IL-8 Levels Research Articles

Investigation of cytokine imbalance in schizophrenia, assessment of the possible role of serum cytokine levels in predicting treatment response, prognosis and psychotic relapses

IntroductionSchizophrenia, a multisystem chronic psychiatric disorder of unknown etiology, is associated with several immune dysfunctions, including abnormal levels of circulating cytokines. Exsisting evidence shows a potential causative role for cytokines in schizophrenia symptom development. Furthermore, disease duration, symptom severity, aggressive behavior, and cognitive deficits are correlated with levels of certain cytokines. Despite the development of new antipsychotics, the negative and cognitive symptoms of schizophrenia often do not respond adequately to pharmacotherapy.ObjectivesResearch questions and hypotheses: 1. Can there be a cytokine or cytokines among the different cytokine levels detected in schizophrenia that can be used as biomarkers of treatment response? 2. Can changes in cytokine levels indicate the occurrence of psychotic relapse? 3. Can changes in the cytokine level play a role in predicting the prognosis of the disease? The secondary objectives of the planned research, in addition to the above, are to clarify the knowledge gathered so far about the relationship between cytokine level changes and the clinical symptoms associated with them.Methods We investigate cytokine levels, blood samples are taken on hospital admission. Based on the publications, we mainly focus on the Il-2, Il-4, Il-6 and Il-10 levels, which can serve as possible predictive biomarkers relating to treatment response. We will also assess the possible role of abnormal cytokine levels and their association with symptoms severity and their potential clinical implications. The severity of the symptoms is monitored with the PANSS.Results15 schizophrenic patients who were hospitalized due to a psychotic relapse have been included. Blood samples were taken to measure cytokine levels, the PANSS scale was recorded during a psychotic relapse. We have included 9 healthy, age- and gender-matched healthy controls in the study, from whom blood samples were taken to measure cytokine levels. Preparation for measurement of cytokine levels is underway. Patient involvement is ongoing.ConclusionsA better understanding of cytokine imbalance in schizophrenia patients can potentially help in early diagnosis, novel therapeutic target indentification and development, patient stratification for choosing the best therapeutic protocol, and predicting prognosis, relapse and treatment response.Disclosure of InterestNone Declared

Studying of anti-inflammatory and antioxidant effects of tectorigenin in ovalbumin-induced asthma mice models.

Allergic asthma is an important respiratory system problem characterized by airway inflammation, breathlessness, and bronchoconstriction. Allergic asthma and its outcomes are triggered by type 2 allergic immune responses. Tectorigenin is a methoxy-isoflavone with anti-inflammatory effects. In this study, we investigated the effects of tectorigenin on the pathophysiology of allergic asthma in an animal model. Asthmatic mice were treated with tectorigenin. Then airway hyperresponsiveness (AHR), eosinophil percentage, levels of interleukin (IL)-33, IL-25, IL-13, IL-5, IL-4, total and ovalbumin (OVA)-specific immunoglobulin (Ig)E, and lung histopathology were evaluated. Tectorigenin significantly (P 〈 0.05) reduced eosinophil infiltration (41 ± 7%) in the broncho-alveolar lavage fluid (BALF), serum IL-5 level (41 ± 5, pg/mL), and bronchial and vascular inflammation (scores of 1.3 ± 0.2 and 1.1 ± 0.3, respectively) but had no significant effects on AHR, serum levels of IL-33, -25, -13, and -4 (403 ± 24, 56 ± 7, 154 ± 11, and 89 ± 6 pg/mL, respectively), total and OVA-specific IgE (2684 ± 265 and 264 ± 19 ng/mL, respectively), goblet cell hyperplasia, and mucus production. Tectorigenin could control inflammation and the secretion of inflammatory mediators of asthma, so it can be regarded as a potential antiasthma treatment with the ability to control eosinophilia-related problems.

Mesenchymal Stem Cells and Curcumin Effectively Mitigate Freund's Adjuvant- induced Arthritis via their Anti-inflammatory and Gene Expression of COX-1, IL-6 and IL-4.

Rheumatoid arthritis (RA) is a type of arthritis that damages joints and can affect the thymus and the spleen. RA is an autoimmune disorder in which the immune system targets the body's own tissues. The causes of RA are unknown, although a genetic link is thought to be involved. The objective of this research was to evaluate the effect of curcumin, mesenchymal stem cells (MSCs), and their combination on the disruption of serum cytokines, ankle joint, thymus and spleen histopathology, and affected genes in complete Freund's adjuvant (CFA)-induced arthritis in male and female Wistar rats. Experimental animals were organized into 16 groups (6 animals for each), eight groups including male rats and the other eight groups including females rats. The groups are normal control, CMC, curcumin, MSCs, CFA, CFA/curcumin, CFA/ MSCs and the arthritic group treated with MSCs and curcumin. One subcutaneous injection of 0.1 mL CFA was given to rats into the right hind leg footpad to induce RA. The arthritic rats were intravenously injected three times with bone marrow-derived MSCs (BM-MSCs) and/or treated orally with curcumin daily (100 mg per kg body weight per day) for 21 days. Curcumin and BM-MSCs work together to dramatically (P < 0.05) restore the high serum PGE2 and IL-17 levels and lower the IL-13 level in arthritic rats to normal levels. Deleterious effects on the spleen and thymus histological structure were counteracted. Gene expression of COX-1 and IL-6 was increased and IL-4 was decreased; these changes were improved by the combination treatment (P < 0.05). Based on these findings, additive therapeutic effects on RA occur from the combined treatment of curcumin and BM-MSCs compared with their individual use (P < 0.05). Thus, it can be said that both curcumin and BM-MSCs are effective at reducing inflammation while also having beneficial effects on the ankle joint, thymus and spleen.

Monosodium iodoacetate induces Cartilage degradation and inflammation in rats in a dose- and time-dependent manner

Monosodium iodoacetate (MIA) is a widely recognized agent for inducing cartilage degradation and inflammation in animal models. In this study, we systematically investigated the dose- and time-dependent effects of MIA on cartilage degradation in the knees of rats. We evaluated knee diameter, knee bend score, and the levels of pro-inflammatory cytokines (IL-1 and IL-8) and cartilage degradation markers (CTX-II) to gain insights into cartilage damage and inflammation progression. Wistar rats were categorized into groups receiving various doses of MIA via intra-articular injection (0.1 mg, 0.5 mg, 1 mg, 3 mg, and 5 mg) and were observed at multiple time intervals (2, 4, 6, 8, 10, and 12 weeks). We measured knee diameter to gauge joint swelling, assigned knee bend scores to assess functional limitations, and analyzed synovial fluid samples for IL-1β, IL-8, and CTX-II levels. Our data show that low-dose MIA increases pain sensitivity, knee diameter, inflammatory cytokines IL-1β and IL-8, and CTX-II levels, which progress slowly over 12 weeks. On the other hand, higher dosages of MIA caused cartilage deterioration after two weeks, followed by an increase in inflammatory cytokines. This research sheds light on MIA administration's dose- and time-dependent effects on cartilage breakdown and inflammation in rat knee joints. Analyzing knee diameter, knee bend score, IL-1β, IL-8, and CTX-II as evaluation parameters provides a multidimensional perspective of cartilage injury and inflammatory dynamics. These findings help us comprehend cartilage-related illnesses and have implications for future research on therapeutic approaches for cartilage disorders.

IL-33/soluble ST2 axis is associated with radiation-induced cardiac injury.

Radiotherapy for treating breast cancer is associated with cardiac damage. This study aimed to investigate the role of the interleukin (IL)-33/soluble receptor ST2 (sST2) axis in radiation-induced cardiac injury. Expressions of IL-33 and sST2 were detected in breast cancer patients following radiotherapy, radiation-induced cardiac damaged mice model, and cardiomyocytes using quantitative real-time PCR (qRT-PCR) and immunohistochemical assay. Cardiac injury was evaluated through an ultrasound imaging system and hematoxylin & eosin staining. The transcriptional factor was assessed using dual-luciferase reporter assay and chromatin immunoprecipitation. The results indicated that IL-33 and sST2 were highly expressed in breast cancer patients, which further elevated post-6 months but reduced after 12 months of radiotherapy. Radiation induces cardiac dysfunction and elevated IL-33 and sST2 levels in a time-dependent manner. However, silencing of IL-33 decreased sST2 expression to alleviate radiation-induced cardiac dysfunction. The IL-33 could be transcriptional activated by TCF7L2 by binding to IL33 promoter sites, which mutation alleviated cardiomyocyte injury caused by radiation. Additionally, radiation treatment resulted in higher levels of TCF7L2, IL-33, and sST2 in cardiomyocytes, and TCF7L2 knockdown reduced IL-33 and sST2 expression. In conclusion, TCF7L2 transcriptional-activated IL-33 mediated sST2 to regulate radiation-induced cardiac damage, providing novel insights into radiotherapy-induced cardiac damage.

Immunomodulation in allergic rhinitis: Insights from Th2 cells and NLRP3/IL-18pathway.

Allergic rhinitis (AR) is characterized by nasal symptoms such as rubbing and sneezing, often triggered by allergen exposure. The purpose of this study is to dissect the roles of NLRP3-mediated immune modulation and macrophage pyroptosis in modulating T cell differentiation within the context of ovalbumin (OVA)-induced AR in mice. OVA-induced AR was established in mice, evaluating nasal symptoms, macrophage infiltration, cytokine levels, and T cell differentiation. Manipulations using NLRP3-/-, ASC-/- mice, clodronate liposome treatment, and NLRP3 inhibitor MCC950 were performed to assess their impact on AR symptoms and immune responses. Following OVA stimulation, increased nasal symptoms were observed in the OVA group along with augmented GATA3 expression and elevated IL-4 and IL-1b levels, indicative of Th2 polarization and cellular pyroptosis involvement. NLRP3-/- and ASC-/- mice exhibited reduced CD3+ T cells post OVA induction, implicating cellular pyroptosis in AR. Macrophage depletion led to decreased IgE levels, highlighting their involvement in allergic responses. Further investigations revealed enhanced macrophage pyroptosis, influencing Th1/Th2 differentiation in AR models. IL-18 released through NLRP3-mediated pyroptosis induced Th2 differentiation, distinct from IL-1b. Additionally, MCC950 effectively mitigated AR symptoms by modulating Th2 responses and reducing macrophage infiltration. This comprehensive study unravels the pivotal role of NLRP3-mediated immune modulation and macrophage pyroptosis in Th1/Th2 balance regulation in OVA-induced AR. Targeting NLRP3 pathways with MCC950 emerged as a promising strategy to alleviate AR symptoms, providing insights for potential therapeutic interventions in AR management.

Characterization of Th17 tissue-resident memory cells in non-inflamed intestinal tissue of Crohn's disease patients

Crohn's disease (CD) is a chronic inflammatory disorder affecting the bowel wall. Tissue-resident memory T (Trm) cells are implicated in CD, yet their characteristics remain unclear. We aimed to investigate the transcriptional profiles and functional characteristics of Trm cells in the small bowel of CD and their interactions with immune cells. Seven patients with CD and four with ulcerative colitis as controls were included. Single-cell RNA sequencing and paired T cell receptor sequencing assessed T cell subsets and transcriptional signatures in lamina propria (LP) and submucosa/muscularis propria-enriched fractions (SM/MP) from small bowel tissue samples. We detected 58,123 T cells grouped into 16 populations, including the CD4+ Trm cells with a Th17 signature and CD8+ Trm clusters. In CD, CD4+ Trm cells with a Th17 signature, termed Th17 Trm, showed significantly increased proportions within both the LP and SM/MP areas. The Th17 Trm cluster demonstrated heightened expression of tissue-residency marker genes (ITGAE, ITGA1, and CXCR6) along with elevated levels of IL17A, IL22, CCR6, and CCL20. The clonal expansion of Th17 Trm cells in CD was accompanied by enhanced transmural dynamic potential, as indicated by significantly higher migration scores. CD-prominent Th17 Trm cells displayed an increased interferon gamma (IFNγ)-related signature possibly linked with STAT1 activation, inducing chemokines (i.e., CXCL10, CXCL8, and CXCL9) in myeloid cells. Our findings underscored the elevated Th17 Trm cells throughout the small bowel in CD, contributing to disease pathogenesis through IFNγ induction and subsequent chemokine production in myeloid cells.

Optimized production of full-length PCV2d virus-like particles in Escherichia coli: A cost-effective and high-yield approach for potential vaccine antigen development

Porcine circovirus type 2 (PCV2) is a globally prevalent infectious pathogen affecting swine, with its capsid protein (Cap) being the sole structural protein critical for vaccine development. Prior research has demonstrated that PCV2 Cap proteins produced in Escherichia coli (E. coli) can form virus-like particles (VLPs) in vitro, and nuclear localization signal peptides (NLS) play a pivotal role in stabilizing PCV2 VLPs. Recently, PCV2d has emerged as an important strain within the PCV2 epidemic. In this study, we systematically optimized the PCV2d Cap protein and successfully produced intact PCV2d VLPs containing NLS using E. coli. The recombinant PCV2d Cap protein was purified through affinity chromatography, yielding 7.5 mg of recombinant protein per 100 ml of bacterial culture. We augmented the conventional buffer system with various substances such as arginine, β-mercaptoethanol, glycerol, polyethylene glycol, and glutathione to promote VLP assembly. The recombinant PCV2d Cap self-assembled into VLPs approximately 20 nm in diameter, featuring uniform distribution and exceptional stability in the optimized buffer. We developed the vaccine and immunized pigs and mice, evaluating the immunogenicity of the PCV2d VLPs vaccine by measuring PCV2-IgG, IL-4, TNF-α, and IFN-γ levels, comparing them to commercial vaccines utilizing truncated PCV2 Cap antigens. The HE staining and immunohistochemical tests confirmed that the PCV2 VLPs vaccine offered robust protection. The results revealed that animals vaccinated with the PCV2d VLPs vaccine exhibited high levels of PCV2 antibodies, with TNF-α and IFN-γ levels rapidly increasing at 14 days post-immunization, which were higher than those observed in commercially available vaccines, particularly in the mouse trial. This could be due to the fact that full-length Cap proteins can assemble into more stable PCV2d VLPs in the assembling buffer. In conclusion, our produced PCV2d VLPs vaccine elicited stronger immune responses in pigs and mice compared to commercial vaccines. The PCV2d VLPs from this study serve as an excellent candidate vaccine antigen, providing insights for PCV2d vaccine research.

A Potential Role of Interleukin-5 in the Pathogenesis and Progression of Amyotrophic Lateral Sclerosis: A New Molecular Perspective.

Cumulative data suggest that neuroinflammation plays a prominent role in amyotrophic lateral sclerosis (ALS) pathogenesis. The purpose of this work was to assess if patients with ALS present a specific peripheral cytokine profile and if it correlates with neurological disability assessed by ALSFRS-R, the rate of disease progression, and the pattern of disease progression (horizontal spreading [HSP] versus vertical spreading [VSP]). We determined the levels of 15 cytokines in the blood of 59 patients with ALS and 40 controls. We identified a positive correlation between levels of pro-inflammatory cytokines (interleukin [IL]-17F, IL-33, IL-31) and the age of ALS patients, as well as a positive correlation between IL-12p/70 and survival from ALS onset and ALS diagnosis. Additionally, there was a positive correlation between the ALSFRS-R score in the upper limb and respiratory domain and IL-5 levels. In our ALS cohort, the spreading pattern was 42% horizontal and 58% vertical, with patients with VSP showing a faster rate of ALS progression. Furthermore, we identified a negative correlation between IL-5 levels and the rate of disease progression, as well as a positive correlation between IL-5 and HSP of ALS. To the best of our knowledge, this is the first study reporting a "protective" role of IL-5 in ALS.

Analysis of Endocrine and Inflammatory Markers in Preserved Ratio Impaired Spirometry.

Chronic Obstructive Pulmonary Disease (COPD) is a disease of the lungs characterized by chronic airflow obstruction. Individuals with preserved ratio impaired spirometry (PRISm) may be at risk for developing COPD. This study aimed to characterize PRISm and COPD patients in terms of their immune response and endocrine profile to identify differences extending beyond lung function. The participants performed the clinical assessment, pulmonary function test, and blood collection to determine serum hormone levels and concentrations of cytokine. Differences were observed in the nutritional status, lung function, and comorbidity. There were no differences in IL-6, IL-8, IL-10, IL-12, and TNF levels between PRISm and COPD groups. Both PRISm and COPD patients have lower dehydroepiandrosterone (DHEA) and dehydroepiandrosterone sulfate (DHEA-S) levels than controls. Correlation analysis of PRISm and COPD patients revealed positive correlations between serum levels of DHEA-S and DHEA, with forced expiratory volume in 1 second (FEV1) and forced vital capacity (FVC), which negatively correlated with IL-8 levels. The results indicated that despite differences in lung function parameters, the PRISm and COPD groups exhibited similarities in endocrine profile alterations. This study represents the first attempt to link endocrine with immune markers and lung function in individuals with PRISm.

Exploration of altered miRNA expression and function in MSC-derived extracellular vesicles in response to hydatid antigen stimulation.

Hydatid disease is caused by Echinococcus parasites and can affect various tissues and organs in the body. The disease is characterized by the presence of hydatid cysts, which contain specific antigens that interact with the host's immune system. Mesenchymal stem cells (MSCs) are pluripotent stem cells that can regulate immunity through the secretion of extracellular vesicles (EVs) containing microRNAs (miRNAs). In this study, hydatid antigens were isolated from sheep livers and mice peritoneal cavities. MSCs derived from mouse bone marrow were treated with different hydatid antigens, and EVs were isolated and characterized from the conditioned medium of MSCs. Small RNA library construction, miRNA target prediction, and differential expression analysis were conducted to identify differentially expressed miRNAs. Functional enrichment and network construction were performed to explore the biological functions of the target genes. Real-time PCR and Western blotting were used for miRNA and gene expression verification, while ELISA assays quantified TNF, IL-1, IL-6, IL-4, and IL-10 levels in cell supernatants. The study successfully isolated hydatid antigens and characterized MSC-derived EVs, demonstrating the impact of antigen concentration on MSC viability. Key differentially expressed miRNAs, such as miR-146a and miR-9-5p, were identified, with functional analyses revealing significant pathways like Endocytosis and MAPK signaling associated with these miRNAs' target genes. The miRNA-HUB gene regulatory network identified crucial miRNAs and HUB genes, such as Traf1 and Tnf, indicating roles in immune modulation and osteogenic differentiation. Protein-protein interaction (PPI) network analysis highlighted central HUB genes like Akt1 and Bcl2. ALP activity assays confirmed the influence of antigens on osteogenic differentiation, with reduced ALP activity observed. Expression analysis validated altered miRNA and chemokine expression post-antigen stimulation, with ELISA analysis showing a significant reduction in CXCL1 expression in response to antigen exposure. This study provides insights into the role of MSC-derived EVs in regulating parasite immunity. The findings suggest that hydatid antigens can modulate the expression of miRNAs in MSC-derived EVs, leading to changes in chemokine expression and osteogenic capacity. These findings contribute to a better understanding of the immunomodulatory mechanisms involved in hydatid disease and provide potential therapeutic targets for the development of new treatment strategies.

Sulforaphane inhibits the migration and invasion of BPDE-induced lung adenocarcinoma cells by regulating NLRP12

This study aims to explore the impact and underlying mechanism of sulforaphane (SFN) intervention on the migration and invasion of lung adenocarcinoma induced by 7, 8-dihydroxy-9, 10-epoxy-benzo (a) pyrene (BPDE). Human lung adenocarcinoma A549 cells were exposed to varying concentrations of BPDE (0.25, 0.50, and 1.00 μM) and subsequently treated with 5 μM SFN. Cell viability was determined using CCK8 assay, while migration and invasion were assessed using Transwell assays. Lentivirus transfection was employed to establish NLRP12 overexpressing A549 cells. ELISA was utilized to quantify IL-33, CXCL12, and CXCL13 levels in the supernatant, while quantitative real-time PCR (qRT-PCR) and Western Blot were used to analyze the expression of NLRP12 and key factors associated with canonical and non-canonical NF-κB pathways. Results indicated an increase in migratory and invasive capabilities, concurrent with heightened expression of IL-33, CXCL12, CXCL13, and factors associated with both canonical and non-canonical NF-κB pathways. Moreover, mRNA and protein levels of NLRP12 were decreased in BPDE-stimulated A549 cells. Subsequent SFN intervention attenuated BPDE-induced migration and invasion of A549 cells. Lentivirus-mediated NLRP12 overexpression not only reversed the observed phenotype in BPDE-induced cells but also led to a reduction in the expression of critical factors associated with both canonical and non-canonical NF-κB pathways. Collectively, we found that SFN could inhibit BPDE-induced migration and invasion of A549 cells by upregulating NLRP12, thereby influencing both canonical and non-canonical NF-κB pathways.

Intra-Articular Injection of Interleukin-8 Neutralizing Monoclonal Antibody Effectively Attenuates Osteoarthritis Progression in Rabbits.

Cytokines are implicated in the pathogenesis of osteoarthritis (OA), and this study aims to assess the therapeutic potential of an IL-8 neutralizing monoclonal antibody (mAb) for OA intervention. The study employed a rabbit model of OA induced by anterior cruciate ligament transection (ACLT) surgery to investigate the effects of an interleukin (IL)-8 neutralizing mAb, with hyaluronic acid (HA) used as a positive control. Primary outcomes assessed in the rabbits included cartilage repair, synovitis, joint effusion, changes in footprints, and lower limb loading conditions. Compared to HA, intra-articular injection of the IL-8 neutralizing mAb demonstrated a more pronounced attenuation of OA progression and enhancement of cartilage repair. We observed a reduction in synovitis and joint effusion, indications of bone marrow edema, as well as improvements in lower limb function. In knees treated with the neutralizing IL-8 mAb, there was a significant decrease in IL-8 levels within the synovial tissues. The IL-8 neutralizing mAb exhibits promising therapeutic potential in the management of OA by attenuating inflammation and facilitating cartilage repair. However, further investigations are warranted to comprehensively elucidate the underlying mechanisms, optimize treatment protocols, and ensure the long-term safety and efficacy of this innovative therapeutic approach.

Role of TRPV1 and TRPA1 in TSLP production in nasal epithelial cells

BackgroundTRP protein is sensitive to external temperature changes, but its pathogenic mechanism in the upper airway mucosa is still unclear. ObjectiveTo investigate the mechanism of TRPV1and TRPA1 in regulating the secretion of inflammatory factors in nasal epithelial cells. MethodsThe expression of TRPV1 and TRPA1 in nasal mucosal epithelial cells was investigated using immunofluorescence assays. Epithelial cells were stimulated with TRPV1 and TRPA1 agonists and antagonists, and changes in Ca2+ release and inflammatory factor secretion in epithelial cells were detected. TSLP secretion stimulated with the calcium chelating agent EGTA was evaluated. The transcription factor NFAT was observed by immunofluorescence staining. ResultsTRPV1 and TRPA1 expression was detected in nasal epithelial cells, and Ca2+ influx was increased after stimulation with agonists. After the activation of TRPV1 and TRPA1, the gene expression of TSLP, IL-25, and IL-33 and the protein expression levels of TSLP and IL-33 were increased, and only TSLP could be inhibited by antagonists and siRNAs. After administration of EGTA, the secretion of TSLP was inhibited significantly, and the expression of the transcription factor NFAT in the nucleus was observed after activation of the TRPV1 and TRPA1 proteins in epithelial cells. ConclusionActivation of TRPV1 and TRPA1 on nasal epithelial cells stimulates the generation of TSLP through the Ca2+/NFAT pathway. It also induces upregulation of IL-25 and IL-33 gene expression levels and increased levels of IL-33 protein, leading to the development of airway inflammation.

IL-4-induced decrease in both the number and CTLA-4 expression of Treg impairs suppression of Th2 type inflammation in severe atopic dermatitis

BackgroundTreg plays a pivotal role in the suppression of Th2 cell and the maintenance of immune homeostasis. The precise molecular mechanism underlying the disruption of Treg suppression of Th2 cell and the promotion of Th2 type inflammation in allergic diseases remains elusive. ObjectiveThis study aims to investigate the molecular mechanism underlying quantitative and functional changes of Treg in AD. MethodsThe molecular mechanism was investigated using flow cytometry, mRNA sequencing, co-culture experiments, co-immunoprecipitation, chromatin immunoprecipitation, and bisulfite sequencing in vitro or in AD mice model and patients with AD. ResultsIncreased proportion of Treg was detected in mild and moderate AD. Conversely, characteristic decrease in both the number and CTLA-4 expression of Treg was relevant to serum IL-4 level in severe AD patients, which was verified under a high concentration of IL-4 treatment in vitro. The underlying mechanism is that IL-4/pSTAT6 pathway recruits DNMT1 and HDAC2 to inhibit transcriptional regulation of Foxp3 and CTLA-4 loci. High level of IL-4 impaired the suppression of Treg against Th2 cell differentiation mediated by CTLA-4, and blockade of IL-4Rα signaling in Treg restored Treg number and suppression of Th2 cell in AD model mice and patients with AD. ConclusionThe number of Treg is relevant to stratification of severity and serum IL-4 level in patients with AD. Abnormal high level of IL-4 epigenetically triggers a decrease in both the number and CTLA-4 expression of Treg. The reduced expression of CTLA-4 on Treg induced by IL-4 impairs suppression of Th2 cell differentiation.

Endoplasmic reticulum stress drives macrophages to produce IL-33 to favor Th2 polarization in the airways.

Interleukin (IL)-33 is a key driver of T helper 2 (Th2) cell polarization. Endoplasmic reticulum (ER) stress plays a role in the skewed T cell activation. The objective of this project is to elucidate the role of IL-33 derived from macrophages in inducing Th2 polarization in the airways. In this study, bronchoalveolar lavage fluids (BALF) were collected from patients with asthma and healthy control subjects. Macrophages were isolated from the BALF by flow cytometry cell sorting. An asthmatic mouse model was established using the ovalbumin/alum protocol. The results showed that increased IL33 gene activity and ER stress-related molecules in BALF-derived M2a macrophages was observed in asthmatic patients. Levels of IL33 gene activity in M2a cells were positively correlated with levels of asthma response in asthma patients. Sensitization exacerbated the ER stress in the airway macrophages, which increased the expression of IL-33 in macrophages of airway in sensitized mice. Conditional ablation of Il33 or Perk or Atf4 genes in macrophages prevented induction of airway allergy in mice. In conclusion, asthma airway macrophages express high levels of IL-33 and at high ER stress status. Inhibition of IL-33 or ER stress in macrophages can effectively alleviate experimental asthma.

Abstract 2363: Enhanced T cell activation and cytotoxicity against AML via targeted anti-CD99-A192 ScFv-ELP nanoparticle treatment

Abstract CD99 is a transmembrane protein overexpressed in acute myeloid leukemia (AML), presenting a potential novel therapeutic target. Our previously developed anti-CD99-A192, comprising scFv and elastin-like polypeptides (ELPs), showed promising anti-leukemic activity by inducing apoptosis in AML cell lines and prolonging survival in AML xenograft models. Considering CD99's expression and role in T cell activation, we proposed that anti-CD99-A192 could have a dual function: targeting leukemic cells while activating T cells. Blood samples were obtained from 20 healthy donors, and peripheral blood mononuclear cells (PBMCs) were isolated and stimulated with PHA and IL-2 cytokines to expand T cells for 72-96 hours. Post-expansion, T cells were treated with either anti-CD99-A192 or control anti-A192. Proliferation was evaluated via cell count and cell trace assays. The percentages of CD4+, CD8+, and CD25+ T cells, and the levels of T cell activation markers CD69 and CD38, were quantified by flow cytometry. Cytokine release profiles were assessed by the Proteome Profiler Human Cytokine Array Kit. The cytotoxic impact of the anti-CD99-A192-activated T cells on MV4-11Luc+ AML cells was measured in co-culture apoptosis and luciferase assays. Statistical comparisons utilized paired and unpaired T-tests with Holm-Sidak correction for multiple comparisons, and P value &amp;lt;0.05 was considered significant. T cells treated with anti-CD99-A192 displayed significant increased proliferation compared with untreated and anti-A192-treated groups at Day 5 (163.3% vs. 97% and 92.4%, p=0.042 and p=0.03, respectively). The percentage of CD4+, CD8+, and CD25+ cells remained unchanged across treated and control groups. Activation marker analysis revealed increase in % CD38+ cells in the anti-CD99-A192 group compared with the control (58.15% vs. 49.15%, p=0.0028). Among the cytokine panel, significant increases in IL-1β/IL-1F2 (11-fold change, p&amp;lt;0.001, adjusted p=0.006) and TNF-α (12-fold change, p=0.002, adjusted p=0.03) were observed, along with a marked decrease in IL-8 (70% reduction, p&amp;lt;0.001, adjusted p=0.001). GM-CSF (20-fold change, p=0.005, adjusted p=0.071) and IL-16 (2-fold change, p=0.003, adjusted p=0.051) levels also increased, though not significantly after adjustment. Co-culture assays showed greater apoptosis in MV4-11 cells at 18 hours (33.6% vs. 41.13%, p=0.015) and a reduction in viable leukemic cells at 18 hours (10.89% vs. 8.05%, p=0.034) and 48 hours (9.69% vs. 6.54%, p=0.065) in the anti-CD99-A192 compared with the control group, results were also validated by the luciferase assay (~32% reduction, p&amp;lt;0.001). Anti-CD99-A192 scFv-ELP nanoparticle stimulates T cell proliferation, elevates activation markers, and increases the release of pro-inflammatory cytokines. Anti-CD99-A192 culminates in heightened cytotoxicity against leukemic cells. Citation Format: Shephali Kadam, Atham Ali, Mateusz Pospiech, Sandra Onyemaechi, Yiting Meng, Kanaka Dhuri, Andrew Mackay, Houda Alachkar. Enhanced T cell activation and cytotoxicity against AML via targeted anti-CD99-A192 ScFv-ELP nanoparticle treatment [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 2363.

Abstract 7652: Plasma markers of hemostasis, fibrinolysis, and angiogenesis prior to androgen deprivation therapy (ADT) in progressive prostate cancer (PC) and associations with long-term thrombotic events and survival

Abstract Background: PC is the third most common malignancy associated with thrombosis, procoagulants may play a role in tumorigenesis and progression, and patients (pts) with PC may have activation of the hemostatic system evident in plasma. The prognostic significance of these markers and their relationship to thrombotic events and overall survival (OS) are under-explored. Methods: Patients &amp;gt;18 years were included if they had progressive PC planned to start ADT, were &amp;gt;6 months from surgery or RT, with no history of prior VTE, unexplained bleeding, current or planned anticoagulation, or an additional active malignancy. Plasma samples were obtained via peripheral venipuncture in tubes containing 3.2% sodium citrate and immediately placed on ice. Samples were analyzed for D-dimer, thrombin-antithrombin complex (TAT), IL-6, IL-8, VEGF, and tissue factor (TF) levels by ELISA. Patients had markers drawn at baseline (prior to initiation of ADT), 1 month, and 6 months after initiation. Results: 40 patients were accrued with a median age of 68 years. At baseline, 29 (73%) had presence of metastases. Baseline median PSA was 15 and median PSA doubling time was 0.32. Baseline median hemoglobin was 13.85, platelets were 218, D-dimer was 322 ng/dL, IL-6 was 0 pg/mL, IL-8 was 0 pg/mL, TAT was 3 ug/L, VEGF was 21 pg/mL, and TF was 26 pg/mL. Median follow-up was 78 months. During follow up, 35 (87.5%) had metastases. 10 (25%) had thrombotic events (6 [15%] venous, 4 [10%] arterial) at median 37 months follow up. Median baseline age of pts with subsequent thrombotic events was 74, vs 67 in those without a thrombotic event. While there was no significant difference in baseline PSA between groups with and without clotting events, pretreatment PSA doubling time was significantly lower in pts with subsequent thrombosis (0.27 vs 0.35, p=0.047). Median baseline D-dimer and TF were higher in patients with subsequent thrombosis (D-dimer: 516 vs 282, p=0.10; TF: 36 vs 24, p=0.12). In univariate analysis, baseline IL-6 and VEGF were associated with OS (IL-6: HR=1.05, 95% CI [1.01,1.09], p=0.025; VEGF: HR=1.02, 95% CI [1.00,1.03], p=0.028). When controlling for presence of metastasis, baseline TAT was associated with OS (HR=1.05, 95% CI [1.00,1.11], p=0.055). Conclusions: Plasma markers of hemostatic activation, fibrinolysis, and angiogenesis prior to initiation of ADT are associated with subsequent thrombotic events and overall survival years later. Further analysis will examine how these markers change over time with hormonal therapy. Citation Format: Jessica S. Palmer, Charlene Thomas, Nicole Jacobs, David Nanus, Scott T. Tagawa. Plasma markers of hemostasis, fibrinolysis, and angiogenesis prior to androgen deprivation therapy (ADT) in progressive prostate cancer (PC) and associations with long-term thrombotic events and survival [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 7652.

Abstract 3435: History of breastfeeding in relation to circulating inflammatory biomarkers

Abstract Background: History of breastfeeding has been associated with reduced risk of various chronic diseases in which chronic inflammation is involved in disease etiology, including cardiovascular disease, type II diabetes, breast cancer, and ovarian cancer. However, few studies have investigated the long-term impact of breastfeeding on systemic inflammation. Thus, we aimed to examine the associations between breastfeeding history and circulating inflammatory biomarkers at midlife. Methods: We conducted a cross-sectional analysis including 3,448 parous women with plasma inflammatory biomarkers in the Nurses’ Health Study and Nurses’ Health Study II. We examined self-reported history of ever breastfeeding (ever vs. never) and lifetime total duration of breastfeeding (never, &amp;lt; 6 months, 7-11 months, 12+ months) prior to the time of blood draw. We calculated the geometric means of eight inflammatory biomarkers [i.e., high sensitivity C-reactive protein (hsCRP), interleukin-6 (IL6], sIL6Rα, IL8, IL10, sIL2-receptor-α(Rα), soluble tumor necrosis factor α receptor 2 (sTNFR2), B-cell activating factor (BAFF), C-X-C motif chemokine ligand 13 (CXCL13)] by self-reported history of breastfeeding status adjusted for age, body mass index (BMI), smoking status, duration of oral contraceptive use, parity, proinflammatory diet, analgesic use, menopausal status, and cohort. Tests for trend were calculated using the median value of each lifetime breastfeeding duration category and modeling as a continuous variable. We also conducted stratified analyses by menopausal status and BMI (&amp;lt; 25kg/m2 vs. ≥ 25kg/m2) at blood draw. Results: Overall, we did not observe statistically significant differences in circulating inflammatory biomarker levels by history of breastfeeding among parous women. When stratifying by menopausal status at blood draw, total breastfeeding duration was inversely associated with sTNFR2 levels among premenopausal women (P-trend = 0.03) but not among postmenopausal women. Plasma IL8 levels were suggestively inversely associated with breastfeeding duration among postmenopausal women (P-trend = 0.07) but not among premenopausal women. Among women with BMI ≥ 25 kg/m2, plasma BAFF levels were lower in parous women who ever breastfed compared to those who never breastfed [1382.5 (95%CI = 1242.0 - 1539.0) pg/ml vs. 1611.9 (95%CI = 1424.9 - 1823.4) pg/ml; p = 0.03], and suggestively inversely associated with total breastfeeding duration (p-trend = 0.07) but no statistically significant association was observed in women with BMI &amp;lt; 25kg/m2. IL10 was inversely associated with total breastfeeding duration among women with BMI ≥25 kg/m2 (P-trend = 0.007), but not in women with BMI &amp;lt; 25kg/m2. Conclusion: Overall, history of breastfeeding was not associated with inflammatory biomarkers at mid-life, however the association may vary by menopausal status and BMI. Citation Format: Nan Lin, Jennifer M. Mongiovi, Leslie V. Farland, Tianyi Huang, Heather Eliassen, Mary K. Townsend, Shelley S. Tworoger, Kathryn L. Terry, Naoko Sasamoto. History of breastfeeding in relation to circulating inflammatory biomarkers [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 3435.

Abstract 1403: The role of Dclk1-expression cells in normal pancreas and in pancreatic tumor progression

Abstract Background: Tuft cells, marked by Doublecortin-like kinase-1 (Dclk1), are rare in the healthy pancreas but increase markedly during pancreatic tumorigenesis. While Group 2 innate lymphoid cells (ILC2s) are known to modulate inflammation and immunity, their specific roles in cancer-related immunity and immunotherapy interactions with pancreatic tuft cells remain unknown. Methods: To investigate tuft cells during early pancreatic neoplasia, we used a Dclk1 reporter mouse model and single-cell RNA-seq to study Dclk1-expressing cells in normal pancreas and neoplasia using Mist1/Kras mouse. We treated mice with IL-13 peptides and anti-IL-13 antibodies to explore the IL-13-ILC2 axis in tuft cell regulation. Results: Within the normal pancreas, Dclk1 expression marked several specialized epithelial cell types, including tuft-like cells, neuroendocrine cells, and a small number of acinar progenitor cells. A notable increase in Dclk1-expressing cells was observed at the onset of early PanIN lesions, with a subsequent decline in advanced PanIN stages and PDAC. Using flow cytometric analysis, we observed an increase in ILC2 cells in the Mist1/Kras pancreas in response to IL33 produced by pancreatic cancer-associated fibroblasts (CAFs). Blocking ILC2 activation with anti-IL33 antibodies reduced Dclk1 cell hyperplasia during PanIN progression. Given the known association between tuft cells, ILC2 cells, and the IL13/IL4 signaling pathways in the intestine, we discovered that Dclk1-expressing cells highly expressed the IL4 receptor (IL4ra). IL-13 levels increased during PanIN progression, and we found that ILC2s comprised the primary source of IL-13 within pancreatic tumors. In vitro, studies showed that the presence of IL-13 maintained the expansion of Dclk1-expressing tuft-like cells in pancreatic organoids derived from Dclk1-DTR-ZsGreen mice. Correspondingly, the in vivo administration of anti-IL-13 resulted in reduced Dclk1 expression. Conclusion: Previous studies have shown that pancreatic tuft cells inhibit pancreatic cancer progression. Our study reveals a novel regulatory mechanism wherein IL-13 secreted by ILC2s modulates Dclk1 tuft cell expression during pancreatic tumorigenesis. The IL13-ILC2 axis appears to be a potential therapeutic target to regulate Dclk1 cell activity, which could influence the course of pancreatic tumorigenesis. Future investigations will further explore the interaction between the IL13/ILC2 axis and Dclk1 in pancreatic cancer development. Citation Format: Giovanni Valenti, Pasquale Laise, Feijing Wu, Hiroki Kobayashi, Quin T. Waterbury, Ryota Takahashi, Tuo Ruan, Zhengyu Jiang, Yosuke Ochiai, Leah B. Zamechek, Timothy C. Wang. The role of Dclk1-expression cells in normal pancreas and in pancreatic tumor progression [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 1403.