Mepolizumab Research Articles

IL-6 and PD-1 antibody blockade combination therapy regulate inflammation and T lymphocyte apoptosis in murine model of sepsis

BackgroundInterleukin-6 (IL-6) plays a central role in sepsis-induced cytokine storm involving immune hyperactivation and early neutrophil activation. Programmed death protein-1 (PD-1) is associated with sepsis-induced immunosuppression and lymphocyte apoptosis. However, the effects of simultaneous blockade of IL-6 and PD-1 in a murine sepsis model are not well understood.ResultsIn this study, sepsis was induced in male C57BL/6 mice through cecal ligation and puncture (CLP). IL-6 blockade, PD-1 blockade, or combination of both was administered 24 h after CLP. Peripheral blood count, cytokine level, lymphocyte apoptosis in the spleen, neutrophil infiltration in the lungs and liver, and survival rate were measured. The mortality rate of the IL-6/PD-1 group was lower, though not statistically significant (p = 0.164), than that of CLP mice (75.0% vs. 91.7%). The IL-6/PD-1 group had lower neutrophil percentage and platelet count compared with the CLP group; no significant difference was observed in other cytokine levels. The IL-6/PD-1 group also showed reduced T lymphocyte apoptosis in the spleen and decreased neutrophil infiltration in the liver and lungs.ConclusionsIL-6/PD-1 dual blockade reduces neutrophil infiltration, lymphocyte apoptosis, and bacterial burden while preserving tissue integrity in sepsis. Although the improvement in survival was not statistically significant, these findings highlight its potential as a therapeutic approach in sepsis.

Long-term antibody responses to COVAXIN and COVISHIELD vaccines in rheumatoid arthritis patients and healthy control population – A cross-sectional study

ABSTRACT Background: Rheumatoid arthritis (RA) is a chronic autoimmune disease that causes inflammation and damage in the joints. It often requires treatment with disease-modifying antirheumatic drugs (DMARDs) to manage symptoms and prevent progression. The study investigates the long-term antibody responses to COVAXIN and COVISHIELD vaccines in RA patients. Methodology: This cross-sectional study (IEC approval no: AlIMS/BBN/IEC/AUG/2021/60-R dated Sept 05, 2022, and Ref No: 799/U/IEC/ESICMC/F490/09/2022 dated Oct 31, 2022) enrolled 103 diagnosed RA patients receiving DMARDs and 183 healthy controls. The participants who completed 1 year after the second dose of vaccination were included, and detailed information on demographic, medical, and vaccination were collected. Laboratory investigations included complete blood count, inflammatory markers, and antispike antibody levels. Statistical analyses assessed differences between COVAXIN and COVISHIELD subgroups, considering DMARDs usage and disease duration. Results: Among RA patients, both COVAXIN and COVISHIELD groups exhibited low disease activity. No significant (P > 0.05) differences were found in IL-6, CRP, or antispike antibody levels between COVAXIN and COVISHIELD subgroups in RA patients and healthy controls. Notably, 89% of female RA patients received COVISHIELD. Co-morbidities, including hypothyroidism (44%), were prevalent in COVISHIELD-received RA patients. Antibody concentration varied significantly among DMARDs usage groups in COVAXIN-vaccinated RA patients, with a notable difference between three-drug and HCQ-alone regimens. However, no such difference was observed in the COVISHIELD group. Disease duration did not significantly impact antispike antibody concentration in either of the vaccination group. Conclusion: RA patients had a decreased antibody response, 1 year after receiving the second dose of the COVID-19 vaccine. Nonetheless, there was no discernible difference in the antispike antibody concentration between the COVISHIELD and COVAXIN vaccination groups. Additionally, immunosuppressive medications significantly impact serological responses to these vaccines.

P94 REX-7117 is a highly potent and selective oral STAT3 inhibitor that has demonstrated potential efficacy and safety differentiation versus JAK/TYK2 targeting in preclinical models of psoriasis

Abstract Inhibition of key cytokine pathways, including IL-23 and IL-17, has translated into transformative efficacy in plaque psoriasis, psoriatic arthritis and other inflammatory diseases. There is a strong unmet need to develop oral medicines which match the efficacy of biologics. Additionally, current small molecule JAK/TYK2 inhibitors are associated with on-target safety signals, including opportunistic infections and dysregulated haematologic homeostasis (anaemia, thrombocytopenia). Oral selective STAT3 inhibitors represent an opportunity to target psoriatic pathogenesis by inhibiting clinically validated inflammatory mediators, such as IL-23 and IL-6 cytokines. The STAT3 SH2 domain mediates both binding to the cytokine receptor and transcriptional activity, and therefore is an attractive therapeutic target. Recludix has developed an innovative and proprietary SH2 domain platform that has enabled the discovery of a new class of small molecule inhibitors of these previously undruggable protein domains. REX-7117 is the first orally available, reversible, SH2 domain-targeting STAT3 inhibitor. REX-7117 demonstrates low nanomolar potency in biochemical and primary human cellular assays, is highly selective across the SH2 family, including other STAT proteins, and inhibits IL-6 and IL-23 driven Th17 cell function. Unlike JAK1/2 and TYK2 inhibitors, REX-7117 does not impair broader immune responses, such as Th1 cell activity, innate interferon-dependent anti-viral immunity, or growth factor signalling critical for haematologic homeostasis. In dogs and mice, REX-7117 achieves deep, durable, and selective STAT3 inhibition at clinically relevant oral doses and exhibits comparable efficacy to an IL-17 antibody in rodent models of plaque psoriasis. Selective STAT3 inhibition has the potential to combine the efficacy of clinically validated biologics with the convenience of oral administration, while avoiding known safety concerns observed with the broader activity of JAK and TYK2 inhibitors.

Secukinumab (Anti-IL-17A Therapeutic Antibody) Improves Clinical Outcome for a Mixed Endotype CRS.

We identified two CRSwNP patients who had previously failed treatment with an anti-IL-4/IL-13 antibody (dupilumab). Based on their clinical characteristics and blood cytokine levels, we considered them mixed Type II/Type III cases and treated them with an anti-IL-17 antibody (secukinumab). Anti-IL-17 antibody secukinumab was superior in reducing their NPS and SNOT-22 values compared to dupilumab. IL-17 could be a promising target for non-type II and mixed endotype CRS treatment.

Cervical Cancer Cells Use the CD95 and IL-2 Pathways to Promote Their Proliferation and Survival.

Cervical cancer is a global health problem; therapies focused on eliminating tumour cells and strengthening different immunotherapies are in development. However, it has been observed that cervical tumour cells can evade cell death mechanisms and generate immune system molecules to promote their proliferation and metastasis. In this context, we analysed the role of the IL-2 and CD95 pathways, essential molecules in activating the immune system and eliminating tumour cells. However, it is important to analyse their role in cervical tumour cells because these cells could be using these pathways to proliferate. In this study, we found that SiHa and HeLa cells respond to treatment, with 10 IU/mL of IL-2 inducing their proliferation and 100 IU/mL of IL-2 decreasing their proliferation. We also observed that they express a high percentage of the CD95 receptor and its ligand (CD95L) and that treatment with CD95 agonist antibodies at low doses increases cell proliferation. Furthermore, simultaneous treatment with high doses of IL-2 plus CD95 agonist antibody positively regulates LC3B accumulation. We did not observe apoptosis under any of the treatments carried out. In conclusion, cervical tumour cells can use the IL-2 and CD95 pathways to induce their proliferation and potentially activate cytoprotective mechanisms for survival.

Comparison of CD30L and OX40L Reveals CD30L as a Promising Therapeutic Target in Atopic Dermatitis.

Blocking IL-13 is highly efficacious in patients with Th2-biased atopic dermatitis (AD), and recent clinical data have highlighted that targeting the T cell costimulatory molecules OX40 and OX40L (TNFSF4) holds promise for future treatment of AD. We asked whether targeting another T cell costimulatory molecule, CD30L (TNFSF8), might also be a possible treatment option in AD. Single-cell RNA-seq data from human AD skin lesions was analyzed to identify pathogenic IL-13- or IL-22-producing T cells and assess expression of CD30 and its ligand in comparison to OX40 and its ligand. Additionally, a murine model of AD with repetitive exposure to house dust mite allergen was used to compare neutralizing antibodies against CD30L with those against IL-13 or OX40L. Analysis of several scRNA-seq datasets from skin lesions of AD patients showed that transcripts for CD30 or CD30L were found expressed with OX40 or OX40L in the primary T cell populations that also expressed mRNA for IL13 and/or IL22. Suggesting that this could be therapeutically relevant, mice treated prophylactically with a blocking CD30L antibody were protected from developing maximal allergen-induced AD features, including epidermal and dermal thickening, immune cell infiltration, and expression of AD-related genes, similar to mice treated with a blocking IL-13 antibody. Moreover, therapeutic neutralization of CD30L in mice with experimental AD also reduced all of the pathological skin lesion features to a comparable extent as blocking OX40L. These data suggest that targeting the CD30-CD30L axis might hold promise as a future therapeutic intervention in human AD, similar to targeting the OX40-OX40L axis.

The role of angiotensin II and endothelin receptor antibodies in COVID19 patients with HFpEF

Abstract Background The infection caused by the coronavirus SARS-COV-2, is associated with an unusual pattern of organ damage in comparison to other viral-induced pathologies. A theory of autoimmunity induced by ACE-2/SARS-COV-2 spike protein complex or its degradation products, was proposed as a possible explanation for the versatile and multiorgan damage in COVID-19. Aim The aim of the study is to determine whether antibodies against angiotensin II receptor type 1 (AT1R) or endothelin-1 receptor type A (ETAR) do exist in plasma of COVID -19 patients with heart failure with preserved ejection fraction and whether they correlate to the severity of the disease. Methods A total of 61 patients with COVID-19 and heart failure with preserved ejection fraction and 48 controls (free of COVID19) with heart failure with preserved ejection fraction were included. Patients were divided into two groups: patients with severe disease (admitted at the intensive care (n=30)), patients with milder clinical presentation that remained in the general therapeutical department (n=31). The plasma levels of IL-6 and auto-antibodies against AT1R or ETAR in peripheral blood were measured and compared between COVID19 patients with severe and milder disease and age matched controls. Clinical and standard laboratory parameters were also collected and their association with the research biomarkers was analysed. Results Auto-antibodies against AT1R were significantly increased in severe disease course (median 19.60 U/mL, IQR 4.8-34.75) compared to milder disease (median 10.95 U/mL, IQR 7.69 – 17.67, p=0.017) and the control group (median 6.79 U/mL, IQR 5.10 – 14.05, p< 0.001). ETAR antibody titers were also significantly increased in severe disease (median 14.52, IQR 6.76-32.94), compared to milder disease course (median 8.73 U/mL, IQR 6.59 –13.45, p=0.023) and to the control group (median 6.64, IQR 6.79 – 17.92, p <0.001). The same was the trend regarding IL-6 antibodies. Severe disease (median 12.11 U/mL, IQR 5.59–19.38); milder disease disease (median 8.64U/mL, IQR 1.84 – 14.13, p=0.039); control group (median 4.28U/mL, IQR 0.11–1.43, p<0.001).Both AT1R and ETAR correlated to IL-6 plasma levels. The results did not show association with the concomitant therapy. Conclusion In conclusion, we establishеd the presence of AT1R and ETAR autoantibodies in COVID-19 patients with heart failure with preserved ejection fraction. They correlated to IL-6 plasma levels (systemic inflammation) and are associated with severe form of the disease.

The diagnostic value of cytokines in chronic glomerulonephritis in children

In modern clinical practice of pediatricians, the problem of timely adequate treatment of chronic glomerulonephritis (CGN) in children is of particular relevance, due to both the high prevalence of the disease and the severity of its course, the complexity of therapy and the ambiguity of the prognosis. Lesions of the urinary system in children are not only common, but also tend to grow, and often at an early age. Deterioration of the environmental background, toxic and allergic effects of drugs lead to damage primarily to the kidneys, which are the eliminating organ. The aim of the study was to study urocytokines to assess the immune response in children with HCG, depending on the association with cytomegalovirus infection. The study included 100 children aged 4-7 years, permanently residing in the Bukhara region of the Republic of Uzbekistan. At the time of the study, the patients were undergoing routine treatment in the Department of Pediatric Nephrology of the Bukhara Regional Children’s Multidisciplinary Medical Center. All children underwent general clinical (general blood test with leukoformula, general urine analysis, biochemical blood test with determination of urea, creatinine and cystatin C, immunological (TNFα, IL-18, MCP-1, IgM and IgG antibodies to cytomegalovirus infection (CMVI) in blood serum, IL-1β and IL-17A in urine) examination methods. Thus, it was found that the concentration of cystatin C in the blood serum was inversely proportional to the glomerular filtration rate in the kidneys – with a decrease in kidney function in sick children, a twofold increase was noted, that is, the accumulation of cystatin C in the blood. An increase in the concentration of IL-18 in serum and IL-1β in urine relative to the control group in CGN was an indicator of the severity of an autoimmune reaction, and a relatively low concentration of cytokines in both blood and urine in CGN with CMVI indicated suppression of the specific antiviral immunity of CMVI. It was found that an increase in serum MCP-1 by 1.4 times in group 1 and 2.9 times in group 2 of CGN with CMVI is an indicator of viral kidney damage. A significantly high concentration of IL-17A in urine in CGN with CMVI indicated local cytokine production in the kidneys and acted as an indicator of the prognosis of the outcome of CGN.

Long-term efficacy of mepolizumab in patients with eosinophilic granulomatosis with polyangiitis: a propensity score matching analysis in the multicenter REVEAL cohort study.

Mepolizumab (MPZ) has demonstrated efficacy in clinical trials for eosinophilic granulomatosis with polyangiitis (EGPA); however, few studies compare the disease course between patients treated with MPZ (MPZ group) and those who were not treated with MPZ (non-MPZ group) in real-world settings. This study aimed to compare the disease course and outcomes between the two groups and assess the long-term efficacy of MPZ in a multicenter cohort in Japan. Methods: We enrolled 113 EGPA patients registered in the cohort until June 2023. Data on clinical characteristics, disease activity, organ damage, treatments, and outcomes were retrospectively collected. To minimize potential confounding factors, we conducted propensity score matching (PSM). After PSM, 37 pairs of matched patients were identified. Clinical characteristics, including age at disease onset, sex, disease duration at last observation, antineutrophil cytoplasmic antibody positivity at disease onset, Birmingham Vasculitis Activity Score (BVAS) at disease onset, and Five-factor score at disease onset, were comparable between the groups. The median BVAS at the last observation was 0 in both groups; however, more cases in the non-MPZ group exhibited elevated BVAS, resulting in a significantly higher BVAS in the non-MPZ group at the last observation (median; MPZ group: 0, non-MPZ group: 0, p=0.028). The MPZ group had significantly lower glucocorticoid (GC) doses at the last observation (median; MPZ group: 4 mg/day, non-MPZ group: 5 mg/day, p=0.011), with a higher proportion achieving a GC dose ≤ 4 mg/day at the last observation (MPZ group: 51.4%, non-MPZ group: 24.2%, p=0.027). Three models of multivariable logistic regression analyses were performed to identify factors associated with GC doses ≤ 4 mg/day at the last observation. In all models, achieving a GC dose ≤ 4 mg/day was positively associated with MPZ administration and inversely associated with asthma at disease onset. Finally, we evaluated the survival rates between the groups, and the 5-year survival rates were significantly higher in the MPZ group compared to the non-MPZ group (MPZ group: 100%, non-MPZ group: 81.3%, p=0.012). Mepolizumab not only contributes to disease activity control but also reduces the GC dose, which may lead to improved survival in EGPA patients.

Evaluation The Role Of Some Immunological Markers In Coronavirus-19 Infection In A Sample Of Iraqi Patients

Background: cells produce proinflammatory cytokines like il-8, which activates neutrophils and stimulates immunological responses. Il-6 is produced quickly and transiently in response to infections and stimulates hematopoiesis and acute phase responses, aiding host defense. Dysregulated il-6 production can lead to chronic inflammation and autoimmunity despite its transcriptional and posttranscriptional regulation. Objective: the study aims to identify immunological markers that could serve as diagnostic factors for covid-19 infections, using rt-pcr for diagnosis and estimation of interleukin-6 and interleukin-8 levels and evaluating their association with infection progression, given the importance of covid-19 in various medical fields. Methods: blood samples collected from al-zafaraniyah general hospital, fatima-alzahra hospital, and al-yarmook teaching hospital in baghdad city identified 40 patients with coronavirus. The study used the enzyme-linked immune sorbent assay (elisa) approach to detect interleukin-6 and interleukin-8 antibodies in the patients ' serum. Results: the study reveals a strong correlation between immunological markers and corona virus-19 infection, with 40 patients showing higher levels of interleukin-6 and interleukin-8 antibodies, indicating a significant association between these markers. Conclusions: the study found a significant association between coronavirus-19 infection patients and immunological markers, with high levels of interleukin-6 and interleukin-8 antibodies in serum highly associated with coronavirus -19 incidence.

Deciphering the mechanisms of the IL-6/CXCL1 and NOD-like receptor signaling pathways in otitis media with effusion in rodents

BackgroundOtitis media with effusion (OME) often leads to pediatric hearing loss and is influenced by innate and adaptive immune responses. Innate immunity serves as the non-specific first line of defense against OME. MethodsWe induced OME in rats using ovalbumin. We administered IL-6 monoclonal antibodies intranasally to inhibit IL-6, and we injected an NF-κB inhibitor intraperitoneally to explore the role of IL-6 in innate immunity and its interaction with the NOD-like receptor signaling pathway. We analyzed RNA-sequencing data with Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathways to assess signaling pathways involved in OME. We also utilized Western blot, quantitative real-time PCR, and immunohistochemistry on middle ear samples and used microscopy to identify immune cells in ear wash fluids. ResultsOur study suggests a pivotal role for IL-6 in the immune pathways of rats with OME via the regulation of CXCL1-mediated pathways. Increased levels of IL-6 and CXCL1 were observed in the middle ear tissues, and activation of the NLRP3 inflammasome in OME rats led to an immune response via NF-κB, thus promoting IL-6 and CXCL1 production, which was reduced by IL-6 antibody treatment. ConclusionsOur findings confirm that IL-6 and CXCL1 play significant roles in the innate immune response in OME in rodents, predominantly via the NOD-like receptor signaling pathway and NLRP3 inflammasome activation. This research sheds light on OME pathogenesis and its immune-related mechanisms.

Optimization of Nanowell-Based Label-Free Impedance Biosensor Based on Different Nanowell Structures.

Nanowell-based impedance-based label-free biosensors have demonstrated significant advantages in sensitivity, simplicity, and accuracy for detecting cancer biomarkers and macromolecules compared to conventional impedance-based biosensors. Although nanowell arrays have previously been employed for biomarker detection, a notable limitation exists in the photolithography step of their fabrication process, leading to a reduced efficiency rate. Historically, the diameter of these nanowells has been 2 μm. To address this issue, we propose alternative geometries for nanowells that feature larger surface areas while maintaining a similar circumference, thereby enhancing the fabrication efficiency of the biosensors. We investigated three geometries: tube, spiral, and quatrefoil. Impedance measurements of the samples were conducted at 10 min intervals using a lock-in amplifier. The study utilized interleukin-6 (IL-6) antibodies and antigens/proteins at a concentration of 100 nM as the target macromolecules. The results indicated that tube-shaped nanowells exhibited the highest sensitivity for detecting IL-6 protein, with an impedance change of 9.55%. In contrast, the spiral, quatrefoil, and circle geometries showed impedance changes of 0.91%, 0.95%, and 1.62%, respectively. Therefore, the tube-shaped nanowell structure presents a promising alternative to conventional nanowell arrays for future studies, potentially enhancing the efficiency and sensitivity of biosensor fabrication.

Abstract 05: Renal Resident Tissue Macrophages Contribute to Salt-Dependent Increases in IL-6

Salt-Sensitive hypertension is associated with an increase in pro-inflammatory cytokines and immune cells. We previously showed an acute increase in systolic blood pressure following IL-6 + high salt (HS) after 3 days, coupled with an immediate (1 day) reduction in sodium excretion. Renal resident tissue macrophages (rRTMs) form an intricate network within the kidney and function to both maintain homeostasis and secrete pro-inflammatory cytokines. We hypothesize that rRTMs are primary contributors to the immediate increases in IL-6 following high salt. To test this hypothesis, male wild-type mice lacking a functional CX 3 CR 1 chemokine receptor (CX 3 CR 1 - EGFP+/+ ) or wild-type (Wt, C57Bl6) were used. CX 3 CR 1 is required for RTM localization to the kidney; thus, these mice are rRTM-depleted. Mice were fed HS (1% saline or 4% HS chow) or normal chow (NC) for 6 days and kidneys harvested. Kidneys were digested and FACS analysis performed on total immune cell populations or used for RT-PCR (n=4-5 Wt & rRTM-depleted). Cells were gated on: Live + CD45 + + MHCII + Lyc6C - + B220 - + CD64 + + F4/F80 hi + CD11b lo. Or Lyc6C hi . Lyc6C hi and rRTMs were pre-stimulated (LPS, 1mg/mL, 3hrs) before staining with intracellular IL-6 antibody. FACS data were analyzed as a percent increase (mean fluorescence intensity, MFI) of IL-6 over control. RT-PCR data were analyzed as DCt/housekeeping. Our data show that rRTMs are the most abundant immune cell in the kidney (23.4% of total CD45 + ) and predominantly reside in the cortex and outer medulla (>99%). HS feeding (4%) significantly increased whole kidney IL-6 mRNA (12.0 ± 0.5 vs . 15.3 ± 0.6 DCt Wt NC, **p<0.01). This increase was completely prevented in rRTM-depleted mice. While intracellular IL-6 levels were generally increased in LPS-stimulated Lyc6C hi inflammatory monocytes, the rRTMs showed a preferential increase in intracellular IL-6 with both hyperosmolar saline (1%, >13% increase) and HS chow (4%, >50%). Here, we show that rRTM-depletion prevents the HS-induced increase in kidney IL-6. We additionally show that rRTMs are preferentially increasing intracellular levels of IL-6 following an acute HS load of hyperosmolar saline (1%) or HS chow (4%). We previously demonstrated that IL-6 contributes to increased blood pressure and sodium retention, and these data suggest that rRTMs preferentially contribute to the increased IL-6.

Non-Invasive Detection of Cytokines in Saliva Towards Asthma Phenotyping Using an Electrochemical Sensor

Introduction: Asthma is a complicated and heterogeneous condition of the lungs which is usually associated with the chronic inflammation of the airways. Traditionally, classification of asthma has been based on clinical attributes such as the symptoms observed and response to treatment. However, with the growing understanding of the heterogeneity of asthma as a spectrum with varying underlying causes, asthma phenotyping has been on the horizon. Cytokines are the chemical messengers that play a crucial role in the immune system. They mediate communication between the cells in the immune system and as such have a direct effect on the inflammation process. The role of some cytokines has been established in the pathogenesis of asthma. Due to the involvement of cytokines in the modulation of asthma, they can be excellent target for asthma phenotyping. This is also helpful because identification and quantification of these inflammatory mediators can allow for more precise medicine for more targeted treatment. To aid the identification and quantification of cytokines in the phenotyping of asthma, we have created a customizable (currently for IL-8 and IL-10) multi-marker electrochemical platform for the detection of cytokines non-invasively in saliva. The developed electrochemical platform can detect cytokines rapidly and non-invasively within reported physiological ranges of the given cytokines. Evaluating the presence and quantity of the cytokines will be useful in the phenotyping and treatment plans adopted by physicians.Materials and Methods: The electrochemical sensor consists of 6 multiplexed gold electrodes on a PCB substrate. IL-8 and IL-10 antibodies were immobilized onto the sensor surface using a gold-thiol linker chemistry. Sensor response was evaluated with non- faradaic Electrochemical Impedance Spectroscopy (nf-EIS). Pooled human saliva was spiked with increasing doses of IL-8 and IL-10 in their respective physiological ranges to develop a dose response curve. Spike and recovery experiments were conducted to validate the efficacy of dose response curve.Results and Discussion: Sensor response to both IL-8 and IL-10 spiked doses in pooled saliva showed a dose-dependent response to increasing concentrations of IL-8 and IL-10 respectively as illustrated by the Nyquist plots in Fig.1. The change in impedance with increasing doses of IL-8 and IL-10 in pooled human saliva lends to credence to the establishment of the presence and quantification of cytokines in saliva that can be used by clinicians and scientists alike in the further development of personalized medicine for severe asthmatics unresponsive to traditional asthmatic treatments.Conclusion: This study presents a noninvasive electrochemical sensor for rapid and early detection of cytokines in pooled human saliva towards asthma phenotyping. The sensor exhibits a dose dependent response to IL-8 and IL-10, markers implicated in pathogenesis of asthma. Sensor capability was demonstrated with spike and recovery experiments illustrating the ability of the developed electrochemical sensor to detect and quantify cytokines in human saliva. Figure 1

IL-2 Complexed With Anti–IL-2 Antibody Expands the Maternal T-Regulatory Cell Pool and Alleviates Fetal Loss in Abortion-Prone Mice

Regulatory T (Treg) cells are essential for immune tolerance of embryo implantation and insufficient Treg cells are implicated in early pregnancy loss. An abortion-prone mouse model was utilized to evaluate the utility of IL-2 complexed with JES6-1 anti-IL-2 antibody (IL-2/JES6-1) to boost uterine Treg cells and improve reproductive success. IL-2/JES6-1 but not IL-2/IgG control administered in the peri-conception phase to CBA/J females mated with DBA/2 males elicited a >2-fold increase in the proportion of CD4+ T cells expressing FOXP3, and an increase in the ratio of FOXP3+ Treg cells to FOXP3- T conventional cells, in the uterus and its draining lymph nodes at embryo implantation that was sustained into mid-gestation. An attenuated phenotype was evident in both thymic-derived and peripheral Treg cells with elevated CTLA4, CD25, and FOXP3 indicating improved suppressive function, as well as increased proliferative marker Ki67. IL-2/JES6-1 treatment reduced fetal loss from 31% to 10%, but this was accompanied by a 6% reduction in late gestation fetal weight, despite comparable placental size and architecture. Similar effects of IL-2/JES6-1 on Treg cells and fetal growth were seen in CBA/J females with healthy pregnancies sired by BALB/c males. These findings show that expanding the uterine Treg cell pool through targeting IL-2 signaling is a strategy worthy of further investigation for mitigating immune-mediated fetal loss.

Au Nanoshell-Based Lateral Flow Immunoassay for Colorimetric and Photothermal Dual-Mode Detection of Interleukin-6.

Interleukin-6 (IL-6) detection and monitoring are of great significance for evaluating the progression of many diseases and their therapeutic efficacy. Lateral flow immunoassay (LFIA) is one of the most promising point-of-care testing (POCT) methods, yet suffers from low sensitivity and poor quantitative ability, which greatly limits its application in IL-6 detection. Hence, in this work, we integrated Aushell nanoparticles (NPs) as new LFIA reporters and achieved the colorimetric and photothermal dual-mode detection of IL-6. Aushell NPs were conveniently prepared using a galvanic exchange process. By controlling the shell thickness, their localized surface plasmon resonance (LSPR) peak was easily tuned to near-infrared (NIR) range, which matched well with the NIR irradiation light. Thus, the Aushell NPs were endowed with good photothermal effect. Aushell NPs were then modified with IL-6 detection antibody to construct Aushell probes. In the LFIA detection, the Aushell probes were combined with IL-6, which were further captured by the capture IL-6 antibody on the test line of the strip, forming a colored band. By observation with naked eyes, the colorimetric qualitative detection of IL-6 was achieved with limit of 5 ng/mL. By measuring the temperature rise of the test line with a portable infrared thermal camera, the photothermal quantitative detection of IL-6 was performed from 1~1000 ng/mL. The photothermal detection limit reached 0.3 ng/mL, which was reduced by nearly 20 times compared with naked-eye detection. Therefore, this Aushell-based LFIA efficiently improved the sensitivity and quantitative ability of commercial colloidal gold LFIA. Furthermore, this method showed good specificity, and kept the advantages of convenience, speed, cost-effectiveness, and portability. Therefore, this Aushell-based LFIA exhibits practical application potential in IL-6 POCT detection.

Prophylactic IL-23 blockade uncouples efficacy and toxicity in dual CTLA-4 and PD-1 immunotherapy

BackgroundImmune-related adverse events (irAEs), characterized by targeted inflammation, occur in up to 60% of patients with melanoma treated with immune checkpoint inhibitors (ICIs). Evidence proved that the baseline peripheral blood...

IL-22/IL-22RA1 Promotes Human Tenon's Capsule Fibroblasts Proliferation and Regulates Fibrosis Through STAT3 Signaling Pathway.

Purpose: Although it is now understood that most antiglaucoma surgeries fail because of scarring of the filtering tract, the underlying mechanism remains to be elucidated. The present study investigated the mechanism by which the interleukin (IL)-22/IL-22 receptor alpha 1 (IL-22RA1) signaling pathway regulates scar formation in glaucoma patients. Method: A total of 31 glaucoma patients who underwent trabeculectomy surgery with uncontrollable intraocular pressure because of scarring and 19 strabismus patients as the control patient group were included in the present study. ELISA was performed to measure the content of IL-22 in peripheral blood. Serum from patients was used to incubate human Tenon's capsule fibroblasts (HTFs) cells and IL-22 antibody rescued the effect of IL-22 on the biological functions. qPCR and Western blot were performed to determine IL-22RA1 mRNA and protein expression levels. Flow cytometry was performed to assess the cell cycle distribution and the Cell Counting Kit-8 assay was used to analyze cell proliferation. Results: The ELISA assay revealed that the serum IL-22 level of glaucoma patients was significantly higher than the healthy group (29.80 ± 5.1 ng/µL vs. 5.21 ± 0.9 ng/µL). After incubation with patient serum, the proliferation and activation of human Tenon fibroblasts (HTFs) were promoted. IL-22 mediated the biological function of HTFs via directly binding IL-22RA1. Moreover, transfection of the siR-IL-22RA1 or IL-22RA1 gene resulted in significant antifibrosis or profibrosis in HTFs. When a signal transducer and activator of transcription (STAT) 3 inhibitor (BAY) was introduced to the IL-22RA1 overexpression group, IL-22-induced proliferation was reduced in HTFs. Additionally, glaucoma patients had increased levels of IL-22 expression following surgery. Conclusions: The IL-22/IL-22RA1/STAT3 signaling pathway promoted fibroblast cell proliferation and alpha-smooth muscle actin, potentially regulating fibrosis in glaucoma filtration tracts. Our results provide hitherto undocumented insights into the pathophysiology of postoperative scarring.

Polymer Micropatches as B-Cell Engagers.

B cells, despite their several unique functionalities, remain largely untapped for use as an adoptive cell therapy and are limited to in vitro use for antibody production. B cells can be easily sourced, they possess excellent lymphoid-homing capabilities, and they can act as antigen-presenting cells (APCs), offering an alternative to dendritic cells (DCs), which have shown limited efficacy in the clinical setting. Soluble factors such as IL-4 and anti-CD40 antibody can enhance the activation, survival, and antigen-presenting capabilities of B cells; however, it is difficult to attain sufficiently high concentrations of these biologics to stimulate B cells in vivo. Micropatches as Cell Engagers (MACE) are polymeric microparticles, surface functionalized with anti-CD40 and anti-IgM, which can attach to B cells and simultaneously engage multiple B-cell receptors (BCR) and CD40 receptors. Stimulation of these receptors through MACE, unlike free antibodies, enhanced the display of costimulatory molecules on the B-cell surface, increased B-cell viability, and improved antigen presentation by B cells to T cells in vitro. B-cell activation by MACE further synergized with soluble IL-4 and anti-CD40. MACE also elicited T-cell chemokine secretion by B cells. Upon intravenous adoptive transfer, MACE-bound B cells homed to the spleen and lymph nodes, key sites for antigen presentation to T cells. Adoptive transfer of MACE-B cells pulsed with the CD4+ and CD8+ epitopes of ovalbumin significantly delayed tumor progression in a murine subcutaneous EG7-OVA tumor model, demonstrating the functional benefit conferred to B cells by MACE.

Evaluation of drug resistance for EGFR-TKIs in lung cancer via multicellular lung-on-a-chip

Drug resistance to irreversible epidermal growth factor receptor tyrosine kinase inhibitors (EGFR-TKIs) is a primary factor affecting their therapeutic efficacy in human non-small cell lung cancer (NSCLC). NSCLC cells can undergo epithelial-mesenchymal transition (EMT) induced by many factors in the tumour microenvironment (TME), which plays a crucial role in tumour drug resistance. In this study, a multicellular lung-on-a-chip that can realise the cell co-culture of the human non-small cell lung cancer cell line HCC827, human foetal lung fibroblasts (HFL-1), and human umbilical vein endothelial cells (HUVECs) is prepared. The TME was simulated on the chip combined with perfusion and other factors, and the drug evaluation of osimertinib was performed to explore the drug resistance mechanism of EGFR-TKIs. In the early stages, a two-dimensional static cell co-culture was achieved by microchip, and the results showed that HFL-1 cells could be transformed into cancer-associated fibroblasts (CAFs), and HCC827 cells could undergo EMT, both of which were mediated by Interleukin-6 (IL-6). Vimentin (VIM) and Alpha Skeletal Muscle Actin (a-SMA) expression of HFL-1 was upregulated, whereas E-cadherin (E-cad) expression of HCC827 was down-regulated. Further, N-cadherin (N-cad) expression of HCC827 was upregulated. In both the static cell co-culture and multicellular lung-on-a-chip, HCC827 cells with CAFs co-culture or IL-6 treatment developed resistance to osimertinib. Further use of the IL-6 antibody inhibitor tocilizumab could reverse EGFR-TKI resistance to a certain extent. Combination therapy with tocilizumab and EGFR-TKIs may provide a novel therapeutic strategy for overcoming EGFR-TKI resistance caused by EMT in NSCLC. Furthermore, the lung-on-a-chip can simulate complex TME and can be used for evaluating tumour resistance and exploring mechanisms, with the potential to become an important tool for personalised diagnosis, treatment, and biomedical research.