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The IGRA (Interferon Gamma Release Assays) test is currently the standard specific test for Mycobacterium tuberculosis infection status. However, a positive test cannot distinguish between active tuberculosis disease (ATBD) and latent tuberculosis infection (LTBI). Developing a test with this characteristic is needed. We conducted longitudinal studies to identify a combination of antigen peptides and cytokines to discriminate between ATBD and LTBI. We studied 54 patients with ATBD disease and 51 with LTBI infection. Cell culture supernatant from cells stimulated with overlapping Mycobacterium tuberculosis novel peptides and 40 cytokines/chemokines were analyzed using the Luminex technology. To summarize longitudinal measurements of analyte levels, we calculated the area under the curve (AUC). Our results indicate that in vitro cell stimulation with a novel combination of peptides (Rv0849-12, Rv2031c-14, Rv2031c-5, and Rv2693-06) and IL-1RA detection in culture supernatants can discriminate between LTBI and ATBD.

BACKGROUNDNasopharyngeal carriage of pathogenic bacteria precedes invasive disease and higher rates are found in low socioeconomic-status (SES) settings. Local immune responses are important for controlling colonization, but whether SES affects these responses is currently unknown.OBJECTIVEExamining bacterial colonization and cytokine response in nasal mucosa of children from high and low SES.METHODSTwenty-five cytokines were measured in nasal fluid. qPCR was performed to determine carriage and density of Haemophilus influenzae (H. influenzae), Streptococcus pneumoniae (S. pneumoniae), Moraxella catarrhalis (M. catarrhalis) and Staphylococcus aureus (S. aureus).RESULTSThe densities of H. influenzae and S. pneumoniae were increased in low compared to the high SES (p=0.006, p=0.026), with respectively 6 and 67 times higher median densities. Densities of H. influenzae and S. pneumoniae were positively associated with levels of IL-1beta (p=0.002, p=0.008) and IL-6 (pp=0.006). After correcting for bacterial density, IL-6 levels were increased in colonized children from high compared to low SES for both H. influenzae and S. pneumoniae (both p=0.039).CONCLUSIONIncreased density of H. influenzae and S. pneumoniae was observed in low SES children, while IL-6 levels associated with colonization were reduced in these children, indicating that immune responses to bacterial colonization were altered by SES.

The task in treating acute nasopharyngitis (ANP) deals with reducing the disease symptoms and the risk of complications. The lack of reliable antiviral drugs makes it important to search for appropriate medicines among other pharmacotherapeutic groups.The study involves a comparative analysis of the efficiency and estimates potential: the recombinant interferon α2b and the compound containing fungal β-D-glucans used in treat ANPThe studies involved patients with ANP from 18 to 55 years old. As many as 152 people were examined including the following: 38 were practically healthy people (group 1); and 114 patients wuth ANP: 38 people (group 2) was subject to a standard therapy (vasoconstrictor nasal drops, nasal cavity irrigation using 0.1% Miramistine solution, gargling using the Furacilin solution); forty people (group 3) were administered application of intranasal interferon α2b of 105IU, it was delivered with a spray into each nasal passage twice a day; 36 people (group 4) were administered an immunotropic drug containing β-D-glucans orally twice a day. The duration of drug administration lasted 7 days. Polymerase chain reaction (PCR) was used to identify the ANP etiological factor. Concentrations of cytokines IL-1β, IL-1ra were estimated using enzyme immunoassay (ELISA) technique. Clinical efficiency was assessed through score approach. The following symptoms were taken into account: general malaise, sore throat, character of nasal discharge, and the difficulty of nasal breathing. The results of the study were analyzed using parametric and nonparametric statistical methods. In 60.0% the nasal secretions of patients revealed RV. The distribution of cytokine concentrations in nasal secretions in group 1 indicated that the concentration of IL-1β was in the range of 20.0-25.0 pg/ml, and the concentration of IL-1ra was about 1250.0-2500.0 pg/ml. Developing ANP stimulated an increase in IL-1β concentration up to 30.0-70.0 pg/ml in nasal secretions of patients without affecting IL-1ra concentrations. On day 7 of treatment, the cytokine concentrations among the patients treated using the immunotropic drugs were the same as in the group of healthy individuals. There were no significant changes in cytokine production on day 7 in the group of patients undergoing the standard treatment. Application of proposed immunobiological medicines to ANP does not result in overproduction of proinflammatory cytokine IL-1β in nasal secretion. This confirms that these drugs are promising in the treating strategy including reduction of the risk of developing complications.

Interleukin-1 antagonism is a proposed biomarker and potential therapy for the delay and/or treatment of type 1 diabetes (T1D). We evaluated the role of circulating interleukin-1 receptor antagonist (IL-1ra) in a prospectively monitored cohort of T1D patients. In order to determine a mechanistic association between IL-1ra and T1D, we performed co-localization analyses between serum IL-1ra protein quantitative trait loci and T1D genome-wide analysis studies. Adjusting for human leukocyte antigen (HLA) genotypes, first degree relative status, gender, and age, serum levels of IL-1ra were lower in subjects who progressed to T1D compared to the controls (p = 0.023). Our results suggest that females have higher levels of IL-1ra compared to males (p = 0.005). The 2q14.1 region associated with serum IL-1ra levels is not associated with a risk of developing T1D. Our data suggest that IL-1 antagonism by IL-1ra is not an effective therapy in T1D, but IL-1ra may be a biomarker for progression to T1D.

BackgroundBiomarkers predicting treatment response may be used to stratify patients with pancreatic ductal adenocarcinoma (PDAC) for available therapies. The aim of this study was to evaluate the association of circulating cytokines with FOLFIRINOX response and with overall survival (OS).MethodsSerum samples were collected before start and after the first cycle of FOLFIRINOX from patients with PDAC (n=83) of all disease stages. Overall, 34 circulating cytokines were analyzed with a multiplex immunoassay. In addition, changes in peripheral blood immune cell counts were determined by flow cytometry to correlate with differences in cytokine levels. Chemotherapy response was determined by CT scans with the RECIST 1.1 criteria, as disease control (n=64) or progressive disease (n=19) within eight cycles of FOLFIRINOX.ResultsPatients with high serum IL-1RA concentrations after one cycle of chemotherapy were less likely to have tumor progression during FOLFIRINOX (OR 0.25,P=0.040). Increase of circulating IL-1RA concentrations correlated with increase of total, classical (CD14+CD16-), and non-classical monocytes (CD14-CD16+), and dendritic cells. In multivariable cox regression, including the variables chemotherapy response outcome and baseline CA19-9 level, serum concentrations of IL-7 (HR 2.14,P=0.010), IL-18 (HR 2.00,P=0.020), and MIP-1β (HR 0.51,P=0.025) after one cycle of FOLFIRINOX showed correlations with OS.ConclusionsCirculating IL-1RA, IL-7, IL-18, and MIP-1β concentrations are biomarkers associated with FOLFIRINOX response in PDAC patients, suggesting an important role for specific immune cells in chemotherapy response and PDAC progression. Cytokine-based treatment might improve patient outcome and should be evaluated in future studies.

Interleukin-1 receptor antagonist (IL-1Ra) is an active pharmaceutical ingredient obtained by the method of gene transformation of Escherichia coli, provides a neuroprotective effect, namely – inhibition of oxidative modification of proteins, normalization of functional activity of mitochondrial energy, mitochondrial energy. Early complex physical-chemical, microbiological, and biopharmaceutical studies have developed a new intranasal formulation, the formulation of which contains: IL-1Ra, sodium carboxymethyl cellulose, D-panthenol, benzalkonium chloride, trilon B, Tween-80, phosphate buffer solution. Given the peculiarities of the technological process, when creating a new gel dosage form, it is necessary to justify the temperature of the carrier base, the introduction of the active substance into the base, as well as to predict the possible chemical interaction of individual components in the dosage form. Therefore, the application of thermogravimetric analysis in pharmaceutical technology is relevant. The aim of the work is to conduct thermogravimetric studies of a new nasal form with an antagonist of interleukin-1β (IL-1Ra). Materials and methods. The objects of thermogravimetric studies were: semi-finished product-solution of the receptor antagonist of interleukin-1 (IL-1Ra). Excipients were: sodium carboxymethylcellulose, tween-80, D-panthenol, trilon B, benzalkonium chloride. To ensure the appropriate pH value used phosphate buffer solution (pH 6.0), according to the recipe to State Pharmacopoeia 2 ed. In addition to the individual components of the finished nasal gel, also prepared nasal gel without IL-1Ra, as well as gel with IL-1Ra. The thermographic analysis was performed on a derivatograph – Shimadzu DTG-60 (Japan) with a platinum-platinum-rhodium thermocouple when heating the samples in aluminum crucibles (from 15 °C to 250 °C). Al2O3 was used as a reference substance. The heating rate was 10 °C per minute. The weight of the test samples ranged from 19.22 mg to 52.91 mg. The derivatograph graphically recorded the obtained data in the form of curves T, DTA, TGA. The T curve on the derivatogram shows the change in temperature, and the TGA curve shows the change in mass of the sample during the study period. The DTA curve reflects the differentiation of thermal effects, contains information about endothermic and exothermic maxima, can be used for qualitative evaluation of the derivatogram. Results. The study consisted of two stages. At the first stage, we studied the derivatograms of experimental samples – individual ingredients of the finished nasal gel - active and excipients, namely: IL-1Ra, sodium carboxymethylcellulose, tween-80, D-panthenol, trilon B, benzalkonium chloride, phosphate buffer solution 6.0. In the second stage, samples of nasal gels were prepared: nasal gel with the required amount of excipients, in addition to the active substance – IL-1Ra, as well as a nasal gel containing IL-1Ra and the required amount of excipients. As a result, we obtained some parameters of thermogravimetric analysis in the form of a graphical image – derivatograms, which were then analyzed. Conclusions. Thermogravimetric studies of individual components of nasal gel with IL-1Ra, as well as nasal gels with active substance and without active substance, were found that the manufacturing process of gel with IL-1Ra should be carried out taking into account thermolabile compounds. First, you need to prepare the base without preservative and IL-1Ra. Then at a temperature not exceeding 40 °C dissolve the preservative, and then when cooled to 20 °C add IL-1Ra. It was found that the developed dosage form of the gel with IL-1Ra is a mixture of active and excipients, the ingredients of which do not interact with each other and can be combined in one dosage form.

Le malattie autoinfiammatorie sistemiche si manifestano quando il sistema immunitario viene attivato in maniera incontrollata. Un sottogruppo cardinale di queste malattie comprende rare febbri periodiche caratterizzate dalla produzione disregolata della citochina proinfiammatoria interleuchina-1 (IL-1). Non esiste una cura per queste condizioni. Anakinra, il farmaco rappresentante la ricombinante dell'antagonista del recettore IL-1 (IL-1RA), è la terapia principale per questi pazienti. Tuttavia, l’anakinra ha una breve emivita e una scarsa distribuzione dei tessuti. I pazienti gravi rispondono in modo inadeguato e i loro sintomi non migliorano. Pertanto, è necessaria una terapia duratura che oltrepassi la somministrazione continua di farmaci e garantisca una risoluzione soddisfacente dell'infiammazione dei tessuti. In questo progetto di dottorato, abbiamo affrontato l'urgenza di sviluppare un trattamento efficace per queste malattie, basato su cellule staminali ematopoietiche che producono costitutivamente la proteina umana IL-1RA utilizzando un approccio di trasferimento genico mediato da vettori lentivirali (LV). Abbiamo dimostrato che le cellue staminali trasdotte producono in maniera efficace I-1RA. Il protocollo di trasduzione associato alla sovraespressione di IL-1RA non hanno alterato la vitalità cellulare, il potenziale clonogenico e l’abilità delle cellule staminali di differenziazione nel topo. Successivamente, abbiamo studiato se le cellule staminali esprimenti IL-1RA, una volta trapiantate nei topi potessero migliorare l'infiammazione acuta e cronica. Sono stati impiegati tre modelli di topi. L'espressione ectopica di IL-1RA da parte delle cellule immunitarie derivate dalle cellule staminali trapiantate ha soppresso il reclutamento dei neutrofili nel sito dell'infiammazione nei topi con peritonite indotta da cristalli di urato monosodico (MSU), noti attivatori del complesso NLRP3 dell’inflammasoma. Questo effetto protettivo era paragonabile a quello ottenuto da anakinra. Successivamente, abbiamo sfruttato un modello murino inducibile esprimente la mutazione dominante Nlrp3A350V associata alle sindromi periodiche associate alla criopirina (CAPS). Il trapianto singenico di cellule staminali trasdotte con IL-1RA in topi Nlrp3A350V+CreT ha efficacemente impedito ai topi l'insorgenza e la progressione della malattia associata a perdita di peso, leucocitosi e livelli sierici elevati dell’interleuchina 6. Infine, i nostri dati preliminari indicano che il nostro approccio di terapia genica potrebbe migliorare il tasso di mortalità e la gravità della malattia in un modello murino di encefalomielite autoimmune sperimentale. Complessivamente, i nostri risultati hanno dimostrato che la somministrazione di IL-1RA mediata da un vettore lentivirale nelle cellule staminali è un approccio sicuro ed efficiente per controllare l'infiammazione mediata da IL-1. Questi risultati hanno posto le basi per studi futuri per valutare la potenziale applicazione clinica della tarapia genica con IL-1RA per le malattie autoinfiammatorie sistemiche mediate da IL-1. Systemic autoinflammatory diseases (SAIDs) delineate a group of diseases that manifest when the immune system is activated uncontrollably. One cardinal subgroup of SAIDs includes rare periodic fevers characterized by the dysregulated production of the proinflammatory cytokine interleukin-1 (IL-1). There is no cure for these conditions. Anakinra, the recombinant form of IL-1 receptor antagonist (IL-1RA), is the mainstay therapy for these patients. However, anakinra has a short half-life and poor tissue distribution. Severe patients respond inadequately and do not experience improvement in symptoms. Therefore, there is a need for a durable therapy that bypasses continuous drug administration and ensures a satisfactory resolution of tissue inflammation. In this PhD project, we addressed the urgency to develop an effective treatment for IL-1 induced SAIDs based on haematopoietic stem and progenitor cells (HSPCs) producing constitutively human IL-1RA using a lentiviral vector (LV)-mediated gene transfer approach. Human and mouse HSPCs transduced with an LV encoding human IL-1RA efficiently released this cytokine. Transduction procedure and IL-1RA over-expression did not alter HSPCs viability, clonogenic and differentiation potential in vivo. Next, we investigated whether, once transplanted in mice, IL-1RA-expressing HSPCs could ameliorate acute and chronic inflammation. Three mouse models were employed. The ectopic expression of IL-1RA by HSPC-derived immune cells suppressed neutrophil recruitment to the site of inflammation in mice with peritonitis induced by monosodium-urate crystals (MSU), well-known activators of the NLRP3-IL-1 axis. This protective effect was comparable to that obtained by anakinra. Next, we exploited an inducible mouse model of the cryopyrin-associated period syndrome (CAPS) carrying the dominant Nlrp3A350V mutation. Syngeneic transplant of IL-1RA-transduced HSPCs in Nlrp3A350V+CreT mice effectively prevented mice from disease onset and progression manifested as weight loss, leucocytosis, and high serum IL-6 level. Finally, preliminary data indicate that our gene therapy approach could improve mortality rate and disease severity in a mouse model of experimental autoimmune encephalomyelitis. Altogether, our results demonstrated that LV-mediated IL-1RA delivery in HSPCs is safe and efficient approach to controlling IL-1-mediated inflammation. These findings set the stage for future studies to evaluate the potential LV-mediated IL-1RA GT clinical application for IL-1-mediated systemic autoinflammatory diseases.

The aim of this study is to evaluate the association of interleukin-4 (IL-4) gene intron 3 and interleukin-1 receptor antagonist (IL-1Ra) gene intron 2 variable number tandem repeat (VNTR) polymorphisms with bladder cancer (BC) susceptibility in Turkish population. A total of 75 BC patients and 126 healthy controls were included in this case-control study. Genotyping for the interested polymorphisms were analyzed through polymerase chain reaction (PCR). The strength of association between both IL-4 and IL-1Ra gene VNTRs and BC susceptibility was estimated utilizing odds ratio (OR) with corresponding 95% confidence interval (CI). In the study, no statistically significant differences were determined in the allele distributions for either in IL-4 gene intron 3 VNTR (OR= 1.33; CI 0.704-2,41, p=0.390) or in IL-1Ra gene intron 2 VNTR polymorphisms (OR= 0.890; CI 0.569-1.394, p=0.346) between BC patients and control groups. The genotype distributions of IL-4 gene were estimated for RP1/RP2 (OR= 1.55; CI 0.11-7.74, p = 0.590) and RP1/RP1 (OR= 2.08; CI 0.48-9.06, p = 0.320), found no difference between BC and control groups. The genotype distributions of IL-1Ra gene were estimated for 2L (OR= 1.401; CI 0.753-2.610, p = 0.287) and 22 (OR=0.908; CI 0.252-3.276, p = 0.883) and found no difference between BC and control groups. This study suggest that there were no statistically significant differences determined either in genotype or allele distributions between BC patients and control groups for both IL-4 intron 3 VNTR and IL-1Ra intron 2 VNTR polymorphisms in Turkish population and therefore there was no association of these variants with BC risk in this population.

Interleukin-1 receptor antagonist is an important acute-phase protein and an immune mediator, and its expression is associated with the development of hepatitis or acute liver failure. The aim of this study was to investigate whether recombinant human interleukin-1 receptor antagonist directly targets and improves cell survival in a carbon tetrachloride-induced hepatocyte injury model in vitro. A human hepatoma cell line and a mouse hepatocyte cell line were used to establish carbon tetrachloride-induced cell injury models in vitro, and cell viability, apoptosis, and reactive oxygen species level were determined to assess the degree of hepatocellular damage. Quantitative real-time polymerase chain reaction was used to analyze the level of interleukin-1β, interleukin-6, and tumor necrosis factor-α mRNA in cells; extracellular regulated protein kinases 1/2 phosphorylation in hepatocytes was analyzed using western blotting. Recombinant human interleukin-1 receptor antagonist could directly target hepatocytes, improve cell survival, and decrease carbon tetrachloride-induced cell apoptosis in vitro. In hepatocytes, recombinant human interleukin-1 receptor antagonist remarkably downregulated expression of interleukin-1β, interleukin-6, and tumor necrosis factor-α in hepatocytes exposed to carbon tetrachloride. It also decreased accumulation of reactive oxygen species and abrogated the suppression of extracellular regulated protein kinases 1/2 phosphorylation induced by carbon tetrachloride. However, stimulation of cells with an extracellular regulated protein kinases 1/2 inhibitor blocked the recombinant human interleukin-1 receptor antagonist-induced upregulation of extracellular regulated protein kinase1/2 activation and abrogated the improvement in hepatocyte survival following carbon tetrachloride treatment. Collectively, these findings provide new insights into the hepatocyte-protective mechanism of recombinant human interleukin-1 receptor antagonist.

Background Dynamically changed levels of serum cytokines might predict the development of active TB from latent tuberculosis infection (LTBI) and monitor preventive treatment effectiveness. The aim of the study was to identify potential serum cytokines associated with LTBI treatment which might predict active disease development in a Chinese population. Methods Based on a randomized controlled trial aiming to explore short-course regimens for LTBI treatment, the dynamic changes of serum cytokines determined by bead-based multiplex assays were investigated for the participants who developed active TB during follow-up and age and gender matched controls stayed healthy. Results Totally, 21 patients diagnosed with active tuberculosis (TB) during the 2-year follow-up (12 from treated groups and 9 from untreated controls) and 42 age and gender matched healthy controls (24 from treated groups and 18 from untreated controls) were included in the study. Before treatment, serum IL-1ra was statistically higher among those who developed active disease during follow-up as compared with those stayed healthy. As for treated participants, the levels of IL-1ra were significantly lower after treatment in comparison with those before treatment both in active TB group (p = 0.002) and non-TB group (p = 0.009). For untreated participants, the levels of IL-1ra were not statistically different between different time points both in active TB group (p = 0.078) and non-TB group (p = 0.265). Conclusion Our results suggested that declined serum level of IL-1ra was associated with LTBI treatment. Further studies are needed to verify whether it could be used to evaluate LTBI treatment and to predict active disease development.

Introduction: It has been recognized for about 20 years that interleukin (IL)-1 signaling is implicated in Multiple Sclerosis (MS), a disabling, chronic, inflammatory and neurodegenerative disease of the central nervous system (CNS). Only recently, multifaceted roles of IL-1 emerged in MS pathophysiology as a result of both clinical and preclinical studies. Notably, drugs that directly target the IL-1 system have not been tested so far in MS.Areas covered: Recent studies in animal models, together with the development of ex vivo chimeric MS models, have disclosed a critical role for IL-1 not only at the peripheral level but also within the CNS. In the present review, we highlight the IL-1-dependent neuropathological aspects of MS, by providing an overview of the cells of the immune and CNS systems that respond to IL-1 signaling, and by emphasizing the subsequent effects on the CNS, from demyelinating processes, to synaptopathy, and excitotoxicity.Expert opinion: Drugs that act on the IL-1 system show a therapeutic potential in several autoinflammatory diseases and preclinical studies have highlighted the effects of these compounds in MS. We will discuss why anti-IL-1 therapies in MS have been neglected to date.

We provide the data on distribution of IL-1β and IL-1ra cytokine concentrations in nasal secretions from the patients with different forms of chronic polypous rhinosinusitis. The control group (group 1) included healthy persons. The second group consisted of patients exhibiting chronic polypous rhinosinusitis (CRS-NP), free of clinical signs of purulent inflammation in paranasal sinuses and/or bronchial asthma. The patients of group 3 were diagnosed with chronic purulent polypous rhinosinusitis (CPRS-NP). Group 4 consisted of the patients with chronic polypoid rhinosinusitis complicated by bronchial asthma (CRS-NP + BA). Cytokine concentration was determined by means of ELISA tests for all the patient groups. We have found that the local concentrations of IL-1ra and IL-1β cytokines were significantly different (p < 0.05) from the group of healthy individuals, dependent on the clinical forms of chronic polyposis. IL-1β concentrations in the group of patients with CRS-NP (No. 2) were increased 5-fold, for group of CPRS-NP patients (No. 3), these levels showed a 15.5-fold increase. In the group of patients with CRS-NP + BA (No. 4), the Il-1 levels were 13fold higher in comparison with healthy controls. The IL-1ra concentration in the group of healthy people (No. 1) was 1825.64+463.70 pg/ml. Upon development of CRS-NP (No. 2), an increase in its values up to 3646.21±263.39 pg/ml was registered. Upon development of CPRS-NP (No. 3), an increase in appropriate values up to 2305.33±282.86 pg/ml was noted, and, in cases of CRS-NP + BA, a decrease in its values to 956.15±213.02 pg/ml was observed.Meanwhile, a conflicting character was revealed for the changes in these cytokines, thus preventing their usage as an indicator of polyposis severity. At the same time, estimation of the CIL-1ra/CIL- 1β ratio in nasal secretions of the patients in groups 1, 2, 3 and 4 based on two-dimensional projection of the spatial CIL-1ra/ CIL-1β distribution provides a basis for discrimination of the patient groups. The CIL-1ra/ CIL-1β ratio as assessed for nasal secretions of patients belonging to groups 1, 2, 3, and 4 shows a general, statistically significant decrease with increasing severity of chronic polyposis (p < 0.001), thus making this parameter indiformative for intensity of the pathological process. The resulting 95% confidence intervals confirm that the cytokine ratios are promising parameter in order to estimate the efficiency of immune therapy.

The monitoring of endocrine and immunologic markers during exercise is of paramount importance to assess and/or maintain the physical well-being of athletes as well as to optimize the athletic performance. This study aimed to investigate the linkage between acute stress response and immune status in Thoroughbred horses competing in an official 1300-m race. From 10 horses blood was collected 1 week before the day of the race (1W-BEFORE), before (REST) and immediately after the race to assess the cortisol, Interleukin-1 receptor antagonist (IL-1Ra), total proteins, white blood cells (WBC), red blood cells (RBC), haemoglobin (Hb) and haematocrit (Hct) concentration. Higher levels of cortisol, Il-1Ra, WBC and erythrocytes indices after exercise was found than 1W-BEFORE and REST (P0.0001). Cortisol concentration was positively correlated with Il-1Ra, WBC, RBC, Hb and Hct. Overall, the findings suggest that submaximal exercise induces an acute stress response and an immune system reaction in athletic horse. Also, the correlation found between cortisol levels and Il-1Ra, WBC and erythrocytes indices open new scenario on the positive role of this hormone on the complex and dynamic physiological adaptation to exercise implemented by the organism to re-establish the homeostatic equilibrium, and, interestingly, to maintain an adequate anti-inflammatory environment after exercise.

Multiple myeloma (MM) is characterized by the malignant proliferation of monoclonal plasma cells in the bone marrow with an elevation in monoclonal paraprotein, renal impairment, hypercalcemia, lytic bony lesions, and anemia. Immune cells and associated cytokines play a significant role in MM growth, progression, and dissemination. While some cytokines and their clinical significance are well described in MM biology, others remain relatively unknown. The present study examines the influence on progression-free survival (PFS) and overall survival (OS) by the serum levels of 27 selected cytokines in 61 newly diagnosed MM patients receiving first-line therapy with bortezomib-based regimens. The measurements were performed using a Bio-Rad Bio-Plex Pro Human Cytokine 27-Plex Assay and a MAGPIX Multiplex Reader, based on the Bio-Plex® 200 System (Bio-Rad). The following levels were determined: IL-1β, IL-1Ra, IL-2, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL-12, IL-13, IL-15, IL-17, Eotaxin, FGF, G-CSF, GM-CSF, IFN-γ, IP-10, MCP-1, MIP-1α, MIP-1β, PDGF-BB, RANTES, TNF-α, and VEGF. Most patients received a VCD chemotherapy regimen (bortezomib, cyclophosphamide, and dexamethasone). In the final multivariate model, IL-13 cytokine level (HR 0.1411, 95% CI: 0.0240–0.8291, p = 0.0302) and ASCT (HR 0.3722, 95% CI: 0.1826–0.7585, p = 0.0065) significantly impacted PFS. Furthermore, ASCT (HR 0.142, 95% CI: 0.046–0.438, p = 0.0007), presence of bone disease at diagnosis (HR 3.826, 95% CI: 1.471–9.949, p = 0.0059), and two cytokine levels—IL-1Ra (HR 1.017, 95% CI: 1.004–1.030, p = 0.0091) and IL-4 (HR 0.161, 95% CI: 0.037–0.698, p = 0.0147)—were independent predictors of OS. Three clusters of MM patients were identified with different cytokine profiles. In conclusion, serum pretreatment levels of IL-13 and IL-4 are predictors of better PFS and OS, respectively, whereas IL-1Ra pretreatment levels negatively impact OS in MM patients treated with bortezomib-based chemotherapy. Cytokine signature profile may have a potential influence on the outcome of patients treated with bortezomib.

Background/Purpose: Although an inflammatory response upon acute injury caused by ultraviolet radiation (UV) can be observed immediately, the influence of long-term, repetitive low-dose UV exposure on the skin cannot be precisely perceived, making early detection of chronic damage difficult. This study investigated bioactive substances in the stratum corneum as a potential early and sensitive indicator of the influence of sun exposure on the skin using receiver operating characteristic (ROC) analysis. Methods: Receiver operating characteristic analysis was performed to assess the responsiveness of cytokines [interleukin (IL)-1α, IL-1 receptor antagonist (IL-1ra), IL-10, tumor necrosis factor (TNF)-α], BCL2-associated protein X (Bax), Toll-like receptor (TLR)3, and TLR4 in the stratum corneum of healthy people exposed (dorsum of the hand) and unexposed (inner arm) to UV. Sunscreen was applied to patients with photodermatosis for 4 weeks to evaluate changes in IL-1ra/IL-1α, TNF-α, Bax, and TLR3 levels after sunscreen application, as these molecules exhibited high responsiveness to sun exposure according to ROC analysis. In addition, IL-1ra, IL-1α, and IL-10 levels were quantified by enzyme-linked immunosorbent assay, and TNF-α, Bax, TLR3, and TLR4 levels were semi-quantitatively assessed by immunocytochemistry. Results: Receiver operating characteristic analysis identified IL-1ra/IL-1α, TNF-α, Bax, and TLR3 in the stratum corneum as highly responsive to sun exposure. Moreover, in participants, including patients with photodermatosis, IL-1ra/IL-1α, TNF-α, and Bax levels decreased significantly after sunscreen application. Conclusion: The results revealed that IL-1ra/IL-1α, TNF-α, and Bax in the stratum corneum represent sensitive indicators of the influence of sun exposure on the skin.

Allergic conjunctivitis (AC) is the most prevalent form of mucosal allergy, and the conditioned medium (CM) from mesenchymal stem cells has been reported to attenuate some allergic diseases. However, the therapeutic effects of CM from different tissue stem cells (TSC-CM) on allergic diseases have not been tested. Here, we studied the effects of topical administration of different human TSC-CM on experimental AC (EAC) mice. Only human amniotic epithelial cell-CM (AECM) significantly attenuated allergic eye symptoms and reduced the infiltration of immune cells and the levels of local inflammatory factors in the conjunctiva compared to EAC mice. In addition, AECM treatment decreased immunoglobulin E (IgE) release, histamine production, and the hyperpermeability of conjunctival vessels. Protein chip assays revealed that the levels of anti-inflammatory factors, interleukin-1 receptor antagonist (IL-1ra) and IL-10, were higher in AECM compared to other TSC-CM. Furthermore, the anti-allergic effects of AECM on EAC mice were abrogated when neutralized with IL-1ra or IL-10 antibody, and the similar phenomenon was for the activation and function of B cells and mast cells. Together, the present study demonstrated that AECM alleviates EAC symptoms by multiple anti-allergic mechanisms mainlyviaIL-1ra and IL-10. Such topical AECM therapy may represent a novel and feasible strategy for treating AC.

Background: Neuroinflammation is a recognized aspect of Alzheimer’s disease (AD) and other neurological illnesses. Interleukin 1 receptor antagonist (IL-1Ra) is an anti-inflammatory molecule, which inhibits inflammatory molecules in different cells including brain cells. However, mechanisms for upregulating IL-1Ra in brain cells are poorly understood. Objective: Since aspirin is a widely available pain reliever that shows promise beyond its known pain-relieving capacity, we examined whether aspirin could upregulate the IL-1Ra in the brain. Methods: We employed PCR, real-time PCR, western blot, immunostaining, chromatin immunoprecipitation (ChIP), and lentiviral transduction in glial cells. 5xFAD mice, an animal model of AD, were treated with aspirin orally via gavage. Results: Aspirin increased the expression of IL-1Ra mRNA and protein in primary mouse astrocytes and mouse BV-2 microglial cells. While investigating the mechanism, we found that the IL-1Ra gene promoter harbors peroxisome proliferator response element (PPRE) and that aspirin upregulated IL-1Ra in astrocytes isolated from peroxisome proliferator-activated receptor-beta knockout (PPARβ–/–), but not PPARα–/–, mice. Moreover, we observed that aspirin bound to tyrosine 314 residue of PPARα to stimulate IL-1Ra and that aspirin treatment also increased the recruitment of PPARα to the IL-1Ra promoter. Accordingly, aspirin increased IL-1Ra in vivo in the brain of wild type and PPARβ–/–, but not in PPARα–/– mice. Similarly, aspirin treatment also increased astroglial and microglial IL-1Ra in the cortex of 5xFAD, but not 5xFAD/PPARα–/– mice. Conclusion: Aspirin may reduce the severity of different neurological conditions by upregulating IL-1Ra and reducing the inflammation.

Psoriasis (PS) is a skin disease with autoimmune features mediated by immune cells, which typically presents inflammatory erythematous plaques, and is associated with many comorbidities. PS exhibits excessive keratinocyte proliferation, and a high number of immune cells, including macrophages, neutrophils, Th1 and Th17 lymphocytes, and mast cells (MCs). MCs are of hematopoietic origin, derived from bone marrow cells, which migrate, mature, and reside in vascularized tissues. They can be activated by antigen-provoking overexpression of proinflammatory cytokines, and release a number of mediators including interleukin (IL)-1 and IL-33. IL-1, released by activated keratinocytes and MCs, stimulates skin macrophages to release IL-36—a powerful proinflammatory IL-1 family member. IL-36 mediates both innate and adaptive immunity, including chronic proinflammatory diseases such as psoriasis. Suppression of IL-36 could result in a dramatic improvement in the treatment of psoriasis. IL-36 is inhibited by IL-36Ra, which binds to IL-36 receptor ligands, but suppression can also occur by binding IL-38 to the IL-36 receptor (IL-36R). IL-38 specifically binds only to IL-36R, and inhibits human mononuclear cells stimulated with IL-36 in vitro, sharing the effect with IL-36Ra. Here, we report that inflammation in psoriasis is mediated by IL-1 generated by MCs—a process that activates macrophages to secrete proinflammatory IL-36 inhibited by IL-38. IL-37 belongs to the IL-1 family, and broadly suppresses innate inflammation via IL-1 inhibition. IL-37, in murine models of inflammatory arthritis, causes the suppression of joint inflammation through the inhibition of IL-1. Therefore, it is pertinent to think that IL-37 can play an inhibitory role in inflammatory psoriasis. In this article, we confirm that IL-38 and IL-37 cytokines emerge as inhibitors of inflammation in psoriasis, and hold promise as an innovative therapeutic tool.

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Background Autophagy induction is an effective approach for OA therapy. IL-1β is one of the major inflammatory cytokines linked to OA pathological progression, and its receptor blockade interrupts OA cartilage destruction. The objective of this study was to decipher the link between autophagy and regulatory mechanism of IL-1β and to investigate the effect of IL-1β receptor blockade by IL-1 receptor antagonist (IL-1Ra) combined with or without an autophagy inducer (TAT-Beclin1) on extracellular matrix (ECM) in OA chondrocytes in vitro and in vivo. Methods IL-1β-treated rat and human OA chondrocytes were cultured in response to IL-1Ra. The expression and distribution of signal molecules regulating ECM synthesis and autophagy were investigated via western blotting, immunoprecipitation, real-time PCR, immunofluorescence, and transmission electron microscope technique. Furthermore, after intra-articular injection of IL-1Ra, TAT-Beclin1, and a combination of both in a rat OA model established by anterior cruciate ligament transection and medial meniscus resection, the morphological changes of cartilage and related signal molecule expression levels were monitored using H.E., Safranin O-Fast green, and immunohistochemistry staining. Results Reduced autophagy by IL-1β contributed to ECM degradation, and blockade of IL-1β by IL-1Ra restored autophagy and attenuated ECM degradation in rat and human OA chondrocytes, as well as in a rat OA model. Akt/mTOR/ULK1, Akt/mTOR/NF-κB, and LC3B deacetylation were involved in autophagy regulated by IL-1β. Intra-articular injection of IL-1Ra combined with TAT-Beclin1 was more effective than IL-1Ra alone. Conclusions IL-1Ra restored autophagy and attenuated ECM degradation, with an implication that blocking IL-1β combined with enhancing autophagy might be a potential therapeutic strategy for OA. Electronic supplementary material The online version of this article (10.1186/s13075-019-1952-5) contains supplementary material, which is available to authorized users.

Summary Kawasaki disease (KD) vasculitis is an acute febrile illness of childhood characterized by systemic vasculitis of unknown origin, and is the most common cause of acquired heart disease among children in the United States. While histological evidence of myocarditis can be found in all patients with acute KD, only a minority of patients are clinically symptomatic and a subset demonstrate echocardiographic evidence of impaired myocardial function, as well as increased left ventricular mass, presumed to be due to myocardial edema and inflammation. Up to a third of KD patients fail to respond to first-line therapy with intravenous immunoglobulin (IVIG), and the use of interleukin (IL)-1 receptor antagonist (IL-1Ra, anakinra) is currently being investigated as an alternative therapeutic approach to treat IVIG-resistant patients. In this study, we sought to investigate the effect of IL-1Ra on myocardial dysfunction and its relation to myocarditis development during KD vasculitis. We used the Lactobacillus casei cell-wall extract (LCWE)-induced murine model of KD vasculitis and investigated the effect of IL-1Ra pretreatment on myocardial dysfunction during KD vasculitis by performing histological, magnetic resonance imaging (MRI) and echocardiographic evaluations. IL-1Ra pretreatment significantly reduced KD-induced myocardial inflammation and N-terminal pro B-type natriuretic peptide (NT-proBNP) release. Both MRI and echocardiographic studies on LCWE-injected KD mice demonstrated that IL-1Ra pretreatment results in an improved ejection fraction and a normalized left ventricular function. These findings further support the potential beneficial effects of IL-1Ra therapy in preventing the cardiovascular complications in acute KD patients, including the myocarditis and myocardial dysfunction associated with acute KD.

Objectives Previously, we reported the improved transfection efficiency of a plasmid DNA-chitosan (pDNA-CS) complex using a phosphorylatable nuclear localization signal-linked nucleic kinase substrate short peptide (pNNS) conjugated to chitosan (pNNS-CS). This study investigated the effects of pNNS-CS-mediated miR-140 and interleukin-1 receptor antagonist protein (IL-1Ra) gene transfection both in rabbit chondrocytes and a cartilage defect model. Methods The pBudCE4.1-miR-140, pBudCE4.1-IL-1Ra, and negative control pBudCE4.1 plasmids were constructed and combined with pNNS-CS to form pDNA/pNNS-CS complexes. These complexes were transfected into chondrocytes or injected into the knee joint cavity. Results High IL-1Ra and miR-140 expression levels were detected both in vitro and in vivo. In vitro, compared with the pBudCE4.1 group, the transgenic group presented with significantly increased chondrocyte proliferation and glycosaminoglycan (GAG) synthesis, as well as increased collagen type II alpha 1 chain (COL2A1), aggrecan (ACAN), and TIMP metallopeptidase inhibitor 1 (TIMP-1) levels. Nitric oxide (NO) synthesis was reduced, as were a disintegrin and metalloproteinase with thrombospondin type 1 motif 5 (ADAMTS-5) and matrix metalloproteinase (MMP)-13 levels. In vivo, the exogenous genes reduced the synovial fluid GAG and NO concentrations and the ADAMTS-5 and MMP-13 levels in cartilage. In contrast, COL2A1, ACAN, and TIMP-1 levels were increased, and the cartilage Mankin score was decreased in the transgenic group compared with the pBudCE4.1 group. Double gene combination produced greater efficacies than each single gene, both in vitro and in vivo. Conclusion This study suggests that pNNS-CS is a good candidate for treating cartilage defects via gene therapy, and that IL-1Ra in combination with miR-140 produces promising biological effects on cartilage defects. Cite this article: R. Zhao, S. Wang, L. Jia, Q. Li, J. Qiao, X. Peng. Interleukin-1 receptor antagonist protein (IL-1Ra) and miR-140 overexpression via pNNS-conjugated chitosan-mediated gene transfer enhances the repair of full-thickness cartilage defects in a rabbit model. Bone Joint Res 2019;8:165–178. DOI: 10.1302/2046-3758.83.BJR-2018-0222.R1.

Interleukin-1 receptor antagonist (IL-1Ra) is an active pharmaceutical ingredient obtained by the method of gene transformation of Escherichia coli, provides a neuroprotective effect, namely – inhibition of oxidative modification of proteins, normalization of functional activity of mitochondrial energy, mitochondrial energy. Early complex physical-chemical, microbiological, and biopharmaceutical studies have developed a new intranasal formulation, the formulation of which contains: IL-1Ra, sodium carboxymethyl cellulose, D-panthenol, benzalkonium chloride, trilon B, Tween-80, phosphate buffer solution. Given the peculiarities of the technological process, when creating a new gel dosage form, it is necessary to justify the temperature of the carrier base, the introduction of the active substance into the base, as well as to predict the possible chemical interaction of individual components in the dosage form. Therefore, the application of thermogravimetric analysis in pharmaceutical technology is relevant. The aim of the work is to conduct thermogravimetric studies of a new nasal form with an antagonist of interleukin-1β (IL-1Ra). Materials and methods. The objects of thermogravimetric studies were: semi-finished product-solution of the receptor antagonist of interleukin-1 (IL-1Ra). Excipients were: sodium carboxymethylcellulose, tween-80, D-panthenol, trilon B, benzalkonium chloride. To ensure the appropriate pH value used phosphate buffer solution (pH 6.0), according to the recipe to State Pharmacopoeia 2 ed. In addition to the individual components of the finished nasal gel, also prepared nasal gel without IL-1Ra, as well as gel with IL-1Ra. The thermographic analysis was performed on a derivatograph – Shimadzu DTG-60 (Japan) with a platinum-platinum-rhodium thermocouple when heating the samples in aluminum crucibles (from 15 °C to 250 °C). Al2O3 was used as a reference substance. The heating rate was 10 °C per minute. The weight of the test samples ranged from 19.22 mg to 52.91 mg. The derivatograph graphically recorded the obtained data in the form of curves T, DTA, TGA. The T curve on the derivatogram shows the change in temperature, and the TGA curve shows the change in mass of the sample during the study period. The DTA curve reflects the differentiation of thermal effects, contains information about endothermic and exothermic maxima, can be used for qualitative evaluation of the derivatogram. Results. The study consisted of two stages. At the first stage, we studied the derivatograms of experimental samples – individual ingredients of the finished nasal gel - active and excipients, namely: IL-1Ra, sodium carboxymethylcellulose, tween-80, D-panthenol, trilon B, benzalkonium chloride, phosphate buffer solution 6.0. In the second stage, samples of nasal gels were prepared: nasal gel with the required amount of excipients, in addition to the active substance – IL-1Ra, as well as a nasal gel containing IL-1Ra and the required amount of excipients. As a result, we obtained some parameters of thermogravimetric analysis in the form of a graphical image – derivatograms, which were then analyzed. Conclusions. Thermogravimetric studies of individual components of nasal gel with IL-1Ra, as well as nasal gels with active substance and without active substance, were found that the manufacturing process of gel with IL-1Ra should be carried out taking into account thermolabile compounds. First, you need to prepare the base without preservative and IL-1Ra. Then at a temperature not exceeding 40 °C dissolve the preservative, and then when cooled to 20 °C add IL-1Ra. It was found that the developed dosage form of the gel with IL-1Ra is a mixture of active and excipients, the ingredients of which do not interact with each other and can be combined in one dosage form., Антагонист рецепторов интерлейкина-1 (IL-1Ra) – активный фармацевтический ингредиент, который получают методом генной трансформации Escherichia coli, обеспечивающий нейропротекторный эффект, а именно торможение процессов окислительной модификации белков, нормализацию функциональной активности митохондрий, энергетического обмена, неврологического статуса в постишемическом периоде. В ходе предварительных комплексных физико-химических, микробиологических и биофармацевтических исследований разработан состав новой интраназальной формы, рецептура которой включает IL-1Ra, натрий КМЦ, D-пантенол, бензалкония хлорид, трилон Б, твин-80, фосфатный буферный раствор – рН 6.0 (ГФУ 2). Учитывая особенности технологического процесса, при создании новой гелевой лекарственной формы необходимо обосновать температурный режим изготовления основы-носителя, введение действующего вещества в основу, а также спрогнозировать возможное химическое взаимодействие отдельных компонентов. Поэтому актуальным является применение термогравиметрического анализа в фармацевтической технологии. Цель работы – проведение термогравиметрических исследований новой назальной формы с антагонистом интерлейкина-1β (IL-1Ra). Материалы и методы. В качестве объектов термогравиметрических исследований использовали полуфабрикат-раствор рецепторного антагониста интерелейкина-1 β (IL-1Ra). Использовали вспомогательные вещества: натрий карбоксиметилцеллюлозу, твин-80, D-пантенол, трилон Б, бензалкония хлорид. Для обеспечения соответствующего значения рН использовали фосфатный буферный раствор (рН 6.0) по рецептуре согласно ГФУ 2 издания. Кроме отдельных компонентов готового назального геля готовили назальный гель без IL-1Ra, а также гель с IL-1Ra. Термографический анализ выполнен на дериватографе Shimadzu DTG-60 (Япония) с платиново-платинородиевый термопарой при нагревании образцов в алюминиевых тиглях (от 15 °С до 250 °С). Как эталонную субстанцию использовали α-Al2O3. Скорость нагрева составляла 10 ºС в минуту. Масса исследуемых образцов – от 19,22 мг до 52,91 мг. Полученные данные дериватограф графически фиксировал в виде кривых T, DTA, TGA. Кривая Т на дериватограмме показывает изменение температуры, кривая TGA – изменение массы образца в период исследования. Кривая DTA отражает дифференцирование тепловых эффектов, содержит информацию об эндотермических и экзотермических максимумах, может быть использована для качественной оценки дериватограммы. Результаты. Исследование состояло из двух этапов. На первом изучали дериватограммы опытных образцов – отдельных ингредиентов готового назального геля (действующие и вспомогательные вещества: IL-1Ra, натрий карбоксиметилцеллюлоза, твин-80, D-пантенол, трилон Б, бензалкония хлорид, фосфатный буферный раствор – рН 6.0)). На втором этапе готовили образцы назальных гелей: назальный гель с необходимым количеством вспомогательных веществ, без действующего вещества – IL-1Ra, а также назальный гель, содержащий IL-1Ra и необходимое количество вспомогательных веществ. В результате получили некоторые параметры термогравиметрического анализа в виде графического изображения – дериватограмм, которые затем анализировали. Выводы. В ходе термогравиметрических исследований отдельных компонентов назального геля с IL-1Ra, а также назальных гелей с действующим веществом и без него установлено: технологический процесс изготовления геля с IL-1Ra целесообразно проводить с учетом термолабильных соединений. Сначала нужно приготовить основу без консерванта и IL-1Ra. Далее при температуре не выше 40 °С растворить консервант, а затем при охлаждении до 20 °С добавить IL-1Ra. Разработанная лекарственная форма геля с IL-1Ra – смесь действующих и вспомогательных веществ, ингредиенты которых не взаимодействуют и могут сочетаться., Антагоніст рецепторів інтерлейкіну-1 (ІL-1Ra) – активний фармацевтичний інгредієнт, який отримують методом генної трансформації Escherichia coli, забезпечує нейропротекторний ефект, як-от гальмування процесів окислювальної модифікації білків, нормалізацію функціональної активності мітохондрій, енергетичного обміну, неврологічного статусу в постішемічному періоді. Впродовж попередніх комплексних фізико-хімічних, мікробіологічних і біофармацевтичних досліджень розробили склад нової інтраназальної форми, рецептура включає ІL-1Ra, натрій КМЦ, D-пантенол, бензалконію хлорид, трилон Б, твін-80, фосфатний буферний розчин – рН 6.0 (ДФУ 2). Враховуючи особливості технологічного процесу, під час створення нової гелевої лікарської форми необхідно обґрунтувати температурний режим виготовлення основи-носія, введення діючої речовини в основу, а також спрогнозувати можливу хімічну взаємодію окремих компонентів. Тому актуальним є застосування термогравіметричного аналізу в фармацевтичній технології. Мета роботи – здійснити термогравіметричне дослідження нової назальної форми з антагоністом інтерлейкіну-1β (IL-1Ra). Матеріали та методи. Як об’єкти термогравіметричних досліджень використовували напівфабрикат-розчин рецепторного антагоніста інтерелейкіну-1 (IL-1Ra). Використали допоміжні речовини: натрій карбоксиметилцелюлозу, твін-80, D-пантенол, трилон Б, бензалконію хлорид. Для забезпечення відповідного значення рН застосовували фосфатний буферний розчин (рН 6.0) за рецептурою згідно з ДФУ 2 видання. Крім окремих компонентів готового назального гелю, готували назальний гель без IL-1RA, а також гель з IL-1Ra. Термографічний аналіз виконали на дериватографі Shimadzu DTG-60 (Японія) з платиново-платинородієвою термопарою при нагріванні зразків в алюмінієвих тиглях (від 15°С до 250°С). Як еталонну субстанцію використовували α-Al2O3. Швидкість нагрівання – 10 ºС за хвилину. Маса зразків – від 19,22 мг до 52,91 мг. Результати дериватограф графічно фіксував як криві T, DTA, TGA. Крива Т на дериватограмі показує зміну температури, крива TGA – зміну маси зразка в період дослідження. Крива DTA показує диференціювання теплових ефектів, містить інформацію про ендотермічні та екзотермічні максимуми, її можна використовувати для якісного оцінювання дериватограми. Результати. Дослідження складалося з двох етапів. На першому вивчали дериватограми дослідних зразків, окремих інгредієнтів готового назального гелю (діюча та допоміжні речовини: IL-1Ra, натрій карбоксиметилцелюлоза, твін-80, D-пантенол, трилон Б, бензалконію хлорид, фосфатний буферний розчин – рН 6.0). На другому етапі готували зразки назальних гелів: назальний гель із необхідною кількістю допоміжних речовин, крім діючої речовини – IL-1Ra, а також назальний гель, що містив IL-1Ra та необхідну кількість допоміжних речовин. У результаті отримали окремі параметри термогравіметричного аналізу у вигляді графічного зображення – дериватограми, які потім аналізували. Висновки. Під час термогравіметричних досліджень окремих компонентів назального гелю з IL-1Ra, а також назальних гелів із діючою речовиною та без неї встановили: технологічний процес виготовлення гелю з IL-1Ra доцільно проводити, враховуючи термолабільні сполуки. Спочатку потрібно приготувати основу без консерванта й IL-1Ra. Далі за температури не більше ніж 40 °С розчинити консервант, а потім при охолодженні до 20 °С додати IL-1Ra. Розроблена лікарська форма гелю з IL-1Ra – суміш діючих і допоміжних речовин, інгредієнти яких не взаємодіють і можуть поєднуватися.

Many alternative treatments aimed at modulating osteoarthritis (OA) progression have been developed in the past decades, including the use of cytokine inhibitors. IL-1β is considered one of the most impactful cytokines in OA disease and therefore, its blockage offers a promising approach for the modulation of OA. Interleukin-1 receptor antagonist (IL-1Ra) is a naturally occurring anti-inflammatory protein belonging to the IL-1 family that competes with IL-1β for occupancy of its receptors, without triggering the same downstream inflammatory response. Because of its natural anti-inflammatory properties, different methods have been proposed to use IL-1Ra therapeutically in OA. Autologous conditioned serum (ACS) and autologous protein solution (APS) are blood-derived products produced with the use of specialized commercial kits. These processes result in hemoderivatives with high concentrations of IL-1Ra and other cytokines and growth factors with potential modulatory effects on OA progression. Several studies have demonstrated potential anti-inflammatory effect of these therapies with promising clinical results. However, as with any hemoderivatives, clinical outcomes may vary. For optimal therapeutic use, further research is warranted for a more comprehensive understanding of the product's composition and interaction of its components in joint inflammation. Additionally, differences between ACS and APS treatments may not be clear for many clients and clinicians. Thus, the objective of this narrative review is to guide the reader in important aspects of ACS and APS therapies, in vitro and in vivo applications and to compare the use of both treatments in OA.

Background Increased perioperative pro-inflammatory biomarkers, renal hypoperfusion and ischemia reperfusion injury (IRI) heighten cardiac surgery acute kidney injury (CS-AKI) risk. Increased urinary anti-inflammatory cytokines attenuate risk. We evaluated whether blood and urinary anti-inflammatory biomarkers, when expressed as ratios with biomarkers of inflammation, hypoperfusion and IRI are increased in CS-AKI patients. Methods Preoperative and 24-h postoperative blood and urinary pro-inflammatory and anti-inflammatory cytokines, blood VEGF and H-FABP (hypoperfusion biomarkers), and MK, a biomarker for IRI, were measured in 401 cardiac surgery patients. Pre- and postoperative concentrations of biomarkers and selected ratios thereof, were compared between non-CS-AKI and CS-AKI patients. Results Compared with non-CS-AKI, blood pro-inflammatory (pre- and post-op TNFα, IP-10, IL-12p40, MIP-1α, NGAL; pre-op IL-6; post-op IL-8, MK) and anti-inflammatory (pre- and post-op sTNFsr1, sTNFsr2, IL-1RA) biomarkers together with urinary pro-inflammatory (pre- and post-op uIL-12p40; post-op uIP-10, uNGAL) and anti-inflammatory (pre- and post-op usTNFsr1, usTNFsr2, uIL-1RA) biomarkers, were significantly higher in CS-AKI patients. Urinary anti-inflammatory biomarkers, when expressed as ratios with biomarkers of inflammation (blood and urine), hypoperfusion (blood H-FABP and VEGF) and IRI (blood MK) were decreased in CS-AKI. In contrast, blood anti-inflammatory biomarkers expressed as similar ratios with blood biomarkers were increased in CS-AKI. Conclusions The urinary anti-inflammatory response may protect against the injurious effects of perioperative inflammation, hypoperfusion and IRI. These finding may have clinical utility in bioprediction and earlier diagnosis of CS-AKI and informing future therapeutic strategies for CS-AKI patients.

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Background: Hydrogen sulfide(H2S) is a bacterial metabolite produced as a result of bacterial growth in subgingival pockets, suggested to partake in the pathogenesis of periodontitis. H2S has previously been shown to induce the secretion of the pro-inflammatory cytokines IL-1β and IL-18 via the NLRP3 inflammasome in monocytes. Objective: To investigate the non-NLRP3 inflammasome-dependent immunological response of human peripheral blood mononuclear cells (PBMCs) of periodontitis patients and healthy controls exposed to H2S in vitro. Methods: PBMCs of periodontitis patients(N = 31) and healthy controls(N = 32) were exposed to 1 mM sodium hydrosulfide (NaHS) at 37°C for 24 h and the secretion of cytokines was compared to resting cells. TNF-α, IFN-γ, IL-6, IL-8, IL-12p40, IL-12p70, IL-17, MCP-1, and IL-1Ra secretions were measured with Bio-Plex Pro™ Human Cytokine Assay. Results: H2S triggered the secretion of the pro-inflammatory IFN-γ, IL-6, IL-17, TNF-α, IL-12p40, and IL-12p70, while the reverse was seen for IL-1Ra. In addition, a higher basal secretion of IFN-γ, IL-6, IL-12p70, IL-17 and MCP-1 was seen from PBMCs of periodontitis patients compared to healthy controls. Conclusion: The bacterial metabolite H2S triggers the secretion of pro-inflammatory cytokines from PBMCs and may thus have a prominent role in the host-bacteria interplay in periodontitis.

International audience; Interleukin-1 beta (IL-1β), a pro-inflammatory cytokine, has been ascribed a role in the expansion of myeloid progenitors in acute myeloid leukemia (AML) and in promoting myeloid cell-induced suppression of lymphocyte-mediated immunity against malignant cells. This study aimed at defining the potential impact of IL-1β in the post-remission phase of AML patients receiving immunotherapy for relapse prevention in an international phase IV trial of 84 patients (ClinicalTrials.gov; NCT01347996). Consecutive serum samples were collected from AML patients in first complete remission (CR) who received cycles of relapse-preventive immunotherapy with histamine dihydrochloride (HDC) and lowdose interleukin-2 (IL-2). Low IL-1β serum levels before and after the first HDC/IL-2 treatment cycle favorably prognosticated leukemia-free survival and overall survival. Serum levels of IL-1β were significantly reduced in patients receiving HDC/IL-2. HDC also reduced the formation of IL-1β from activated human PBMCs in vitro. Additionally, high serum levels of the IL-1 receptor antagonist IL-1RA were associated with favorable outcome, and AML patients with low IL-1β along with high IL-1RA levels were strikingly protected against leukemic relapse. Our results suggest that strategies to target IL-1β might impact on relapse risk and survival in AML.

Although the range of immune responses to severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is variable, cytokine storm is observed in a subset of symptomatic individuals. To further understand the disease pathogenesis and, consequently, to develop an additional tool for clinicians to evaluate patients for presumptive intervention, we sought to compare plasma cytokine levels between a range of donor and patient samples grouped by a COVID-19 Severity Score (CSS) based on the need for hospitalization and oxygen requirement. Here we utilize a mutual information algorithm that classifies the information gain for CSS prediction provided by cytokine expression levels and clinical variables. Using this methodology, we found that a small number of clinical and cytokine expression variables are predictive of presenting COVID-19 disease severity, raising questions about the mechanism by which COVID-19 creates severe illness. The variables that were the most predictive of CSS included clinical variables such as age and abnormal chest x-ray as well as cytokines such as macrophage colony-stimulating factor, interferon-inducible protein 10, and interleukin-1 receptor antagonist. Our results suggest that SARS-CoV-2 infection causes a plethora of changes in cytokine profiles and that particularly in severely ill patients, these changes are consistent with the presence of macrophage activation syndrome and could furthermore be used as a biomarker to predict disease severity.

Background: Circulating markers are attractive molecules for prognosis and management of cancer that allow sequential monitoring of patients during and after treatment. Based on previous protein profiling data, circulating interleukin 1 receptor antagonist (IL-1Ra) was evaluated as a potential diagnostic and prognostic marker for squamous cell carcinomas of the head and neck (SCCHN). In this study, we aimed at confirming the clinical relevance of plasma IL-1Ra in SCCHN and exploring its potential as a prediction marker for SCCHN. Methods: Plasma from 87 patients with SCCHN, control plasma from 28 healthy individuals and pre-diagnostic plasma from 44 patients with squamous cell carcinoma of the oral tongue (SCCOT) and 88 matched controls were analysed with IL-1Ra electrochemiluminescence immunoassays from mesoscale diagnostics. Results: Plasma IL-1Ra was found to be up-regulated in patients with oral tongue, gingiva and base of tongue tumours compared to healthy individuals (p

Sequence variants in genes involved in the immune system have previously been linked to neutropenia as well as infections in cancer patients. Sequence variants in genes coding for TLR4, MBL, and IL-1Ra were investigated in relation to clinical utility of identifying severe episodes of febrile neutropenia (FN) in a cohort of children undergoing treatment for acute lymphoblastic leukemia. The study included 122 children, where data on FN and microbiological findings were retrospectively collected from medical records. Sequence variants in genes coding for MBL, TLR4, and IL-1Ra were identified by pyrosequencing, TaqMan SNP genotyping assay, and gel electrophoresis. A total of 380 episodes of FN were identified and in 139 episodes, there was a microbiological defined infection. Age and treatment intensity were all associated with the risk of developing FN. No sequence variant was associated to increased numbers of FN episodes. Two sequence variants in the TLR4 gene increased the risk of viral infection, whilst sequence variants in the IL-1Ra gene were associated to a decreased risk of bacterial blood-stream infection (BSI). The investigated sequence variants did not associate with increased risk for FN or to severe infections, as to why the clinical utility as a risk-stratification tool is low. Most episodes of FN were classified as fever with unknown origin, emphasizing the need for improved microbial detection methods. Funding Agencies|Swedish Childhood Cancer Foundation; Swedish Society of Medicine

(1) Background: Hospital mortality in patients suffering from SARS-CoV-2 infection has been associated with thrombocytopenia. The present study was conducted to establish the correlation of thrombocytopenia and the severity of infection. The impact of IL-1Ra gene polymorphism on the incidence and severity of thrombocytopenia was also studied. (2) Methods: Various biochemical parameters measured in all the 1200 enrolled patients included full blood counts, renal and liver function tests, iron study, inflammatory markers, and coagulation assays. A further 70 patients each were selected from the severe thrombocytopenic and non-thrombocytopenic patient groups to study the IL-1Ra gene polymorphism by RCR. (3) Results: Out of 1200 patients, 436 (36.3%) had thrombocytopenia. Among these patients, 118 (27.1%), 75 (17.2%), and 42 (9.6%) had mild, moderate, severe, and very severe thrombocytopenia, respectively. Severe cases mostly resulted from peripheral consumption (73.5%), hemo-phagocytosis (15.4%), and bone marrow suppression (11.11%). A statistically significant correlation was found between the occurrence and severity of thrombocytopenia with perturbated levels of inflammatory markers and the presence of comorbidities. The IL-1Ra∗3 variant was found to be significantly associated with thrombocytopenia. The IL-1Ra∗2 variant was significantly seen among controls. (4) Conclusions: The present study revealed a significant correlation between thrombocytopenia and the severity of COVID-19 disease. Moreover, the IL-1Ra∗3 variant of IL-1Ra gene was associated with thrombocytopenia.

Sequence variants in genes involved in the immune system have previously been linked to neutropenia as well as infections in cancer patients. Sequence variants in genes coding for TLR4, MBL, and IL-1Ra were investigated in relation to clinical utility of identifying severe episodes of febrile neutropenia (FN) in a cohort of children undergoing treatment for acute lymphoblastic leukemia. The study included 122 children, where data on FN and microbiological findings were retrospectively collected from medical records. Sequence variants in genes coding for MBL, TLR4, and IL-1Ra were identified by pyrosequencing, TaqMan SNP genotyping assay, and gel electrophoresis. A total of 380 episodes of FN were identified and in 139 episodes, there was a microbiological defined infection. Age and treatment intensity were all associated with the risk of developing FN. No sequence variant was associated to increased numbers of FN episodes. Two sequence variants in the TLR4 gene increased the risk of viral infection, whilst sequence variants in the IL-1Ra gene were associated to a decreased risk of bacterial blood-stream infection (BSI). The investigated sequence variants did not associate with increased risk for FN or to severe infections, as to why the clinical utility as a risk-stratification tool is low. Most episodes of FN were classified as fever with unknown origin, emphasizing the need for improved microbial detection methods.

Background Many laboratory parameters have been associated with morbidity and mortality in SARS‐CoV‐2 (COVID‐19), which emerged in an animal market in Wuhan, China in December 2019 and has infected over 20 million people. This study investigated the relationship between serum interleukin (IL)‐18, IL‐1 receptor antagonist (IL‐1Ra), and alpha defensin levels and the clinical course and prognosis of COVID‐19. Materials and Methods This study included 100 patients who were admitted to the chest diseases department and intensive care unit of our hospital and diagnosed with COVID‐19 by real‐time polymerase chain reaction (PCR) of nasopharyngeal swab samples between March 24 and May 31, 2020. The control group consisted of 50 nonsymptomatic health workers with negative real‐time PCR results in routine COVID‐19 screening in our hospital. Results Serum alpha defensin, IL‐1Ra, and IL‐18 levels were significantly higher in patients who developed macrophage activation syndrome (MAS) and acute respiratory distress syndrome (ARDS) compared to patients who did not (p < .001 for all). Alpha defensin, IL‐1Ra, and IL‐18 levels were significantly higher in COVID‐19 patients with and without MAS or ARDS when compared to the control group (p < .001 for all). When the 9 patients who died were compared with the 91 surviving patients, IL‐1Ra and IL‐18 levels were found to be significantly higher in the nonsurvivors (p < .001). Conclusion Our findings of correlations between alpha defensin and levels of IL‐1Ra and IL‐18, which were previously shown to be useful in COVID‐19 treatment and follow‐up, indicates that it may also be promising in treatment.

International audience; Despite early antiretroviral therapy (ART), treatment interruption is associated with viral rebound, indicating early viral reservoir (VR) seeding and absence of full eradication of human immunodeficiency virus type 1 (HIV-1) that may persist in tissues. Herein, we address the contributing role of monocytes in maintaining VRs under ART, since these cells may represent a source of viral dissemination due to their ability to replenish mucosal tissues in response to injury. To this aim, monocytes with classical (CD14+), intermediate (CD14+ CD16+), and nonclassical (CD16+) phenotypes and CD4+ T cells were sorted from the blood, spleen, and intestines of untreated and early-ART-treated simian immunodeficiency virus (SIV)-infected rhesus macaques (RMs) before and after ART interruption. Cell-associated SIV DNA and RNA were quantified. We demonstrated that in the absence of ART, monocytes were productively infected with replication-competent SIV, especially in the spleen. Reciprocally, early ART efficiently (i) prevented the establishment of monocyte VRs in the blood, spleen, and intestines and (ii) reduced systemic inflammation, as indicated by changes in interleukin-18 (IL-18) and IL-1 receptor antagonist (IL-1Ra) plasma levels. ART interruption was associated with a rebound in viremia that led to the rapid productive infection of both CD4+ T cells and monocytes. Altogether, our results reveal the benefits of early ART initiation in limiting the contribution of monocytes to VRs and SIV-associated inflammation.IMPORTANCE Despite the administration of antiretroviral therapy (ART), HIV persists in treated individuals and ART interruption is associated with viral rebound. Persistent chronic immune activation and inflammation contribute to disease morbidity. Whereas monocytes are infected by HIV/SIV, their role as viral reservoirs (VRs) in visceral tissues has been poorly explored. Our work demonstrates that monocyte cell subsets in the blood, spleen, and intestines do not significantly contribute to the establishment of early VRs in SIV-infected rhesus macaques treated with ART. By preventing the infection of these cells, early ART reduces systemic inflammation. However, following ART interruption, monocytes are rapidly reinfected. Altogether, our findings shed new light on the benefits of early ART initiation in limiting VR and inflammation.

Category: Ankle Arthritis; Basic Sciences/Biologics Introduction/Purpose: Post-traumatic osteoarthritis (PTOA) is a frequent cause of disability. The most common predisposing factor for ankle PTOA is intra-articular ankle fracture. It has been hypothesized that an early inflammatory response after intra- articular injury could lead to cartilage damage. Our group recently demonstrated significantly elevated inflammatory cytokines in the synovial fluid fracture hematoma (SFFH) immediately after intra-articular ankle fracture. The negative effect that this inflammatory environment has on cartilage is unknown but may be the reason for PTOA development. The purpose of this study was to determine if cartilage occurred when exposed to intra-articular SFFH and if anti-inflammatory agents could attenuate this damage. Methods: SFFH was obtained from 52 intra-articular ankle fractures at the time of surgery. Samples were divided into three groups based on time from fracture: acute (0-2 days), intermediate (3-9 days), late (>=10 days). A biopsy punch was used to create 5mm cartilage discs from the medial and lateral shoulders of fresh human talus cartilage. The cartilage discs were weighed and cultured in standard chondrocyte media (SCM) for 3 days. Next, the discs were randomly assigned to one of 4 groups (n=10 each) and cultured as follows for 6 days: A=control, SCM only; Group B=acute SFFH+SCM; Group C=intermediate SFFH+SCM; Group D=late SFFH+SCM. After 6 days of culture, the discs were rinsed and transferred to new wells and cultured for 3 more days in SCM only, to determine inflammatory cytokine and MMP release from the cartilage discs (cartilage damage). The post SFFH spent culture media was collected. The initial and post SFFH culture spent media were analyzed for inflammatory cytokines, MMPs, CTXII (cartilage breakdown marker), and sGAG. Safranin-O staining was then performed on the discs. The initial and post SFFH media from groups A, B, C, and D were compared to determine if there was a difference in damage caused by SFFH based on time from fracture. Based on the results, Group B (early) was felt to cause the most damage. Therefore, the following treatment groups were created and cultured exactly as before: Group E=same as Group B + IL-1Ra treatment; Group F=same as Group B + doxycycline treatment. Groups B, E, and F were compared to determine if the addition of IL-1Ra or doxycycline (known MMP inhibitor) attenuated the damage. Results: Early, intermediate, and late SFFH all caused some degree of cartilage damage in the form of significantly elevated inflammatory cytokine and MMP production from the cartilage discs ( Figure 1 ). The early SFFH (Group B) was noted to cause significantly elevated production of IL-6, IL-8, CTXII, and decreased safranin-O staining. Therefore, it was chosen to compare with the therapeutic anti-inflammatory agents. The addition of IL-1Ra or doxycycline significantly reduced the production of IL-6, IL-8, CTXII, and MMPs 3, 9, and 10 ( Figure 2 ). Conclusion: This is the first study to show that intra-articular hematoma immediately after ankle fracture causes cartilage damage that can be partially attenuated by current clinically available therapeutic agents. Additionally, further research should be performed regarding the safety and efficacy of anti-inflammatory agents to prevent ankle PTOA.

Although most bone fractures typically heal without complications, a small proportion of patients (≤10%) experience delayed healing or potential progression to non-union. Interleukin-1 (IL-1β) plays a crucial role in fracture healing as an early driver of inflammation. However, the effects of IL-1β can impede the healing process if they persist long after the establishment of a fracture hematoma, making it a promising target for novel therapies. Accordingly, the overall objective of this study was to develop a novel gene-based therapy that mitigates the negative effects of IL-1β-driven inflammation while providing a structural template for new bone formation. A collagen-hydroxyapatite scaffold (CHA) was used as a platform for the delivery of nanoparticles composed of pDNA, encoding for IL-1 receptor antagonist (IL-1Ra), complexed to the robust non-viral gene delivery vector, polyethyleneimine (PEI). Utilizing pDNA encoding for Gaussia luciferase and GFP as reporter genes, we found that PEI-pDNA nanoparticles induced a transient gene expression profile in rat bone marrow-derived mesenchymal stromal cells (BM-MSCs), with a transfection efficiency of 14.8 ± 1.8% in 2D. BM-MSC viability was significantly affected by PEI-pDNA nanoparticles as evaluated using CellTiter Blue; however, after 10 days in culture this effect was negligible. Transfection with PEI-pIL-1Ra nanoparticles led to functional IL-1Ra production, capable of antagonizing IL-1β-induced expression of secreted embryonic alkaline phosphatase from HEK-Blue-IL-1β reporter cells. Sustained treatment with IL-1β (0.1, 1, and 10 ng/ml) had a dose-dependent negative effect on BM-MSC osteogenesis, both in terms of gene expression (Alpl and Ibsp) and calcium deposition. BM-MSCs transfected with PEI-IL-1Ra nanoparticles were found to be capable of overcoming the inhibitory effects of sustained IL-1β (1 ng/ml) treatments on in vitro osteogenesis. Ultimately, IL-1Ra gene-activated CHA scaffolds supported mineralization of BM-MSCs under chronic inflammatory conditions in vitro, demonstrating potential for future therapeutic applications in vivo.

Summary: Background: Screening for pancreatic ductal adenocarcinoma (PDAC) in populations at high risk is recommended. Individuals with new-onset type 2 diabetes mellitus (NOD) are the largest high-risk group for PDAC. To facilitate screening, we sought biomarkers capable of stratifying NOD subjects into those with type 2 diabetes mellitus (T2DM) and those with the less prevalent PDAC-related diabetes (PDAC-DM), a form of type 3c DM commonly misdiagnosed as T2DM. Methods: Using mass spectrometry- and immunoassay-based methodologies in a multi-stage analysis of independent sample sets (n=443 samples), blood levels of 264 proteins were considered using Ingenuity Pathway Analysis, literature review and targeted training and validation. Findings: Of 30 candidate biomarkers evaluated in up to four independent patient sets, 12 showed statistically significant differences in levels between PDAC-DM and T2DM. The combination of adiponectin and interleukin-1 receptor antagonist (IL-1Ra) showed strong diagnostic potential, (AUC of 0.91; 95% CI: 0.84-0.99) for the distinction of T3cDM from T2DM. Interpretation: Adiponectin and IL-1Ra warrant further consideration for use in screening for PDAC in individuals newly-diagnosed with T2DM. Funding: North West Cancer Research, UK, Cancer Research UK, Pancreatic Cancer Action, UK.

Extracellular interleukin 1 alpha (IL-1α) released from keratinocytes is one of the endpoints for in vitro assessments of skin irritancy. Although cells dying via primary skin irritation undergo apoptosis as well as necrosis, IL-1α is not released in apoptotic cells. On the other hand, active secretion has been identified in interleukin-1 receptor antagonist (IL-1ra), which was discovered to be a common, upregulated, differentially-expressed gene in a microarray analysis performed with keratinocytes treated using cytotoxic doses of chemicals. This study examined whether and how IL-1ra, particularly extracellularly released IL-1ra, was involved in chemically-induced keratinocyte cytotoxicity and skin irritation. Primary cultured normal adult skin keratinocytes were treated with cytotoxic doses of chemicals (hydroquinone, retinoic acid, sodium lauryl sulfate, or urshiol) with or without recombinant IL-1ra treatment. Mouse skin was administered irritant concentrations of hydroquinone or retinoic acid. IL-1ra (mRNA and/or intracellular/extracellularly released protein) levels increased in the chemically treated cultured keratinocytes with IL-1α and IL-1β mRNAs and in the chemically exposed epidermis of the mouse skin. Recombinant IL-1ra treatment significantly reduced the chemically-induced apoptotic death and intracellular/extracellularly released IL-1α and IL-1β in keratinocytes. Collectively, extracellular IL-1ra released from keratinocytes could be a compensatory mechanism to reduce the chemically-induced keratinocyte apoptosis by antagonism to IL-1α and IL-1β, suggesting potential applications to predict skin irritation.

Previously, we reported that electroporation-mediated (EP) delivery of the FER gene improved survival in a combined trauma-pneumonia model. The mechanism of this protective effect is unknown. In this paper, we performed a pneumonia model in C57/BL6 mice with 500 CFU of Klebsiella pneumoniae. After inoculation, a plasmid encoding human FER was delivered by EP into the lung (PNA/pFER-EP). Survival of FER-treated vs. controls (PNA; PNA/EP-pcDNA) was recorded. In parallel cohorts, bronchial alveolar lavage (BAL) and lung were harvested at 24 and 72 h with markers of infection measured. FER-EP-treated animals reduced bacterial counts and had better 5-day survival compared to controls (80 vs. 20 vs. 25%; p

Introduction Among the broad spectrum of cytokines, interleukin 1-β (IL-1β) has been implicated in induction and subsequent aggravation of skin lesions in atopic dermatitis (AD). A considerable body of evidence suggests that vitamin D status also influences the risk and/or severity of AD. Material and methods Fifty-seven children suffering from mild to severe AD were enrolled in the study. The control group consisted of 33 matched healthy children. In all the children serum concentrations of IL-1β/IL-1F2 and the interleukin-1 receptor antagonist IL-Ra/1F3 were measured. Serum 25(OH)D concentration was obtained for 49 patients with AD and all healthy children. Results In children with AD 59.2% of children had insufficiency, 24.5% had deficiency and 16.3% had a sufficient serum 25(OH)D level. In the control group 26.5%, 52.9% and 20% of participants had insufficiency/deficiency/sufficiency of 25(OH)D, respectively. The severity of AD was positively correlated with total IgE level, percentage and absolute count of eosinophils and IL-1Ra. IL-1β correlated with IL-1Ra. Conclusions In children with AD the serum vitamin D level was lower than in healthy children. The correlation between severity of AD and IL-1Ra may prove that inflammasome-dependent IL-1β is involved in immunopathogenesis of the disease. Further studies are needed on a larger population of children to confirm the role of this cytokine in development of AD.

This study was aimed to give a better understanding of the mechanisms of early immune response to trauma by assessing the concentration of cytokines in peripheral blood. The study group comprised 32 patients admitted to the Emergency Department due to injury. Depending on the magnitude of the Injury Severity Score (ISS) trauma patients were divided into two groups. In group A (ISS ≥ 20), 13 patients had complications, and five died, while in group B (ISS < 20) only three patients had complications (e.g. respiratory failure and infections). Depending on the extent of the injury, significant differences were observed in the concentrations of cytokines in the treatment groups. The highest levels of IL-6 and IL-1Ra in both groups were recorded in the third hour of hospitalisation and were considerably higher in group A compared to the concentration of these cytokines in group B (p = 0.001). In patients with complications, IL-6 and IL-1Ra concentrations were significantly higher compared to those without complications. Spearman’s rho-correlation showed a statistically significant positive correlation between baseline concentrations of IL-6 (r = 0.64, p < 0.001) and IL-1Ra (r = 0.37, p = 0.042) and the values of the ISS. A high diagnostic sensitivity calculated from ROC curves was found for IL-6 concentrations. In summary, our findings suggest that elevated levels of the cytokines tested, determined in the peripheral blood shortly after injury, may be significantly associated with the occurrence of severe complications, which in some patients can lead to death. Monitoring the levels of these cytokines in patients with a high risk of serious complications should be used routinely.

Background Mast cells are resident immune effector cells, often studied in the context of allergic disease. Found in substantial numbers at sites of potential infection they are increased at sites of angiogenesis and can be pivotal for the sensing and clearance of a variety of pathogens. Interferons (IFNs) are cytokines that are critical for host defence against intracellular pathogens. Increased levels of IFNs are observed during viral infection and in autoimmune diseases. IFNs are also widely used therapeutically and have been examined in the therapy of severe asthma. Objective To define the selective human mast cell cytokine and chemokine response following activation with type I or type II IFN's. Methods The ability of both IFNα2 and IFNγ to induce cytokine production by human cord blood‐derived mast cells was examined in vitro. Cytokine and chemokine production at 6 and 24 h was assessed by multiplex protein analysis. Degranulation was assessed by β‐hexosaminidase release. Mast cells were also treated with reovirus or respiratory syncytial virus and their production of CXCL10, IL‐1 receptor antagonist (IL‐1Ra), and vascular endothelial growth factor (VEGF) examined after 24 h. Results In addition to increased expression of classical IFN response genes, such as CXCL10, small but significant increases in CCL5 and IL‐17 production were observed following IFN activation. Notably, human mast cells produced both VEGF and IL‐1Ra in a dose dependent manner. These responses occurred in the absence of mast cell degranulation by a mechanism consistent with classical IFN signaling. Both reovirus and respiratory syncytial virus infection of mast cells, were also associated with IFN‐dependent IL‐1Ra expression. Conclusion and Clinical Relevance Our findings demonstrate that IFNs have profound impact on cytokine and chemokine expression by human mast cells, alone or in the context of viral infection. Mast cell VEGF and IL‐1Ra responses to IFNs could impact the regulation of local inflammatory responses and subsequent tissue remodeling.

Osteoarthritis (OA) is a progressive and degenerative disease, which may result in significant pain and decreased quality of life. Recent updates in our understanding of OA have demonstrated that it is a whole joint disease that has many similarities to an unhealed wound containing inflammatory cytokines. The nSTRIDE Autologous Protein Solution (APS) Kit is a medical device under development for the treatment of OA. The APS Kit processes a patient's own blood at the point of care to contain high concentrations of anti-inflammatory cytokines and anabolic growth factors. This study assessed the safety and treatment effects of a single intra-articular injection of APS. Eleven patients were enrolled in this study. Sufficient blood could not be drawn from one patient who was subsequently withdrawn, leaving 10 patients treated. Minor adverse events (AEs) were experienced by seven subjects (63.6%). There was one serious AE (diverticulitis) unrelated to the device or procedure. One subject experienced AEs that were judged “likely” to be procedure related (arthralgia/musculoskeletal discomfort) and all resolved within 6 days of injection. All other AEs were unrelated to the device or procedure. Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) pain scores improved significantly over time (ANOVA, p

Purpose This article investigates the relationship between cancer cells and stromal cells in carcinoma cell living microenvironment and elucidates the mechanism that interleukin-1 receptor antagonist (IL-1RA) blocks metastatic potential in colon cancer. Methods Western blot and RT-PCR assay were used to determine the expression of hepatocyte growth factor (HGF) and IL-1α in colon carcinoma cells and stromal cells. Effect of IL-1RA and HGF on metastatic potential of colon cancer cells were examined by proliferation, invasion, and angiogenesis assays. The interactional role of IL-1RA and HGF were detected by ELISA assay, invasion, and angiogenesis assay making up a co-culture system consisting of stromal and colon cancer cells in cells living microenvironment. Results IL-1α was expressed in human umbilical vein endothelial cells (HUVECs) and HT-29 and WiDr (colon cancer cell lines with higher liver metastatic potential). HGF was expressed only in fibroblast. HGF secretion from fibroblasts was significantly inhibited by IL-1RA (P

The contributions of the innate immune system to the development of pancreatic cancer are still ill defined. Inflammatory macrophages can initiate metaplasia of pancreatic acinar cells to a duct-like phenotype (acinar-to-ductal metaplasia [ADM]), which then gives rise to pancreatic intraepithelial neoplasia (PanIN) when oncogenic KRas is present. However, it remains unclear when and how this inflammatory macrophage population is replaced by tumor-promoting macrophages. Here, we demonstrate the presence of interleukin-13 (IL-13), which can convert inflammatory into Ym1+ alternatively activated macrophages, at ADM/PanIN lesions. We further show that Ym1+ macrophages release factors, such as IL-1ra and CCL2, to drive pancreatic fibrogenesis and tumorigenesis. Treatment of mice expressing oncogenic KRas under an acinar cell-specific promoter with a neutralizing antibody for IL-13 significantly decreased the accumulation of alternatively activated macrophages at these lesions, resulting in decreased fibrosis and lesion growth.

Dengue virus (DENV) infection causes several diseases ranging from dengue fever to life-threatening dengue hemorrhagic fever and dengue shock syndrome characterized by endothelial dysfunction, vascular leakage, and shock. Here, we identify a potential mechanism by which DENV induces tissue injury and vascular leakage by promoting the activation of interleukin (IL)-1β. DENV facilitates IL-1β secretion in infected patients, mice, human peripheral blood mononuclear cells (PBMCs), mouse bone marrow-derived macrophages (BMDMs), and monocyte-differentiated macrophages (THP-1) via activating the NLRP3 inflammasome. The accumulated data suggest that IL-1β probably induces vascular leakage and tissue injury in interferon-alpha/beta receptor 1 deficient C57BL/6 mice (IFNAR -/- C57BL/6), whereas IL-1 receptor antagonist (IL-1RA) alleviates these effects of IL-1β. Finally, administration of recombinant IL-1β protein results in vascular leakage and tissue injury in C57BL/6 mice. Together, the accumulated results demonstrate that IL-1β contributes to DENV-associated pathology and suggest that IL-1RA acts as a potential agent for the treatment of DENV-associated diseases.

Background: Blockade of the cytokine interleukin-1 (IL-1) with IL-1 receptor antagonist (IL-1Ra) is a candidate treatment for stroke entering phase II/III trials, which acts by inhibiting harmful inflammatory responses. Infection is a common complication after stroke that significantly worsens outcome and is related to stroke-induced deficits in systemic immune function thought to be mediated by the sympathetic nervous system. Therefore, immunomodulatory treatments for stroke, such as IL-1Ra, carry a risk of aggravating stroke-associated infection. Our primary objective was to determine if factors associated with antibody-mediated antibacterial defences were further compromised in patients treated with IL-1Ra after stroke. Methods: We assessed plasma concentrations of immunoglobulin isotypes and complement components in stroke patients treated with IL-1Ra or placebo and untreated non-stroke controls using multiplex protein assays. Activation of the sympathetic nervous system (SNS) was determined by measuring noradrenaline, a major SNS mediator. Results: There were significantly lower plasma concentrations of IgM, IgA, IgG1 and IgG4 in stroke-patients compared to non-stroke controls, however there were no differences between stroke patients treated with placebo or IL-1Ra. Concentrations of complement components associated with the classical pathway were increased and those associated with the alternative pathways decreased in stroke patients, neither being affected by treatment with IL-1Ra. Noradrenaline concentrations were increased after stroke in both placebo and IL-1Ra-treated stroke patients compared to non-stroke controls. Conclusion: These data show treatment with IL-1Ra after stroke does not alter circulating immunoglobulin and complement concentrations and is therefore unlikely to further aggravate stroke-associated infection susceptibility through altered availability of these key anti-microbial mediators.

Background Interleukin‐1 promotes tumor angiogenesis through VEGF production. The interleukin‐1 receptor antagonist can suppress tumors by blocking this effect. Methods Immunohistochemistry, WB, and gene sequencing were used to analyze the expression of IL‐1RA in esophageal cancer patients. WB was used to detect the expression of IL‐1RA and interleukin‐1α in esophageal cancer cells. Stable ESCC cell models overexpressing the IL‐1RA were constructed. Their cell functions were tested, and their effects on VEGF were examined. Results IL‐1RA is downregulated in primary EC tumors, and this downregulation of IL‐1RA is closely related to TNM staging and survival prognosis. The overexpression of IL‐1RA increased the proliferation of KYSE410 EC cells, which have a high level of IL‐1α expression. Overexpression of IL‐1RA in KYSE410 cells promotes a decrease in the expression of VEGF‐A. However, IL‐1RA expression did not cause any changes in EC9706 cells with low IL‐1α expression. Conclusion IL‐1RA acts as a tumor suppressor, and its deletion promotes tumor progression by increasing VEGF‐A expression in ESCC.