Central European Journal of Immunology最新文献

Introduction: Neutrophil autophagy and neutrophil extracellular trap (NET) formation are closely related to asthma pathogenesis. Src homology domain 2-containing protein tyrosine phosphatase 2 (SHP2) is an important regulatory factor in airway remodeling in asthma. This study aimed to explore the molecular mechanisms of SHP2 in neutrophils.

Material and methods: Peripheral blood samples were collected from healthy individuals and asthma patients. A dimethylsulfoxide-induced HL-60-driven neutrophil-like cell model was established. Neutrophil-like cells were treated with rapamycin to activate autophagy. Neutrophil-like cells or neutrophils were transfected with oe-SHP2, si-SHP2, oe-ERK5 or their negative controls.

Results: There was an abnormal increase of neutrophils in the peripheral blood of asthma patients. Neutrophil autophagy gradually decreased with the severity of asthma while the NET formation increased. Pearson's correlation analysis revealed that SHP2 was negatively correlated with BECN1 and LC3 and positively correlated with p62 and MPO. Moreover, SHP2 inhibited autophagy in neutrophil-like cells. Overexpression of ERK5 partially counteracted the inhibitory effect of interfering with SHP2 expression on NET formation in neutrophil-like cells. After interfering with SHP2 expression in neutrophils, the expression of BECN1 and LC3 were significantly increased, while dsDNA levels, MPO activity, and the expression levels of p62, cit-H3, MPO, ELANE, PADI4 and ERK5 were decreased.

Conclusions: Down-regulation of SHP2/ERK5 promoted neutrophil autophagy and inhibited NET formation. SHP2 could be a new indicator of asthma.

Esophageal cancer is considered one of the most significant challenges to public health worldwide. While various therapeutic options exist for esophageal cancer, including chemotherapy, radiotherapy, and surgery, several adverse effects of these medications have been reported. Therefore, a new generation of therapeutic lines should be applied to minimize complications. In this regard, immunotherapy is a novel approach that aims to kill tumor cells directly by targeting them. Specifically, monoclonal antibodies can target specific markers of esophageal cancer tumor cells, keeping other normal cells safe. Multiple monoclonal antibodies optimized for esophageal cancer, such as pembrolizumab, ramucirumab, trastuzumab, nivolumab, and ipilimumab, are available. On the other hand, esophageal cancer tumor cells express a specific inhibitory ligand and its receptor called programmed cell death, which can suppress T cell immune responses. This receptor provides an inhibitory signal, causing the highest expression of the PD-L1 ligand on tumor cells. The outcomes of this interaction lead to the suppression of the activation and function of T lymphocytes. Therefore, immunotherapy for esophageal cancer targeting the PD-1/PD-L1 pathway has shown a remarkable correlation with cancer care. This study presents a comprehensive review of the latest findings related to immunotherapy in esophageal cancer.

Introduction: Diabetic encephalopathy (DE) is a central nervous complication of type 2 diabetes (T2D). Swertiamarin (SW) is a secoiridoid glycoside reported to have anti-hyperglycemic properties in T2D animal models. Nonetheless, the precise function of SW in T2D-induced DE remains unclarified.

Material and methods: A T2D rat model was established by high-fat diet feeding plus streptozotocin injection, followed by SW administration. Fasting blood glucose and insulin levels were determined. The Morris water maze test was implemented to evaluate rat cognitive function. Hematoxylin-eosin staining was performed for hippocampal morphological observation. Hippocampal p-tau level was detected using immunofluorescence staining. ELISA was utilized to determine inflammatory cytokine production. Western blotting was performed to estimate PI3K/Akt/GSK3 β signaling-related protein levels.

Results: Swertiamarin treatment improved spatial learning and memory and reduced fasting blood glucose as well as insulin levels in T2D rats. SW ameliorated hippocampal morphological changes, reduced tau phosphorylation, and attenuated the inflammatory response in T2D rat hippocampal tissues. SW restored PI3K/Akt/GSK3 β signaling in diabetic rat hippocampus.

Conclusions: Swertiamarin exerts anti-diabetic and anti-inflammatory effects possibly by activating PI3K/Akt/GSK3 β signaling, thereby ameliorating cognitive impairment in T2D rats.

Introduction: Alternative in vitro tests that can be used instead of animal experiments are those that can most closely evaluate the biological activity of the drug of interest. For testing the potency of antivenom, these are the methods used to assess cytotoxicity. The aim of this study was to evaluate the most commonly used cytotoxicity methods for determining the protective potency of the antivenom Viekvin, which neutralizes Vipera ammodytes venom.

Material and methods: The selected methods are based on different biological mechanisms: MTT assay, based on the activity of cell oxidoreductase enzymes; crystal violet staining, based on the degree of cell adhesion; trypan blue staining, based on cell membrane permeability, and propidium iodide staining, based on measurement of nucleic acids of dead cells. The pro-apoptotic effect of the venom was also determined with annexin V staining.

Results: The IC₅₀ value of V. ammodytes venom obtained by these methods was very similar, while the EC₅₀ values differed significantly.

Conclusions: We concluded that the choice of the method used to measure the anticytotoxic anti-venom potency depends on the immunogenicity of the venom components that cause cell death; for each venom/antivenom pair, it is necessary to select the appropriate assay separately, and at present, none of the standard cytotoxic methods can be universally applied to determine antivenom potency.

Introduction: The low-grade inflammation occurring in obese individuals leads to many diseases, including cardiovascular disease (CVD). Dietary patterns, food groups or nutrients in a well-balanced diet may reduce the level of pro-inflammatory markers and the risk of obesity-related morbidities. Our study aims to describe three cytokines in obese patients in relation to dietary habits, lifestyle and body composition.

Material and methods: Serum samples were collected from 84 obese adult volunteer subjects [body mass index (BMI) ≥ 30 kg/m²] to analyze the concentrations of interleukin 6 (IL-6), tumor necrosis factor α (TNF-α) and interferon γ (IFN-γ). The subjects were tested by bioelectrical impedance analysis (BIA) and completed a three-day food diary and original questionnaire with the FFQ-6 food consumption frequency questionnaire.

Results and conclusions: Higher serum levels of IL-6 and IFN-γ were found in patients with atherosclerosis, but the group was too small for a reliable correlation. Subcutaneous but not visceral adipose tissue correlated positively with IL-6 levels. Dietary factors such as amount of sugars, including galactose and sucrose, in the diet and the frequency of consumption of sweet flavored dairy products correlated positively with the levels of IL-6 and TNF-α, while the frequency of alcohol consumption negatively correlated with the level of IL-6. The greater the frequency of sports, the higher was the level of IL-6. In obese individuals, the level of pro-inflammatory cytokines could predispose to atherosclerosis and is associated with dietary factors and lifestyle.

Introduction: Inflammatory bowel disease (IBD) is an inflammatory pathological condition for which effective drugs are currently lacking. The objective of this study was to reveal the regulatory mechanisms of growth arrest specific transcript 5 (GAS5) in IBD.

Material and methods: To mimic IBD in vitro, human fetal colon (FHC) cells were exposed to lipopolysaccharide (LPS) to induce inflammation. The MTT method and flow cytometry were utilized to detect cell viability and apoptosis, respectively. Enzyme-linked immunosorbent assay was performed to determine cytokine concentrations. A luciferase reporter kit was utilized to confirm the binding between GAS5 and the microRNA miR-23a-3p, and between vascular epicardial substance (BVES) and miR-23a-3p.

Results: GAS5 and BVES were lowly expressed in the colonic mucosal tissues obtained from patients with IBD, while miR-23a-3p was abundantly expressed. Both GAS5 upregulation and miR-23a-3p inhibition promoted proliferation, impeded apoptosis and abolished inflammatory cytokine release in FHC cells. The expression levels of miR-23a-3p and GAS5 and those of BVES and miR-23a-3p in the colonic mucosa of IBD patients were negatively correlated. GAS5 decreased the level of miR-23a-3p via direct targeting. BVES was targeted and suppressed by miR-23a-3p. Lastly, GAS5 promoted FHC cell proliferation, impeded apoptosis and inhibited cytokine release by upregulating BVES.

Conclusions: GAS5 promoted cell viability, impeded apoptosis, and inhibited inflammation in colonic mucosal cells exposed to LPS by targeting miR-23a-3p and then promoting BVES expression. These findings imply that GAS5 could be further explored as a target for IBD.

Introduction: In adult-onset asthma, two major endotypes have been proposed: T2 with eosinophilia and non-T2 characterised by neutrophils and interleukin (IL)-17. The objective of the study was to examine the endotype marker profile in patients with severe asthma who were hospitalized for exacerbations, with a focus on differentiating between viral and non-viral triggers.

Material and methods: Forty-nine patients with asthma, admitted for exacerbations, and 51 healthy controls (HCs) were recruited. We further categorized the exacerbated asthma patients into two groups: non-viral infected (n = 38) and viral infected (n = 11) groups. Blood was drawn and a nasopharyngeal swab taken at the time of admission and eosinophil numbers, eosinophil cationic protein (ECP), immuno- globulin E (IgE), tryptase and viral infection were determined. Additionally, levels of IL-17, IL-33 and IL-31 were assessed.

Results: The majority of patients had adult onset asthma (age of diagnosis, 42.8 ±16.1) with a duration of 7.7 ±10.8 years, 24.5% being atopic. Patients had higher levels of eosinophils, ECP and IgE than healthy controls (eosinophils, p = 0.003; ECP and IgE, p = 0.0001). Immunohistochemistry confirmed eosinophils as a source of ECP. Tryptase (p = 0.0001), IL-17 (p = 0.0005), IL-31 (p = 0.0001) and IL-33 (p = 0.0002) were also higher in patients than controls. ECP correlated with tryptase (r = 0.08, p = 0.62). IL-17 showed the best correlation with other mediators, including ECP (r = 0.35, p = 0.24), tryptase (r = 0.69, p = 0.0001), IgE (r = 0.50, p = 0.0001), IL-33 (r = 0.95, p = 0.0001) and IL-31 (r = 0.89, p = 0.0001). IgE, IL-17, and IL-31 had a high AUC when differentiating those with severe and non-severe asthma. The group with exacerbated viral infection showed elevated levels of serum IL-17 and IL-31 compared to the non-infected group.

Conclusions: Patients with asthmatic exacerbations were found to have higher levels of both T2 and non-T2 inflammatory markers than healthy controls. In the study, levels of IgE, IL-17, and IL-31 differentiated between patients with severe and non-severe asthma. The last two cytokines were also able to distinguish between exacerbated asthma caused by viral infection and exacerbated asthma caused by non-viral infection.

Introduction: STK11 mutation is common in lung adenocarcinoma (LUAD), but the molecular mechanism of STK11 regulation in LUAD remains uncharacterized. This study intended to explore the effect of STK11 mutation on activity and proliferation of CD4⁺ T cells in LUAD.

Material and methods: qRT-PCR experiments verified the STK11 level in different cell models. Cell Counting Kit-8 (CCK-8) and colony formation experiments evaluated proliferation ability. CCK-8 detected activity of CD4⁺ T cells. Immunohistochemistry detected levels of related genes. Immunofluorescence assayed levels of CD4⁺ T cell infiltration.

Results: STK11 mutation could accelerate proliferation of LUAD cells and impact activity of CD4⁺ T cells. Further research found that STK11 mutation affected tumor proliferation by impacting CD4⁺ T cell activity in LUAD.

Conclusions: This study revealed the regulatory mechanism of STK11 mutation affecting tumor proliferation by impacting CD4⁺ T cell activity in LUAD. It suggested that STK11 may be a possible biological target for LUAD patients, and inhibiting STK11 mutation or cutting off its regulatory pathway for immune function may be an effective strategy for STK11-mutated tumor patients.

Introduction: The study was designed to determine whether and how sevoflurane (Sev) regulates tumor associated macrophage (TAM) polarization and cervical cancer (CC) cell progression.

Material and methods: The M2 polarized THP-1 was treated with 3%Sev. The culture supernatant of M2 polarized THP-1 was co-cultured with the CC cell line Hela. The NF-κB activity was determined by luciferase reporter assay. The key genes dis-regulated by 3%Sev were determined by RNA sequencing (RNA-seq) followed by real-time reverse transcription PCR (qRT-PCR) assay. Luciferase reporter assay was used to analyze the function of 3%Sev based on N6-methyladenosine (m6A) site activity on MAP3K7 3 ' untranslated regions (3 ' UTRs). RNA immunoprecipitation (IP) using an anti-m6A antibody (anti-m6A RNA-IP) was performed to determine the m6A levels at MAP3K7 3 ' UTR.

Results: 3%Sev treatment significantly up-regulated the M2 polarization markers and down-regulated the NF-κB activity of THP-1. Meanwhile, 3%Sev treated macrophages could enhance the migratory and invasive potential of CC cells. Further, 3%Sev significantly regulated the NF-κB pathway, including MAP3K7 inhibition. MAP3K7 overexpression reversed the 3%Sev-regulated NF-κB activity and M2 polarization. 3%Sev treatment increased m6A levels in the 3 ' UTR of MAP3K7. Mutational analysis of potential m6A sites within MAP3K7 3 ' UTR revealed that these sites were required for 3%Sev regulation. In conclusion, the m6A pattern of MAP3K7 mediates the effects of 3%Sev on macrophage M2 polarization and cervical cancer progression.

Conclusions: 3%Sev enhanced TAMs M2 polarization through regulating the m6A pattern of MAP3K7, and therefore enhanced the stimulatory effect of M2 TAMs on the migration and invasion of CC cells.