Search

Urinary tract infection (UTI) is a disease of extremely high incidence in both community and nosocomial settings. UTIs cause significant morbidity and mortality, with approximately 150 million cases globally per year. Uropathogenic Escherichia coli (UPEC) is the primary cause of UTI and is generally treated empirically. However, the rapidly increasing incidence of UTIs caused by multidrug-resistant UPEC strains has led to limited available treatment options and highlights the urgent need to develop alternative treatment and prevention strategies. In this study, we performed a comprehensive analysis to define the regulation, structure, function, and immunogenicity of recently identified UPEC vaccine candidate C1275 (here referred to as IrmA). We showed that the irmA gene is highly prevalent in UPEC, is cotranscribed with the biofilm-associated antigen 43 gene, and is regulated by the global oxidative stress response OxyR protein. Localization studies identified IrmA in the UPEC culture supernatant. We determined the structure of IrmA and showed that it adopts a unique domain-swapped dimer architecture. The dimeric structure of IrmA displays similarity to those of human cytokine receptors, including the interleukin-2 receptor (IL-2R), interleukin-4 receptor (IL-4R), and interleukin-10 receptor (IL-10R) binding domains, and we showed that purified IrmA can bind to their cognate cytokines. Finally, we showed that plasma from convalescent urosepsis patients contains high IrmA antibody titers, demonstrating the strong immunogenicity of IrmA. Taken together, our results indicate that IrmA may play an important role during UPEC infection., IMPORTANCE Uropathogenic E. coli (UPEC) is the primary cause of urinary tract infection (UTI), a disease of major significance to human health. Globally, the incidence of UPEC-mediated UTI is strongly associated with increasing antibiotic resistance, making this extremely common infection a major public health concern. In this report, we describe the regulatory, structural, functional, and immunogenic properties of a candidate UPEC vaccine antigen, IrmA. We demonstrate that IrmA is a small UPEC protein that forms a unique domain-swapped dimer with structural mimicry to several human cytokine receptors. We also show that IrmA binds to IL-2, IL-4, and IL-10, is strongly immunogenic in urosepsis patients, and is coexpressed with factors associated with biofilm formation. Overall, this work suggests a potential novel contribution for IrmA in UPEC infection.

Zhen-Yi Long,1,* Yi-Feng Zhou,2,* Hao Yuan,1 Ya-Meng Peng,1 Si-Xian Wu,1 Fang Peng1 1Department of Clinical Laboratory, Hunan Provincial People’s Hospital, The First Affiliated Hospital of Hunan Normal University, Changsha, 410005, People’s Republic of China; 2Operating Room, Hunan Provincial People’s Hospital, The First Affiliated Hospital of Hunan Normal University, Changsha, 410005, People’s Republic of China*These authors contributed equally to this workCorrespondence: Hao YuanDepartment of Clinical Laboratory, Hunan Provincial People’s Hospital, The First Affiliated Hospital of Hunan Normal University, Changsha, 410005, People’s Republic of ChinaTel +86-13677366519Fax +86-731-82278012Email yuanhao666@hunnu.edu.cnPurpose: We explored the expression levels of IgG4 and interleukin (IL)-21 in the serum and ankle joints of collagen-induced arthritis (CIA) rats at different disease stages.Materials and Methods: Wistar rats were randomly divided into normal and model groups, and the latter group was administered bovine type II collagen to induce arthritis. Enzyme-linked immunosorbent assay was performed at 21, 28, 35, and 42 days to detect IgG4 and IL-21 in the serum, followed by histological and immunohistochemical analyses of IgG4 and IL-21r in the ankle joint of rats.Results: The contents of IgG4 and IL-21 in the serum of the CIA model group were positively correlated and increased with disease progression. The expression of IgG4 and IL-21 receptors in the ankle joint of the CIA model group was significantly higher than that in the control group. These proteins were closely related to the pathological score. The serum IL-21 level in the model group was closely related to the level of IL-21 receptor in the ankle joint.Conclusion: IL-21 may promote the occurrence and development of rheumatoid arthritis by combining with IL-21r to regulate the content of IgG4.Keywords: rheumatoid arthritis, immunoglobulin, interleukin, interleukin receptor

Objectives: This is a protocol for a Cochrane Review (intervention). The objectives are as follows: To assess the clinical benefits and harms of interleukin‐receptor antagonists and tumor necrosis factor (TNF) inhibitors for the primary and secondary prevention of atherosclerotic cardiovascular diseases. To determine whether interleukin‐receptor antagonists or TNF inhibitors, provided to people both with and without pre‐existing cardiovascular disease, are: useful in preventing adverse cardiovascular outcomes; and safe. post-print 2743 KB

Sterile alpha and toll/interleukin receptor (TIR) motif–containing protein 1 (SARM1) is a neuronally expressed NAD+ glycohydrolase whose activity is increased in response to stress. NAD+ depletion triggers axonal degeneration, which is a characteristic feature of neurological diseases. Notably, loss of SARM1 is protective in murine models of peripheral neuropathy and traumatic brain injury. Herein, we report that citrate induces a phase transition that enhances SARM1 activity by ~2000-fold. This phase transition can be disrupted by mutating a residue involved in multimerization, G601P. This mutation also disrupts puncta formation in cells. We further show that citrate induces axonal degeneration in C. elegans that is dependent on the C. elegans orthologue of SARM1 (TIR–1). Notably, citrate induces the formation of larger puncta indicating that TIR–1/SARM1 multimerization is essential for degeneration in vivo. These findings provide critical insights into SARM1 biology with important implications for the discovery of novel SARM1-targeted therapeutics.

Sterile alpha and toll/interleukin receptor (TIR) motif–containing protein 1 (SARM1) is a neuronally expressed NAD+glycohydrolase whose activity is increased in response to various stressors. The consequent depletion of NAD+triggers axonal degeneration (i.e., Wallerian degeneration), which is a characteristic feature of neurological diseases, including peripheral neuropathies and traumatic brain injury. Notably, SARM1 knockout mice show minimal degeneration in models of peripheral neuropathy and traumatic brain injury, making SARM1 a promising therapeutic target. However, the development of SARM1 inhibitors has been challenging as the purified enzyme is largely inactive. Herein, we report that SARM1 activity is increased ∼2000–fold by a liquid-to-solid phase transition. These findings provide critical insights into SARM1 biochemistry with important implications for the situationin vivo. Moreover, they will facilitate the discovery of novel SARM1–targeted therapeutics.Graphical Abstract

Background Acknowledging the importance of type I interferon (IFN) in Systemic Lupus Erythematosus (SLE), we asked if RNA containing immune complexes (RNA-IC), which trigger the IFN-α synthesis by plasmacytoid dendritic cells (pDCs), also activate type III IFN (IFN-λ1-3) production, and how this is regulated. Methods Peripheral blood mononuclear cells (PBMCs) were isolated from SLE patients and healthy individuals and depleted of monocytes. Immune cells were isolated from healthy PBMCs. Cells were stimulated with RNA-IC. Cytokines were measured by immunoassays, a microarray of pDCs, NK and B cells, as well as single-cell RNA-sequencing of pDCs was performed. Results Type III IFN mRNA was induced in RNA-IC stimulated pDC-NK and pDC-B cell co-cultures, type III IFN was produced in pDC and pDC-NK cell co-culture supernatants. A small subset (3%) of RNA-IC activated pDCs expressed both IFNs type III and type I. Priming with IFN-λ2, IFN-α2b, interleukin (IL)-3, IL-6 and granulocyte-macrophage colony stimulating factor (GM-CSF) significantly enhanced IFN-λ1/3 production by 2–5 fold. In pDC-NK cell co-cultures from SLE patients, IFN-α2b and GM-CSF increased the proportion of RNA-IC responding IFN-λ1/3 producing individuals from 9% to 36%. Hydroxychloroquine as well as an interleukin receptor 1 associated kinase 4 inhibitor (IRAK4i) significantly inhibited the RNA-IC-triggered IFN-λ1/3 production by pDCs and pDC-NK cell co-cultures by >90%. Conclusions Type III IFN production in a small subset of pDCs can be triggered by RNA containing IC, enhanced by NK cells and several pro-inflammatory cytokines, and inhibited by blocking the TLR-MyD88 pathway, resembling the regulation of type I IFN. Thus, our results support a contributing role for both type I and type III IFN in SLE, which needs to be considered when targeting the IFN system in this disease.

Interleukin 33 (IL-33) is among the earliest-released cytokines in response to allergens that orchestrate type 2 immunity. The prolyl cis-trans isomerase PIN1 is known to induce cytokines for eosinophil survival and activation by stabilizing cytokines mRNAs, but the function of PIN1 in upstream signaling pathways in asthma is unknown. Here we show that interleukin receptor associated kinase M (IRAK-M) is a PIN1 target critical for IL-33 signaling in allergic asthma. NMR analysis and docking simulations suggest that PIN1 might regulate IRAK-M conformation and function in IL-33 signaling. Upon IL-33-induced airway inflammation, PIN1 is activated for binding with and isomerization of IRAK-M, resulting in IRAK-M nuclear translocation and induction of selected proinflammatory genes in dendritic cells. Thus, the IL-33-PIN1-IRAK-M is an axis critical for dendritic cell activation, type 2 immunity and IL-33 induced airway inflammation., IL-33 orchestrates type 2 immunity in allergic asthma. Here the authors show, using biochemical, structural and patient data, that upon IL-33 or allergic challenge, the isomerase Pin1 modifies IRAK-M to control the production of pro-inflammatory cytokines in the setting of airway inflammation.

Sterile alpha and toll/interleukin receptor (TIR) motif–containing protein 1 (SARM1) plays a pivotal role in triggering the neurodegenerative processes that underlie peripheral neuropathies, traumatic brain injury, and neurodegenerative diseases. Importantly, SARM1 knock down or knock out prevents degeneration, thereby demonstrating that SARM1 is a promising therapeutic target. Recently, SARM1 was shown to promote neurodegeneration via its ability to hydrolyze NAD(+), forming nicotinamide and ADP ribose (ADPR). Herein, we describe the initial kinetic characterization of full length SARM1, as well as the truncated constructs corresponding to the SAM(1−2)TIR and TIR domains, highlighting the distinct challenges that have complicated efforts to characterize this enzyme. Moreover, we show that bacterially expressed full length SARM1 (k(cat)/K(M) = 6000 ± 2000 M(−1)s(−1)) is at least as active as the TIR domain alone (k(cat)/K(M) = 1500 ± 300 M(−1)s(−1)). Finally, we show that the SARM1 hydrolyzes NAD(+) via an ordered uni–bi reaction in which nicotinamide is released prior to ADPR.

Objective Patients with systemic lupus erythematosus (SLE) have an ongoing interferon (IFN) production due to an activation of plasmacytoid dendritic cells (pDCs), which can be triggered to type I IFN synthesis by RNA containing immune complexes (RNA-IC). Considering emerging data suggesting a role of type III IFN in the SLE disease process, we asked if RNA-IC can induce type III IFN production in pDC and how this production can be regulated. Methods Peripheral blood mononuclear cells (PBMCs) or immune cell subsets were isolated from healthy blood donors or SLE patients and stimulated with IC containing U1 snRNP and SLE-IgG (RNA-IC). Hydroxychloroquine (HCQ) and an interleukin receptor 1-associated kinase 4 inhibitor (IRAK4i) were added to cell cultures. Cytokine mRNA levels were determined with a microarray and protein levels with immunoassays. Single-cell RNA sequencing of pDCs using ddSEQ technology was performed. Results Type III IFN mRNA and protein was induced in RNA-IC-stimulated pDC-NK and pDC-B cell co-cultures. A subset of activated pDCs (3%) expressed both type III and type I IFN mRNA. IFN-λ2, IFN-α2b, interleukin (IL)-3, IL-6, or granulocyte-macrophage colony-stimulating factor (GM-CSF) enhanced IFN-λ1/3 production 2–5-fold. HCQ and an IRAK4i blocked the RNA-IC-triggered IFN-λ1/3 production (p Conclusions Type III IFN production is triggered by RNA-IC in pDCs in a TLR-MyD88-dependent manner, enhanced by NK and B cells as well as several pro-inflammatory cytokines. These results support a contributing role for both type I and type III IFNs in SLE, which needs to be considered when targeting the IFN system in this disease.

Background: The type I Interferons (IFNs) are the most important drivers of the IFN gene signature in Systemic Lupus Erythematosus (SLE). However, both type II and type III IFNs (IFN-λ1-3) can be measured in a proportion of patients with SLE and contribute to the IFN signature. The exact role of type III IFNs in SLE is not completely clear, but serum levels of type III IFN correlate with disease activity and specific organ manifestations, such as arthritis, nephritis and anti-dsDNA antibodies. Type III IFN can be induced in pDCs by TLR9 agonist Oligodinucleotide (ODN) 2216 and many viruses. Whether type III IFN can also be induced in pDCs by nucleic acid containing immune complexes (IC), has, to our knowledge, not been investigated before. Objectives: We asked if RNA containing immune complexes (RNA-IC), which trigger the synthesis of large amounts of IFN-α by plasmacytoid dendritic cells (pDCs), can act as stimuli for type III IFN production, and how this production is regulated by Natural Killer (NK) cells and different cytokines. We also investigated if the type III IFN production could be blocked by hydroxychloroquine (HCQ) and an interleukin receptor 1 associated kinase 4 inhibitor (IRAK4i). Methods: Peripheral blood mononuclear cells (PBMCs) from SLE patients or healthy individuals were used to isolate PDCs and natural killer (NK) cells, or were depleted of monocytes. Cells were stimulated with RNA-IC, and cytokines were measured by immunoassays. mRNA expression in RNA-IC stimulated pDCs and NK cells cells was analyzed with a microarray. The effect of HCQ and IRAK4i on the IFN-λ1/3 production was investigated in pDCs and NK cells from healthy individuals. Results: Type III IFN mRNA expression was strongly upregulated in co-cultures of pDC-NK cells stimulated with RNA-IC. High levels of IFN-λ1/3 and IFN-λ2 (medians 2000 pg/ml and 100 pg/ml) were detected in supernatants from RNA-IC stimulated pDC-NK cell co-cultures. IFN-λ2 enhanced IFN-λ1/3 and IFN-α production by purified pDCs. Interleukin (IL) -3, IL-6, and GM-CSF significantly enhanced IFN-λ1/3 production (4-5 fold) by RNA-IC stimulated pDCs. Monocyte depleted PBMCs and pDC-NK cell co-cultures from 15% and 9% of SLE patients produced IFN-λ1/3 in response to RNA-IC stimulation. Exogenous IFN-α2b and GM-CSF in pDC-NK cell co-cultures increased the proportion of patients responding to RNA-IC stimulation from 9 to 36%. IFN-λ1/3 production by RNA-IC-stimulated pDCs and pDC-NK cells was significantly inhibited by HCQ (by 99% and 93% respectively) and an IRAK4i (by 98% and 96% respectively). Conclusion: pDCs produce both type I and type III IFN in response to RNA containing immune complexes. This is promoted by activated NK cells as well as a number of pro-inflammatory cytokines, including IFN type I and type III, considered important in SLE. Consequently, in order to achieve a proper control the IFN driven autoimmune process in SLE, both type I and type III IFN need to be targeted. In this system of stimulated, co-cultivated pDCs and NK cells, HCQ and an IRAK4 inhibitor blocked the type III IFN production. Disclosure of Interests: None declared

Background:Interleukin receptor associated kinases (IRAK) 1 and 4 are kinases involved in Toll-Like Receptor (TLR) and Interleukin-1 Receptor (IL-1R) signaling pathways, which regulate innate immunity and inflammation. Dysregulation of IRAK1/4 signaling can lead to a variety of inflammatory conditions including rheumatoid and gouty arthritis. As a result, IRAK1/4 are promising therapeutic targets for rheumatic diseases (1). We have identified a potent and selective IRAK1/4 inhibitor, R835, that substantially suppressed the elevation of LPS (TLR4 agonist)-induced serum cytokines in healthy human volunteers in a recently completed phase 1 study.Objectives:The aim of our study was to investigate the effect of IRAK1/4 selective inhibition as a potential therapeutic approach for rheumatological diseases. We evaluated the inhibition by our clinical candidate, R835, on TLR-, IL-1R- and NLRP3 inflammasome-induced cytokine production, as well as in preclinical models of arthritis.Methods:The effect of R835 on TLR- or IL-1R-induced cytokine production was evaluated in vitro using THP-1, human primary endothelial cells and human primary dendritic cells. The activity of R835 on the NLRP3 inflammasome was also tested in vitro using THP-1 cells. The pharmacokinetic-pharmacodynamic relationship of R835 was evaluated in a mouse model of IL-1b-induced cytokine release. Mice were pre-treated orally with vehicle or R835 prior to challenge; serum cytokine and plasma compound levels were determined. The efficacy of IRAK1/4 inhibition by R835 in rodent models of joint inflammation was evaluated in a mouse model monosodium (MSU)-induced peritonitis, in rat model of MSU-induced gouty arthritis and in a rat model of collagen-induced arthritis (CIA).Results:In human cells, R835 blocked proinflammatory cytokine production in response to TLR, IL-1R and NLRP3 inflammasome activation. In mice, R835 dose-dependently decreased serum cytokines in response to administration of IL-1b. Mice pre-treated with R835 demonstrated dose-dependent reductions in MSU crystal-induced serum and peritoneal cytokine levels, as well as neutrophil influx in the peritoneal cavity. Prophylactic and therapeutic treatment with R835 also resulted in significant inhibition of MSU crystal-induced knee edema and pain in a rat model of human gouty arthritis. In the rat model of CIA, R835 blocked both onset and progression of disease, by reducing inflammation, cartilage degeneration and synovial inflammation.Conclusion:R835 is a promising clinical candidate for the treatment of a range of cytokine-driven rheumatological diseases. R835 has proven to have desirable pharmacokinetic properties, was well tolerated and suppressed LPS-induced serum cytokines in healthy volunteers in a recent phase 1 study.References:[1]Bahia M S, Kaur M, Silakari P, Silakari O. Interleukin-1 receptor associated kinase inhibitors: Potential therapeutic agents for inflammatory- and immune-related disorders. Cellular Signalling 27 (2015) 1039–1055.Disclosure of Interests:Chrystelle Lamagna Shareholder of: Rigel Pharmaceuticals, Employee of: Rigel Pharmaceuticals, Meagan Chan Shareholder of: Rigel Pharmaceuticals, Employee of: Rigel Pharmaceuticals, Ernest Tai Shareholder of: Rigel Pharmaceuticals, Employee of: Rigel Pharmaceuticals, Stacey Siu Shareholder of: Rigel Pharmaceuticals, Employee of: Rigel Pharmaceuticals, Roy Frances Shareholder of: Rigel Pharmaceuticals, Employee of: Rigel Pharmaceuticals, Sothy Yi Shareholder of: Rigel Pharmaceuticals, Employee of: Rigel Pharmaceuticals, Chi Young Shareholder of: Rigel Pharmaceuticals, Employee of: Rigel Pharmaceuticals, Vadim Markovtsov Shareholder of: Rigel Pharmaceuticals, Employee of: Rigel Pharmaceuticals, Yan Chen Shareholder of: Rigel Pharmaceuticals, Employee of: Rigel Pharmaceuticals, Lu Chou Shareholder of: Rigel Pharmaceuticals, Employee of: Rigel Pharmaceuticals, Gary Park Shareholder of: Rigel Pharmaceuticals, Employee of: Rigel Pharmaceuticals, Esteban Masuda Shareholder of: Rigel Pharmaceuticals, Employee of: Rigel Pharmaceuticals, Vanessa Taylor Shareholder of: Rigel Pharmaceuticals, Employee of: Rigel Pharmaceuticals

Objective To develop a combinatorial panel of salivary cytokines that manifests the presence of non-small cell lung cancer (NSCLC) that will eventually improve prognosis by facilitating the early diagnosis and management of this common cancer. Methods We performed a case-control study comparing salivary cytokine profiles of 35 adult subjects with NSCLC with those of 35 matched, healthy nonsmokers. Multiplex bead array assays were used to quantify 27 cytokines in saliva, serum, and oral mucosal transudate samples. Logistic regression analysis was used to develop an informative cytokine panel. Receiver operating characteristic (ROC) curves were generated to evaluate the discriminant ability of the panel. Results A combinatorial 12-cytokine panel (interleukin receptor antagonist [IL1RN], IL1B, IL6, IL7, IL8, IL10, C-C motif chemokine ligand 11 [CCL11], tumor necrosis factor, C-X-C motif chemokine ligand 10 [CXCL10], C-C motif chemokine ligand 3, C-C motif chemokine ligand 4, and platelet-derived growth factor-BB) distinguished patients with NSCLC from healthy controls. Further, ROC analysis revealed that a cytokine panel comprising IL10 (odds ratio, 1.156) and CXCL10 (odds ratio, 1.000) discriminated NSCLC with a sensitivity of 60.6% and specificity of 80.8% (area under the ROC curve, 0.701). Conclusion A combinatorial panel of select salivary cytokines indicates the presence of NSCLC., Article, JOURNAL OF INTERNATIONAL MEDICAL RESEARCH.46(9):3570-3582(2018)

Nod-like Receptor Protein3 (NLRP3) inflammasome in macrophages infected with Leishmania sp. enhances the secretion of IL-1β. Excess IL-1β production is linked to disease severity in patients with cutaneous leishmaniasis (CL) caused by L. mexicana. Blockade of the NLRP3 inflammasome in cell cultures from skin biopsies of patients with CL caused by L. braziliensis inhibited the release of IL-1β. We hypothesized that common single nucleotide polymorphisms in the IL1B and in its receptor antagonist IL1RN genes may be predictive of CL caused by L. guyanensis. The SNPs –511T/C (rs16944) and +3954C/T (rs1143634) of the IL1B and IL1RN VNTR (rs2234663) were assessed in 881 patients with CL and 837 healthy controls by PCR-RFLP and direct PCR respectively. Plasma cytokines levels were also assayed. The plasma levels of IL-1β were higher in patients compared to control subjects. In contrast, increased plasma levels of IL-1Ra were observed in controls. The rs16944 C/C genotype was more common among the patients (OR = 1.5 [95%CI 1.1–2.0]; P = 0.004) and the C allele suggests susceptibility to CL (OR = 1.2 [95%CI 1.1–1.4]; P = 0.003). The rs16944 C/C genotype shows a tendency to correlate with lower levels of the IL-1Ra cytokine. Low levels of IL-1Ra cytokine and rs16944 C/C genotype seem to confer susceptibility to L. guyanensis-infection in the Amazonas.

Background All known biological functions of the pro-inflammatory cytokine interleukin-1β (IL-1β) are mediated by type 1 interleukin receptor (IL-1R1). IL-1β–IL-1R1 signaling modulates various neuronal functions including spinal pain processing. Although the role of IL-1β in pain processing is generally accepted, there is a discussion in the literature whether IL-1β exerts its effect on spinal pain processing by activating neuronal or glial IL-1R1. To contribute to this debate, here we investigated the expression and cellular distribution of IL-1R1 in the superficial spinal dorsal horn in control animals and also in inflammatory pain. Methods Experiments were performed on rats and wild type as well as IL-1R1-deficient mice. Inflammatory pain was evoked by unilateral intraplantar injection of complete Freund adjuvant (CFA). The nociceptive responsiveness of control and CFA-treated animals were tested daily for withdrawal responses to mechanical and thermal stimuli before and after CFA injection. Changes in the expression of 48 selected genes/mRNAs and in the quantity of IL-1R1 protein during the first 3 days after CFA injection were measured with the TaqMan low-density array method and Western blot analysis, respectively. The cellular localization of IL-1R1 protein was investigated with single and double staining immunocytochemical methods. Results We found a six times and two times increase in IL-1R1 mRNA and protein levels, respectively, in the dorsal horn of CFA-injected animals 3 days after CFA injection, at the time of the summit of mechanical and thermal allodynia. Studying the cellular distribution of IL-1R1, we found an abundant expression of IL-1R1 on the somatodendritic compartment of neurons and an enrichment of the receptor in the postsynaptic membranes of some excitatory synapses. In contrast to the robust neuronal localization, we observed only a moderate expression of IL-1R1 on astrocytes and a negligible one on microglial cells. CFA injection into the hind paw caused a remarkable increase in the expression of IL-1R1 in neurons, but did not alter the glial expression of the receptor. Conclusion The results suggest that IL-1β exerts its effect on spinal pain processing primarily through neuronal IL-1R1, but it can also interact in some extent with IL-1R1 expressed by astrocytes.

Anakinra is an interleukin (IL) receptor antagonist that works by blocking the biological activity of IL-1 by competitively inhibiting binding of IL-1 to the type 1 interleukin receptor. IL-1 production is induced in response to inflammatory stimuli and mediates various physiological mechanisms, including inflammation and immunological reactions. Patients with neonatal onset multisystem inflammatory disease (NOMID) produce excess IL-1 β, a major proinflammatory cytokine that regulates innate immune responses. Anakinra binds competitively and this results in a rapid reduction in disease severity. NOMID, also known as chronic infantile neurologic, cutaneous, articular syndrome, is the most severe clinical phenotype in the spec- trum of cryopyrin-associated periodic syndromes. It is characterized by cutaneous symptoms, arthropathy, and central nervous system involvement. Extensive studies in patients with NOMID have led to advances in characterizing the extent of organ-specific involvement and damage that occurs with chronic overproduction of IL-1β. NOMID is caused predominantly by mutations in the NLRP3/CIAS1 gene that encodes for the protein cryopyrin, leading to activation of the "NLRP3 inflammasome complex". This in turn regulates the maturation and secretion of the inflammatory cytokine, IL-1 β. The clinical value of IL-1β has been demonstrated by the positive response of patients after treatment with anakinra, with rapid improvement in clinical symptoms, markers of inflammation, and a significant decrease in major organ manifestations.

Objective: To investigate the serum expressions of chemokines CCL2 and CCL3 in patients with rheumatoid arthritis (RA) who were treated with recombinant human interleukin 1 (IL-1) receptor antagonist (IL-1Ra). Material and methods: Serum CCL2 and CCL3 were determined using an enzyme-linked immunosorbent assay in 54 active RA patients before and after treatment with IL-1Ra or a placebo, as well as 36 healthy controls. Results: Compared with the healthy controls, all the 54 RA patients exhibited higher serum CCL2 and CCL3 before and after treatment (p < 0.05). However, patients who had a good response to IL-1Ra treatment had significantly lower mean changes in the serum CCL2 and CCL3 levels from baseline to the last injection than IL-1Ra non-responders (p < 0.01). Conclusions: CCL2 and CCL3 may be useful efficacy markers of IL-1Ra treatment.

E-mail: psy@ssu.ac.krReceived May 8, 2012, Accepted May 31, 2012Key Words : TIR-domain containing Adaptor Protein (TIRAP), MyD88-Adaptor Like protein (MAL), Tollsignalling, Toll/Interleukin Receptor (TIR)Toll-like receptors (TLRs) and interleukin-1 receptors (IL-1Rs) have integral roles in host immune and inflammatoryresponse. In TLR signalling, pathogens bind to the receptor,and stimulate the cytoplasmic Toll/interleukin-1 receptor(TIR)-domain to recruit other TIR-domain containing pro-teins. Recruitment of one or more of these TIR-domainproteins, such as MyD88, TIRAP/MAL, TRIF/TICAM-1and TIRP/TRAM/TICAM-2, to the receptors ultimatelyresults in the downstream activation of transcription factors(e.g. NF-κB), which induce genes necessary for the hostdefense.

Background. Interleukin receptor alpha 1 (IL-13RA1) is a component of a heterodimeric IL-13 receptor that is shared with IL-4 and it is ubiquitously expressed in normal tissues and also tumors. In contrast, IL-13RA2 is a monomeric receptor which is overexpressed in various solid tumors, but is virtually absent in normal tissues. Ligands specifically targeting IL-13RA2 but not IL-13RA1, such as Pep-1L and IL-13.E13K.D2.Cys, have been developed and validated [1,2]. Objective. To assess the expression of IL-13RA2 in pancreatic ductal adenocarcinoma and metastatic melanoma specimens and to develop tracers for selective recognition of the tumors. Methods. The expression of IL-13RA2in patient specimens, patient-derived xenografts (PDX) and established cell lines was assessed using Western blot, immunochemistry or immunofluorescence. The ligands were conjugated to a chelator and labeled with Yttrium 86. Ligand specificity was confirmed using IL-13RA2 positive and negative cells and blocking with a “cold” ligand. Biodistribution studies and PET/CT imaging were conducted to assess distribution of the tracer in IL-13RA2 positive tumors models. Results. IL-13RA2 receptor is overexpressed in melanoma and pancreatic cell lines and tumor tissues. Western blot analysis of 56 patients tumor tissue lysates obtained from pancreatic cancer PDX models showed the presence of IL-13RA2 in over 60% of patients. Similarly, tissue microarray analysis of metastatic melanoma samples identified IL-13RA2 receptor expression in 40% of tumor specimens. Uptake study performed in receptor positive and negative cells confirmed the specificity of radiotracer binding. In vivo experiments indicated fast clearance of 86Y- Pep1L ligand with glomerular filtration as a primary mechanism of the tracer removal. Mice bearing IL-13RA2 positive tumors had up taken the tracer with “tumor to muscle” ratio of 5.6 1 h after probe administration. Yttrium 86-labelled IL-13-E13K.D2.Cys was also taken up by IL-13 RA2 positive tumors. Conclusion. IL-13RA2 is an attractive target for the development of comprehensive theranostic strategies for melanoma and pancreatic cancer. References 1. Pandya, H., et al., An interleukin 13 receptor alpha 2-specific peptide homes to human Glioblastoma multiforme xenografts. Neuro Oncol, 2012. 14(1): p. 6-18. 2. Nguyen, V., et al., A novel ligand delivery system to non-invasively visualize and therapeutically exploit the IL13Ralpha2 tumor-restricted biomarker. Neuro Oncol, 2012. 14(10): p. 1239-53. Citation Format: Rafal Zielinski, Izabela Tworowska, Stanislaw Skora, Aleksandra Rusin, Radjendirane Venugopal, Arumugam Jayakumar, Izabela Fokt, Yaan Kang, Jason Fleming, Yue Huang, Denise Herpai, Waldemar Debinski, Waldemar Priebe. Targeting IL-13RA2 in melanoma and pancreatic cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 1927.

When activated by its ligand, the interleukin receptor type I (IL-1RI) transduces signals in cooperation with the IL-1 receptor accessory protein (IL-1RacP). In contrast, IL-1RII functions as a decoy receptor without participating in IL-1 signalling. Brain astrocytes are cellular targets of IL-1 and play a pivotal role in brain responses to inflammation. The regulation of IL-1 receptors on astrocytes by anti-inflammatory cytokines such as IL-4 and IL-10 has not been studied, despite its importance for understanding the way these cells respond to IL-1. Using RT-PCR, we first showed that the expression of IL-1RI and IL-1RII, but not IL-1RacP, mRNAs are up-regulated by IL-1 beta in a time-dependent manner. Using a radioligand binding technique, we then showed that astrocytes display an equivalent number of IL-1RI and IL-1RII. IL-1 beta decreases the number of IL-1RI binding sites, whereas it increases those of IL-1RII. IL-4 and IL-10 both up-regulate IL-1RII IL-1 beta-induced, but only IL-4 does so for IL-1RI. At the protein level, IL-4 and IL-10 dramatically reverse the ability of IL-1 beta to inhibit expression of IL-1RI but neither affects the ability of IL-1 beta to enhance the number of IL-1RII. Collectively, these results establish the existence of receptor cross-talk between pro- and anti-inflammatory cytokines on a critical type of cell that regulates inflammatory events in the brain.

SUMMARYMarsupials are born in a relatively underdeveloped state and develop during a period of intensive maturation in the postnatal period. During this period,the young marsupial lacks a competent immune system, but manages to survive despite the potential of exposure to environmental pathogens. Passive immune transfer via the milk is one well-recognised strategy to compensate the neonate, but there also may be innate immune mechanisms in place. In this study, CD14 and Toll-like receptor 4 (TLR4), integral molecular components of pathogen recognition, were identified and characterised for the first time in a marsupial, the tammar wallaby (Macropus eugenii). Functional motifs of tammar CD14 and the toll/interleukin receptor (TIR) domain of TLR4 were highly conserved. The lipopolysaccharide (LPS) binding residues and the TLR4 interaction site of CD14 were conserved in all marsupials. The TIR signalling domain had 84% identity within marsupials and 77% with eutherians. Stimulation of adult tammar leukocytes resulted in the induction of a biphasic pattern of CD14 and TLR4 expression, and coincided with increased production of the pro-inflammatory cytokine TNF-α. Differential patterns of expression of CD14 and TLR4 were observed in tammar pouch young early in development,suggesting that early maturation of the innate immune system in these animals may have developed as an immune survival strategy to protect the marsupial neonate from exposure to microbial pathogens.

Loss of kidney graft function with tubular atrophy (TA) and interstitial fibrosis (IF) causes most kidney allograft losses. We aimed to identify the molecular pathways involved in IF/TA progression. Kidney biopsies from normal kidneys (n = 24), normal allografts (n = 6), and allografts with IF/TA (n = 17) were analyzed using high-density oligonucleotide microarray. Probe set level tests of hypotheses tests were conducted to identify genes with a significant trend in gene expression across the three groups using Jonckheere-Terpstra test for trend. Interaction networks and functional analysis were used. An unsupervised hierarchical clustering analysis showed that all the IF/TA samples were associated with high correlation. Gene ontology classified the differentially expressed genes as related to immune response, inflammation, and matrix deposition. Chemokines (CX), CX receptor (for example, CCL5 and CXCR4), interleukin, and interleukin receptor (for example, IL-8 and IL10RA) genes were overexpressed in IF/TA samples compared with normal allografts and normal kidneys. Genes involved in apoptosis (for example, CASP4 and CASP5) were importantly overexpressed in IF/TA. Genes related to angiogenesis (for example, ANGPTL3, ANGPT2, and VEGF) were downregulated in IF/TA. Genes related to matrix production-deposition were upregulated in IF/TA. A distinctive gene expression pattern was observed in IF/TA samples compared with normal allografts and normal kidneys. We were able to establish a trend in gene expression for genes involved in different pathways among the studied groups. The top-scored networks were related to immune response, inflammation, and cell-to-cell interaction, showing the importance of chronic inflammation in progressive graft deterioration.

Objective. To compare multiple serum markers for their ability to detect active disease in patients with GCA and in those with PMR.Methods. Twenty-six markers related to immune cells that may be involved in GCA and PMR were determined by ELISA and multiplex assay in the serum of 24 newly diagnosed, untreated GCA/PMR patients, 14 corticosteroid (CS)-treated GCA/PMR patients in remission and 13 healthy controls. Receiver operating characteristic analysis with area under the curve and Spearman's correlation coefficients were performed.Results. Serum B-cell activating factor (BAFF), CXCL9 and IL-6 were increased in newly diagnosed GCA and PMR patients. Serum CCL2, CCL11, IL-10 and sIL-2R were modulated in GCA patients only and CXCL10 in PMR patients only. BAFF, CXCL9 and IL-6 accurately distinguished newly diagnosed GCA and PMR patients from healthy controls, as shown by area under the curve> 0.80. Upon CS-induced remission, serum BAFF and IL-6 decreased significantly in both GCA and PMR patients, whereas CXCL9 remained high. Serum BAFF and IL-6 correlated strongly with ESR and CRP in GCA and PMR patients.Conclusion. Among the serum markers tested, BAFF and IL-6 showed the strongest association with disease activity in both GCA and PMR patients. The diagnostic value of these markers should be evaluated in larger, longitudinal studies with GCA and PMR patients, and in patients with infections or other inflammatory conditions.

Background: The interleukins IL-4 and IL-13 play a key role in the pathophysiology of asthma. The interleukin receptor IL-13Rα2 is believed to act as a decoy receptor, but until now, the functional significance of IL-13Rα2 remains vague. Methods: Bronchial reactivity was quantified in murine lung slices by digital video microscopy and acetylcholine (ACH)-induced Ca2+ signaling was measured in human airway smooth muscle cells (ASMC) using fluorescence microscopy. Results: IL-4 or IL-13 up to 50 ng/ml induced bronchial hyperreactivity. But after incubation with 100 ng/ml this effect was lost and bronchial responsiveness was again comparable to the control level. The effects of IL-4 and IL-13 on bronchial reactivity were paralleled by the effects on ASMC proliferation. Fifty nanograms per milliliter of IL-4 and IL-13 increased the Ca2+ response of human ASMC to ACH. At 100 ng/ml, however, the effects of the cytokines on the Ca2+ response were no longer evident. The expression of IL-13Rα2 increased with increasing concentrations of IL-4 or IL-13, reaching its maximum at 100 ng/ml. Blocking IL-13Rα2, the loss of the effect of IL-4 and IL-13 at 100 ng/ml on human ASMC proliferation and the ACH-induced Ca2+ response were no longer present. Conclusions: IL-4 and IL-13 induce bronchial hyperreactivity by changing the Ca2+ homeostasis of ASMC. These effects are counteracted by IL-13Rα2. The biological significance of IL-13Rα2 might be a protective function by regulating IL-13- and IL-4-mediated signal transduction and thereby limiting pathological alterations in Th2-mediated inflammatory diseases.

To identify novel cytokine-related genes, we searched the set of 60,770 annotated RIKEN mouse cDNA clones (FANTOM2 clones), using keywords such as cytokine itself or cytokine names (such as interferon, interleukin, epidermal growth factor, fibroblast growth factor, and transforming growth factor). This search produced 108 known cytokines and cytokine-related products such as cytokine receptors, cytokine-associated genes, or their products (enhancers, accessory proteins, cytokine-induced genes). We found 15 clusters of FANTOM2 clones that are candidates for novel cytokine-related genes. These encoded products with strong sequence similarity to guanylate-binding protein (GBP-5), interleukin-1 receptor-associated kinase 2 (IRAK-2), interleukin 20 receptor α isoform 3, a member of the interferon-inducible proteins of the Ifi 200 cluster, four members of the membrane-associated family 1-8 of interferon-inducible proteins, one p27-like protein, and a hypothetical protein containing a Toll/Interleukin receptor domain. All four clones representing novel candidates of gene products from the family contain a novel highly conserved cross-species domain. Clones similar to growth factor-related products included transforming growth factor β-inducible early growth response protein 2 (TIEG-2), TGFβ-induced factor 2, integrin β-like 1, latent TGF-binding protein 4S, and FGF receptor 4B. We performed a detailed sequence analysis of the candidate novel genes to elucidate their likely functional properties.

Rheumatoid arthritis is a debilitating disease that has significant impact on quality of life. Biological and non biological DMARDs improved the outcome in RA patients. New agents are still required in inadequate responders or intolerant patients. FDA has recently approved a new drug Sarilumab, a monoclonal antibody for the treatment of moderate to severe form of RA which acts by binding to Interleukin receptor 6 and interrupts the resultant cytokine-mediated inflammatory signaling.

Ulcerative colitis (UC) is one of the main types of inflammatory bowel disease, which is caused by dysregulated immune responses in genetically predisposed individuals. Several genetic factors, including interleukin and interleukin receptor gene polymorphisms and other inflammation-related genes play central role in mediating and modulating the inflammation in the human body, thereby these can be the main cause of development of the disease. It is clear these data are very important for understanding the base of the disease, especially in terms of clinical utility and validity, but summarized literature is exiguous for challenge health specialist that can used in the clinical practice nowadays. This review summarizes the current literature on inflammation-related genetic polymorphisms which are associated with UC. We performed an electronic search of Pubmed Database among publications of the last 10 years, using the following medical subject heading terms: UC, ulcerative colitis, inflammation, genes, polymorphisms, and susceptibility.

The main aim of the present study was a search for a characteristic serum marker of inflammatory activity in the airways of asthmatics with difficult-to-control disease. Therefore, serum levels of interleukin-4 (IL-4), serum low-affinity Fc Epsilon Receptor II (sFcER II), Interferon-gamma (INF-gamma) Immunoglobulin-E (IgE), Interleukin-2 (IL-2), serum Interleukin Receptor 2 (sIL-2R) and Intercellular Adhesion Molecule-1 (sICAM-1) were measured in 2 groups of asthmatics: 1-26 patients with difficult-to-control asthma (DTCA), 2-22 asthmatics, minimally symptomatic (MSA). Results: No significant difference in either measured parameters between the DTCA and MSA group in peripheral blood samples was found. Conclusion: The above mentioned serum markers of T- and B-cell activation as well as the serum ICAM-1 level are not sensitive enough to determine the type, activity and severity of the inflammatory process in the asthmatic airways.

Background— Concentrations of soluble (s)ST2 predict prognosis in heart failure. We recently found changing doses of β-blocker (BB) may affect sST2 concentrations. It remains unclear whether sST2 concentrations identify benefit of BB therapy, however. Methods and Results— A total of 151 subjects with heart failure attributable to left ventricular systolic dysfunction were examined in this post hoc analysis; >96% were taking BB at enrollment. Medication regimen and sST2 values were obtained during 10 months. Cardiovascular events were examined as a function of baseline sST2 status (low ≤35 versus high >35 ng/mL) and final achieved BB dose (high ≥50 versus low P =0.001), and lowest cumulative hazard ( P =0.003). Those with low sST2/low-dose BB, or high sST2/high-dose BB had intermediate outcomes (0.92 and 1.19 events). Patients with high sST2 treated with low-dose BB had the highest cardiovascular event rate (2.08 events) and the highest cumulative hazard. Compared with low sST2/high-dose BB, those with high sST2 treated with low-dose BB had an odds ratio of 6.77 ( P P =0.18 and 0.02). Similar results were found for heart failure hospitalization and cardiovascular death. Conclusions— Although BB therapy exerted dose-related benefits across all study participants, sST2 measurement identifies patients with chronic heart failure who may particularly benefit from higher BB doses. Clinical Trial Registration— URL: http://www.clinicaltrials.gov . Unique identifier: NCT00351390.

IntroductionThe influence of the immune deregulation on the risk and psychopathology of schizophrenia is increasingly recognized in the literature.AimTo assess the association between serum IL-1RA on schizophrenia psychopathology.MethodsWe recruited 88 schizophrenia patients (38 males and 49 females, mean age 38.12 ± 12.67 years) and 88 healthy adult control subjects (68 males, 20 females, mean age 40.63 ± 7.99 years). Lifetime psychopathology was evaluated using Operational Criteria for Psychotic Illness (OPCRIT) checklist, while current psychopathology was assessed using Positive and Negative Syndrome Scale (PANSS). Serum samples were stored in aliquots at –80 °C. Serum levels of IL1-RA were measured using Immunoassay (ELISA).ResultsThere were statistically significant differences between schizophrenia patients and healthy controls (median ± interquartile range: 350,81 ± 227.04 and 888.74 ± 762.63, respectively [pg/ml]) (U Mann–Whitney test, Z = –7.99, P < 0.0001). There were no differences in serum IL1-RA levels between male and female among patients with schizophrenia (U Mann–Whitney test, Z = –0.22, P = 0.82) nor among healthy control subjects (U Mann–Whitney test, Z = –0.17, P = 0.86). Among schizophrenia patients, there was a trend-level association between IL-1RA serum level with negative symptoms (Spearman correlation coefficient, r = –0.23, P = 0.056), positive symptoms (Spearman correlation coefficient r = –0.22, P = 0.066), and on a statistically significant level with general symptoms (Spearman correlation coefficient r = –0.28, P = 0.018).ConclusionSerum IL1-RA level is higher in schizophrenia patients in comparison to healthy controls and it is associated with schizophrenia psychopathology.Disclosure of interestThe authors have not supplied their declaration of competing interest.

Intravenous immunoglobulin (IVIG)-resistant Kawasaki disease (KD) patients comprise at least 20% of treated patients and are at higher risk of coronary artery abnormalities. If identified early in the course of the disease, such patients may benefit from additional anti-inflammatory therapy. The aim of this study was to compare the transcript abundance between IVIG-resistant and – responsive KD patients to identify biomarkers that might differentiate between these two groups and to generate new targets for therapies in IVIG-resistant KD patients. We compared the transcript abundance profiles of whole blood RNA on Agilent arrays from acute and convalescent KD subjects and age-similar, healthy controls. KD subjects were stratified as IVIG-resistant or – responsive based on response to initial IVIG therapy. Transcript abundance was higher for IL-1 pathway genes (IL-1 receptor, interleukin receptor associated kinase, p38 mitogen-activated protein kinase), and MMP-8. These findings point to candidate biomarkers that may predict IVIG-resistance in acute KD patients. The results also underscore the importance of the IL-1 pathway as a mediator of inflammation in KD and suggest that IL-1 or its receptor may be reasonable targets for therapy, particularly for IVIG-resistant patients.

IL-25, a member of the IL-17 cytokine family, is known to enhance Th2-like responses associated with increased serum levels of IgE, IgG1, IgA, blood eosinophilia, and eosinophilic infiltrates in various tissues. However, IL-25 also abrogates inflammatory responses driven by Th17 cells. However, the cell types that respond to IL-25 and the mechanisms by which IL-25 differentially regulates immune reactions are not well explored. To identify potential targets of IL-25, we initially examined IL-25 receptor (IL-25R) in human peripheral blood cells. IL-25R was predominantly expressed by CD14+ cells. We next assessed the functional role of IL-25 in modulating the response of CD14+ cells to various inflammatory signals. CD14+ cells responded to IL-25 by down-regulating the synthesis of inflammatory cytokines induced by toll-like receptor (TLR) ligands and inflammatory cytokines. Inhibition of cytokine response by IL-25 occurred via a p38 Map kinase–driven Socs-3–dependent mechanism. In vivo, IL-25 inhibited monocyte-derived cytokines and protected against LPS-induced lethal endotoxemia in mice. These data indicate that IL-25 is a negative regulator of monocyte proinflammatory cytokine responses, which may have therapeutic implications.

Genetic variation in immune-regulating components such as cytokines may lead to interindividual differences in immunosuppression response and susceptibility to melanoma. We evaluated the associations between genetic variants in five interleukins (IL) and IL receptor genes (IL-4, IL-4R, IL-6, IL-6R, and IL-10) and the risk of melanoma. Twenty-five single nucleotide polymorphisms (SNPs) were selected that are functionally relevant or tagging SNPs of each gene. We conducted a nested case-control study of 219 female patients and 219 matched controls within the Nurses' Health Study. We observed that in the IL-6R gene, four SNPs in linkage disequilibrium were associated with an increased risk of melanoma. Three are located in introns (rs6684439, rs4845618, and rs4845622), and one is a nonsynonymous SNP in exon 9 [rs8192284 (Asp358Ala)]. An elevated risk of melanoma was observed in the heterozygous groups of these SNPs with odds ratio of 1.74 [(95% confidence interval, 1.07, 2.81) for rs6684439, 1.72 (1.04, 2.84) for rs4845618, 1.69 (1.03, 2.75) for rs4845622, and 1.68 (1.04, 2.73) for rs8192284]. But these associations were not observed in the homozygous variant group with odds ratios ranging from 0.93 to 1.03. We did not find significant results for the SNPs in the other four genes. These data suggest the involvement of IL-6R in melanoma development. Further studies are needed to confirm these findings.

Objectives: Traumatic injury to cartilage has been shown to lead to Post-Traumatic Osteoarthritis (PTOA). The acute phase of PTOA is characterized with increased expression of aggrecanases and inflammatory cytokines in the injured cartilage. Early intervention therapies aim to be administered during the acute phase for the prevention of PTOA development. Our objective was to determine the effect of Interleukin Receptor Antagonist Protein (IRAP), Hyaluronan (HA), and Mesenchymal Stem Cell (MSC) treatment as early intervention strategies by examining the changes in microRNA (miRNA) and mRNA expression in cartilage at 8 hours after impact injury. Methods: Custom impact device was used to create replicable injury ex-vivo to intact porcine knee joint. Injury was caused by dropping a 10kg weight one time from 1m directly above the knee in extension. One hour after impact 20µg/mL IRAP, 15mg/mL HA (MW 1.9 MDa), or 5x106 P4 MSCs in 1mL saline was intra-articularly injected. Control legs (no injury) and injury legs (injury, no treatment) received saline injection. At 8 hours post-injury, cartilage samples were harvested for genetic expression analysis. Genetic expression of miR-140 (regulates ADAMTS-5) miR-125b (regulates ADAMTS-4), ADAMTS-4, ADAMTS-5, MMP-3, IL-1β, and TNF-α were analyzed by RT-PCR. Groups were compared by one-way analysis of variance followed by Tukey's post-hoc test. A P-value Results: After IRAP treatment, expressions of ADAMTS-4, ADAMTS-5, IL-1β, and TNF-α in cartilage were significantly down-regulated from injury group (all PConclusion: The results show that IRAP, HA, and MSC treatment administered during acute phase of cartilage injury each have a distinct effect on catabolic and inflammatory regulation. HA and MSC treatment did not have significant effect on the inflammatory and catabolic response of injured cartilage within the first 8 hours of injury. However, the beneficial effects of HA may take place beyond the acute time frame as suggested by increased miR-125b and miR-140 expressions after treatment. IRAP treatment increased expressions of miR-140, -125b, and -27b in cartilage, indicating increased inhibition of their respective enzymes, as shown by reduced aggrecanase and inflammatory genetic expressions. Clinically, these findings support the potential of IRAP and HA treatment as early intervention strategies for the prevention of cartilage degeneration after impact injury.

The activation of a macrophage (Mphi)-dependent proinflammatory cascade following thermal injury plays an important role in the development of immunosuppression and increased susceptibility to subsequent sepsis in burn patients. In contrast, although interleukin (IL)-10, an anti-inflammatory cytokine that can downregulate M phi activity, has also been implicated in postburn immune dysfunction, its role in the regulation of M phi function postburn remains unclear. To study this, C57BL/6 female mice were subjected to a 25% total body surface area third-degree scald burn, and splenic Mphis were isolated 7 days later. Lipopolysaccharide (LPS)-stimulated IL-10, IL-6, tumor necrosis factor (TNF)-alpha, and nitric oxide (NO) production were significantly increased in the burn group compared with shams. Blockade of endogenous IL-10 activity enhanced IL-6 and TNF-alpha release, but not NO release, in both groups. The addition of exogenous IL-10 to the M phi cultures dose dependently suppressed production of these inflammatory mediators in both groups. The timing of IL-10 addition to the cultures in relation to LPS stimulation, however, was critical. The suppressive effect of exogenous IL-10 was attenuated in both groups when the cells were exposed to IL-10 at 4-6 h after LPS stimulation; however, Mphis from injured mice were significantly better able to maintain inflammatory mediator-productive capacity. The resistance of Mphis from injured mice to IL-10-mediated suppression correlated with decreased IL-10 receptor (IL-10R) expression and increased CD11b expression. These findings suggest that Mphis, following thermal injury, display resistance to suppression by IL-10 due in part to downregulation of IL-10R expression.

Interleukin-1 (IL-1) is induced immediately after brain imjury and elevated levels of IL-1 have been strongly implicated in the neurodegeneration that accompanies stroke, Alzheimer's disease and Multiple Sclerosis. Antagonizing IL-1 reduces cell death; however, the basis for this protection has not been elucidated. Here we analyzed the response to penetrating brain injury in mice lacking the type 1 interleukin receptor (IL-1R1) to determine which cellular and molecular mediators of tissue damage require IL-1 signaling. At the cellular level fewer amoeboid microglia/macrophages appeared adjacent to the injured brain tissue in IL-1R1 null mice, and those microglia present at early postinjury intervals retained their resting morphology. Astrogliosis also was mildly abrogated. At the molecular level, cyclooxygenase 2 and IL-6 expression were depressed and delayed. Interestingly, basal levels of cyclooxygenase 2, IL-1 and IL-6 were significantly lower in the IL-1R1 null mice. Additionally, stimulation of VCAM-1 mRNA was depressed in the IL-1R1 null mice, and correspondingly, there was reduced migration of peripheral macrophages into the IL-1R1 null brain after injury. This observation correlated with a reduced number of cyclooxygenase 2+ amoeboid phagocytes adjacent to the injury. By contrast, the production of nerve growth factor was only mildly affected. Since antagonizing IL-1 protects neural cells in experimental models of stroke and multiple sclerosis, our data suggest that cell preservation is achieved by abrogating microglial/macrophage activation and the subsequent self-propagating cycle of inflammation. Acknowledgements: Supported by NMSS Award #RG 3837.

Rheumatoid arthritis (RA) is an autoimmune disease, resulting in a chronic, systemic inflammatory disorder. It may affect many tissues and organs, but it primarily affects the flexible joints. In clinical practice patient care generates many questions about diagnosis, prognosis, and treatment. It is challenging for health care specialists to keep up to date with the medical literature. This review summarizes the pathogenesis, the polymorphisms of interleukin and interleukin genes and the standard available and possible future immunologic targets for RA treatment. The identification of disease-associated interleukin and interleukin receptor genes can provide precious insight into the genetic variations prior to disease onset in order to identify the pathways important for RA pathogenesis. The knowledge of the complex genetic background may prove useful for developing novel therapies and making personalized medicine based on the individual’s genetics.