Cytokine Pattern Research Articles

Pulmonary and serum cytokine profiles of patients with anti-ARS and anti-MDA5 antibodies.

The present study aimed to determine the pulmonary cytokine profiles of patients with anti-RNA synthetase (ARS) and anti-melanoma differentiation-associated protein 5 (MDA5) antibodies. The study included patients with ARS and MDA5 whose serum or bronchoalveolar fluid (BALF) was available. Sandwich enzyme-linked immunoassay microarray multiplex assay was used to measure 18 cytokine levels in serum and BALF. The cytokine patterns were investigated using factor and cluster analyses. Pulmonary cytokine production was examined using the BALF/Seum cytokine ratio. Forty participants were enrolled in the study: 19 with ARS and 21 with MDA5. All patients had interstitial lung disease (ILD). BALF was collected from 10 patients with ARS and 6 with MDA5. Serum type 1 IFN, IP-10, MCP-1, and TNF-α were elevated in both ARS and MDA5. IL-6, IL-10, and IL-15 were elevated in MDA5. Serum cytokine patterns differed between ARS and MDA5. In BALF, IFN-α, IP-10, MCP-1, and ferritin were increased in both ARS and MDA5. Higher levels of IFN-α, IL-6, and ferritin were observed in MDA5. One patient with severe MDA5-ILD showed higher levels of multiple cytokines, including IL-6 and IFN-α. BALF cytokine patterns were similar in ARS and MDA5 cases except the one with severe MDA5-ILD. IL-6, IP-10, IL-15, MCP-1, and ferritin were produced in the lungs in ARS and MDA5 and IFN-α in MDA5. In conclusion, IFN-α and pulmonary macrophage activation play important roles in ILD development in both ARS and MDA5-ILD. MDA5-ILD could be characterized by higher production of multiple cytokines and macrophage activation, particularly in severe cases.

Association of increased serum I-309 with phenotypes, disease activity, and cytokine pattern in primary Sjögren's syndrome.

The aim of this study was to determine serum I-309 levels in primary Sjögren's syndrome (pSS) patients, as well as the association with disease phenotype, systemic activity, and T helper cell-related cytokines.A total of 58 pSS patients and 30 healthy controls (HC) were enrolled in this study. The concentrations of serum I-309, interleukin-4 (IL-4), IL-6, IL-9, IL-13, IL-17, IL-22, IL-23, tumor-necrosis factor-α (TNF-α), interferon-γ (IFN-γ), IFN-α, and IFN-β were measured with multiplex immunoassay. The relationships between I-309 and various clinical and laboratory variables were analyzed.The serum concentrations of I-309 were significantly increased (median, IQR, 14.24, 10.99--20.35, pg/ml) in pSS patients compared with HC (median, IQR, 8.27, 6.74--9.62, pg/ml) (P < 0.001).Serum I-309 is increased in pSS, and may be associated with systemic inflammation, Th1-, Th17,- and Th9 cells, type Key Points • Serum I-309 levels are increased in pSS patients. • Increased I-309 may be associated with systemic rather than local manifestations in pSS. • Increased I-309 may be associated with Th1, Th17 and Th9 other than Th2 in pSS. • There may be close relationships between I-309 and type land type II IFNs in pSS.

P0015 The local intestinal interstitial cytokine profile provides an insight into inflamed bionaive patients with Ulcerative Colitis

Abstract Background Proinflammatory cytokines identified in the mucosal microenvironment at the site of inflammation are involved in the pathophysiology of chronic inflammatory bowel diseases such as ulcerative colitis (UC). We hypothesize that the local cytokine production is significantly altered in inflamed regions of the intestine and cannot be predicted by plasma measurements. Our goal is to identify personalized cytokine signatures in UC patients' interstitial space depending on their disease progression. This can significantly advance our understanding of UC and support the development of more personalized treatment strategies as cytokine signaling is already a target of current therapies. Methods We isolated interstitial fluid (IF) from the intestinal mucosa using the tissue elution method in bionaive UC patients (mild activity (UCEIS 1-4) n = 6; inflamed (UCEIS 5-8) n = 6) and healthy controls (n = 25). We took biopsies from 4 different intestinal segments (rectum, descending colon, ascending colon and ileum) with an average weight of 13,4 mg in 400 mg NaCl for 2h elution in 4°C. Obtained IF was used for multiplex immunoassays of 18 proinflammatory cytokines in the same way as plasma cytokines. Results Cytokines such as VEGF, IL-16 and IL-12p40 are present in healthy controls and inflamed patients in the IF. Depending on the disease's severity, more cytokines can be detected. Cytokine concentrations decrease in the more aboral localizations of the colon with the rectum showing a unique cytokine pattern. Cytokines that can be detected in plasma show much lower concentrations compared to IF. Inflamed patients can be distinguished from mild activity in local interstitial profiles. In plasma, different cytokines appear to drive inflammation – those can be identified by linear discriminant analysis, and their composition differs from that in the rectum. Moreover, plasma and rectal interstitial mucosa cytokine patterns correlate well with endoscopic and clinical UC scores. Conclusion Isolation of IF is a valuable approach to assessing inflammation in UC patients, improving our understanding of mild residual activity and more severe conditions compared to traditional plasma measurements. We can identify cytokines that drive inflammation in UC and characterize the inflammatory cytokine profile in the intestinal interstitial space associated with a healthy gut. Future targeted therapies should also consider the unique cytokine profiles of individual patients being expressed in the rectum. Comparing bionaive cytokine profiles with profiles from patients treated with biologicals, will allow us to demonstrate how the network changes in response to immunosuppression.

Tattooed human in vitro skin model for testing the biocompatibility of tattoo inks and healing progression after tattooing

Tattoos are widespread in the population. Tattoo inks, which contain a variety of ingredients among them hazardous compounds such as polyaromatic hydrocarbons, heavy metals and nanoparticles and that are made for injection into the skin, are not dermatologically tested. New testing systems for evaluation of biocompatibility of tattoo inks as composite products and the tattooing process itself are needed. This paper describes an in vitro 3D human skin model that was tattooed with black and red ink. Biocompatibility including analysis of cytotoxicity, cytokine release, and gene expression patterns of proinflammatory cytokines, proliferation markers, growth factors and structural components was investigated over a period of 7 days. Tattooing of the 3D skin model resulted in a strong inflammatory reaction comparable to in vivo observations that subsided 4 days after treatment. The subsequent healing phase was detectable in the gene expression patterns. Tattooing with two different tattoo inks resulted in distinguishable inflammatory reactions. The described 3D skin model is a useful tool for evaluation of the biocompatibility of tattoo inks and the tattooing process itself and for characterizing the healing process after tattooing.

Distinct clinical outcomes based on multiple serum cytokine and chemokine profiles rather than autoantibody profiles and ultrasound findings in rheumatoid arthritis: a prospective ultrasound cohort study

ObjectivesTo evaluate the potential of clinical factors, ultrasound findings, serum autoantibodies, and serum cytokine and chemokine profiles as predictors of clinical outcomes in rheumatoid arthritis (RA).Patients and methodsWe included 200...

Destructive and protective effects and therapeutic targets of IL-36 family cytokines in dry eye disease.

To explore the destructive and protective effects and therapeutic targets of IL-36 cytokines in dry eye disease using a murine dry eye model. A dry eye model was established in C57BL/6 mice exposed to desiccating stress (DS) with untreated mice as controls. A topical challenge model was performed in normal mice with exogenous rmIL-36α, rhIL-38 and 2% ectoine, or PBS vehicle. IL-36 cytokine expression was assessed by RT-qPCR and immunofluorescent (IF) staining. Corneal epithelial damage was evaluated by corneal smoothness score, Oregon Green Dextran (OGD) fluorescent staining, and tight junction barrier. All members of the IL-36 family were expressed by murine ocular surface epithelium. The expression of IL-36α and IL-36γ was upregulated while IL-38 and IL-36RN were down regulated in ocular surface of dry eye mice. A topical challenge of rmIL-36α directly destructed corneal surface with distorted smoothness, increased OGD uptake and IF intensity, and disrupted tight junction proteins ZO-1 and occludin. Co-application with rhIL-38 prevented all these corneal damages by rmIL-36α. Ectoine treatment reversed the pathological expression pattern of IL-36 cytokines, protected corneal epithelium from defects, and restored the tight junction barrier in DS mice, and even prevented corneal damage by rmIL-36α. Our findings demonstrate the upregulated pro-inflammatory agonists IL-36α and IL-36γ with downregulated antagonists IL-38 and IL-36RA in dry eye model, which provides a previously unknown mechanism and therapeutic targets in dry eye disease. The therapeutic efficacy of ectoine may be through reversing the pathological alteration of IL-36 cytokines in dry eye mice.

Neuroinflammation as an Integral Component of Neurodegeneration in Parkinson's Disease

Parkinson's disease (PD) is a progressively advancing neurodegenerative disorder, the pathogenetic mechanisms of which remain poorly understood. The disease is characterized by the degeneration of dopaminergic neurons in the substantia nigra of the midbrain. Given the improvement in the quality of medical care provided to the population, it is projected that the total number of patients diagnosed with PD worldwide will rise to 8.7 million by 2030. This review addresses the fundamental aspects of neuroinflammation in the context of PD pathogenesis. There is no doubt that pro-inflammatory immunological mechanisms play a critical role in the onset and progression of the disease. Neuronal-derived cells, such as microglia and astrocytes, act as inducers of neuroinflammation, affecting the permeability of the blood-brain barrier to peripheral immune-competent cells. Furthermore, cytokine patterns of the immune response in PD appear to exist. Potential therapeutic approaches for mitigating neuroinflammation in PD, which have been studied in experimental and in vitro models, are also discussed.

Characterization of Serum Cytokine Patterns in Frequent-Exacerbation Asthma: Implications for Phenotyping and Management.

(1) Background: Asthma exacerbations represent significant clinical events, however, the underlying inflammatory mechanisms and cytokine profiles in patients with frequent exacerbations remain incompletely understood; (2) Methods: In this prospective, cross-sectional study of 120 stable asthma patients, we compared the serum concentrations of eight key cytokines (IL-4, IL-12, IL-13, IL-17, IFN-α, IFN-γ, TNF-α, and IL-1β) between two groups: 60 patients with frequent exacerbations (≥ 2 events per year) and 60 matched controls with few exacerbations (1 event per year); (3) Results: Patients with frequent exacerbations showed significantly higher serum concentrations of IL-4 and IL-13 (p < 0.05), along with an increased prevalence of allergic history and comorbidities (chronic rhinosinusitis, GERD, OSA; all p < 0.05). The IgE levels correlated positively with IFN-α (rh = 0.26) and TNF-α (rh = 0.29), while the FeNO levels correlated with IL-17 (rh = 0.26) and IL-1β (rh = 0.33) (all p < 0.05); (4) Conclusions: Our findings identify a distinct cytokine signature in frequent exacerbators characterized by elevated IL-4 and IL-13 levels. The correlations between specific cytokines and established biomarkers suggest potential mechanisms underlying exacerbation susceptibility, which may inform targeted therapeutic strategies for this high-risk population.

Bacillus Subtilis-Derived Exopolysaccharide Halts Depigmentation and Autoimmunity in Vitiligo.

Vitiligo has a complex multifactorial etiology involving a T-cell-mediated autoimmune response to cutaneous melanocytes. Microbial dysbiosis has been assigned a contributing role in vitiligo etiology. Treating vitiligo can be a challenging task, and finding novel treatment approaches is crucial. In this study, we tested exopolysaccharides (EPSs) isolated from Bacillus subtilis as a microbiome-based therapy. Vitiligo-prone h3TA2 mice were treated by weekly intraperitoneal EPS injection for 18 weeks. Depigmentation was evaluated over time, measuring immune responses at end point. EPS treatment significantly limited the rate of depigmentation. The abundance of cutaneous T cells, specifically CD8+ cytotoxic T cells, was reduced, whereas regulatory T cells were more abundant in the skin of treated mice than in untreated mice. Moreover, EPS treatment was associated with increased numbers of splenic M2 macrophages, elevated splenic indoleamine 2,3-dioxygenase expression, and a systemic cytokine shift toward a type 2 pattern of cytokines. Importantly, splenocytes retrieved from EPS-treated mice were less responsive to cognate tyrosinase peptide, as demonstrated by limited release of IFN-γ and other inflammatory cytokines. In summary, EPS isolated from Bsubtilis interfered with T-cell-mediated depigmentation in the h3TA2 mouse model of vitiligo, suggesting that Bsubtilis EPS could serve as a novel treatment entity for vitiligo.

From glia to cytokine signatures across the Alzheimer’s spectrum

AbstractBackgroundEarly neuroinflammation is involved in pathophysiology of Alzheimer’s Disease (AD) and contributes to faster clinical decline. Thus, neuroinflammation has emerged as a promising therapeutic target for dementia. However, a better understanding of the interaction between central and peripheral inflammation in human disease and in vivo biomarkers are required for successful clinical trials. Here we assess inflammatory patterns of serum cytokines and their relationship with central inflammation as quantified by TSPO PET (PK11195) in participants on the aging‐AD spectrum.MethodBlood samples were obtained from 56 participants, including30 controls, 22 patients with MCI and 14 with AD) Serum cytokines were quantified using the MesoScale Discovery V‐Plex‐Human Cytokine panel. We applied a Principal Component Analysis (PCA) across all participants to identify inflammatory profiles, which were compared across groups with ANOVA and post hoc tests. Individual scores of the resulting components were correlated with regional TSPO PET binding potentials, as index of microglial activation, and cognitive impairment (MMSE scores).ResultAnalyses on 30 identified 2 components (explaining 19.4 % and 10.8% of the variance, respectively). Both components were strongly represented by pro‐inflammatory cytokines (Figure 1A). One‐way analyses of variance on the first component could not detected statistically significant differences across the groups (χ2(2)=1.69, p=0.193), while individual scores of Component 2 (χ2(2)=8.46, p&lt;0.001) were statistically higher in patients with AD as compared to patients with MCI and to controls (Figure 1B). Correlation analyses with regional TSPO PET values identified positive widespread associations with individual Component 2 scores in frontal, temporal and parietal regions (r &gt; 0.250, p &lt; 0.05; while associations with Component 1 where limited to the anterior frontal/temporal regions (Figure 1C). Individual scores of Component 2 but not Component 1 were significantly associated with worse cognitive performance (lower MMSE; r = ‐0.315, p = 0.01).ConclusionOur approach identified proinflammatory blood signatures that relate to central inflammation and cognitive impairment across the aging‐AD spectrum. These findings encourage the combination of TSPO imaging and blood‐based markers to clarify the interaction between peripheral and central inflammation in AD, and to identify promising therapeutic targets.

The Pattern of Cytokines, Chemokines, and Growth Factors of the Maxillary and Mandibular Periosteum After Exposure to Titanium Fixations-Ti6Al4V.

Objectives: Titanium miniplates and screws are commonly used in the surgical management of dentofacial deformities. Despite the opinion of the biocompatibility of these bone fixations, some patients experience symptoms of chronic inflammation around titanium implants even many years after their application. The aim of this study was to examine the levels of cytokines, chemokines, and growth factors released from the maxilla and mandible periosteum surrounding titanium fixations 11 months after the implantation procedure. Methods: From the study group (n = 20) consisting of patients with maxillofacial defects who underwent bimaxillary osteotomy, fragments of the periosteum of the maxilla and mandible adjacent to the titanium miniplates and screws were taken during routine bone fixation removal procedures. From the control group subjects (n = 20), fragments of healthy maxillary and mandibular periosteum were taken prior to surgical treatment of dentofacial deformities. The examination of cytokines, chemokines, and growth factors levels released from the periosteum of jaws was performed using the Bio-Plex Pro Human Cytokine Screening Panel (48-Plex). Results: The study group was characterized by a significant increase in the concentration of most of the tested-for proinflammatory cytokines/chemokines/growth factors compared to the control group, with greater amounts of inflammatory factors released from the periosteum covering the titanium implants in the mandible than from the periosteal cells surrounding the titanium implants in the maxilla. Conclusions: Prolonged exposure to titanium miniplates and screws leads to a disturbance of immune homeostasis in the periosteal cells of the maxilla and mandible. The data obtained indicate the need to remove fixations after the bone fragments have healed.

The diurnal pattern of cytokines, chemokines and growth factors in human saliva—a pilot study

BackgroundUnderstanding of possible periodicity of cytokines, chemokines and growth factors is of great interest and provide valuable information for research into pathophysiological mechanism of inflammatory disease and chronic pain. Significant efforts have been made to identify different analytes in saliva. For precision and accuracy in measurement and interpretation of results, it is crucial to know the source of variability, especially the circadian variation for the analytes.ObjectiveThe study aimed to analyze circadian variation in 71 inflammatory markers in both unstimulated and stimulated saliva, as well as plasma, from a sample of healthy individuals.MethodsTen young adults participated. Unstimulated and stimulated whole saliva were collected at 3-h intervals between between 7:30 am and 7:30 pm. Blood samples were drawn in connection with the first and last saliva collection. All samples were analyzed using the U-PLEX 71-Plex assay.ResultsThe analysis showed distinct clustering of the 71 inflammatory mediators between plasma and saliva. Furthermore, differences were also observed between stimulated and unstimulated saliva. The proteins were clustered into three groups that expressed different circadian rhythms. These clusters were stable over time in stimulated saliva but showed significant variability in unstimulated saliva (P &amp;lt; 0.05).ConclusionsThese results suggest that time of the day could influence the detection and interpretation of inflammatory markers and collecting saliva samples at consistent times across participants will help control for the natural fluctuations in salivary composition. The results encourage further exploration of salivary diagnostics, particularly in understanding circadian rhythms and localized immune responses.

The Microbiome Modifies Manifestations of Hemophagocytic Lymphohistiocytosis in Perforin-Deficient Mice.

Primary hemophagocytic lymphohistiocytosis (HLH) is a life-threatening hyperinflammatory syndrome caused by inborn errors of cytotoxicity. Patients with biallelic PRF1 null mutations (encoding perforin) usually develop excessive immune cell activation, hypercytokinemia, and life-threateningimmunopathology in the first 6 months of life, often without an apparent infectious trigger. In contrast, perforin-deficient (PKO) mice only develop HLH after systemic infection with lymphocytic choriomeningitis virus (LCMV). We hypothesized that restricted microbe-immune cell interactions due to specific pathogen-free (SPF) housing might explain the need for this specific viral trigger in PKO mice. To investigate the influence of a "wild" microbiome in PKO mice, we fostered PKO newborns with Wildling microbiota ('PKO-Wildlings') and monitored them for signs of HLH. PKO-Wildlings survived long-term without spontaneous disease. Also, systemic infection with vaccinia virus did not reach the threshold of immune activation required to trigger HLH in PKO-Wildlings. Interestingly, after infection with LCMV, PKO-Wildlings developed an altered HLH pattern. This included lower IFN-γ serum levels along with improved IFN-γ-driven anemia, but more elevated levels of IL-17 and increased liver inflammation compared with PKO-SPF mice. Thus, wild microbiota alone is not sufficient to trigger HLH in PKO mice, but host-microbe interactions shape inflammatory cytokine patterns, thereby influencing manifestations of HLH immunopathology.

Cytokine Patterns for Secondary HLH: Experiences from a Single Center

Cytokine Patterns for Secondary HLH: Experiences from a Single Center

A Defined Diet Combined with Sonicated Inoculum Provides a High Incidence, Moderate Severity Form of Experimental Autoimmune Encephalomyelitis (EAE).

Myelin oligodendrocyte glycoprotein 35-55 (MOG35-55)-peptide induced experimental autoimmune encephalomyelitis (EAE) is a model for inflammation of the brain and spinal cord. However, its severity and incidence vary within and between laboratories. Severe scores can lead to premature termination and are both unnecessary for readouts and detrimental to animal welfare. Ideally, the model would have high incidence, moderate severity, and low interindividual variability to fulfill the "Refine" aspect of the 3R concept. Nevertheless, most efforts to increase incidence also increase the severity. When the effects of potential therapies are tested, moderate severity is sufficient to detect useful drug effects as long as variation is low. Low variation can also reduce group sizes, which supports the "Reduce" aspect of 3R approaches in disease modeling. We set out to reduce variation and control severity by assessing the effects of mouse age, dietary fiber, antigen emulsion, and the dose of MOG and pertussis toxin on incidence, variability, and severity in the MOG-EAE model. We compared 14- and 33-week-old female C57BL/6 mice and varied the diet and inoculum in two studies. We measured disease signs in vivo as well as gene expression in the brain and spinal cord and histology by immunofluorescence. Ordinary one-way ANOVA was used for multiple comparisons. The most reliable induction conditions were with a low-fermentative/fiber diet (AIN 93M) combined with a sonicated emulsion of the MOG35-55-peptide. High-dose pertussis toxin increased EAE severity and incidence in 14-week-old mice (25% survival) while being more moderate in mature mice (100% survival). Varying all parameters suggests that duration of prefeeding defined diet, emulsion quality, and mouse maturity were factors that increase uniformity of response allowing incidence to reach 100% without excess severity. Microglia and astrocyte-associated markers were upregulated proportionally to score consistent with known EAE pathology. A defined fiber/high-sugar diet with sonicated inoculum provides for a moderate severity, high incidence, and less variable EAE. The resulting uniformity in animal response and associated cytokine patterns, and the strong link to a defined diet, suggest that this may be a more clinically translatable protocol for the induction of EAE. This is consistent with reported effects of low-fermentable diets on immune modulation in human patients with autoimmune diseases.

Cooling lactating sows exposed to early summer heat wave alters circadian patterns of behavior and rhythms of respiration, rectal temperature, and saliva melatonin.

Heat stress (HS) exerts detrimental effects on animal production, with lactating sows being particularly vulnerable. Understanding the mechanisms involved in HS response could aid in developing effective strategies against the negative impacts on livestock. Recent genome wide association studies identified two core circadian clock genes as potential candidates in mediating HS response. The study aimed to investigate how cooling lactating sows under natural heat stress conditions impacted circadian patterns of respiration rate (RR), rectal temperature (RT), behavior, salivary melatonin and cortisol levels, and diurnal patterns of cytokines in saliva. Mixed parity lactating sows were assigned to one of two treatment groups: electronic cooling pad (C; n = 9) and heat-stressed (H; n = 9). The experiment spanned two 48 h periods of elevated ambient temperatures due to summer heat wave. In the first 48 h period, RR was recorded every 30 min, RT every 60 min, and behaviors (eating, standing, sitting, laying, sleeping, drinking, and nursing) every 5 min. In the second 48 h period, saliva samples were collected every 4 h. Cooling reduced RR and RT and altered circadian patterns (P < 0.05). Cooling did not affect amount of time engaged in any behavior over the 48 h period (P > 0.05), however, daily patterns of eating, standing and laying differed between the treatments (P < 0.05), with altered eating behavior related to RT increment in H sows (P < 0.05). Cooling increased and altered the circadian pattern of salivary melatonin (P < 0.05). Cooling also influenced the diurnal pattern of saliva cytokines. Cooling had no impact on saliva cortisol levels. In conclusion, cooling HS sows impacted circadian rhythms of physiology and behavior, supporting the need for further research to understand if circadian disruption underlies decreased production efficiency of HS animals.

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.

Plasma Cytokines Pattern as a Prognostic Marker for Esophageal Squamous Cell Carcinoma via Unsupervised Clustering Analyses

Introduction: Cytokines such as tumor necrosis factor-alpha (TNF-α), interleukin 6 (IL6), interferon-gamma (IFN-γ), interleukin 17-alpha (IL17-α), and interleukin 33 (IL33) play critical roles in immune responses and may impact cancer prognosis in future. However, few studies have simultaneously explored the prognostic impact of these cytokines for cancer. In this study, we aim to apply the unsupervised clustering analysis to approach the correlation between the expression of these cytokines and the subsequent prognosis of patients with esophageal squamous cell carcinoma (ESCC). Methods: A robust clustering algorithm was used, the Gaussian mixture method (GMM), through the mclust R package to group patients based on the expression of their cytokines in plasma or tumors. The 324 NTU patients were grouped into 4 clusters, and the 179 GSE53625 patients were grouped into 3 clusters based on expression in plasma and tumors, respectively. Five- and 3-year overall survival (OS) and progression-free survival (PFS) curves of each cluster were compared. Univariate and multivariate Cox regression analyses were also performed. Results: We successfully distinguished the multimodal distribution of cytokines through GMM clustering and discovered the relationship between cytokines and clinical outcomes. We observed that NTU-G3 and NTU-G4 subgroups showed most variation in 5-, 3-year OS and 5-, 3-year PFS with NTU-G3 being associated with poorer prognosis compared to NTU-G4 (p = 0.016, 0.0052, 0.0575, and 0.0168, respectively). NTU-G3 was characterized with higher TNF-α (median = 3.855, N = 78) and lower IL33 (median = 0.000, N = 78), while NTU-G4 showed lower TNF-α (median = 1.76, N = 51) and higher IL33 (median = 1.070, N = 51). The difference was statistically significant for TNF-α and IL33, with p = 0.0002 and p < 0.0001, respectively. A multivariate Cox-regression analysis revealed that GMM clustering and T/N stage were independent factors for prognosis, suggesting that the prognosis might be dependent on these cytokines. Conclusions: Our data suggest that expression patterns of IL33 and TNF-α in plasma might serve as a convenient marker to predict the prognosis of ESCC in the future.

A novel encapsulation approach to enhance the delivery and antitumor activity of docetaxel in breast cancer therapy

Docetaxel (DTX) is one of the most potent anticancer drugs but its extensive side effects necessitate innovative formulations. In this study, we aimed to investigate the expression pattern of apoptotic proteins, cell cycle arrest, and apoptosis induction after treatment with encapsulated DTX in alginate-chitosan nanoparticles in both breast cancer cells (MCF-7) and peripheral blood mononuclear cells (PBMCs). The characterization of the nanoparticles revealed a spherical shape with a size <50 nm, a hydrodynamic diameter of 200 nm, a Polydispersity Index of 0.5, and an encapsulation efficiency of 98.75 %. The free drug was released completely within 11 h while encapsulated DTX was released only 34 % in 96 h. The encapsulated drug indicated higher cytotoxicity on MCF-7 cells and the half inhibitory concentration (IC50) value was 2 µg/ml after 72 h. Quantitative real-time PCR demonstrated a significant increase in cell death as the expression of apoptosis regulatory protein (Bcl-2) was downregulated with no impact on Bax in the MCF-7 cells. A notable decrease in the expression pattern of pro-inflammatory cytokine (IL-1β) in PBMCs indicated less inflammation induction. Flow cytometry analysis revealed that the newly formulated drug induced less opoptosis in PBMCs than the free DTX. Cell cycle arrest in the sub-G1 phase was observed for the free drug while the encapsulated drug exhibited no significant changes. Our results suggest the high toxicity of the formulated drug in contrast to the free DTX on the MCF-7 cell line, minimal blood cell side effects, and no inflammation positioning it as a promising alternative to free docetaxel.

From Womb to World: Immunological Pathways Influencing Stunting and 2030 Sustainability

Stunting affects 150 million children globally, indicating an existing inequality in nutrition and health; despite focusing on nutrient deficiencies, the immunological continuum from prenatal to postnatal needs to be better described. This study synthesized the literature analyzing the role of immune functions in stunting from prenatal to early childhood and its implications for the 2030 Sustainable Development Goals. The database search was performed on PubMed, Scopus, Web of Science, and Google Scholar, in accordance with the PRISMA 2020 guidelines. The objective of the current narrative review was to determine immunological factors and stunting using peer-reviewed English research published from 2000 to 2024. The review also reveals that stunting is a complex process involving immune and nutritional factors, but the significant determinants of growth are prenatal immune priming and early-life infection. Some findings are related to the knowledge of the interrelation between nutritional deficiencies and immune system dysfunctions, cytokine patterns, and gut microbiota dynamics. Thus, this study provides a general definition of stunting and recommends additional research in immunology and the need for public health measures involving nutritional and immunological interventions. The assessment provides recommendations for preventing stunting and ensures that the actions are aligned with the 2030 Sustainable Development Goals for improving children’s well-being.