Human Interleukin-1β Research Articles

The Methylation Level of a CpG Site in the Human Interleukin-1β Promoter Reflects Both Current and Past Inflammation

The Methylation Level of a CpG Site in the Human Interleukin-1β Promoter Reflects Both Current and Past Inflammation

Ligandability Assessment of IL-1β by Integrated Hit Identification Approaches.

Human interleukin-1β (IL-1β) is a pro-inflammatory cytokine that plays a critical role in the regulation of the immune response and the development of various inflammatory diseases. In this publication, we disclose our efforts toward the discovery of IL-1β binders that interfere with IL-1β signaling. To this end, several technologies were used in parallel, including fragment-based screening (FBS), DNA-encoded library (DEL) technology, peptide discovery platform (PDP), and virtual screening. The utilization of distinct technologies resulted in the identification of new chemical entities exploiting three different sites on IL-1β, all of them also inhibiting the interaction with the IL-1R1 receptor. Moreover, we identified lysine 103 of IL-1β as a target residue suitable for the development of covalent, low-molecular-weight IL-1β antagonists.

Exploring the anti-inflammatory potential of the polyphenolic compounds in Moringa oleifera leaf: in silico molecular docking

ABSTRACT A recent study conducted an in silico model to investigate the potential of Moringa oleifera’s ethanolic leaf extract as a natural anti-inflammatory agent. The HPLC analysis identified 7 phenolic acids (benzoic acid 28%, syringic acid 23%, ellagic acid 19%, chlorogenic acid 12%, catechol 11%, gallic acid 5%, and caffeic acid 2%) and 5 flavonoids (naringenin 31%, kaempferol 30%, apigenin 14%, rutin 13%, and quercetin 12%) that are known to have anti-inflammatory activities in vitro tested on human interleukin-1β (IL-1β) and interleukin-6 (IL-6) in a previously prepared study (Hamdy, 2023) The Moringa polyphenol compounds were docked against IL-1β, IL-6, and the human interleukin-1 receptor (IL1RA) using Auto-Dock Vina. Caffeic acid, chlorogenic acid, and kaempferol interacted with amino acid residues in IL-1β by creating hydrogen bonds and hydrophobic interactions. The co-crystalized ligand attached to IL-1β displayed an affinity score of −8.96 kcal/mol. Caffeic acid, chlorogenic acid, quercetin, rutin, and syringic acid had unique interaction patterns with IL-6, forming hydrogen bonds and being hydrophobic. The interactions’ binding energies ranged from −5.97 to 5.26 kcal/mol. Moringa’s polyphenolic compounds also docked with IL1RA, where rutin and chlorogenic acid exhibited binding energies of −8.96 kcal/mol and −7.12 kcal/mol, respectively, and created hydrophobic interactions and hydrogen bonds. In conclusion, Moringa oleifera polyphenols can interact with IL-1β, IL-6, and IL-1RA to reduce inflammation, making it a potential treatment strategy for improving recovery.

Discovery of a selective and biologically active low-molecular weight antagonist of human interleukin-1β

Human interleukin-1β (hIL-1β) is a pro-inflammatory cytokine involved in many diseases. While hIL-1β directed antibodies have shown clinical benefit, an orally available low-molecular weight antagonist is still elusive, limiting the applications of hIL-1β-directed therapies. Here we describe the discovery of a low-molecular weight hIL-1β antagonist that blocks the interaction with the IL-1R1 receptor. Starting from a low affinity fragment-based screening hit 1, structure-based optimization resulted in a compound (S)-2 that binds and antagonizes hIL-1β with single-digit micromolar activity in biophysical, biochemical, and cellular assays. X-ray analysis reveals an allosteric mode of action that involves a hitherto unknown binding site in hIL-1β encompassing two loops involved in hIL-1R1/hIL-1β interactions. We show that residues of this binding site are part of a conformationally excited state of the mature cytokine. The compound antagonizes hIL-1β function in cells, including primary human fibroblasts, demonstrating the relevance of this discovery for future development of hIL-1β directed therapeutics.

Multivalent Nanobody Conjugate with Rigid, Reactive Oxygen Species Scavenging Scaffold for Multi-Target Therapy of Alzheimer's Disease.

Efficient therapeutic strategies that concurrently target both Aβ aggregation and oxidative stress in the Alzheimer'sdisease(AD)microenvironment emerge as a cutting-edge tool to combat the intricate pathogenesis of AD. Here, a multivalent nanobody conjugate with rigid, reactive oxygen species (ROS) scavenging scaffold is developed to achieve simultaneous Aβ amyloidogenesis mitigation, ROS elimination, and Aβ plaque clearance. Grafting Aβ segment (33-GLMVGGVVIA-42) into the third complementary-determining region of a parent nanobody generates an engineered nanobody NB that can recognize Aβ and inhibit its aggregation through homotypic interactions. NB is further genetically modified with a fragment of human interleukin-1β (163-VQGEESNDK-171), so that the obtained fusion nanobody NBIL can also facilitate the Aβ clearance by microglia. Linking NBIL covalently onto a rigid, ROS scavenging scaffold poly(deca-4,6-diynedioic acid) (PDDA) creates the multivalent nanobody conjugate PNBIL, which not only boosts the binding affinity between NBIL and Aβ aggregates for nearly 100 times but also possesses a long-term capability of oxidative stress alleviation, inflammation reduction, and neuron protection. PNBIL has significantly attenuated symptoms on two AD mouse models through amyloidogenesis inhibition and AD microenvironment modulation, validating that the multivalent nanobody conjugate design based on combinatory nanobody and molecular engineering is a promising approach of multi-target therapeutic strategies.

Human Interleukin-1β Profile and Self-Reported Pain Monitoring Using Clear Aligners with or without Acceleration Techniques: A Case Report and Investigational Study.

There is a growing demand for more aesthetic, comfortable, and faster orthodontic treatments, and clear aligners emerged as a solution to fulfill this need. However, the effectiveness of clear aligners to treat complex malocclusions is yet contentious. The use of acceleration methods could improve the efficacy of clear aligners by stimulating cells' mechanobiology through numerous pathways, but this hypothesis is still poorly explored. We aimed to monitor the release profile of an inflammatory marker-the interleukin-1β-and to evaluate its relationship with self-reported pain scores with and without the use of acceleration techniques during an orthodontic treatment requiring difficult tooth movements with clear aligners. Case Report. Here, we report a case of a 46-year-old female patient who presented functional and aesthetic complaints. Intraoral examination revealed a diminished overjet and overbite, rotation of teeth 45 and 24, absence of teeth 25, 35, and 36, buccolingual dislocation of tooth 21, a tendency to a Class III malocclusion, and a 2 mm left deviation of the lower midline. This study is divided into three stimulation phases: no stimulation, mechanical vibration stimulation, and photobiomodulation. Interleukin-1β levels in gingival crevicular fluid samples from the pressure side of six selected teeth were evaluated at four time points after the orthodontic treatment onset. Pain monitoring in those teeth was performed using a visual analogue scale at the same time points. Interleukin-1β protein production peaked 24 h after treatment onset. Complex movements were associated with increased self-reported pain. Clear aligners show limitations in solving complex tooth movements, even when combined with acceleration. The development of customized and programmable stimulation microdevices integrated into "smart aligners," which could be designed to specifically stimulate the direction of movement and stimulation parameters and could constitute a solution to optimize the orthodontic tooth movement with clear aligners.

A host-vector toolbox for improved secretory protein overproduction in Bacillus subtilis

Target proteins in biotechnological applications are highly diverse. Therefore, versatile flexible expression systems for their functional overproduction are required. In order to find the right heterologous gene expression strategy, suitable host-vector systems, which combine different genetic circuits, are useful. In this study, we designed a novel Bacillus subtilis expression toolbox, which allows the overproduction and secretion of potentially toxic enzymes. This toolbox comprises a set of 60 expression vectors, which combine two promoter variants, four strong secretion signals, a translation-enhancing downstream box, and three plasmid backbones. This B. subtilis toolbox is based on a tailor-made, clean deletion mutant strain, which is protease and sporulation deficient and exhibits reduced autolysis and secondary metabolism. The appropriateness of this alternative expression platform was tested for the overproduction of two difficult-to-produce eukaryotic model proteins. These included the sulfhydryl oxidase Sox from Saccharomyces cerevisiae, which forms reactive hydrogen peroxide and undesired cross-linking of functional proteins, and the human interleukin-1β, a pro-inflammatory cytokine. For the best performing Sox and interleukin, overproducing and secreting variants of these new B. subtilis toolbox fermentation strategies were developed and tested. This study demonstrates the suitability of the prokaryotic B. subtilis host-vector system for the extracellular production of two eukaryotic proteins with biotechnological relevance.Key points• Construction of a versatile Bacillus subtilis gene expression toolbox.• Verification of the toolbox by the secretory overproduction of two difficult-to-express proteins.• Fermentation strategy for an acetoin-controlled overproduction of heterologous proteins.

Inhibition of Autophagy at Different Stages by ATG5 Knockdown and Chloroquine Supplementation Enhances Consistent Human Disc Cellular Apoptosis and Senescence Induction rather than Extracellular Matrix Catabolism

The intervertebral disc is the largest avascular organ. Autophagy is an important cell survival mechanism by self-digestion and recycling damaged components under stress, primarily nutrient deprivation. Resident cells would utilize autophagy to cope with the harsh disc environment. Our objective was to elucidate the roles of human disc cellular autophagy. In human disc cells, serum deprivation and pro-inflammatory interleukin-1β (IL-1β) stimulation increased autophagy marker microtubule-associated protein 1 light chain 3 (LC3)-II and decreased autophagy substrate p62/sequestosome 1 (p62/SQSTM1), indicating enhanced autophagy. Then, RNA interference (RNAi) of autophagy-related gene 5 (ATG5), essential for autophagy, showed decreases in ATG5 protein (26.8%–27.4%, p < 0.0001), which suppressed early-stage autophagy with decreased LC3-II and increased p62/SQSTM1. Cell viability was maintained by ATG5 RNAi in serum-supplemented media (95.5%, p = 0.28) but reduced in serum-free media (80.4%, p = 0.0013) with IL-1β (69.9%, p = 0.0008). Moreover, ATG5 RNAi accelerated IL-1β-induced changes in apoptosis and senescence. Meanwhile, ATG5 RNAi unaffected IL-1β-induced catabolic matrix metalloproteinase release, down-regulated anabolic gene expression, and mitogen-activated protein kinase pathway activation. Lysosomotropic chloroquine supplementation presented late-stage autophagy inhibition with apoptosis and senescence induction, while catabolic enzyme production was modest. Disc-tissue analysis detected age-related changes in ATG5, LC3-II, and p62/SQSTM1. In summary, autophagy protects against human disc cellular apoptosis and senescence rather than extracellular matrix catabolism.

Anti-Inflammatory and Antioxidant Activity of Hydroxytyrosol and 3,4-Dihydroxyphenyglycol Purified from Table Olive Effluents.

New liquid effluents based on the use of acetic acid in the table olive industry make it easier to extract bioactive compounds to be used for food, cosmetic, and pharmaceutical purposes. The use of water acidified with acetic acid or in brine with or without acetic acid for storing the table olive enhances the extraction of two more active phenolic compounds: hydroxytyrosol (HT) and 3,4-dihydroxyphenylglycol (DHPG). This work has two aims: (1) measure the solubilization of phenolics controlled for two years using more than thirty olive varieties with different ripeness index as a potential source of HT and DHPG, and (2) evaluate the anti-inflammatory activity of the purified phenolics. The effluents with a higher concentration of phenolics were used for the extraction of HT and DHPG in order to evaluate its antioxidant and anti-inflammatory activity in vitro by the determination of pro-inflammatory cytokines such as Human Tumor Necrosis Factor-α (TNF), Interleukin-6 (IL-6), and Interleukin-1β (Il-1β). The anti-inflammatory activity of these phenolic extracts was demonstrated by studying the expression of cytokines by qPCR and the levels of these proteins by enzyme-linked immunosorbent assay (ELISA).

Correlation between inflammatory cytokines and liver cancer stem cell markers in DEN-induced liver cancer rats.

To study the association of inflammatory factors and hepatocarcinoma stem cells of induced liver cancer rats. 30 SD male healthy rats were selected. 10 rats were given water as normal control group. 10 rats only were implemented laparotomy as sham operation group. The remaining 10 rats were the liver cancer model group and treated with diethylnitrosamine (DEN) to induce liver cancer. Real-time quantitative PCR was used to detect the related inflammatory factors in HCC tissues, including interleukin-6 (IL-6), monocyte chemoattractant protein-1 (MCP-1), transforming growth factor-β1 (TGF-β), human interleukin-1α (IL-1α), human interleukin 1β (IL-1β) and levels of hepatocarcinoma stem cells indicators CD90, CD133, Alpha-fetoprotein (AFP). Correlation analysis was used to analyze the correlation between inflammatory factors and hepatocarcinoma stem cells markers CD90 and CD133. The expression levels of IL-6, MCP-1 and TGF-β of HCC tissues in liver cancer model group were significantly higher than those in the control group and the sham operation group. The expression levels of CD90 and CD133 of tissues in the liver cancer model group were significantly higher than those in the control group and the sham operation group. The differences were statistically significant (p<0.001). By inhibiting related inflammatory factors, the growth, migration and invasion of liver cancer cells were significantly inhibited, and apoptosis was promoted. Correlation analysis results showed that the expression changes of IL-6, MCP-1 and TGF-β were significantly positively correlated with CD90 up-regulation (p<0.05), while the expression changes of IL-6, MCP-1 and TGF-β were significantly positively correlated with CD133 up-regulation (p<0.05). The inflammatory factors IL-6, MCP-1 and TGF-β are closely related to hepatocarcinoma stem cells, which play an important role in promoting the occurrence and deterioration of liver cancer.

Salvianolic acid B decreases interleukin-1β-induced colitis recurrence in mice.

BackgroundDegree of mucosal recovery is an important indicator for evaluating the therapeutic effects of drugs in treatment of inflammatory bowel disease (IBD). Increasing evidences has proved that tight junction (TJ) barrier dysfunction is one of the pathological mechanisms of IBD. The aim of this study was to observe whether enhancement of TJ can decrease colitis recurrence.MethodsEighty C57BL/6 mice were randomly divided into four groups including normal group, colitis group, sulfasalazine (SASP) treated group, and traditional Chinese drug salvianolic acid B (Sal B) treated group. Colitis was established in mice by free drinking water containing dextran sulfate sodium, after treatments by SASP and Sal B, recombinant human interleukin-1β (IL-1β) was injected intraperitoneally to induce colitis recurrence.ResultsCompared with sham control, cell apoptosis in colitis group was increased from 100.85 ± 3.46% to 162.89 ± 11.45% (P = 0.0038), and TJ dysfunction marker myosin light chain kinase (MLCK) was also significantly increased from 99.70 ± 9.29% to 296.23 ± 30.78% (P = 0.0025). The increased cell apoptosis was reversed by both SASP (125.99 ± 8.45% vs. 162.89 ± 11.45%, P = 0.0059) and Sal B (104.27 ± 6.09% vs. 162.89 ± 11.45%, P = 0.0044). High MLCK expression in colitis group was reversed by Sal B (182.44 ± 89.42% vs. 296.23 ± 30.78%, P = 0.0028) but not influenced by SASP (285.23 ± 41.04% vs. 296.23 ± 30.78%, P > 0.05). The recurrence rate induced by recombinant human IL-1β in Sal B-treated group was significantly lower than that in SASP-treated group.ConclusionsThese results suggested a link between intestinal mucosal barrier dysfunction, especially TJ barrier dysfunction, and colitis recurrence. The TJ barrier dysfunction in remission stage of colitis increased the colitis recurrence. This study might provide potential treatment strategies for IBD recurrence.

Modern Condition and Prospects for the Development of Medicines towards Prevention and Early Treatment of Radiation Damage

An analysis of the current status and future directions of developing medicines for the prevention and early treatment of radiation damage has been conducted. Among radioprotectors, only B-190 (Indralin) is approved for use in the Russian Federation; the main radioprotector in foreign countries is amifostine (Ethyol). Some prospects in the development of new radioprotectors are associated with inhibitors of NO-synthases from the class of N-acyl-S-alkyl-substituted isothiourea derivatives, among which chemical compounds with a pronounced radioprotective effect are found. A recombinant human interleukin-1β (Betaleukin) has been registered in Russia as a radiomitigator intended for use in the early period after accidental irradiation. The US Food and Drug Administration (FDA) has approved the investigational new drug status for seven radiomitigators, including 5-androstenediol (Neumune), genistein (BIO 300), CBLB502 (Entolimod), a kinase inhibitor ON01210 (Ex-RAD), recombinant human interleukin-12 (HemaMax), beclomethasone (OrbeShield), and a granulocyte colony-stimulating factor (G-CSF, Neupogen). Ondansetron hydrochloride dihydrate (Latran) was registered as an antiemetic drug for prevention and therapy of the primary response to radiation in Russia, and granisetron (Kytril) was registered as an antiemetic drug upon irradiation abroad. Potassium iodide, potassium ferric hexacyanoferrate (Ferrocin), diethylenetriaminepentaacetate calcium trisodium (Pentacin, calcium trisodium pentetate), and 2,3-dimercaptopropansulfonate sodium (Unithiol) are used in Russia for the prevention and treatment of damage from irradiation via incorporated radionuclides. Potassium iodide (ThyroShield), diethylenetriaminepentaacetate zinc trisodium (Zn-DTPA, Pentetate zinc trisodium), diethylenetriaminepentaacetate calcium trisodium (Ca-DTPA, Pentetate calcium trisodium), and ferric hexacyanoferrate (Prussian blue, Radiogardase) are used as decorporation agents abroad. Cytokines, vitamins, low-molecular compounds, inhibitors of apoptosis, etc., are considered promising means for the prevention and early treatment of radiation damage.

The cost-effectiveness of canakinumab for secondary prevention of cardiovascular disease: The Australian healthcare perspective

BackgroundCanakinumab is a fully human monoclonal antibody targeting interleukin-1β. It is currently indicated for use in those with rheumatologic disorders due to its anti-inflammatory properties, and was recently shown to be beneficial for the secondary prevention of cardiovascular disease (CVD). However, the cost-effectiveness of canakinumab used to treat CVD is unknown. MethodsA Markov state transition model was developed and populated with a hypothetical sample of 1000 individuals profiled on the Canakinumab Antiinflammatory Thrombosis Outcome Study (CANTOS); with a history of myocardial infarction (MI) and blood concentrations of high-sensitivity C-reactive protein (hsCRP) of >2 mg/L. With each annual cycle, individuals could have a recurrent non-fatal CVD event (MI or stroke), or die from a CVD event or die from other causes based on data from CANTOS. Individuals continued to cycle through the model for 20 years or until death. Cost and utility data was applied. Outcomes were discounted (5% annually). ResultsOver a 20-year time horizon, canakinumab is predicted to prevent 40 recurrent cardiovascular events and save 287 (discounted) years of life and 239 (discounted) quality-adjusted life years (QALYs) in 1000 individuals. At an annual cost of AUD36,049 (USD25,590, GBP19,662) per person, canakinumab would not be considered cost-effective within the Australian healthcare system, with an incremental cost-effectiveness ratio (ICER) of AUD1,221,170 per QALY gained. ConclusionsCanakinumab is an attractive treatment option to reduce recurrent CVD among patients with high hsCRP. It would be considered cost-effective in this treatment setting within the perspective of the Australian public healthcare system if its annual costs do not exceed AUD1500 (USD1065, GBP818) per person.

Evaluation of the radioprotective properties of recombinant flagellin when used alone or in combination with interleukin-1 beta

Research objective. To evaluate the radioprotective effectiveness of recombinant flagellin when used alone or in combination with interleukin-1 beta for prophylactic or therapeutic effect on animals. Materials and methods. The effect of hybrid flagellin FliC Salmonella typhimurium and human interleukin-1β (Institute of Highly Pure Biopreparations, Saint Petersburg, Russia) on the 30-day survival of male mice exposed to lethal doses of X-ray radiation was studied. Survival of irradiated animals was analyzed by Kaplan-Meier method. Results. The preventive use of flagellin had a protective effect on the 30-day survival of mice irradiated with lethal doses of X-rays. Compared with the irradiated control, the administration of flagellin (1 mg/kg or 2 mg/kg) prior to X-ray exposure (7.5 Gy, 8.0 Gy or 8.5 Gy) increased the animal survival rate to 67-87%. Complex preventive administration of flagellin (1 mg/kg) and interleukin-1 beta (50 μg/kg) provided a 100% survival rate of the irradiated mice. Separate use of drugs was also effective; 92.8% of mice survived when flagellin was administered prior to irradiation and interleukin after. Conclusion. Recombinant flagellin is a promising candidate product for designing new generation of domestic radiation countermeasures, including combinations with interleukin-1 beta.

Proposed 3-D Structure of a Critical Protein-Protein Interaction Required for Transcription of the IL1B Gene Coding for Human Interleukin 1β

Abstract We previously reported that expression of the human IL1B gene coding for Interleukin 1β in LPS-activated macrophages requires long-range chromatin looping mediated by a mutual DNA binding domain (DBD) interaction between two transcription factors, Spi-1/PU.1 (Spi1) at the promoter and C/EBPβ at a far-upstream enhancer. We have now combined preexisting X-ray structures and advanced computational techniques with additional biochemical and cell-based studies in order to further investigate the structure of this critical interaction. In vitro binding was used to generate a surface interaction map for the two proteins that suggested an association similar to that of a C/EBPβ·Myb X-ray structure reported by others. The distinction is that, in addition to a C/EBPβ leucine zipper interaction similar to that with the Myb DBD, there is an interaction between a carboxyl-tail extension of one of the C/EBPβ leucine zipper monomers with Spi1. This is consistent with our previous report that the carboxyl-tail of C/EBPβ is critical for association with arginine 232 within a pocket at one end of the Spi1 DNA recognition helix. L-arginine, a known anti-inflammatory, was computationally identified to bind in this interaction pocket. In vivo studies of LPS-activated THP-1 macrophages treated with L-arginine decreased IL1B transcription in parallel with a reduction in C/EBPβ association with Spi1 on the IL1B promoter. No significant change in direct binding of Spi1 and C/EBPβ to DNA was observed. These results suggest that L-arginine inhibits IL1B gene expression by competitive inhibition of C/EBPβ binding to Spi1 in the absence of any effect on direct DNA binding for either factor, and support the predicted structure of the C/EBPβ·Spi1 interaction.

L‐Arginine Inhibits a Critical Structural Contact Required for Transcription of the IL1B Gene Coding for Human Interleukin 1β

Interleukin 1β (IL‐1β) is a proinflammatory cytokine that is normally expressed and released by monocytes in response to infection and injury. Chronic expression of IL‐1β has been associated with numerous inflammatory diseases. We previously reported chromatin looping for the IL1B gene that is mediated by a mutual DNA‐binding domain (DBD) interaction between two transcription factors, Spi‐1/PU.1 (Spi‐1) at the promoter and C/EBPβ at a distal enhancer. The nature of this critical interaction between the Spi‐1 wHTH domain and the C/EBPβ leucine zipper was investigated by using the existing X‐ray structures of these two proteins bound to DNA as a guide in designing mutants for in vitro protein‐protein interaction studies. This generated surface interaction maps for the two proteins that in concert with computational techniques supported an association mode similar to, but distinct from, the C/EBPβ·c‐Myb X‐ray structure reported by others. The distinction was that, in addition to the C/EBPβ leucine zipper interaction with the c‐Myb wHTH domain, Spi‐1 utilizes an additional pseudo‐symmetrical bidentate interaction between the carboxyl‐tail extensions of each of the C/EBPβ leucine zipper monomers with the two ends of the Spi‐1 DNA recognition helix. This is consistent with our previous report that the carboxyl‐tail of C/EBPβ is critical for Spi‐1 association, suggesting that pocket‐like surfaces at the ends of the Spi‐1 DNA recognition helix might be targets for inhibition. Candidate small molecules were generated using a virtual screening approach. L‐arginine, a known anti‐inflammatory molecule, was identified from this approach as having potential to bind to each end of the Spi‐1 recognition helix, possibly inhibiting IL1B gene expression. Studies in 2.5 h LPS‐treated human THP‐1 macrophages in the presence of L‐arginine revealed a concentration‐dependent blunting of transcription in parallel with a decrease in C/EBPβ association with Spi‐1 on the IL1B gene promoter, while Spi‐1 binding to DNA was unaffected. In the same experiment, there was no significant change in direct binding of C/EBPβ to DNA at a well‐characterized site in the IL6 gene. In conclusion, these studies are consistent with an L‐arginine‐dependent decrease in IL1B gene expression that is due to its ability to inhibit the protein‐protein interaction between C/EBPβ and Spi‐1 in the absence of any effect on direct DNA binding for either factor. This is consistent with the predicted structure of the C/EBPβ·Spi‐1 interaction.

Respiratory Syncytial Virus (RSV) Pulmonary Infection in Humanized Mice Induces Human Anti-RSV Immune Responses and Pathology.

Respiratory syncytial virus (RSV) is a leading cause of lower respiratory tract disease, which causes high rates of morbidity and mortality in infants and the elderly. Models of human RSV pulmonary disease are needed to better understand RSV pathogenesis and to assess the efficacy of RSV vaccines. We assessed the RSV-specific human innate, humoral, and cellular immune responses in humanized mice (mice with a human immune system [HIS mice]) with functional human CD4(+) T and B cells. These mice were generated by introduction of HLA class II genes, various human cytokines, and human B cell activation factor into immunodeficient NOD scid gamma (NSG) mice by the use of an adeno-associated virus vector, followed by engraftment of human hematopoietic stem cells. During the first 3 days of infection, HIS mice lost more weight and cleared RSV faster than NSG mice. Human chemokine (C-C motif) ligand 3 (CCL3) and human interleukin-1β (IL-1β) expression was detected in the RSV-infected HIS mice. The pathological features induced by RSV infection in HIS mice included peribronchiolar inflammation, neutrophil predominance in the bronchioalveolar lavage fluid, and enhanced airway mucus production. Human anti-RSV IgG and RSV-neutralizing antibodies were detected in serum and human anti-RSV mucosal IgA was detected in bronchioalveolar lavage fluid for up to 6 weeks. RSV infection induced an RSV-specific human gamma interferon response in HIS mouse splenocytes. These results indicate that human immune cells can induce features of RSV lung disease, including mucus hyperplasia, in murine lungs and that HIS mice can be used to elicit human anti-RSV humoral and cellular immunity. Infections with respiratory syncytial virus (RSV) are common and can cause severe lung disease in infants and the elderly. The lack of a suitable animal model with disease features similar to those in humans has hampered efforts to predict the efficacy of novel anti-RSV therapies and vaccines for use in humans. A murine model consisting of mice with a human immune system (HIS mice) could be useful for assessment of RSV disease and anti-RSV responses specific to humans. This study investigates an HIS mouse model to imitate human RSV disease and immune responses. We found that RSV lung infection in HIS mice results in an RSV-specific pathology that mimics RSV disease in humans and induces human anti-RSV immune responses. This model could be useful for better understanding of human RSV disease and for the development of RSV therapies.

Anterior chamber interleukin 1β, interleukin 6 and prostaglandin E2 in patients undergoing femtosecond laser-assisted cataract surgery

PurposeTo evaluate the interleukin (IL) 1β, IL-6 and prostaglandin E2 (PGE2) concentrations in aqueous humour in patients undergoing femtosecond laser-assisted cataract surgery.MethodsIn 27 eyes of 27 patients undergoing femtosecond laser-assisted...

СAPSULAR ANTIGEN OF YERSINIA PESTIS

Plague is a zoonosis caused by gram-negative bacteria Yersinia pestis, which, as a rule, is transmitted to humans from septicemic rodents by the bites of infected fleas. This microbe killed more people than all of the wars in the human history. Y. pestis circulation in the natural plague foci is ensured by the whole number of pathogenicity factors with differing functional orientation. This review is devoted to one of them, Y. pestis capsular antigen (F1 or Caf1). The history of its discovery and studying of its genetic control, biosynthesis, isolation and purification, and physicochemical properties are reviewed. Its roles in plague pathogenesis and its application as a main component of plague vaccines are also discussed. Y. pestis capsule under light microscopy is visually amorphous, while high-resolution electron microscopy displays the structure formed from separate fimbria-like cords up to 200 nm long, diverging from the bacterial surface in different directions. At 37°C Y. pestis produce 800–1000 times more capsular antigen than at 28°C. Genes coding for 17.6-kD Caf1 protein, which contains 170 amino acids, are located in caf1 operon of pFra plasmid. Analysis of caf1 operon nucleotide sequence testified its close phylogenetic relationship with the gene clusters coding for pilus adhesins that were secreted with the help of chaperone/usher systems in enterobacteria including six additional adhesins in Y. pestis. Y. pestis multiplication within macrophages is the obligatory stage of plague pathogenesis, and the plague pathogen virulence correlates not with resistance to phagocyte ingesting but with bacterial ability to survive and multiply within phagolysosomes of phagocytes due to neutralization of antibacterial functions of eukaryotic cells. The capsule formed out of the Caf1 aggregates protects Y. pestis from ingestion by naïve host’s phagocytes and prevents from initiation of the alternative pathway of the complement system. Molecular usher Caf1A responsible for capsular antigen anchoring on the surface of bacterial cell has a high affinity to human interleukin 1β. Caf1 can compete with interleukins 1α, 1β, and 1ra in binding to receptors on lymphoid cells preventing development of adequate immune response. Immunodiagnosis of plague is based on detection of Caf1 or anti-Caf1 antibodies since this Y. pestis antigen is species specific. Covering bacterial surfaces capsular antigen is also the paramount component of all modern plague vaccines. Its leading role in induction of intense immunity in mice, rats, monkeys, and men was shown clearly. However, non-capsulated (Caf1–) variants of Y. pestis keeping their virulence at the level of the wild-type strains might be selected and accumulated in immune animals. This indicates inadmissibility of application of monoantigen plague vaccines and necessity for design of immunoprophylactic preparations aimed at two or three molecular targets.

CFTR Knockdown induces proinflammatory changes in intestinal epithelial cells.

BackgroundHyperinflammation is a hallmark feature of cystic fibrosis (CF) airways. However, inflammation has also been documented systemically and, more recently, in extrapulmonary CF-affected tissues such as the pancreas and intestine. The pathogenesis of CF-related inflammation and more specifically the role of the cystic fibrosis transmembrane conductance regulator (CFTR) in that respect are not entirely understood. We have tested the hypothesis that genetic depletion of CFTR will affect the inflammatory status of human intestinal epithelial cell lines.MethodsCFTR expression was genetically depleted from Caco-2/15 and HT-29 cells using short hairpin RNA interference (shRNAi). Inflammatory conditions were induced by the addition of human recombinant tumor necrosis factor (TNF) or Interleukin-1β (IL-1β) for various periods of time. Gene expression, mRNA stability and secreted levels of interleukin (IL)-6, −8 and 10 were assessed. Analysis of pro- and anti-inflammatory signaling pathways including mitogen-activated protein kinases (p38, ERK 1/2 and JNK), nuclear factor of kappa light polypeptide gene enhancer in B-cells inhibitor alpha (IκBα), and nuclear factor-kappa B (NF-κB) was also performed. Eosinophils were counted in the jejunal mucosa of Cftr−/− and Cftr+/+ mice.ResultsCFTR gene and protein knockdown caused a significant increase in basal secretion of IL-8 as well as in IL-1β-induced secretion of IL-6 and −8. Release of the anti-inflammatory cytokine, IL-10, remained unaffected by CFTR depletion. The enhanced secretion of IL-8 stems in part from increased IL8 mRNA levels and greater activation of ERK1/2 MAPK, IκBα and NF-κB in the CFTR knockdown cells. By contrast, phosphorylation levels of p38 and JNK MAPK did not differ between control and knockdown cells. We also found a higher number of infiltrating eosinophils in the jejunal mucosa of Cftr −/− females, but not males, compared to Cftr +/+ mice, thus providing in vivo support to our in vitro findings.ConclusionCollectively, these data underscore the role played by CFTR in regulating the intestinal inflammatory responses. Such findings lend support to the theory that CFTR exerts functions that may go beyond its role as a chloride channel whereby its disruption may prevent cells to optimally respond to exogenous or endogenous challenges. These observations are of particular interest to CF patients who were found to display alterations in their intestinal microbiota, thus predisposing them to pathogens that may elicit exaggerated inflammatory responses.Electronic supplementary materialThe online version of this article (doi:10.1186/s12950-015-0107-y) contains supplementary material, which is available to authorized users.