Central European Journal of Immunology最新文献

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: 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.

Introduction: The main pathological feature of osteoarthritis (OA) is chondrocyte injury. LncRNA mitochondrial RNA processing endoribonuclease (RMRP) has been shown to be a chondrogenic differentiation factor. This study aimed to explore the role of RMRP in chondrocyte injury.

Material and methods: Cell counting kit-8 (CCK-8) and TUNEL assays were used to determine lipopolysaccharide (LPS)-induced chondrocyte viability and apoptosis, respectively. The interaction between RMRP and FOXC1 was analyzed by RIP and RNA pull-down. Dual luciferase reporter and ChIP were employed to analyze the interaction between FOXC1 and RBP4. The levels of RMRP, FOXC1, RBP4, apoptosis-related and extracellular matrix (ECM)-related genes were detected by RT-qPCR and western blot. ELISA assay was used for detection of inflammatory cytokines in LPS-induced chondrocytes.

Results: The levels of RMRP, FOXC1 and RBP4 were significantly upregulated in OA cartilage tissues and LPS-induced chondrocytes. Knockdown of RMRP inhibited chondrocyte apoptosis and inflammation under LPS. RMRP interacted with FOXC1 and promoted RBP4 expression. FOXC1 could upregulate RBP4 and promote LPS-induced chondrocyte apoptosis and inflammation. Similarly, RMRP combined with FOXC1 and aggravated apoptosis and inflammation in LPS-treated chondrocytes.

Conclusions: RMRP promoted upregulation of RBP4 and activation of the JNK signaling pathway by binding to FOXC1, thereby accelerating LPS-induced apoptosis and inflammation in chondrocytes.

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: Chimeric antigen receptor (CAR)-NK cells are considered safer than CAR-T cells due to their short lifetime and production of lower toxicity cytokines. By virtue of unlimited proliferative ability in vitro, NK-92 cells could be utilized as the source for CAR-engineered NK cells. CD22 is highly expressed in B cell lymphoma. The goal of our study was to determine whether CD22 could become an alternative target for CAR-NK-92 therapy against B cell lymphoma.

Material and methods: We first generated m971-BBZ NK-92 that expressed a CAR for binding CD22 in vitro. The expression of CAR was assessed by flow cytometric analysis as well as immunoblotting. The cytotoxicity of the m971-BBZ NK-92 cells towards target lymphoma cells was determined by the luciferase-based cytolysis assay. The production of cytokines in CAR NK-92 cells in response to target cells was evaluated by ELISA assay. Lastly, the cytolytic effect was evaluated by the cytolysis assay mentioned above following irradiation. The level of inhibitory receptor of CAR-expressing cells was assessed by flow cytometry.

Results: CD22-specific CAR was expressed on m971-BBZ NK-92 cells successfully. m971-BBZ NK-92 cells efficiently lysed CD22-expressing lymphoma cells and produced large amounts of cytokines after coculture with target cells. Meanwhile, irradiation did not apparently influence the cytotoxicity of m971-BBZ NK-92 cells. Inhibitory receptor detection exhibited a lower level of PD-1 in m971-BBZ NK-92 cells than FMC-63 BBZ T cells after repeated antigen stimulation.

Conclusions: Our data show that adoptive transfer of m971-BBZ NK-92 could serve as a promising strategy for immunotherapy of B cell lymphoma.