Changes In Cytokine Profile Research Articles

The Association between Gut Microbiota and Serum Biomarkers in Children with Atopic Dermatitis.

Background. Currently, it is known that the gut microbiota plays an important role in the functioning of the immune system, and a rebalancing of the bacterial community can arouse complex immune reactions and lead to immune-mediated responses in an organism, in particular, the development of atopic dermatitis (AD). Cytokines and chemokines are regulators of the innate and adaptive immune response and represent the most important biomarkers of the immune system. It is known that changes in cytokine profiles are a hallmark of many diseases, including atopy. However, it remains unclear how the bacterial imbalance disrupts the function of the immune response in AD. Objectives. We attempted to determine the role of gut bacteria in modulating cytokine pathways and their role in atopic inflammation. Methods. We sequenced the 16S rRNA gene from 50 stool samples of children aged 3-12 years who had confirmed atopic dermatitis, and 50 samples from healthy children to serve as a control group. To evaluate the immune status, we conducted a multiplex immunofluorescence assay and measured the levels of 41 cytokines and chemokines in the serum of all participants. Results. To find out whether changes in the composition of the gut microbiota were significantly associated with changes in the level of inflammatory cytokines, a correlation was calculated between each pair of bacterial family and cytokine. In the AD group, 191 correlations were significant (Spearman's correlation coefficient, p ≤ 0.05), 85 of which were positive and 106 which were negative. Conclusions. It has been demonstrated that intestinal dysbiosis is associated with alterations in cytokine profiles, specifically an increase in proinflammatory cytokine concentrations. This may indicate a systemic impact of these conditions, leading to an imbalance in the immune system's response to the Th2 type. As a result, atopic conditions may develop. Additionally, a correlation between known AD biomarkers (IL-5, IL-8, IL-13, CCL22, IFN-γ, TNF-α) and alterations in the abundance of bacterial families (Pasteurellaceae, Barnesiellaceae, Eubacteriaceae) was observed.

Systemic pro-inflammatory response following bronchoscopic lung volume reduction using endobronchial valves

RationaleBronchoscopic lung volume reduction treatment using endobronchial valves (EBV) is an effective treatment for severe COPD patients by improving lung function and quality of life. However, little is known about its effects on systemic inflammation. Therefore, the aim of our study was to investigate whether EBV treatment impacts the inflammatory cytokine profile.MethodsThis study was a predefined substudy of the SoLVE trial (NCT03474471) that investigated the combination of EBV treatment with pulmonary rehabilitation (PR). The substudy included the collection of blood samples with assessment of 10 inflammatory markers prior to EBV treatment and 6 months after EBV or EBV+PR treatment.ResultsIn 66 patients, six months after treatment a pro-inflammatory cytokine profile was observed, with an increase in all pro-inflammatory markers and a decrease in the anti-inflammatory cytokine IL-10. The changes in plasma cytokine profile were not associated with changes in clinical outcomes such as lung function or exercise capacity.DiscussionIn conclusion, our study demonstrated an elevation in systemic pro-inflammatory cytokine levels following successful EBV-treatment, which was not associated with adverse clinical outcomes. It would be interesting to further explore, whether this increase is attributed to a foreign body response or if other factors contribute to this phenomenon.

In Vivo Biocompatibility of Synechococcus sp. PCC 7002-Integrated Scaffolds for Skin Regeneration.

Cyanobacteria, commonly known as blue-green algae, are prevalent in freshwater systems and have gained interest for their potential in medical applications, particularly in skin regeneration. Among these, Synechococcus sp. strain PCC 7002 stands out because of its rapid proliferation and capacity to be genetically modified to produce growth factors. This study investigates the safety of Synechococcus sp. PCC 7002 when used in scaffolds for skin regeneration, focusing on systemic inflammatory responses in a murine model. We evaluated the following three groups: scaffolds colonized with genetically engineered bacteria producing hyaluronic acid, scaffolds with wild-type bacteria, and control scaffolds without bacteria. After seven days, we assessed systemic inflammation by measuring changes in cytokine profiles and lymphatic organ sizes. The results showed no significant differences in spleen, thymus, and lymph node weights, indicating a lack of overt systemic toxicity. Blood cytokine analysis revealed elevated levels of IL-6 and IL-1β in scaffolds with bacteria, suggesting a systemic inflammatory response, while TNF-α levels remained unaffected. Proteome profiling identified distinct cytokine patterns associated with bacterial colonization, including elevated inflammatory proteins and products, indicative of acute inflammation. Conversely, control scaffolds exhibited protein profiles suggestive of a rejection response, characterized by increased levels of cytokines involved in T and B cell activation. Our findings suggest that Synechococcus sp. PCC 7002 does not appear to cause significant systemic toxicity, supporting its potential use in biomedical applications. Further research is necessary to explore the long-term effects and clinical implications of these responses.

72 Impact of microbiome modulation on serum cytokine profile in a randomized, phase I trial of cabozantinib/nivolumab (cabo/nivo) with or without CBM588 in pts with metastatic renal cell carcinoma (mRCC)

Abstract Background We have previously demonstrated that the orally administered, live bacterial product CBM588 can augment clinical outcomes with cabo/nivo in pts with mRCC (Ebrahimi et al ASCO 2023). As it is postulated this effect may be mediated through changes in systemic immunity, we sought to characterize changes in serum cytokine profile induced by CBM588. Methods Pts with previously untreated mRCC with clear cell, papillary or sarcomatoid histology were enrolled. Key eligibility criteria included Karnofsky performance status ≥ 70% and measurable disease. Pts were randomized to receive cabo/nivo on a standard schedule alone or the same regimen with CBM588 at 80 mg bid. Pts had blood collection performed at baseline and at weeks 9, 13, 17 and 25. Samples underwent processing within 4-6 hrs of collection, and a total of 30 cytokines and growth factors were assessed using the Luminex Flexmap 3D system (Biotechne). Changes in circulating cytokine levels between baseline and week 13 (± 7 days) were examined across the treatment arms to investigate the impact of CBM588 on the immune system. The Wilcoxon matched-pairs test was used to compare the levels of cytokines at the two timepoints. Results A total of 30 pts were enrolled; the majority were male (67%) and had intermediate- or poor-risk disease (60%). Most pts had clear cell histology (87%); 5 pts (17%) had sarcomatoid features. As previously reported, pts receiving cabo/nivo/CBM588 had a significantly higher response rate as compared to pts receiving cabo/nivo alone (74% vs 20%; P=0.01). Across baseline and week 13, a total of 53 samples were available for assessment. In pts receiving cabo/nivo/CBM588, there was a significant change in the level of IL-12, IL-13, eotaxin, interferon-γ, and granulocyte-macrophage colony stimulating factor (GM-CSF); this was not observed in the cabo/nivo alone arm. Additional flow cytometric analyses characterizing relevant CD4- and CD8-positive subsets will be presented. Conclusions Taken together with our previous study evaluating nivolumab/ipilimumab with CBM588 (Dizman et al Nat Med 2022), these results suggest that the augmented clinical outcome seen with the addition of CBM588 to cabo/nivo may be related to systemic immunomodulation. Combined analyses of the cohorts across studies are planned.

Astaxanthin Supplementation Does Not Alter Training-Related Changes in Inflammatory Cytokine Profile in Arabian Racing Horses.

This study aimed to evaluate the oral supplementation of astaxanthin (ATX) on inflammatory markers in 3-year-old Arabian racehorses. Despite the recognized antioxidant and anti-inflammatory properties of ATX observed in vitro in rodent models and in human athletes, the effects in equine subjects remain unknown. This study involved a controlled trial with 14 horses receiving either ATX (six horses) or a placebo (eight horses), monitored over four months of race training. Inflammatory cytokines: TNFα, IFNγ, IL-6, IL-10, and prostaglandin E (PGE), were measured monthly to assess the impact of ATX on the inflammatory response. The results indicated no significant differences in measured parameters between the ATX and the control group during the study. However, a significant time-dependent decrease in TNFα and IFNγ levels (p = 0.001) was observed in both groups, suggesting that regular training naturally modulates inflammatory responses. Moreover, positive correlations were noted between TNFα and IFNγ (p < 0.001) in the early phase of the study and between IL-6 and IL-10 (p = 0.008) in the later phase. Hematological parameters remained stable and within reference ranges, indicating no adverse effects of ATX supplementation. Performance metrics, including the number of races completed and wins, showed no significant differences between groups, suggesting that ATX did not enhance athletic performance under the study conditions. Overall, while ATX supplementation affected neither cytokine levels nor performance in Arabian racehorses, the natural anti-inflammatory effects of regular training were evident. Further research is needed to explore potential benefits of ATX supplementation under different conditions, such as in horses with subclinical inflammation or varying training regimens, to fully clarify its role and applications in equine sports medicine.

Heligmosomoides bakeri and Toxoplasma gondii co-infection leads to increased mortality associated with changes in immune resistance in the lymphoid compartment and disease pathology.

Co-infections are a common reality but understanding how the immune system responds in this context is complex and can be unpredictable. Heligmosomoides bakeri (parasitic roundworm, previously Heligmosomoides polygyrus) and Toxoplasma gondii (protozoan parasite) are well studied organisms that stimulate a characteristic Th2 and Th1 response, respectively. Several studies have demonstrated reduced inflammatory cytokine responses in animals co-infected with such organisms. However, while general cytokine signatures have been examined, the impact of the different cytokine producing lymphocytes on parasite control/clearance is not fully understood. We investigated five different lymphocyte populations (NK, NKT, γδ T, CD4+ T and CD8+ T cells), five organs (small intestine, Peyer's patches, mesenteric lymph nodes, spleen and liver), and 4 cytokines (IFN©, IL-4, IL-10 and IL-13) at two different time points (days 5 and 10 post T. gondii infection). We found that co-infected animals had significantly higher mortality than either single infection. This was accompanied by transient and local changes in parasite loads and cytokine profiles. Despite the early changes in lymphocyte and cytokine profiles, severe intestinal pathology in co-infected mice likely contributed to early mortality due to significant damage by both parasites in the small intestine. Our work demonstrates the importance of taking a broad view during infection research, studying multiple cell types, organs/tissues and time points to link and/or uncouple immunological from pathological findings. Our results provide insights into how co-infection with parasites stimulating different arms of the immune system can lead to drastic changes in infection dynamics.

Immune Response Dynamics and Biomarkers in COVID-19 Patients.

The immune response dynamics in COVID-19 patients remain a subject of intense investigation due to their implications for disease severity and treatment outcomes. We examined changes in leukocyte levels, eosinophil activity, and cytokine profiles in patients hospitalized with COVID-19. Serum samples were collected within the first 10 days of hospitalization/confirmed infection and analyzed for eosinophil granule proteins (EGP) and cytokines. Information from medical records including comorbidities, clinical symptoms, medications, and complete blood counts were collected at the time of admission, during hospitalization and at follow up approximately 3 months later. Serum levels of eotaxin, type 1 and type 2 cytokines, and alarmin cytokines were elevated in COVID-19 patients, highlighting the heightened immune response (p < 0.05). However, COVID-19 patients exhibited lower levels of eosinophils and eosinophil degranulation products compared to hospitalized controls (p < 0.05). Leukocyte counts increased consistently from admission to follow-up, indicative of recovery. Attenuated eosinophil activity alongside elevated chemokine and cytokine levels during active infection, highlights the complex interplay of immune mediators in the pathogenesis COVID-19 and underscores the need for further investigation into immune biomarkers and treatment strategies.

Changes of aqueous humor cytokine profiles of patients with high intraocular pressure after PPV for retinal detachment

High intraocular pressure (IOP) is one of the early complications after pars plana vitrectomy (PPV), which may cause glaucoma and poor visual prognosis secondary to surgery. Proliferative vitreoretinopathy (PVR) is one of the complications of retinal detachment (RD) and is the main reason for the poor prognosis, which is related to different kinds of cytokines. It’s essential for the basic mechanism to analyze the relative aqueous humor cytokine profiles with IOP after PPV for RD. In this study, we have collected the aqueous humor of 16 patients and qualified 27 cytokines using Luminex and compared biomarkers with the high IOP group and the normal group. As a result, the concentrations of VEGF, IL-6, FGF2, and G-CSF upregulated significantly (P < 0.05), while VEGFR2 downregulated significantly (P < 0.05) in the high IOP group. IL-6 was positively correlated with high IOP (r = 0.561, P = 0.041). Meanwhile, the concentrations of IL-6 (r = 0.543, P = 0.03), IL-5 (r = 0.576, P = 0.019), IL-15 (r = 0.614, P = 0.011), IL-4 (r = 0.517, P = 0.04), ICAM-1 (r = 0.611, P = 0.012), and G-CSF (r = 0.636, P = 0.008) were significantly associated with preoperative PVR classification, and the aqueous humor levels of IL-4 (r = 0.567, P = 0.022), HGF (r = 0.701, P = 0.005), and MCP-1 (r = 0.565, P = 0.035) are significant relative to laser points. Hence, cytokines might potentially be the therapeutic target of high IOP after PPV.

P159 Safety, clinical effectiveness, and changes in systemic cytokine profiles in solid organ transplant recipients post 6 months of elexacaftor/tezacaftor/ivacaftor therapy. A case series

P159 Safety, clinical effectiveness, and changes in systemic cytokine profiles in solid organ transplant recipients post 6 months of elexacaftor/tezacaftor/ivacaftor therapy. A case series

Understanding the dynamics of TKI-induced changes in the tumor immune microenvironment for improved therapeutic effect

BackgroundThe dynamic interplay between tyrosine kinase inhibitors (TKIs) and the tumor immune microenvironment (TME) plays a crucial role in the therapeutic trajectory of non-small cell lung cancer (NSCLC). Understanding the...

Kinetics of IFNγ-Induced Cytokines and Development of Immune-Related Adverse Events in Patients Receiving PD-(L)1 Inhibitors.

Immune checkpoint inhibitors (ICI) have the potential to trigger unpredictable immune-related adverse events (irAEs), which can be severe. The underlying mechanisms of these events are not fully understood. As PD-L1 is upregulated by IFN, the heightened immune activation resulting from PD-1/PD-L1 inhibition may enhance the IFN response, triggering the expression of IFN-inducible genes and contributing to irAE development and its severity. In this study, we investigated the interplay between irAEs and the expression of IFN-inducible chemokines and cytokines in 134 consecutive patients with solid tumours treated with PD-(L)1 inhibitors as monotherapy or in combination with chemotherapy or other immunotherapy agents. We compared the plasma levels of IFN-associated cytokines (CXCL9/10/11, IL-18, IL-10, IL-6 and TGFβ) at various time points (at baseline, at the onset of irAE and previous to irAE onset) in three patient groups categorized by irAE development and severity: patients with serious irAEs, mild irAEs and without irAEs after PD-(L)1 inhibitors. No differences were observed between groups at baseline. However, patients with serious irAEs exhibited significant increases in CXCL9/10/11, IL-18 and IL-10 levels at the onset of the irAE compared to baseline. A network analysis and correlation patterns highlighted a robust relationship among these chemokines and cytokines at serious-irAE onset. Combining all of the analysed proteins in a cluster analysis, we identified a subgroup of patients with a higher incidence of serious irAEs affecting different organs or systems. Finally, an ROC analysis and a decision tree model proposed IL-18 levels ≥ 807 pg/mL and TGFβ levels ≤ 114 pg/mL as predictors for serious irAEs in 90% of cases. In conclusion, our study elucidates the dynamic changes in cytokine profiles associated with serious irAE development during treatment with PD-(L)1 inhibitors. The study's findings offer valuable insights into the intricate IFN-induced immune responses associated with irAEs and propose potential predictive markers for their severity.

390 Modelling Glioma Associated Macrophages Using Human Embryonic Stem Cell Derived Microglia

INTRODUCTION: Glioma associated macrophages/microglia (GAMs) are the most abundant immune cell type in glioblastoma and play a key role in immunotherapeutic resistance. The tumor microenvironment (TME) is thought to modulate GAM phenotype and function leading to a more pro-tumorigenic immune milieu however the exact nature of this crosstalk is unclear. Existing systems to study human GAMs are suboptimal as primary human GAMs are difficult to culture. There is therefore a need to develop human model systems of sufficient fidelity to probe GAM function. Here we report on the development of a human embryonic stem cell (hES)-based platform to derive microglia without recourse to immortalizing oncogenic transduction. This technology can assess the impact of tumor derived factors on the microglial phenotype and represents a novel, scalable model system to investigate GAM function. METHODS: hES-microglia were generated using a stepwise cytokine-based differentiation protocol. Upon exposure to conditioned media obtained from cultures of a panel of patient-derived glioblastoma cell lines (n=6 biological replicates), hES-GAMs were analyzed using flow cytometry, single cell RNA sequencing (scRNA). Functional assays included phagocytosis of pH rhodo particles, wound scratch migration assays and intracellular proteome analysis. RESULTS: Exposure to tumor media leads to transcriptional changes similar to those seen in primary GAMs with increased chemotactic genes such as CCL2(MCP1), CCL3 and CCL4, with corresponding changes in cytokine profile at the protein level. Other upregulated cytokines of note include VEGF, OSM, CCL4 and IL10RB. Functional impact of co-culture with tumor media included a dramatic increase in GAM motility (&gt;30 fold) and a statistically significant decrease in phagocytic ability (29.1% p &lt; 0.001). CONCLUSIONS: The hES-derived GAM model is a promising novel platform to dissect the impact of the TME on tumor-derived GAMs.

Serum cytokines profile changes in amyotrophic lateral sclerosis

BackgroundAmyotrophic lateral sclerosis (ALS) is a devastating neurodegenerative disorder, characterized by progressive limb weakness, dysphagia, dysphonia, and respiratory failure due to degeneration of upper and lower motor neurons. The pathogenesis of ALS is still unclear. Neuroinflammation has been found to be involved in its development and progression. Cytokines play a significant role in the inflammatory process. This study aims to identify novel biomarkers that may assist in the diagnosis of ALS. MethodsIn Fujian Medical University Union Hospital and Huashan Hospital Fudan University, two independent centers, we prospectively recruited 50 ALS patients, and 41 healthy controls (25 ALS and 26 controls in the first stage and 25 ALS and 15 controls in the validation stage). An 18-plex Luminex kit was used to screen the serum cytokines levels in the first stage. Commercial ELISA kits were used to measure the levels of target cytokines in the validation stage. A single-molecule array HD-X platform was applied to assess the levels of serum neurofilament light chain (NFL). ResultsThe levels of serum IL-18 were markedly increased in patients with ALS in the first stage (p = 0.016). The ROC curve showed an area under the curve at 0.695 (95% CI 0.50–0.84) in distinguishing ALS patients from healthy controls. The IL-21 was decreased in elderly patients when grouped by 55 years old (the medium age). Furthermore, the IL-5, IL-13, IL-18, and NFL had a positive relationship with the disease progression of ALS. We also found that serum IL-18 was markedly increased in ALS patients in the validation stage (167.67 [148.25–175.59] vs 116.44 [102.43–122.19]pg/ml, p < 0.0015). ConclusionIn this study, we identified systemic cytokine profile changes in the serum of ALS patients, especially the elevated IL-18, as well as the decreased IL-21 in elder patients. These changes in serum cytokine profiles may shed new light on an in-depth understanding of the immunopathogenic characteristics of ALS.

The impact of age and sex on the inflammatory response during bone fracture healing.

Inflammation is thought to be dysregulated with age leading to impaired bone fracture healing. However, broad analyses of inflammatory processes during homeostatic bone aging and during repair are lacking. Here, we assessed changes in inflammatory cell and cytokine profiles in circulation and in bone tissue to identify age- and sex-dependent differences during homeostasis and repair. During homeostatic aging, male mice demonstrated accumulation of CD4+ helper T cells and CD8+ cytotoxic T cells within bone while both pro-inflammatory "M1" and anti-inflammatory "M2" macrophage numbers decreased. Female mice saw no age-associated changes in immune-cell population in homeostatic bone. Concentrations of IL-1β, IL-9, IFNγ, and CCL3/MIP-1α increased with age in both male and female mice, whereas concentrations of IL-2, TNFα, TNFR1, IL-4, and IL-10 increased only in female mice - thus we termed these "age-accumulated" cytokines. There were no notable changes in immune cell populations nor cytokines within circulation during aging. Sex-dependent analysis demonstrated slight changes in immune cell and cytokine levels within bone and circulation, which were lost upon fracture injury. Fracture in young male mice caused a sharp decrease in number of M1 macrophages; however, this was not seen in aged male mice nor in female mice of any age. Injury itself induced a decrease in the number of CD8+ T cells within the local tissue of aged male and of female mice but not of young mice. Cytokine analysis of fractured mice revealed that age-accumulated cytokines quickly dissipated after fracture injury, and did not re-accumulate in newly regenerated tissue. Conversely, CXCL1/KC-GRO, CXCL2/MIP-2, IL-6, and CCL2/MCP-1 acted as "fracture response" cytokines: increasing sharply after fracture, eventually returning to baseline. Collectively, we classify measured cytokines into three groups: (1) age-accumulated cytokines, (2) female-specific age-accumulated cytokines, and (3) fracture response cytokines. These inflammatory molecules represent potential points of intervention to improve fracture healing outcome.

Abatacept Enhances Natural Killer Cell Cytotoxicity Following Haploidentical HCT: A Novel Platform to Dissociate GVL from Gvhd

Background: Abatacept (Abata, CTLA4 Ig), has been explored for the past 3 decades for blockade of CD28-B7 costimulation pathway in organ transplantation. One of the disadvantages of Abata has been its sparing effect on natural killer (NK) cells, resulting in NK-mediated rejection in preclinical models. However, we explored this caveat of abatacept to augment NK cell mediated anti-leukemia effect without invoking T cell mediated GVHD, where unmodified donor lymphocyte infusions (DLI) were administered 6 hours following administration of abatacept in-vivo (Abata-DLI) to patients with advanced malignancies undergoing haploidentical (Haplo)-HCT on days +7, +21 and +35. An early and rapid proliferation of mature NK cells was observed, which correlated with an improved relapse-free survival, without an increase in GVHD. To explore the mechanistic pathway behind this phenomenon, we studied the effect of Abata on cytotoxicity as well as cytokine profile of NK cells both in-vitro as well as in-vivo (CTRI:REF/2021/08/046552). Methods: NK cells and monocytes were isolated from PBMC of healthy donors. NK cells ± monocytes, were incubated with IL2, IL15 and IL12/15/18 with or without Abata for 72 hours and 7 days. They were assessed for cytotoxicity and IFN-g and perforin expression against K562 cell lines at effector: target (E:T) ratio of 5:1 and 1:1 . In HCT recipients, samples were obtained at day +20 (baseline), +30, +45, +60, and 6, 12 and 18 months. The samples were cryopreserved and both cytotoxicity and cytokine assays were carried out in batches after thawing. Cytokine profile was analysed in total NK cells and in NKG2A+ and NKG2C+ subsets. Results: The study was carried out on 11 healthy donors and 20 haplo-HCT recipients (15 with malignant diseases receiving Abata-DLI and 5 with non-malignant diseases receiving Abata without DLI). NK cells from healthy donors showed significant increase in cytotoxicity at 72 hours in presence of Abata with IL2 and IL15 (Figure1, p&amp;lt;0.01) at E:T ratio of 5:1 as well as 1:1 at both 72 hours as well as 7 days, but not with IL12/15/18. The effect was independent of monocytes. Perforin tended to be upregulated at both 72 hours and 7 days with Abata and IL15. In patients receiving Abata-DLI, cytotoxicity of NK cells was significantly increased at all time points compared to baseline at E:T ratio of 5:1 and 1:1 (Fig 2, P &amp;lt; 0.01). NK-cytotoxicity was significantly higher than the corresponding donor samples at days +45 and +60 (p&amp;lt;0.05). This was particularly so in 11 patients without any event compared to 4 patients with relapse/GVHD. To evaluate if Abata-DLI had greater impact on NK-cytotoxicity, the data was compared with 5 patients who received Abata alone. This showed significantly greater cytotoxicity at days +45 and +60 in the Abata-DLI group, which was more pronounced, when only those without events were analysed from the Abata-DLI group (p&amp;lt;0.01). There was no significant change in cytokine profile in those receiving Abata alone, but in Abata-DLI group, IFN-g was significantly upregulated in the NKG2C+ subset at day +60 at 1:1 ratio. Perforin expression was significantly higher at days+45 and +60 at both 5:1 and 1:1 in the Abata-DLI cohort, for both NKG2C+ and NKG2A+ subsets. In the Abata-DLI group NKG2C+ subset showed significantly higher IFN-g at days+45 and +60 and upregulation of perforin at all time points. 3 patients transplanted with active leukemia, assessed beyond day +60, showed greater cytotoxicity compared to the corresponding donors at 6-18 months. Conclusion: In this study, Abata was shown to increase cytotoxicity of NK cells, independent of monocytes, in the presence of IL2 and IL15, but not with IL12/15/18. This was observed at both high as well as low E:T ratios, with significant upregulation of perforin. In Haplo-HCT with Abata-DLI, there was marked increase in NK-cytotoxicity with upregulation of both IFN-g and Perforin, when compared to baseline as well as in comparison to those receiving Abata without DLI. Our findings suggest that Abatacept directly augments NK-cytotoxicity and provides rationale for exploring Abata-augmented NK cell therapy as a novel approach to augment NK-mediated GVL effect without invoking T cell mediated GVHD

Role of altered amniotic fluid cytokine profile in in development of fetal membrane insufficiency

Studies on pathogenesis of fetal membrane insufficiency in preterm pregnancy should expand the opportunities of predicting prenatal rupture of amniotic fluid and improve the strategy of anticipant gestation management in cases of premature rupture of the fetal membranes. The clinical significance of studying this obstetric problem is unquestionable due to the high risk of complications and perinatal losses caused by preterm birth. This research is devoted to studying the changes in cytokine profile of amniotic fluid during prenatal rupture of amniotic fluid at 22 to 34 weeks of gestation. The levels of pro-inflammatory (TNFα, IL-6, IL-1β, IL-2, IL-6, IL-8) and anti-inflammatory (IL-4, IL-10) cytokines in amniotic fluid were determined in 30 patients, whose premature pregnancy was complicated by early rupture of the membranes. For reference, the level of these cytokines in the amniotic fluid was studied in 25 pregnant women with a physiological course of pregnancy at full-term gestation. The ELISA technique was used with a test system produced by JSC Vector-Best. The study was carried out on the basis of the Perinatal Center of the Saratov Region. We have found that premature rupture of the membranes is preceded by increased levels of pro-inflammatory cytokines in amniotic fluid. The latter result suggests a significant alteration in feto-placental complex, which may be the starting point both for damage to the fetal membranes, as well as for development of labor activity.

Early Plasma Levels of IL7 and IL15 Predict Response to Chimeric Antigen Receptor T Cell Therapy (CAR-T) in Large B-Cell Lymphoma (LBCL) Patients

Introduction Anti-CD19 chimeric antigen receptor T cells (CAR-T) have rapidly shifted the treatment paradigm for relapsed/refractory large B-cell lymphoma (LBCL) but produce durable complete response (CR) in only 30-40% of patients. In addition, CAR-T-mediated toxicities such as cytokine release syndrome (CRS) and immune effector cell-associated neurotoxicity syndrome (ICANS) remain a major concern. To investigate the relationships of circulating cytokines and chemokines on CAR-T outcomes, we applied specialized comprehensive cytokine panels to peripheral blood samples serially collected from LBCL patients undergoing treatment with anti-CD19 CAR-T cells. Methods LBCL patients were consented to our IRB-approved study and longitudinally followed for sample collection at pre-specified timepoints: prior to lymphodepleting chemotherapy (LD chemo), day 0 (day of [and prior to] CAR-T infusion), and days 3, 7, 10, 14, 21, and 90. MSD R-PLEX and U-PLEX assays were used to detect 47 cytokines and chemokines in plasma; vasoactive intestinal peptide (VIP) levels were quantified via a VIP-specific enzyme immunoassay. Mann-Whitney U test, Spearman's correlation coefficient, and a LASSO (least absolute shrinkage and selection operator) model were utilized to identify key cytokines contributing to critical clinical outcomes. Results Peri-CAR-T blood samples were collected from 24 LBCL patients, including one patient who received 2 infusions of an investigational allogeneic anti-CD19 product; all other patients received commercially available products, each associated with their distinct LD chemo dosing regimens. With median follow-up of 11.2 months, 54% patients achieved CR as best response, 50% experienced CRS (maximum Grade 2 in 25%), and 29% experienced ICANS (Grade ≥3 in 8%). Most cytokines demonstrated characteristic peak patterns over time that were not dependent on whether cytokines were produced by hematopoietic cells ( Table). Among most cytokines with peak values at Day 0, the mean percentage change from Pre-LD Chemo to Day 0 increased as chemo intensity increased (i.e., fludarabine dose from 25 to 30 mg/m 2, and cyclophosphamide dose from 250 to 500 mg/m 2). Notably, levels of the non-hematopoietic cytokines IL7 and IL15 were highest at day 0, reflecting effects of LD chemo. Using statistical models to filter relevant values, we developed a signature of 14 key cytokines that could be used to predict clinical response at 3 months. Half of these were stromal-produced cytokines from early timepoints: Cytokines positively correlated with CR or partial response (PR) included IL7 at Day 0 (Mann-Whitney U p&amp;lt;0.001) and Day 7 (p=0.044), and IL15 at Day 0 (p=0.016). Negatively correlated cytokines included IL4 at Day 3 (p=0.041), IL10 at Day 7 (p=0.027), CCL19 at Day 10 (p=0.017), and MIP1α at Day 14 (p=0.03). Intriguingly, several Type 1 T helper cell (Th1)-related cytokines with peaks at Day 3 or Day 7 (e.g., IL2, IFN-γ, TNF-α, and IL18) were not associated with response. Strong associations of timepoint-specific cytokine levels with patient outcomes are exemplified in the Figure: all patients achieving CR had IL7 levels above average on Day 0, while most patients (9/10) who failed to achieve CR had IL7 levels below average on Day 0. This same striking pattern was observed for IL15 at Day 0. Similar analyses gauged which timepoint-specific cytokines were associated with CRS and ICANS. Of 14 values associated with CRS, 13 were from Day 3 or later, likely reflecting sequelae of CRS rather than representing predictive biomarkers. Pre-LD Chemo CCL18 was the exception, positively correlated with CRS (p=0.035). Only 5 cytokine values were associated with development of ICANS, 3 from Day 10 or later; IL7 at Day 0 (p=0.011) and Day 3 (p&amp;lt;0.001) was positively correlated with ICANS . Conclusions Comprehensive cytokine profiling revealed that early measurements of several stromal-produced cytokines can predict response in LBCL patients who receive CAR-T-most notably IL7 and IL15 levels on Day 0, prior to CAR-T infusion. Conversely, few early cytokine measurements were helpful in predicting CRS and ICANS, limited to CCL18 and IL7. Analyses are ongoing to characterize associations of patient characteristics, product differences, and circulating immune cell subsets with changes in cytokine profile over time. Further studies are warranted to validate our findings in a larger data set.

Effect of immunology biomarkers associated with hip fracture and fracture risk in older adults

Osteoporosis is a skeletal disease that can increase the risk of fractures, leading to adverse health and socioeconomic consequences. However, current clinical methods have limitations in accurately estimating fracture risk, particularly in older adults. Thus, new technologies are necessary to improve the accuracy of fracture risk estimation. In this observational study, we aimed to explore the association between serum cytokines and hip fracture status in older adults, and their associations with fracture risk using the FRAX reference tool. We investigated the use of a proximity extension assay (PEA) with Olink. We compared the characteristics of the population, functional status and detailed body composition (determined using densitometry) between groups. We enrolled 40 participants, including 20 with hip fracture and 20 without fracture, and studied 46 cytokines in their serum. After conducting a score plot and two unpaired t-tests using the Benjamini-Hochberg method, we found that Interleukin 6 (IL-6), Lymphotoxin-alpha (LT-α), Fms-related tyrosine kinase 3 ligand (FLT3LG), Colony stimulating factor 1 (CSF1), and Chemokine (C-C motif) ligand 7 (CCL7) were significantly different between fracture and non-fracture patients (p < 0.05). IL-6 had a moderate correlation with FRAX (R2 = 0.409, p < 0.001), while CSF1 and CCL7 had weak correlations with FRAX. LT-α and FLT3LG exhibited a negative correlation with the risk of fracture. Our results suggest that targeted proteomic tools have the capability to identify differentially regulated proteins and may serve as potential markers for estimating fracture risk. However, longitudinal studies will be necessary to validate these results and determine the temporal patterns of changes in cytokine profiles.

Changes in serum cytokine profile and deficit severity in patients with relapsing-remitting multiple sclerosis.

In experimental autoimmune encephalomyelitis, neurological deficit correlates with axonal loss and the CD8+ T cells are a likely mediator of axonal damage. In relapsing-remitting multiple sclerosis, there is a correlation of the immune inflammatory activity in the lesion foci with the axon transection.

Gut microbiota and changes in cytokine profile in animals with experimental acute disseminated peritonitis on the background of diabetes

In the pathogenesis of acute widespread peritonitis and accompanying diabetes, a vital link is an endogenous intoxication caused by the translocation of microorganisms and their toxins from the intestine into the blood, metabolic changes, and immunological reactivity of the body. Our work aimed to investigate the microbial composition in the parietal intestinal biotope and the features of the blood cytokine profile in animals with acute disseminated peritonitis on the background of streptozotocin-induced diabetes. The study was conducted on 56 sexually mature non-linear white male rats. Diabetes mellitus was modeled by a single intraperitoneal injection of streptozotocin (60 mg/kg). On the 14th day of the development of streptozotocin-induced diabetes mellitus, a 10% filtered fecal suspension (0.5 mL) was injected into the abdominal cavity of animals, and acute generalized peritonitis was initiated. Sowing on nutrient media was carried out for bacteriological research to isolate a pure culture of microorganisms and their identification. The concentration of TNF-α, IL-1β, and IL-6 was studied by solid-phase enzyme immunoassay. The research results demonstrate an imbalance of cytokines in the dynamics of experimental acute disseminated peritonitis against the background of diabetes and quantitative and qualitative changes in the microbiota of the parietal intestinal biotope. A decrease in the number of Escherichia coli strains isolated in monoculture and an increase in the number of two-component and three-component microbial associations were revealed, among which Enterobacter aerogenes, Escherichia coli, Bacteroides spp., Proteus mirabilis, Klebsiella spp. and Candida species prevailed.