Mepolizumab Research Articles

Predictors of a weak antibody response to COVID-19 mRNA vaccine in systemic lupus erythematosus.

To characterize the antibody response to COVID-19 mRNA vaccination in patients with Systemic Lupus Erythematosus (SLE) and identify predictors of poor response. SLE patients who are followed at the Beth Israel Deaconess Medical Center Lupus Cohort (BID-LC) were enrolled. SARS-CoV-2 IgG Spike antibody was measured in patients who received two doses of either the BNT162b2 (Pfizer-BioNTech) or the mRNA-1273 (Moderna) COVID-19 vaccine (n = 62). We defined non-responders as patients with an IgG Spike antibody titer less than two-fold (< 2) the index value of the test and responders as patients with antibody levels greater or equal to two-fold (≥ 2). A web-based survey was used to collect information regarding immunosuppressive medication use and SLE flares after vaccination. In our cohort of lupus patients, 76% were vaccine responders. The use of two or more immunosuppressive drugs was associated with being a non-responder (Odds Ratio 5.26; 95% CI 1.23-22.34, p = 0.02). Both Belimumab use and higher Prednisone dose were associated with vaccine non-response (p = 0.04 and p = 0.04). The non-responder group had higher mean levels of serum IL-18 than the responder group (p = 0.04) as well as lower C3 levels (p = 0.01). Lupus flares and breakthrough infections were uncommon post-vaccination. Immunosuppressive medications have a negative impact on vaccine humoral response in SLE individuals. We observed a trend towards vaccine no-response in BNT162b2 recipients and a relationship between IL-18 and impaired antibody response that merits further investigation.

Copper-free click chemistry assisted antibodies for immunodetection of interleukin-10 in saliva

Interleukin-10 (IL-10) is an anti-inflammatory cytokine that is secreted in response to an acute phase inflammation in patients who are suffering from heart failure (HF). The aim of this work was to develop an electrochemical biosensor for determining salivary IL-10 levels. Biofunctionalization strategy was improved through the use of copper-free click chemistry for the developed sensor due to its advantages, leading to high quantitative yields of stable triazoles, rapid reaction, no cytotoxic Cu(I) catalyst requirement, and high specificity of cyclooctynes toward azides. The approach involved in binding of dibenzocyclooctyne acid (DBCO-COOH) to thiol-azide assembled gold microelectrodes, later capturing the monoclonal IL-10 antibody (IL-10 mAb), and ultimately allowing direct detection of IL-10 antigen. Fourier transform infrared spectroscopy (FTIR) and nanoplotter associated with fluorescence microscopy methods have been employed to analyze and prove the biofunctionalization of the gold microelectrodes. Moreover, the electrochemical impedance spectroscopy (EIS) technique was used for detecting IL-10 antigen. The developed immunosensor showed a semi-logarithmic linear range, from 0.1 pg/mL to 5 pg/mL with R2 = 0.9815 and a limit of quantitation (LOQ) of 0.1 pg/mL with relative standard deviation (RSD) of 10.67%. The specificity of the immunosensor was evaluated using an inflammatory cytokine, and none of it generated detectable EIS signals. Finally, the successful analysis of saliva samples from a healthy volunteer without Coronavirus (COVID-19) infection demonstrated the usefulness of the developed immunosensor.

Akkermansia muciniphila, which is enriched in the gut microbiota by metformin, improves cognitive function in aged mice by reducing the proinflammatory cytokine interleukin-6

BackgroundMetformin, a type 2 diabetes treatment, improves the cognitive function of aged mice; however, whether the protective effects of metformin on cognitive function in aged mice are associated with the gut microbiome is poorly understood. Although some studies suggest that the gut microbe composition influences cognitive function and that manipulating the gut microbiota might protect against age-related cognitive dysfunction, there is no direct evidence to validate that the gut microbiota mediates the effect of metformin on cognitive improvement.ResultsIn this study, we show that the gut microbiota is altered by metformin, which is necessary for protection against ageing-associated cognitive function declines in aged mice. Mice treated with antibiotics did not exhibit metformin-mediated cognitive function protection. Moreover, treatment with Akkermansia muciniphila, which is enriched by metformin, improved cognitive function in aged mice. Mechanistically, A. muciniphila decreased pro-inflammatory-associated pathways, particularly that of the pro-inflammatory cytokine interleukin (IL)-6, in both the peripheral blood and hippocampal profiles, which was correlated with cognitive function improvement. An IL-6 antibody protected cognitive function, and an IL-6 recombinant protein abolished the protective effect of A. muciniphila on cognitive function in aged mice. ConclusionThis study reveals that A. muciniphila, which is mediated in the gut microbiota by metformin, modulates inflammation-related pathways in the host and improves cognitive function in aged mice by reducing the pro-inflammatory cytokine IL-6.D24d6SVgEDjFieya1hSRMmVideo Graphical

Development of a specific, active human IL-18 assay for assessing inflammasome activation

Abstract Interleukin-18 (IL-18) was first identified for its ability to induce interferon-γ expression. IL-18 is a proinflammatory cytokine and its release along with IL-1β is a primary output of inflammasome activation. We have developed convenient tools for monitoring inflammasome activation, including assays for caspase-1 activity and IL-1β release. While both IL-1β and IL-18 are expressed as inactive zymogens requiring processing to active forms, they differ in expression and regulation. IL-1β requires upregulation for expression and is primarily restricted to myeloid cells, whereas IL-18 is constitutively expressed more broadly in epithelial cells, as well as myeloid cells. IL-18 is also uniquely regulated by an IL-18 binding protein (IL-18bp). The IL-18bp has a very high affinity for active IL-18 and effectively blocks receptor activation. IL-18-stimulated interferon-γ release stimulates IL-18bp release in a negative feedback loop. The Lumit™ technology is a Nanoluc®-based platform that enables direct, no wash assays for analyte detection using standard luminometers. For Lumit IL-18 assays, we either NanoBiT®-labeled two IL-18 antibodies or NanoBiT®-labeled the IL-18bp as well as one IL-18 antibody. These resulted in, respectively, one assay that measures both IL-18 and proIL-18 and another assay specific for active IL-18. The specific assay was used to measure active IL-18 release from canonical inflammasome-activated THP-1 cells and PBMCs, as well as non-canonical inflammasome-activated lung A549 epithelial cells. Enabling direct quantitation of unbound, active IL-18 is essential for elucidating IL-18’s role in inflammation and disease. A Lumit™ IL-18 assay adds to the toolbox for monitoring inflammasome activation.

B cell intrinsic Toll-like receptor 9 signaling regulates class switching to IgE

Abstract Chronic allergic diseases such as allergic asthma are driven by excessive production of allergen specific IgE. B cells produce IgE by class switch recombination (CSR) in response to T cell help and Th2 cytokines. However, development of allergic diseases can also be influenced by factors such as early-life exposure to microbial products. In vivo, stimulation of Toll-like receptor 9 (TLR9) with microbial DNA reduces IgE levels, largely thought to be a response to Th1 cytokines. However, the directeffects of TLR9 signaling on B cell class switching to IgE remain unclear. To delineate the B cell intrinsic effects of TLR9 stimulation, we treated mouse splenic B cells in vitrowith IL-4 and anti-CD40 antibody with or without a synthetic TLR9 agonist, CpG oligodeoxynucleotides (CpG). Compared to stimulation with IL-4 and anti-CD40 alone, the addition of CpG decreased IgE production. Using B cells from reporter mice that track germline transcription, a prerequisite for CSR, we found that CpG inhibited Iɛ but increased Iγ1 germline transcription. By Western blot analyses, we probed the expression of transcription factors known to negatively regulate Iɛ and positively regulate Iγ1 transcription. These transcription factors were synergistically upregulated in the presence of CpG with IL-4 and anti-CD40. Together, these data illustrate a molecular mechanism by which microbial products directly perturb the production of IgE and influence the development of allergic disease. R01 HL165120

Clinical features and biomarkers of acute encephalitis with claustrum sign (P5-5.027)

<h3>Objective:</h3> To determine clinical features and biomarkers of acute encephalitis with claustrum sign on brain MRI and to clarify the pathogenetic mechanism. <h3>Background:</h3> The claustrum sign has been described as reversible bilateral claustrum MRI hyperintensity on a T2-weighted MRI image. This finding has been reported in acute encephalitis, including acute necrotizing encephalopathy, febrile infection–related epilepsy syndrome, a subcategory of new-onset refractory status epilepticus. <h3>Design/Methods:</h3> We retrospectively analyzed the clinical features and cerebrospinal fluid (CSF) findings of three patients with acute encephalitis with claustrum sign. <h3>Results:</h3> All patients had prior symptoms suspected of infection, including fever, cough, and general fatigue. Two patients had meningeal signs and refractory status epilepticus. One patient had pyramidal signs and tremor. Claustrum sign was noted at 8, 28, and 52 days after the onset. In CSF, a glial fibrillary acidic protein (GFAP) antibody was detected in one patient with pyramidal signs and tremor. The other two patients had no antineuronal antibodies detected by tissue-based immunofluorescence assay using frozen rat brain sections. Compared with GFAP antibody-negative patients, GFAP antibody-positive patients had increased levels of CSF cell counts (antibody-positive vs antibody-negative; 127/μL vs 13 and 32/μL), protein concentrations (157 mg/dL vs 39 and 58 mg/dL), and tumor necrosis factor α (25.3 pg/mL vs 2.82 and 2.95 pg/mL). Interleukin 6 (IL-6) in CSF with GFAP antibody-negative patients was significantly higher than those with antibody-positive patients (195 and 609 pg/mL vs. 19.2 pg/mL). All patients required ventilator management and received immunotherapies. Two GFAP antibody-negative patients had improved, but a GFAP antibody-positive patient had died due to pneumonia. <h3>Conclusions:</h3> Acute encephalitis with claustrum sign is associated with at least increased levels of IL6 and GFAP antibody (i.e., GFAP astrocytopathy). In addition to CSF cytokine measurements, it is considered that GFAP antibody measurements should be performed in this disorder. <b>Disclosure:</b> Dr. Shimohata has nothing to disclose. Akira Takekoshi has nothing to disclose. Kimihiko Kaneko has nothing to disclose. Juichi Fujimori has nothing to disclose. Dr. Ioku has nothing to disclose. Keisuke Imai has nothing to disclose. Dr. Kimura has nothing to disclose.

A Subset of Breg Cells, B10, Contributes to the Development of Radiation-Induced Pulmonary Fibrosis

Radiation-induced pulmonary fibrosis (RIPF) is a serious side effect of radiotherapy, but the underlying mechanisms are unknown. B10 cells, as negative B regulatory cells, play important roles in regulating inflammation and autoimmunity. However, the role of B10 cells in RIPF progression is unclear. The aim of this study was to determine the role of B10 cells in aggravating RIPF and the underlying mechanism. The role of B10 cells in RIPF was studied by constructing mouse models of RIPF and depleting B10 cells with an anti-CD22 antibody. The mechanism of B10 cells in RIPF was further explored through cocultivation of B10 cells and MLE-12 or NIH3T3 cells and administration of an IL-10 antibody to block IL-10. B10 cell numbers increased significantly during the early stage in the RIPF mouse models compared with the controls. In addition, depleting B10 cells with the anti-CD22 antibody attenuated the development of lung fibrosis in mice. Subsequently, we confirmed that B10 cells induced EMT and the transformation of myofibroblasts via activation of STAT3 signaling in vitro. After blockade of IL-10, it was verified that IL-10 secreted by B10 cells mediates the EMT of myofibroblasts, thereby promoting RIPF. Our study uncovers a novel role for IL-10-secreting B10 cells that could be a new target of research for relieving RIPF.

Abstract 5016: IL-6 influences PIM1 expression in renal cell carcinoma

Abstract Renal cell carcinoma (RCC) is the most common kidney neoplasm, accounting for over 430,000 new cases worldwide and 179,000 deaths annually. The overexpression of the proviral integration site for moloney murine leukemia virus 1 (PIM1) kinase is associated with poor clinical outcomes in patients with RCC. PIM1 is a constitutively active effector serine/threonine kinase with roles in tumor progression including cell proliferation, apoptosis, invasion, and migration. Yet, the mechanisms underlying PIM1 expression and its function in RCC are not fully delineated. IL-6 is a pleiotropic cytokine involved in the activation of the JAK/STAT signaling cascade. High serum IL-6 levels are implicated in the poor prognosis of RCC patients and are hypothesized to contribute to RCC invasion and metastasis. PIM1 is shown to be transcriptionally regulated downstream of JAK/STAT signaling and an IL-6/STAT3/PIM1 axis exists in pancreatic cancer. We thus hypothesize that this pathway also regulates expression of PIM1 in RCC. Here, we explore the levels of PIM1 in RCC and identify potential signaling pathways influencing PIM1 expression. Our results show five RCC cell lines, 769-P, 786-O, ACHN, CAKI-1, and RCC-4 cells overexpress PIM1 relative to normal renal proximal tubule epithelial cells. In RCC cell lines, IL-6 secretion correlates with PIM1 protein levels. Treatment with ruxolitinib, a JAK1/2 inhibitor, leads to a dose and concentration dependent decrease in PIM1 levels. We then tested whether incubation with an IL-6 neutralizing antibody is sufficient to regulate PIM1 protein levels. Indeed, use of an IL-6 antibody resulted in decreased pJAK2 protein and PIM1 protein. Our initial studies suggest that differential expression of PIM1 among RCC cell lines may be linked to autocrine IL-6 signaling. Interestingly, 769-P cells demonstrate increased PIM1 levels but do not secrete IL-6 and neither ruxolitinib treatment nor incubation with an IL-6 neutralizing antibody appear to affect PIM1 levels. Therefore, an IL-6/JAK-independent pathway resulting in increased PIM1 protein levels likely exists in 769-P cells. Further investigation is required to elucidate the contexts of PIM1 regulatory pathways in order to develop novel targeted anti-cancer strategies in RCC. Citation Format: Kimberly S. Meza, Sheldon L. Holder. IL-6 influences PIM1 expression in renal cell carcinoma. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 5016.

Interleukin-17 is involved in neuropathic pain and spinal synapse plasticity on mice.

Neuropathic pain seriously affects people's life, but its mechanism is not clear. Interleukin-17 (IL-17) is a proinflammation cytokine and involved in pain regulation. Our previous study found that IL-17 markedly enhanced the excitatory activity of spinal dorsal neurons in mice spinal slices. The present study attempts to explore if IL-17 contributes to neuropathic pain and spinal synapse plasticity. A model of spared nerve injury (SNI) was established in C57BL/6J mice and IL-17a mutant mice. The pain-like behaviors was tested by von Frey test and dynamic mechanical stimuli, and the expression of IL-17 and its receptor, IL-17RA, was detected by immunohistochemical staining. C-fiber evoked field potentials were recorded in vivo. In the spinal dorsal horn, IL-17 predominantly expressed in the superficial spinal astrocytes and IL-17RA expressed mostly in neurons and slightly in astrocytes. The SNI-induced static and dynamic allodynia was significantly prevented by pretreatment of neutralizing IL-17 antibody (intrathecal injection, 2μg/10μL) and attenuated in IL-17a mutant mice. Post-treatment of IL-17 neutralizing antibody also partially relieved the established mechanical allodynia. Moreover, spinal long-term potentiation (LTP) of C-fiber evoked field potentials, a substrate for central sensitization, was suppressed by IL-17 neutralizing antibody. Intrathecal injection of IL-17 recombinant protein (0.2μg/10μL) mimicked the mechanical allodynia and facilitated the spinal LTP. These data implied that IL-17 in the spinal cord played a crucial role in neuropathic pain and central sensitization.

Cadherin-11-Interleukin-6 Signaling between Cardiac Fibroblast and Cardiomyocyte Promotes Ventricular Remodeling in a Mouse Pressure Overload-Induced Heart Failure Model

Heart failure is a serious and life-threatening disease worldwide. Cadherin-11 (Cad-11) is highly expressed in the heart and closely associated with inflammation. There is currently limited understanding on how Cad-11 contributes to cardiac remodeling and its underline molecular mechanism. We found an increased expression of Cad-11 in biopsy heart samples from heart failure patients, suggesting a link between Cad-11 and heart failure. To determine the role of Cad-11 in cardiac remodeling, Cad-11-deficient mice were used in a well-established mouse transverse aortic constriction (TAC) model. Loss of Cad11 greatly improved pressure overload-induced LV structural and electrical remodeling. IL (interleukin)-6 production was increased following TAC in WT mice and this increase was inhibited in cadherin-11-/- mice. We further tested the effect of IL-6 on myocyte hypertrophy and fibrosis in a primary culture system. The addition of hCad-11-Fc to cultured cardiac fibroblasts increased IL-6 production and fibroblast cell activation, whereas neutralizing IL-6 with an IL-6 antibody resulted in alleviating the fibroblast activation induced by hCad-11-Fc. On the other hand, cardiomyocytes were promoted to cardiomyocyte hypertrophy when cultured in condition media collected from cardiac fibroblasts stimulated by hCad-11-Fc.Similarly, neutralizing IL-6 prevented cardiomyocyte hypertrophy. Finally, we found that MAPKs and CaMKII-STAT3 pathways were activated in both hCad-11-Fc stimulated fibroblasts and cardiomyocytes treated with hCad-11-Fc stimulated fibroblast condition medium. IL-6 neutralization inhibited such MAPK and CaMKII-STAT3 signaling activation. These data demonstrate that Cad-11 functions in pressure overload-induced ventricular remodeling through inducing IL-6 secretion from cardiac fibroblasts to modulate the pathophysiology of neighboring cardiomyocytes.

Interleukin-6 promotes skeletal muscle catabolism by activating tryptophan-indoleamine 2,3-dioxygenase 1-kynurenine pathway during intra-abdominal sepsis.

Inflammatory cytokine interleukin-6 (IL-6) plays a pivotal role in skeletal muscle degradation after intra-abdominal sepsis (IAS), with mechanism remained to be elucidated. Indoleamine 2,3-dioxygenase 1 (IDO-1), a key enzyme in converting tryptophan into kynurenine, could be activated by IL-6, and kynurenine has been shown to be involved in muscle degradation. We hypothesized that IL-6 could promote muscle degradation via tryptophan-IDO-1-kynurenine pathway in IAS patients. Serum and rectus abdominis (RA) were obtained from IAS or non-IAS patients. Mouse model of IAS-induced muscle wasting was generated by caecal ligation and puncture (CLP) and lipopolysaccharide (LPS) injection. IL-6 signalling was blocked by anti-mouse IL-6 antibody (IL-6-AB), and the IDO-1 pathway was blocked by navoximod. To elucidate the role of kynurenine in muscle mass and physiology, kynurenine was administered to IAS mice treated with IL-6-AB. Compared to non-IAS patients, kynurenine levels in serum (+2.30-fold vs. non-IAS, P<0.001) and RA (+3.11-fold vs. non-IAS, P<0.001) were elevated, whereas tryptophan levels in serum (-53.65% vs. non-IAS, P<0.01) and RA (-61.39% vs. non-IAS, P<0.01) were decreased. Serum IL-6 level of the IAS group was significantly higher compared to non-IAS patients (+5.82-fold vs. non-IAS, P=0.01), and muscle cross-sectional area (MCSA) was markedly reduced compared to non-IAS patients (-27.73% vs. non-IAS, P<0.01). In animal experiments, IDO-1 expression was up-regulated in the small intestine, colon and blood for CLP or LPS-treated mice, and there was correlation (R2 =0.66, P<0.01) between serum and muscle kynurenine concentrations. Navoximod significantly mitigated IAS-induced skeletal muscle loss according to MCSA analysis (+22.94% vs. CLP, P<0.05; +23.71% vs. LPS, P<0.01) and increased the phosphorylated AKT (+2.15-fold vs. CLP, P<0.01; +3.44-fold vs. LPS, P<0.01) and myosin heavy chain (+3.64-fold vs. CLP, P<0.01; +2.13-fold vs. LPS, P<0.01) protein expression in myocytes. In the presence of anti-IL-6 antibody, a significantly decreased IDO-1 expression was observed in the small intestine, colon and blood in CLP or LPS mice (all P<0.01), whereas the decrease of MCSA was alleviated (+37.43% vs. CLP+IgG, P<0.001; +30.72% vs. LPS+IgG, P<0.001). In contrast, additional supplementation of kynurenine decreased the MCSA in septic mice treated with IL-6-AB (both P<0.01). This study provided novel insights into the tryptophan-IDO-1-kynurenine-dependent mechanisms that underlie inflammatory cytokine-induced skeletal muscle catabolism during intra-abdominal sepsis.

Regulatory T cell expansion promotes white matter repair after stroke

Recent research highlights the function of regulatory T cells (Tregs) in white matter integrity in CNS diseases. Approaches that expand the number of Tregs have been utilized to improve stroke recovery. However, it remains unclear if Treg augmentation preserves white matter integrity early after stroke or promotes white matter repair. This study evaluates the effect of Treg augmentation on white matter injury and repair after stroke. Adult male C57/BL6 mice randomly received Treg or splenocyte (2 million, iv) transfer 2 h after transient (60 min) middle cerebral artery occlusion (tMCAO). Immunostaining showed improved white matter recovery after tMCAO in Treg-treated mice compared to mice received splenocytes. In another group of mice, IL-2/IL-2 antibody complexes (IL-2/IL-2Ab) or isotype IgG were administered (i.p) for 3 consecutive days starting 6 h after tMCAO, and repeated on day 10, 20 and 30. The IL-2/IL-2Ab treatment boosted the number of Tregs in blood and spleen and increased Treg infiltration into the ischemic brain. Longitudinal in vivo and ex vivo diffusion tensor imaging analysis revealed an increase in fractional anisotropy 28d and 35d, but not 14d, after stroke in IL-2/IL-2Ab-treated mice compared to isotype-treated mice, suggesting a delayed improvement in white matter integrity. IL-2/IL-2Ab also improved sensorimotor functions (rotarod test and adhesive removal test) 35d after stroke. There were correlations between white matter integrity and behavior performance. Immunostaining confirmed the beneficial effects of IL-2/IL-2Ab on white matter structures 35d after tMCAO. IL-2/IL-2Ab treatment starting as late as 5d after stroke still improved white matter integrity 21d after tMCAO, suggesting long-term salutary effects of Tregs on the late-stage tissue repair. We also found that IL-2/IL-2Ab treatment reduced the number of dead/dying OPCs and oligodendrocytes in the brain 3d after tMCAO. To confirm the direct effect of Tregs on remyelination, Tregs were cocultured with lysophosphatidyl choline (LPC)-treated organotypic cerebella. LPC exposure for 17 h induced demyelination in organotypic cultures, followed by gradual spontaneous remyelination upon removal of LPC. Co-culture with Tregs accelerated remyelination in organotypic cultures 7d after LPC. In conclusion, Boosting the number of Tregs protects oligodendrocyte lineage cells early after stroke and promotes long-term white matter repair and functional recovery. IL-2/IL-2Ab represents a feasible approach of Treg expansion for stroke treatment.

The impact of curcumin supplementation on systemic lupus erythematosus and lupus nephritis: A systematic review.

Curcumin is the active ingredient in the curry spice turmeric. It has anti-inflammatory properties due to the inhibition of transcription factors and inflammatory mediators such as nuclear factor-κβ (NF-κβ), cyclooxygenase-2 (COX2), lipoxygenase (LOX), tumor necrosis factoralpha (TNF-alpha), and interleukin-1 (IL-1) and 6 (IL-6). This review examines the literature regarding the efficacy of curcumin on systemic lupus erythematosus disease activity. A search was conducted following guidelines in the Preferred Reporting Items for Systematic reviews and Meta-Analyses (PRISMA) using the PubMed, Google Scholar, Scopus, and MEDLINE electronic databases to retrieve relevant studies assessing the impact of curcumin supplementation on SLE. The initial search yielded three double-blind, placebo-controlled, randomized clinical trials, three human in vitro studies, and seven mouse-model studies. In human trials, curcumin decreased 24-h and spot proteinuria, but the trials were small, ranging from 14 to 39 patients, with varied curcumin doses and different study durations ranging from 4 to 12 weeks. There was no change in C3, dsDNA, or the Systemic Lupus Erythematosus Disease Activity (SLEDAI) scores even in the longer trials. The mouse-model trials yielded more data. NF-κβ activation was suppressed along with inducible nitric oxide synthase (NOS) species expression when 1mg/kg/day of curcumin was administered for 14 weeks, leading to significant decreases in dsDNA, proteinuria, renal inflammation, and IgG subclasses. Another study suggested that curcumin reduced B cell-activating factor (BAFF) when used for up to 8 weeks at 50mg/kg/day. A reduction in pro-inflammatory Th1 and Th17 percentages, IL-6 and anti-nuclear antibody (ANA) levels were reported. The doses used in the murine models were much higher than those used in human trials, with 12.5mg-200mg/kg/day used for over 16 weeks; highlighting that the optimal time for an immunological effect to be observed may require 12-16 weeks of curcumin use. Despite the wide use of curcumin in everyday life, its molecular and anti-inflammatory use has only been partially explored. Current data show a potential benefit on disease activity. Still, no uniform dose can be advised because long-duration, large-scale randomized trials using defined dosing are needed in different subsets of SLE, including lupus nephritis patients.

Transplantation of Umbilical Cord-Derived Mesenchymal Stem Cells Attenuates Surgical Wound-Induced Blood-Brain Barrier Dysfunction in Mice.

Blood-brain barrier (BBB) is the most important component of central nervous system (CNS) to keep toxins and pathogens from CNS. Although our studies demonstrated that using interleukin-6 antibodies (IL-6-AB) reversed the increased permeability of BBB, IL-6-AB is limited in their application that only could be used a few hours before surgery and seemed delayed the surgical wounds healing process, which urges us to find another more effective method. In this study, we employed the C57BL/6J female mice to investigate the potential effects of umbilical cord-derived mesenchymal stem cells (UC-MSCs) transplantation on BBB dysfunction induced by surgical wound. Compared to IL-6-AB, the transplantation of UC-MSCs more effectively decreased the BBB permeability after surgical wound evaluated by dextran tracer (immunofluorescence imaging and luorescence quantification). In addition, UC-MSCs can largely decrease the ratio of proinflammatory cytokine IL-6 to the anti-inflammatory cytokine IL-10 in both serum and brain tissue after surgical wound. Moreover, UC-MSCs successfully increased the levels of tight junction proteins (TJs) in BBB such as ZO-1, Occludin, and Claudin-5 and extremely decreased the level of matrix metalloproteinase-9 (MMP-9). Interestingly, UC-MSCs treatment also had positive effects on wound healing while protecting the BBB dysfunction induced by surgical wound compared to IL-6-AB treatment. These findings suggest that UC-MSCs transplantation is a highly efficient and promising approach on protecting the integrity of BBB which caused by peripheral traumatic injuries.

Rheumatoid arthritis and the proinflammatory cytokine IL-17.

Rheumatoid arthritis (RA) is the most common inflammatory joint disease. Various proinflammatory cytokines are involved in the pathogenesis of this chronic disorder. It is characterized by the presence of autoantibodies, such as rheumatoid factor and antibodies against citrullinated peptides. The present study focuses on investigation of possible association between the proinflammatory cytokine interleukin 17 and anti-CCP, anti-MCV, and anti-CarP antibodies seropositivity in RA patients. To assess serum levels of interleukin 17 (IL-17) in patients with rheumatoid arthritis and healthy controls (HC) and to investigate the relationship between IL-17 and anti-cyclic citrullinated protein (anti-CCP) antibodies, antimutated citrullinated vimentin (anti-MCV) antibodies, and anti-carbamylated protein (anti-CarP) antibodies in patients with RA. Forty-seven patients diagnosed with rheumatoid arthritis and 44 healthy controls were included in the study. Serum IL-17 levels were examined in all participants. Anti-CCP, anti-MCV, and anti-CarP antibodies were tested in the group of RA patients. The mean serum level of IL-17 in RA patients was higher (12.8pg/ml) than that in healthy controls (7.9pg/ml), but the difference was not statistically significant (p=0.276). No significant correlation was observed between anti-CCP (+/-) and IL-17 (rs=0.162, p=0.380), and between anti-MCV (+/-) and IL-17 (rs=0.157, p=0.340). A significant positive correlation of moderate value was reported between anti-CarP (+/-) and IL-17 (rs=0.388, p=0.015). The present study demonstrated that the IL-17 serum levels in RA patients were increased compared to healthy controls. No correlation was found between ACPA immunological markers and IL-17 levels in patients with RA. A positive correlation was found between anti-CarP antibodies and IL-17 in the patients' group. The increased level of IL-17 is suggestive of its possible role in the pathogenesis of CarP positive RA patients.

Single dose tocilizumab for COVID-19 associated cytokine storm syndrome: Less is more.

We aim to evaluate the clinical pharmacokinetics of a single dose interleukin-6 (IL-6) antibody tocilizumab (TCZ) in methylprednisolone (MP)-treated COVID-19 patients with cytokine storm syndrome (CSS). MP pre-treated patients with COVID-19-associated CSS, defined as at least two elevations of C-reactive protein (CRP) >100 mg/L, ferritin >900 μg/L or D-dimers >1500 μg/L, received intravenous TCZ (8mg/kg, max. 800 mg) upon clinical deterioration. A nonlinear-mixed effects model was developed based on TCZ serum concentrations and dosing information. Population pharmacokinetic parameters were estimated and concentration-time profiles were plotted against individual predicted values. Fixed dose simulations were subsequently performed based on the final model. In total 40 patients (mean [SD] age: 62 [12] years, 20% female, body weight: 87 [17] kg) with COVID-19 induced CSS were evaluated on pharmacokinetics and laboratory parameters. A biphasic elimination of TCZ serum concentration was described by a homogeneous population pharmacokinetic model. Serum TCZ concentrations above the 1μg/L target saturation threshold were covered for 16 days in all evaluated patients treated with a single dose of 8mg/kg. In a simulation with TCZ 400 mg fixed dose, this condition of full IL-6 receptor occupancy at minimum serum concentration was also met. A single dose (8mg/kg, max. 800 mg) is sufficient to cover a period of 16 days of IL-6-mediated hyperinflammation in COVID-19-induced CSS in MP-treated patients. Based on body weight PK simulations, a fixed-dose tocilizumab of 400 mg should be considered to prevent overtreatment, future drug shortage and unnecessary drug expenditure.

A long-acting anti-IL-8 antibody improves inflammation and fibrosis in endometriosis.

Current pharmacological treatments for endometriosis are limited to hormonal agents that can relieve pain but cannot cure the disease. Therefore, the development of a disease-modifying drug for endometriosis is an unmet medical need. By studying human endometriotic samples, we found that the progression of endometriosis was associated with the development of inflammation and fibrosis. In addition, IL-8 expression was highly up-regulated in endometriotic tissues and closely correlated with disease progression. We created a long-acting recycling antibody against IL-8 (AMY109) and evaluated its clinical potency. Because rodents do not produce IL-8 and do not experience menstruation, we analyzed the lesions in cynomolgus monkeys that spontaneously developed endometriosis and in a surgically induced endometriosis monkey model. Both spontaneously developed and surgically induced endometriotic lesions demonstrated pathophysiology that was highly similar to that of human endometriosis. Once-a-month subcutaneous injection of AMY109 to monkeys with surgically induced endometriosis reduced the volume of nodular lesions, lowered the Revised American Society for Reproductive Medicine score as modified for monkeys, and ameliorated fibrosis and adhesions. In addition, experiments using cells derived from human endometriosis revealed that AMY109 inhibited the recruitment of neutrophils to endometriotic lesions and the production of monocyte chemoattractant protein-1 from neutrophils. Thus, AMY109 may represent a disease-modifying therapy for patients with endometriosis.

A novel electrochemical IL-6 sensor based on Au nanoparticles-modified platinum carbon electrode.

Introduction: Interleukin-6 (IL-6) is a multifunctional polypeptide cytokine composed of two glycoprotein chains, which plays an important role in many cellular reactions, pathological processes, diagnosis and treatment of diseases and so on. The detection of IL-6 plays a promising role in the cognition of clinical diseases. Methods: 4-mercaptobenzoic acid (4-MBA) was immobilized on the gold nanoparticles modified platinum carbon (PC) electrode with the linker IL-6 antibody, and finally formed an electrochemical sensor that specifically recognized IL-6. Through the highly specific antigen-antibody reaction, the IL-6 concentration of the samples to be detected. The performance of the sensor was studied by cyclic voltammetry (CV) and differential pulse voltammetry (DPV). Results: The experimental results showed that the linear detection range of the sensor for IL-6 was 100pg/mL-700pg/mL and the detection limit was 3pg/mL. In addition, the sensor had the advantages of high specificity, high sensitivity, high stability and reproducibility under the interference environment of bovine serum albumin (BSA), glutathione (GSH), glycine (Gly) and neuron specific enolase (NSE), which provided a prospect for specific antigen detection sensor. Discussion: The prepared electrochemical sensor successfully detected the content of IL-6 in standard and biological samples, showing excellent detection performance. No significant difference was found between the detection results of the sensor and that of ELISA. The sensor showed a very broad prospect in the application and detection of clinical samples.

Driving the Chimeric Antigen Receptor: The Road Ahead for Cell Therapy

Driving the Chimeric Antigen Receptor: The Road Ahead for Cell Therapy

Evaluation of the Immunogenicity of Recombinant Espb, Espc Proteins from Mycobacterium Tuberculosis and the Fusion Espc/Espb Protein in BALB/C Mice.

Two newly identified proteins, EspB and EspC are involved in the pathogenesis of Mycobacterium tuberculosis. The objective of the present study was to evaluate the immunogenicity of recombinant EspC, EspB, and EspC/EspB fusion proteins in mice. BALB/c mice were immunized subcutaneously with recombinant EspC, EspB, and fusion EspC/EspB proteins, three times with along with Quil-A as an adjuvant. The cellular and humoral immune responses were evaluated by quantifying IFN-γ, IL-4, IgG, IgG1, and IgG2a antibodies against the antigens. The results showed that the mice immunized with recombinant EspC, EspB, and EspC/EspB proteins did not produce IL-4, whereas IFN-γ was secreted in response to all three proteins. EspC/EspB group produced significant amounts of IFN-γ in response to stimulation with all the three recombinant proteins (P<0.001). In mice immunized with EspC, high levels of IFN-γ were detected in response to EspC/EspB, and EspC (P<0.0001); while mice immunized with EspB produced lower levels of IFN-γ in response to EspC/EspB, and EspB (P<0.05).Mice immunized with recombinant EspC, EspB, and EspC/EspB proteins exhibited significantly high levels of IgG and IgG2a/IgG1 ratio (P< 0.001). Moreover, high levels of IgG and IgG2a were detected in the sera of mice immunized with EspC/EspB fusion protein. All the three recombinant proteins induced Th1-type immune responses in mice against EspB and EspC; however, EspC/EspB protein is more desirable due to the presence of epitopes from both EspC and EspB proteins and the production of immune responses against both.