. T-cell associated cytokines were in general expressed at extremely low levels, and did not differ between the two infections. The inflammatory markers IL-6, IL-1α and IL1-β were expressed at similar levels in both infections at early times, while TNFα was preferentially present in S. mitis infections. In EP infected chambers, reciprocal changes in levels of IL-6 and IL-1α were observed at later times suggesting a switch in the inflammatory response. Analysis of the cytokine response to infection with the individual species from the EP mix suggests that P. intermedia drives this inflammatory switch. Together these results show a surprising level of divergence of the host response to pathogenic and non-pathogenic organisms associated with oral infections, and supports a dominant effect of P. intermedia in polymicrobial endodontic infections.

OBJECTIVE

The aim of the current study was to investigate whether Smad2 overexpression in JE cells induced alveolar bone loss, and to understand the mechanisms regulating the bone loss.

METHODS

A mouse line was created that used a cytokeratin 14 (K14) promoter to overexpress Smad2 in the epithelium of the transgenic mice (K14-Smad2). Micro CT radiographs (μCT) were used to assess bone loss, bone volume, and bone density. The expression of Tnfα, Il1-β, Ifγ, Rankl, and Opg were assessed by RT-PCR. Western blots were used to detect the protein levels of TNF-α and IL1-β. Tartrate-resistant acid phosphatase (TRAP) was used as a marker for osteoclasts. Wild type (WT) mice were used as controls in all steps of the current study.

RESULTS

K14-Smad2 mice had 52.5% (±4.2) root exposed compared to 32.4%(±3.2) in the WT mice. There was a significant difference in alveolar bone volume in the K14-Smad2 mice when compared to WT mice 2.65mm3 (±0.3) and 4.3mm3 (±0.35) respectively. K14-Smad2 mice also had reduced bone density 696.8mg/cc (±70) at 12 months when compared to WT mice 845.9mg/cc(±10). The mRNA levels of Tnfα and Rankl increased by 3.26- and 2.5-fold respectively in the K14-Smad2 mice when compared to controls. The protein level of TNF-α was also significantly increased to 2.8-fold in K14-Smad2 mice when compared to WT mice. Smad2 overexpression increased the total numbers of osteoclasts in K14-Smad2 mice (3.4±0.2)-fold when compared to WT mice.

CONCLUSIONS

Smad2 overexpression induces alveolar bone loss and increases the numbers of osteoclasts. Also, Smad2 overexpression up-regulates TNF-α and RANKL.