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To study the relationship between peripheral blood Toll like receptor 4 (TLR4), interleukin-1β (IL-1β) and neutrophil-to-lymphocyte ratio (NLR) and the progression and prognosis of acute pancreatitis (AP). Methods A total of 250 patients with AP were divided into the mild group (121 cases), the moderately severe group (89 cases) and the severe group (40 cases) according to the disease severity. Patients were divided into the poor prognosis group (33 cases) and the good prognosis group (217 cases) based on the prognosis after 5 days of treatment. Peripheral blood levels of TLR4, IL1β and NLR were compared at different time points, different conditions and different prognosis in patients. The correlation of peripheral blood levels of TLR4, IL-1β and NLR, Ranson score and amylase were analyzed by Pearson correlation analysis. The value of each indicator in the diagnosis of severe AP was analyzed using ROC curve, and multivariate Logistic regression analysis was conducted to identify the prognostic risk factors. Results TLR4, IL-1β and NLR levels in AP patients were higher 24 h after admission than 48 h and 72 h after admission (P＜0.05). Peripheral blood TLR4 and IL-1β levels in AP patients at 24 h after admission increased with the condition became worse. NLR level was higher in the severe group than that in the moderately group or the mild group (P＜0.05). Peripheral blood TLR4, IL-1β and NLR levels in AP patients 24 h after admission were positively correlated with Ranson score at 48 h after admission and amylase level at 24 h after admission (P＜0.05). The area under ROC curve, sensitivity and specificity of the combination of TLR4, IL-1β and NLR at 24 h after admission for predicting severe AP were 0.895, 84.53% and 81.69%. Peripheral blood TLR4, IL-1β and NLR levels 24 h after admission were higher in the poor prognosis group than those in the good prognosis group (P＜0.05). Multivariate Logistic regression analysis showed that elevated TLR4, IL-1β and NLR at 24 h after admission were independent risk factors for poor prognosis of patients with AP (P＜0.05). Conclusion Peripheral blood TLR4, IL-1β and NLR in patients with AP are closely related to disease progression and prognosis. Therefore, they can be taken as important monitoring indicators. [ABSTRACT FROM AUTHOR]

BACKGROUND: M2 macrophages have the function of reducing inflammatory factors and promoting tissue healing. Therefore, how to regulate M2 polarization of macrophages has been a hot research topic in recent years, and some miRNAs have been found to have this function. OBJECTIVE: To investigate the effects of miR-378a on the polarization of the Raw264.7 macrophage cell line. METHODS: The M1 polarization of macrophages was induced by lipopolysaccharide and interferon-γ. Interleukin-4 induced M2 polarization and the expression of endogenous miR-378a in each cell type was detected using qRT-PCR to verify whether miR-378a was involved in the polarization of macrophages. By transfection with lentivirus as the vector of overexpression of miR-378a, the stable expression of miR-378a cell lines was screened. Macrophage M1 polarization was induced synergically by lipopolysaccharide and interferon-γ. Macrophage M2 polarization was induced by interleukin-4. The levels of M1/M2 polarizationrelated cytokines in the supernatant of the macrophage culture medium were determined by enzyme-linked immunosorbent assay. qRT-PCR was used to detect the polarization characteristics of M1/M2-type macrophages and the mRNA expression levels of related cytokines. RESULTS AND CONCLUSION: (1) The expression level of endogenous miR-378a in Raw264.7 cells of each group increased after macrophage polarization. (2) Compared with the non-transfected group, the expressions of proinflammatory cytokine-induced nitric oxide synthase, tumor necrosis factor-α, interleukin-6 and interleukin-1β in macrophage M1 induced polarization were significantly decreased in the miR-378a transfection group (P < 0.05); the levels of inducible nitric oxide synthase, tumor necrosis factor-α and interleukin-6 in cell supernatant were also significantly decreased (P < 0.05). (3) Compared with the nontransfected group, the expressions of CD206, interleukin-10 and arginase-I in macrophage M2 induced polarization were significantly increased (P < 0.05); the levels of CD206 and interleukin-10 in cell supernatant were also significantly increased (P < 0.05) in the miR-378a transfection group. (4) It is indicated that overexpression of miR-378a promotes the M2 polarization of macrophages and inhibits the M1 polarization of macrophages. [ABSTRACT FROM AUTHOR]

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

Interleukin-4 (IL4) is a Th2 cytokine that can signal through two different receptors, one of which—the type II receptor—is overexpressed by various cancer cells. Previously, we have shown that type II IL4 receptor signaling increases proliferation and metastasis in mouse models of breast cancer, as well as increasing glucose and glutamine metabolism. Here, we expand on those findings to determine mechanistically how IL4 signaling links glucose metabolism and histone acetylation to drive proliferation in the context of triple-negative breast cancer (TNBC). We used a combination of cellular, biochemical, and genomics approaches to interrogate TNBC cell lines, which represent a cancer type where high expression of the type II IL4 receptor is linked to reduced survival. Our results indicate that type II IL4 receptor activation leads to increased glucose uptake, Akt and ACLY activation, and histone acetylation in TNBC cell lines. Inhibition of glucose uptake through the deletion of Glut1 ablates IL4-induced proliferation. Additionally, pharmacological inhibition of histone acetyltransferase P300 attenuates IL4-mediated gene expression and proliferation in vitro. Our work elucidates a role for type II IL4 receptor signaling in promoting TNBC progression, and highlights type II IL4 signaling, as well as histone acetylation, as possible targets for therapy. [ABSTRACT FROM AUTHOR]

Background: The preservation of retinal ganglion cells (RGCs) and the facilitation of axon regeneration are crucial considerations in the management of various vision-threatening disorders. Therefore, we investigate the efficacy of interleukin-4 (IL-4), a potential therapeutic agent, in promoting neuroprotection and axon regeneration of retinal ganglion cells (RGCs) as identified through whole transcriptome sequencing in an in vitro axon growth model. Methods: A low concentration of staurosporine (STS) was employed to induce in vitro axon growth. Whole transcriptome sequencing was utilized to identify key target factors involved in the molecular mechanism underlying axon growth. The efficacy of recombinant IL-4 protein on promoting RGC axon growth was validated through in vitro experiments. The protective effect of recombinant IL-4 protein on somas of RGCs was assessed using RBPMS-specific immunofluorescent staining in mouse models with optic nerve crush (ONC) and N-methyl-D-aspartic acid (NMDA) injury. The protective effect on RGC axons was evaluated by anterograde labeling of cholera toxin subunit B (CTB), while the promotion of RGC axon regeneration was assessed through both anterograde labeling of CTB and immunofluorescent staining for growth associated protein-43 (GAP43). Results: Whole-transcriptome sequencing of staurosporine-treated 661 W cells revealed a significant upregulation in intracellular IL-4 transcription levels during the process of axon regeneration. In vitro experiments demonstrated that recombinant IL-4 protein effectively stimulated axon outgrowth. Subsequent immunostaining with RBPMS revealed a significantly higher survival rate of RGCs in the rIL-4 group compared to the vehicle group in both NMDA and ONC injury models. Axonal tracing with CTB confirmed that recombinant IL-4 protein preserved long-distance projection of RGC axons, and there was a notably higher number of surviving axons in the rIL-4 group compared to the vehicle group following NMDA-induced injury. Moreover, intravitreal delivery of recombinant IL-4 protein substantially facilitated RGC axon regeneration after ONC injury. Conclusion: The recombinant IL-4 protein exhibits the potential to enhance the survival rate of RGCs, protect RGC axons against NMDA-induced injury, and facilitate axon regeneration following ONC. This study provides an experimental foundation for further investigation and development of therapeutic agents aimed at protecting the optic nerve and promoting axon regeneration. [ABSTRACT FROM AUTHOR]

Interleukin (IL)− 33, a nuclear factor and pleiotropic cytokine of the IL-1 family, is gaining attention owing to its important role in chronic inflammatory and autoimmune diseases. This review extends our knowledge of the effects exerted by IL-33 on target cells by binding to its specific receptor serum stimulation-2 (ST2). Depending on the tissue context, IL-33 performs multiple functions encompassing host defence, immune response, initiation and amplification of inflammation, tissue repair, and homeostasis. The levels and activity of IL-33 in the body are controlled by complex IL-33-targeting regulatory pathways. The unique temporal and spatial expression patterns of IL-33 are associated with host homeostasis and the development of immune and inflammatory disorders. Therefore, understanding the origin, function, and processes of IL-33 under various conditions is crucial. This review summarises the regulatory mechanisms underlying the IL-33/ST2 signalling axis and its potential role and clinical significance in immune and inflammatory diseases, and discusses the current complex and conflicting findings related to IL-33 in host responses. [Display omitted] • IL-33 is a dual identity alarmin with abundant roles and wide tissue distribution. • Host exists a complex and sophisticated regulatory mechanism targeting the IL-33-ST2 signalling axis. • Immunomodulation, tissue homeostasis, and metabolic regulation are the main roles of the IL-33-ST2 signalling axis. • IL-33 involves in many diseases pathophysiological process and has clinical implications. [ABSTRACT FROM AUTHOR]

BACKGROUND: Interleukin-4 can promote the osteogenic effect of bone substitute materials, but its molecular mechanism is not yet clear. Further elucidating the mechanism of interleukin-4 promoting osteogenic effect can help find safe, economical, and effective methods for the regeneration treatment of alveolar bone defects in patients. OBJECTIVE: To explore the effect of interleukin-4 intervention on polarization transformation of macrophages and osteogenic differentiation of bone marrow mesenchymal stem cells and its possible mechanism. METHODS: RAW264.7 cells in the M1 group were induced with interferon gamma + lipopolysaccharide for 24 hours. RAW264.7 cells in the interleukin-4+M1 group were induced with interferon gamma + lipopolysaccharide for 24 hours and then interleukin-4 was added for 24 hours. RAW264.7 cells in the interleukin4+AG+M1 group were induced with interferon gamma + lipopolysaccharide for 24 hours, and then interleukin-4 and AG-490, a JAK/STAT pathway inhibitor, were added for 24 hours. After intervention, immunofluorescence staining was used to analyze the expression of inducible nitric oxide synthase and CD206, the phenotypic marker protein of macrophages. ELISA kit was used to detect the expression of interleukin-10 and tumor necrosis factor-α in the supernatant of cell culture. The gene expressions of nodular receptor protein-3 (NLRP3), interleukin-1β, and caspase-1 were detected by RT-qPCR. The expression levels of tyrosine protein kinase 1 (JAK1)/ phosphorylated tyrosine protein kinase 1 (p-JAK1), signal transduction and transcription activator 6 (STAT6)/phosphorylated signal transduction and transcription activator 6 (p-STAT6), NLRP3, pro-interleukin-1β and pro-caspase-1 were detected by western blot assay. Then, RAW264.7 cells in the above four groups were indirectly co-cultured with bone marrow mesenchymal stem cells by transwell for 24 hours, followed by alkaline phosphatase staining and alizarin red staining. The mRNA expressions of alkaline phosphatase, collagen type I, and osteocalcin were detected by RT-qPCR. RESULTS AND CONCLUSION: (1) Immunofluorescence and ELISA results showed that interleukin-4 intervention could promote the expression of CD206 and interleukin-10 in M2 macrophages, and inhibit the secretion of inducible nitric oxide synthase and tumor necrosis factor-α. (2) RT-qPCR results showed that interleukin-4 could suppress the expression of NLRP3, interleukin-1β, and caspase-1 mRNAs. (3) Western blot assay showed that interleukin-4 could promote the expression of JAK1/p-JAK1, STAT6/p-STAT6 and NLRP3 proteins. (4) The alkaline phosphatase staining and alizarin red staining of bone marrow mesenchymal stem cells co-cultured with the interleukin-4+M1 group were significantly enhanced, and the mRNA expressions of alkaline phosphatase, collagen type I, and osteocalcin were significantly increased. It is concluded that interleukin-4 may inhibit the activation of NLRP3 by up-regulating JAK1/STAT6 pathway, thus promoting the transformation of macrophages from M1 polarization to M2 polarization, and finally enhancing the osteogenic differentiation ability of bone marrow mesenchymal stem cells. [ABSTRACT FROM AUTHOR]

Microglia/macrophages are major contributors to neuroinflammation in the central nervous system (CNS) injury and exhibit either pro- or anti-inflammatory phenotypes in response to specific microenvironmental signals. Our latest in vivo and in vitro studies demonstrated that curcumin-treated olfactory ensheathing cells (aOECs) can effectively enhance neural survival and axonal outgrowth, and transplantation of aOECs improves the neurological outcome after spinal cord injury (SCI). The therapeutic effect is largely attributed to aOEC anti-inflammatory activity through the modulation of microglial polarization from the M1 to M2 phenotype. However, very little is known about what viable molecules from aOECs are actively responsible for the switch of M1 to M2 microglial phenotypes and the underlying mechanisms of microglial polarization. Herein, we show that Interleukin-4 (IL-4) plays a leading role in triggering the M1 to M2 microglial phenotype, appreciably decreasing the levels of M1 markers IL‑1β, IL‑6, tumour necrosis factor-alpha (TNF-α) and inducible nitric oxide synthase (iNOS) and elevating the levels of M2 markers Arg-1, TGF-β, IL-10, and CD206. Strikingly, blockade of IL-4 signaling by siRNA and a neutralizing antibody in aOEC medium reverses the transition of M1 to M2, and the activated microglia stimulated with the aOEC medium lacking IL-4 significantly decreases neuronal survival and neurite outgrowth. In addition, transplantation of aOECs improved the neurological function deficits after SCI in rats. More importantly, the crosstalk between JAK1/STAT1/3/6-targeted downstream signals and NF-κB/SOCS1/3 signaling predominantly orchestrates IL-4-modulated microglial polarization event. These results provide new insights into the molecular mechanisms of aOECs driving the M1-to-M2 shift of microglia and shed light on new therapies for SCI through the modulation of microglial polarization. [ABSTRACT FROM AUTHOR]

Atopic dermatitis (AD) is a persistent inflammatory skin condition that impacts individuals of various age groups, including both children and adults. Its pathophysiology involves allergens penetrating a disrupted epidermal barrier, triggering the dermal cells to produce pro-inflammatory cytokines and eliciting a T-cell-mediated immune response. Notably, interleukins (ILs), particularly interleukin 4 (IL-4) and interleukin 13 (IL-13), play a key role in AD pathogenesis. Therapies directed at inflammatory mechanisms, including Dupilumab, have demonstrated notable effectiveness in enhancing skin lesions, alleviating subjective symptoms, and improving the overall quality of life for individuals with AD. Despite therapeutic advances, assessing AD severity remains challenging. The commonly used tools, such as the SCORAD and DLQI scores, rely on subjective patient responses. Paraclinically, the search for universal biomarkers continues, with efforts to identify reliable indicators reflecting disease severity and treatment response. Various biomarkers, including Th2-related chemokines and cytokines, have been explored, but none have gained universal recognition for routine clinical use. This study aims to investigate the dynamics of the plasma levels of IL-4 and IL-13 during Dupilumab treatment and establish correlations between these ILs and disease severity, as measured using the SCORAD and DLQI scores. The ultimate endpoint is to determine whether IL-4 and IL-13 can serve as reliable biomarkers, assessing their correlation with patient-reported feelings and disease activity and potentially influencing their inclusion or exclusion as diagnostic elements in routine clinical practice. [ABSTRACT FROM AUTHOR]

Pruritic papular eruption is a dermatosis characterized by pruritic symptoms in patients with human immunodeficiency virus/acquired immunodeficiency syndrome (HIV/AIDS). Similarly, nodular prurigo is an itchy dermatosis with lesions and histopathological features that resemble those of pruritic papular eruption. Both conditions share a common etiopathogenesis, which involves the cytokines produced by T helper 2 (Th2) cells. Nodular prurigo and pruritic papular eruption are chronic and frequently recalcitrant, thus posing challenges in treatment. The use of biological agents represents a treatment development for chronic and recalcitrant dermatoses. This study aimed to determine the difference in the mean percentage of interleukin-4 (IL-4) and interleukin-5 (IL-5) expressions between nodular prurigo and pruritic papular eruption lesions, which may establish a basis for further biological agent therapy. A cross-sectional study was conducted using paraffin block preparations of the skin lesions of patients diagnosed with nodular prurigo (n=16) and pruritic papular eruption (n=16). Each paraffin block preparation involved immunohistochemical staining using IL-4 and IL-5 monoclonal antibodies. The expressions of IL-4 and IL-5 were assessed through ImageJ for Windows, version 1.53 (National Institutes of Health and the Laboratory for Optical and Computational Instrumentation, University of Wisconsin, USA) by pathologists. The data were analyzed using an unpaired t-test with a significance level of p<0.05. The analytical results indicated that data on the average age of the two groups, disease duration, and storage sample duration followed a normal distribution (p>0.05). The mean percentage of IL-4 expression was significantly different between the nodular prurigo and pruritic papular eruption groups (p=0.000). However, the mean percentage of IL-5 expression was not significantly different between the two groups (p=0.060). In conclusion, the expression of IL-4 was higher in the nodular prurigo group in comparison to the pruritic papular eruption group. Nonetheless, the expression of IL-5 was comparably high in both the nodular prurigo and pruritic papular eruption groups. [ABSTRACT FROM AUTHOR]

Tuberculosis (TB) is a global health problem caused by Mycobacterium tuberculosis (MTB), a bacterium that can evade the host immune system and keep at in a latent state. Drug-resistant strains of MTB, such as multi-drug resistant TB (MDR-TB), pose a significant challenge toward the TB control efforts. This study aimed to investigate the role of two MTB virulence genes, pknF and fbpA, and two host cytokines, Interleukin-4 and Interleukin-6, in the pathogenesis of TB. The expression levels of pknF and fbpA genes were measured by qPCR in (12) sensitive TB and (12) MDR-TB isolates. IL-4 and IL-6 were measured by ELISA in serum samples from (24) healthy controls, (12) patients having sensitive TB, and 12 patients having MDR-TB. The results revealed a significant difference in fbpA gene expression between MDR-TB (2.59± SE 0.36) and sensitive TB (1.01± SE0.037) isolates (P=0.0003), whereas pknF gene expression did not vary significantly across the two groups (2.53±0.62 in MDR-TB and 1.72±0.40 in sensitive TB) (p=0.289). IL-4 levels were markedly elevated in patients with MDRTB (1091.967± SE 108.793 pg/ml) compared to the control group (105.3358 ± SE 5.543 pg/ml) (p<0.0001), but not significantly different from patients with sensitive TB (1054.763 ± SE 71.482 pg/ml). IL-6 levels were significantly higher in both MDR-TB and sensitive TB patients than in the control group (9.253 ± SE 0.456 pg/ml). However, MDR-TB patients showed a non-significant lower ratio of IL-6 (38.5851 ± Se 4.601 pg/ml) than sensitive TB patients (42.458 ± SE 1.809). A significant negative correlation were observed between fbpA gene expression and IL-4 levels in both MDR-TB and sensitive TB patients (r = -0.375; p < 0.0001 and r = - 0.165; p < 0.0001, respectively), and a positive correlation between fbpA gene expression and IL-6 levels in both groups (r = 0.1006; p < 0.0001 and r = 0.466; p < 0.0001, respectively). These findings suggest that pknF and fbpA genes may play a role in the virulence of MTB, especially in drug-resistant strains, and that IL-4 and IL-6 may be involved in the host immune response to MTB infection. These potential biomarkers could be used to develop targeted therapies for MDR-TB and improve TB control efforts globally. [ABSTRACT FROM AUTHOR]

Neuroinflammation, a key pathological feature following subarachnoid hemorrhage (SAH), can be therapeutically targeted by inhibiting microglia M1 polarization and promoting phenotypic transformation to M2 microglia. Interleukin-4 (IL-4) is a pleiotropic cytokine known to its regulation of physiological functions of the central nervous system (CNS) and mediate neuroinflammatory processes. However, its specific role in neuroinflammation and microglia responses following SAH remains unexplored. In this investigation, we established both in vivo and in vitro SAH models and employed a comprehensive array of assessments, including ELISA, neurofunctional profiling, immunofluorescence staining, qRT-PCR, determination of phagocytic capacity, and RNA-Seq analyses. The findings demonstrate an elevated expression of IL-4 within cerebrospinal fluid (CSF) subsequent to SAH. Furthermore, exogenous administration of IL-4 ameliorates post-SAH neurofunctional deficits, attenuates cellular apoptosis, fosters M2 microglia phenotype conversion, and mitigates neuroinflammatory responses. The RNA-Seq analysis signifies that IL-4 governs the modulation of neuroinflammation in microglia within an in vitro SAH model through intricate cascades of signaling pathways, encompassing interactions between cytokines and cytokine receptors. These discoveries not only augment comprehension of the neuropathogenesis associated with post-SAH neuroinflammation but also present novel therapeutic targets for the management thereof. [ABSTRACT FROM AUTHOR]

Introduction: Atopic dermatitis (AD) patients have an increased risk of herpes zoster (HZ). The relationship of dupilumab, tralokinumab, upadacitinib, and abrocitinib to HZ incidence in AD patients remains unclear. Aim: To elucidate whether patients with AD have an increased risk of HZ infections when given one of the aforementioned immunomodulatory therapies. Material and methods: Systematic searches were conducted in Ovid Medline and Embase. The primary outcome was incidence of HZ in patients with moderate to severe AD receiving placebo or the aforementioned treatments. A frequentist random-effects NMA was conducted with odds ratio. Results: Our search identified 16 trials comprising 10,689 patients. Upadacitinib was associated with a dose-dependent increase in the incidence of HZ compared to placebo (OR = 2.55 [1.09, 5.95] and (OR = 4.29 [1.89, 9.74], respectively) and compared to various dupilumab doses (OR = 4.48 [1.29, 15.57], 3.61 [1.28, 10.18] and 7.54 [2.21, 25.68], 6.09 [2.24, 16.52], respectively). Upadacitinib 30 mg was associated with a higher incidence of HZ when compared to upadacitinib 15 mg (OR = 1.68 [1.19, 2.38]). Abrocitinib 200 mg was associated with a higher increase in HZ compared to placebo (OR = 3.34 [1.34, 8.31]). According to SUCRA ranks, both JAK-1 inhibitors had a higher cumulative incidence of HZ compared to dupilumab. Conclusions: JAK-1 inhibitors are associated with a significantly higher incidence of HZ compared to dupilumab and placebo. Our results suggest that HZ vaccination should be highly considered for all adult patients prior to starting oral JAK-1 inhibitors. [ABSTRACT FROM AUTHOR]

Although calcineurin inhibitors are very effective as immunosuppressants in organ transplantation, complete graft acceptance remains as a challenge. Transfer of genes with immunosuppressant functions could contribute to improving the clinical evolution of transplantation. In this sense, hydrodynamic injection has proven very efficacious for liver gene transfer. In the present work, the hIL-10 gene was hydrofected 'ex vivo' to pig livers during the bench surgery stage, to circumvent the cardiovascular limitations of the procedure, in a model of porcine orthotopic transplantation with a 10-day follow-up. We used IL-10 because human and porcine proteins can be differentially quantified and for its immunomodulatory pleiotropic functions. Safety (biochemical parameters and histology), expression efficacy (RNA transcription and blood protein expression), and acute inflammatory response (cytokines panel) of the procedure were evaluated. The procedure proved safe as no change in biochemical parameters was observed in treated animals, and human IL-10 was efficaciously expressed, with stationary plasma protein levels over 20 pg/mL during the follow-up. Most studied cytokines showed increments (interferon-α, IFN-α; interleukin-1β, IL-1β; tumor necrosis factor α, TNFα; interleukin-6, IL-6; interleukin-8, IL-8; interleukin-4, IL-4; and transforming growth factor-β, TGF-β) in treated animals, without deleterious effects on tissue. Collectively, the results support the potential clinical interest in this gene therapy model that would require further longer-term dose–response studies to be confirmed. [ABSTRACT FROM AUTHOR]

Dendritic cells (DCs), professional antigen-presenting cells that process and deliver antigens using MHC II/I molecules, can be enhanced in numerous ways. Exosomes derived from heat‐ shocked tumor cells (HS‐TEXs) contain high amounts of heat-shock proteins (HSPs). HSPs, as chaperons, can induce DC maturation. This study aimed to investigate whether HS‐TEXs can promote DC maturation. To generate DC, bone marrow-derived cells were treated with Interleukin-4 and GM-CSF. Exosomes were isolated from heat-treated CT-26 cells. The expression level of HSP in exosomes was checked by western blot and the increase in the expression of this protein was observed. Then, HS‐TEXs were co-cultured with iDCs to determine DC maturity, and then DCs were co-cultured with lymphocytes to determine DC activity. Our results showed that DCs treated with HS‐TEXs express high levels of molecules involved in DC maturation and function including MHCII, CD40, CD83, and CD86. HS‐TEXs caused phenotypic and functional maturation of DCs. In addition, flow cytometric results reflected a higher proliferative response of lymphocytes in the iDC / Tex + HSP group. HS‐TEXs could be used as a strategy to improve DC maturation and activation. [ABSTRACT FROM AUTHOR]

[Display omitted] Macrophages are known to depict two major phenotypes: classically activated macrophages (M1), associated with high production of pro-inflammatory cytokines, and alternatively activated macrophages (M2), which present an anti-inflammatory function. A precise control over M1-M2 polarization is a promising strategy in therapeutics to modulate both tissue regeneration and tumor progression processes. However, this is not a simple task as macrophages behave differently depending on the microenvironment. In agreement with this, non-consistent data have been reported regarding macrophages response to magnetic iron oxide nanoparticles (MNPs). To investigate the impact of both tissue microenvironment and MNPs properties on the obtained macrophage responses, single-core (SC) and multi-core (MC) citrate coated MNPs, are synthesized and, afterwards, loaded with a macrophage polarization trigger, IL-4. The developed MNPs are then tested in macrophages subjected to different stimuli. We demonstrate that macrophages treated with low concentrations of MNPs behave differently depending on the polarization stage independently of the concentration of iron. Moreover, we find out that MNPs size and morphology determines the effect of the IL-4 loaded MNPs on M1 macrophages, since IL-4 loaded SC MNPs favor the polarization of M1 macrophages towards M2 phenotype, while IL-4 loaded MC MNPs further stimulate the secretion of pro-inflammatory cytokines. [ABSTRACT FROM AUTHOR]

Recently, scientists have focused on pro-inflammatory cytokines and immunological dysregulation in major depressive disorder (MDD). Some research suggests pro-inflammatory cytokines' role in MDD development, whereas anti-inflammatory studies are sparse. There is no systematic investigation of Bangladeshi MDD patients' pro- and anti-inflammatory cytokines. This study examines the blood levels of IL-12 and IL-4 in Bangladeshi patients and healthy controls (HCs) to determine the diagnostic accuracy of these cytokines to identify MDD patients from those without MDD. A total of 110 people with MDD from the department of psychiatry of a teaching hospital in Dhaka and 107 HCs from Dhaka participated in this case–control study. Depression and illness severity were gauged using DSM-5 criteria and Ham-D scores. Commercially marketed ELISA kits were used in accordance with manufacturer guidelines to measure the levels of IL-12 and IL-4 in peripheral blood, allowing a comparison of the patient and control groups. In comparison to HCs, MDD patients (5333.00 ± 307.40 pg/ml) showed noticeably higher levels of IL-12 than in HCs (2331.00 ± 207.40 pg/ml). The increased levels were positively correlated with Ham-D scores (male: r = 0.351, p < 0.050; female: r = 0.389, p < 0.050), suggesting a possible relationship to disease progression. Additionally, compared to HCs (272.81 ± 23.94 pg/ml), MDD patients had significantly higher peripheral blood levels of IL-4 (876.35 ± 66.73 pg/ml) (p < 0.001). Also, there was a positive correlation between IL-4 serum levels and Ham-D scores (male: r = 0.361, p < 0.050; female: r = 0.398, p < 0.050). Therefore, we observed increased levels of these serum cytokines and their association with the severity of depression. The results of this study demonstrate the possibility of IL-12 and IL-4 blood levels as distinct markers capable of differentiating between MDD patients and HCs, possibly acting as markers of MDD susceptibility. To ascertain the diagnostic effectiveness of these two cytokines, more research is necessary. [ABSTRACT FROM AUTHOR]

Fumonisins (FBs), particularly fumonisin B1 (FB1) and fumonisin B2 (FB2) produced mainly by Fusarium verticillioide and Fusarium proliferatum, are common contaminants in animal feed and pose a serious threat to both animal and human health. The use of microbial enzymes to efficiently and specifically convert fumonisins into non-toxic or low-toxic metabolites has emerged as the most promising approach. However, most of the available enzymes have only been evaluated in vitro and lack systematic evaluation in vivo. In this study, the detoxification efficacy of two carboxylesterases, FumD (FUMzyme®) and FumDSB, was evaluated comparatively in piglets. The results show that feeding piglets 4.4 mg/kg FBs-contaminated diets for 32 days did not significantly affect the average daily gain, organ indices, and immunoglobulins of the piglets. However, a significant reduction (21.2%) in anti-inflammatory cytokine interleukin-4 was observed in the FBs group, and supplementation with FUMzyme® and FumDSB significantly increased interleukin-4 by 62.1% and 28.0%, respectively. In addition, FBs-contaminated diets resulted in a 3-fold increase in the serum sphinganine/sphingosine (Sa/So) ratio, which is a specific biomarker that has been used to accurately reflect fumonisin levels. The serum Sa/So ratio was significantly reduced by 48.8% after the addition of FUMzyme®, and was insignificantly reduced by 8.2% in the FumDSB group. These results suggested that FUMzyme was more effective than FumDSB in mitigating FBs toxicity in piglets by down-regulating the Sa/So ratio. [ABSTRACT FROM AUTHOR]

Objective: Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2), the virus responsible for Coronavirus Disease 2019 (COVID-19), elicits a strong immune response similar to that seen in other viral infections. The predominant cell type in this immune response also influences the disease prognosis. This study, conducted between 2022 and May 2023, aimed to evaluate the transcription factors and cytokine expressions of Th (helper T) cell subsets at the time of diagnosis and after discharge in patients with nonsevere COVID-19. Materials and Methods: Forty-eight patients with non-severe COVID-19 were included in the study. Transcription factor and cytokine expressions of Th cell subsets were evaluated using the quantitative polymerase chain reaction (qPCR) method, and the results were compared at the time of diagnosis and after discharge. Results: It was determined that the cytokines and transcription factors of T helper 1 (Th1) cells (T-box expressed in T cells [T-bet], 2.71-fold, p<0.001; Interferon-gamma [IFN-γ], 1.42- fold, p=0.010) and T helper 17 (Th17) cells (RAR-related orphan receptor gamma [RORγt], 1.06-fold, p=0.946; Interleukin-22 [IL-22], 1.01-fold, p=0.599) decreased, whereas the expression of T helper 2 (Th2) cells (GATA binding protein 3 [GATA3], 2.56-fold, p<0.001; Interleukin-4 [IL-4], 1.34-fold, p=0.012; Interleukin-5 [IL-5], 1.02-fold, p=0.649; Interleukin-13 [IL-13], 2.06-fold, p=0.0119) and regulatory T (Treg) cells (Forkhead box P3 [FoxP3], 3.56-fold, p<0.001; Transforming growth factor-beta [TGF-β], 1.03-fold, p=0.670; Interleukin-10 [IL-10], 1.40-fold, p=0.010) increased. Conclusion: Our study in non-severe COVID-19 patients demonstrated significant changes in the transcription factor and cytokine expressions of Th cell subsets at the time of diagnosis compared to discharge. We think that even if the patients do not exhibit severe clinical and laboratory findings, Th cell immune responses may be strong, warranting careful consideration. [ABSTRACT FROM AUTHOR]

Objective To observe the proportion of peripheral blood T follicular helper (Tfh) cell subsets and serum cytokine levels in patients with acute lymphoblastic leukemia (ALL), and to analyze the relationship between the two and their relationship with the prognosis of patients. Methods Thirty newly treated ALL patients (observation group, includ‑ ing13 patients with good prognosis and 17 patients with poor prognosis according to the risk stratification criteria for ALL patients) and 20 healthy subjects (control group) were selected. Flow cytometry was used to detect the proportion of Tfh cell subgroups (Tfh, Tfh1, Tfh2, Tfh17) in the peripheral blood of the two groups. Serum cytokine levels [interferon-γ (IFN-γ), interleukin (IL）-4, IL-17] were detected by ELISA, and the correlation between the proportion of Tfh subsets in peripheral blood and serum cytokine level in the observation group was analyzed by Pearson correlation analysis. Results Compared with the control group, the proportions of Tfh and Tfh2 in peripheral blood increased, and the proportion of Tfh1 decreased in the observation group (all P<0. 05). Compared with those with poor prognosis, the proportions of Tfh and Tfh2 decreased in the observation group with good prognosis (both P<0. 05). Compared with the control group, the serum IFN-γ level decreased and serum IL-4 level increased in the observation group (both P<0. 05). Compared with those with poor prognosis, the serum IFN-γ level increased and serum IL-4 level decreased in patients with good prognosis in the observation group (both P<0. 05). The proportions of Tfh and Tfh2 in the observation group were positively correlated with the serum IL-4 level of ALL patients (r=0.860 and 0. 965, respectively; both P<0. 05), and the proportion of Tfh was negatively correlated with the serum IFN-γ level (r =–0. 289, P<0. 05). Conclusion In ALL patients, the proportions of Tfh and Tfh2 in peripheral blood increase, the proportion of Tfh1 decreases, the level of serum IFN-γ decreases, and the level of IL-4 increases, which are related to the poor prognosis of patients, and they may participate in the immune dysfunction of ALL, enhance the immunosuppressive effect, and participate in the disease progression. [ABSTRACT FROM AUTHOR]

BACKGROUND: Acupoint catgut embedding can alleviate airway inflammation in asthma, which is related to regulating the imbalance of Th1/Th2 in the p38 MAPK pathway. Whether the above process is affected by the functional state of type II alveolar epithelial cells at the same time remains to be further studied. OBJECTIVE: To observe the effect of acupoint catgut embedding on p38 MAPK pathway, interleukin-4, γ-interferon and type II alveolar epithelial cells in lung tissue of asthma rats. METHODS: Forty male Wistar rats were randomly divided into blank control group, asthma model group, dexamethasone group and acupoint catgut embedding group (n=10 per group). Asthma models of asthma were prepared by intraperitoneal injection of ovalbumin and aluminum hydroxide suspension on days 1 and 8 and aerosol inhalation of ovalbumin solution from day 15 was given to induce symptoms of airway inflammatory responses, once a day for 2 weeks, in the latter three groups. Meanwhile, the dexamethasone group began to receive intraperitoneal injection of dexamethasone on day 15, once a day for 2 weeks; and the acupoint catgut embedding group received embedding treatment at Feishu, Dingchuan, and Danzhong acupoints on day 15. At the end of aerosol inhalation, the relative content of eosinophils in the alveolar lavage fluid was counted by Richter-Kimsa staining, the ultrastructural changes of type II alveolar epithelial cells in lung tissue were observed by transmission electron microscopy, the positive expression of interleukin-4 and γ-interferon in lung tissue was detected by immunohistochemistry, and the protein and gene expression of p-p38 MAPK, interleukin-4 and γ-interferon in lung tissue were detected by western blot and PCR respectively. RESULTS AND CONCLUSION: Compared with the asthma model group, the relative content of eosinophils in the bronchoalveolar lavage fluid was significantly reduced in the dexamethasone group and acupoint catgut embedding group (P < 0.01), while compared with the dexamethasone group, the relative content of eosinophils in the bronchoalveolar lavage fluid was significantly higher in the acupoint catgut embedding group (P < 0.05). Under the transmission electron microscopy, damage to lamellar vesicles and mitochondria in type II alveolar epithelial cells was alleviated in the dexamethasone and acupoint catgut embedding groups compared with the asthma model group. Immunohistochemical detection showed that compared with the asthma model group, the expression of interleukin-4 in lung tissue was decreased (P < 0.01) and that of γ-interferon was increased (P < 0.01) in the dexamethasone and acupoint catgut embedding group; compared with the dexamethasone group, the expression of interleukin-4 in lung tissue was increased (P < 0.01) and that of γ-interferon was decreased (P < 0.01) in the acupoint catgut embedding group. Compared with the asthma model group, the protein and mRNA expressions of p-p38 MAPK and interleukin-4 in the lung tissue of the dexamethasone and acupoint catgut embedding groups were decreased (P < 0.01, P < 0.05), while the protein and mRNA expressions of γ-interferon were increased (P < 0.01). To conclude, acupoint catgut embedding may upregulate γ-Interferon and downregulate interleukin-4 by inhibiting the p38 MAPK signal pathway, thereby reducing stress damage in type II alveolar epithelial cells and alleviating the airway inflammatory response in asthma. [ABSTRACT FROM AUTHOR]

BACKGROUND: Studies have shown that a certain dose of interleukin-4 intervention can yield the appropriate ratio of M1/M2 macrophage profile to generate a microenvironment conducive to bone healing and promoting bone regeneration. OBJECTIVE: To investigate the effect of interleukin-4 on NLRP3 inflammasome activation and osteoclast differentiation during bone tissue regeneration guided by bone replacement materials. METHODS: Forty-eight 6-8-week-old male SD rats were selected to establish a 5-mm diameter cranial bone defect model on the left and implanted with BioOss bone replacement material at the same time, and the periosteum was sutured. Rat models were randomly divided into control and experimental groups (n=24) at postoperative 3 days. Interleukin-4 and PBS were injected locally into the cranial bone defect area for the experimental and control groups, once a day, for 5 consecutive days. SD rats were executed at 1 and 2 weeks after surgery and skull samples were taken. Immunohistochemical staining was performed to detect the expression of cleaved caspase-1 and interleukin-1β protein. Immunofluorescence staining was performed to observe the expression of inducible nitric oxide synthase (M1 surface marker) and cleaved caspase-1 (NLRP3 indicator). Real-time fluorescence quantitative PCR was performed to detect the expression of related inflammatory factors caspase-1, interleukin-1β and osteoclast factor histone K gene. The differentiation and number of osteoclasts were observed by anti-tartrate acid phosphatase staining. At 6 and 12 weeks after surgery, micro-CT and hematoxylin-eosin staining were performed to observe the osteogenesis of the skull. RESULTS AND CONCLUSION: Immunofluorescence staining showed that the number of inducible nitric oxide synthase and cleaved caspase-1 doublestained cells in the experimental group was significantly lower than that in the control group at 1 and 2 weeks (P < 0.05). Immunohistochemical staining showed that the expression of cleaved caspase-1 and interleukin-1β in the experimental group was significantly lower than that in the control group at 1 week (P < 0.05). RT-qPCR suggested that the expression of caspase-1 mRNA in the experimental group was lower than that in the control group at 1 week (P < 0.05). The expression intensity of interleukin-1β and osteoclast factor histone K mRNA was significantly lower in the experimental group than that in the control group at postoperative 1 and 2 weeks (P < 0.05). Anti-tartrate acid phosphatase staining showed that the number of osteoclasts was significantly lower in the experimental group at 1 and 2 weeks than that in the control group (P < 0.05). Micro-CT results showed that the bone volume fraction and bone mineral density in the bone defect area were significantly higher in the experimental group than those in the control group at 6 and 12 weeks after injection (P < 0.05). Hematoxylin-eosin staining showed that 12 weeks after injection, multiple ossification centers were formed in the center of the defect in the experimental group, with scattered mature bone cells and lacunae, and a large number of osteoblasts around the material arranged into a single layer to participate in the formation of bone matrix. The results suggest that interleukin-4 may be involved in downregulating NLRP3 inflammasome expression, inhibiting the activation of caspase-1 and thus reducing the secretion of interleukin-1β, reducing the inflammatory state of the local microenvironment, as well as inhibiting osteoclast differentiation and playing a role in promoting new bone production during bone tissue regeneration guided by bone replacement materials. [ABSTRACT FROM AUTHOR]

Introduction: Atopic dermatitis is a relapsing, chronic, inflammatory dermatosis. So far, treatment options for more severe forms of the disease have been limited. The prospect has changed with the advent of biological drug registrations. Dupilumab is a monoclonal antibody targeting the a subunit of the IL-4 receptor and is responsible for blocking the signalling of interleukin (IL) 4 (IL-4) and IL-13. Clinical trials conducted for over 10 years have confirmed the efficacy and safety of dupilumab's treatment for atopic dermatitis. Aim: Evaluating the efficacy of dupilumab treatment in patients with moderate and severe atopic dermatitis in real life. Material and methods: We retrospectively evaluated medical records of patients treated with dupilumab for atopic dermatitis at the Department of Dermatology, Venereology and Allergology in Gdansk. Results: Ten patients in total were studied. They received dupilumab with standard dosing. The mean percentage reduction in SCORAD score was 52.16% in 8 weeks. Dupilumab was generally well tolerated and did not cause serious side effects. The most common adverse event was conjunctivitis. Conclusions: Dupilumab is an effective disease-modifying drug for patients with moderate to severe atopic dermatitis. The effects of treatment in real life are consistent with those demonstrated in clinical trials. [ABSTRACT FROM AUTHOR]

Objective(s) The prognostic value of systemic cytokine profiles and inflammatory markers in colorectal cancer were explored by several studies. We want to know more about inflammatory biomarkers in colorectal adenoma and early cancer. Method The level of 38 inflammatory markers in the plasma of 112 adenoma patients, 72 Tis-T1 staging of colorectal carcinoma patients, 34 T2–T4 staging of colorectal carcinoma patients and 53 normal subjects were detected and compared. Result(s) Eight inflammatory biomarkers (Eotaxin, GCSF, IL-4, IL-5, IL-17E, MCP-1, TNF-α and VEGF-A) have higher plasma concentrations in colorectal adenoma and cancer patients compared with normal participants over 50 years old. Conclusion(s) Inflammatory markers may have the prognostic value for colorectal adenoma and early-stage carcinoma. [ABSTRACT FROM AUTHOR]

Drug-induced liver injury (DILI) remains a challenging diagnosis due to an absence of specific biomarkers. DILI due to volatile anaesthetics (VA-DILI) is characterised by trifluoroacetyl and CYP2E1 antibodies, but may not be seen for weeks after injury. Interleukin-4 (IL-4) may be involved in the production of these antibodies and may serve as a clinically useful early biomarker of VA-DILI. To prospectively compare serum IL-4 levels between patients who develop VA-DILI and controls following exposure to the volatile anaesthetic. A nested case-control study of patients exposed to VA during surgery was conducted. Thirteen DILI cases were identified from the original cohort, and 26 controls were matched according to age, sex and VA agent. Serum samples were collected before and 48-96 h after VA exposure, and analysed for IL-4 using quantitative enzyme-linked immunosorbent assay techniques. There was a statistically significant difference in serum IL-4 in post-VA samples between DILI cases and controls (control: 0.030 pg/mL, IQR: 0.030 − 0.030 pg/mL vs DILI: 0.044 pg/mL, IQR: 0.030 − 0.061 pg/mL; p = 0.039). A greater proportion of DILI cases had post-VA IL-4 levels above the assay lower limit of detection compared to controls (control: 23% vs DILI: 69%; p = 0.013). IL-4 is a potential biomarker of DILI. Clinical diagnosis and understanding of DILI disease mechanisms may be improved by further investigation of novel biomarkers, and this IL-4 signal in serum is important as proof of concept for prospective study designs. [ABSTRACT FROM AUTHOR]

BACKGROUND: Previous studies had shown that hair follicle epidermal neural crest stem cells have the great potential in facilitating peripheral nerve rehabilitation, but the role in regulating inflammation after facial nerve injury had rarely been reported. OBJECTIVE: To investigate whether epidermal neural crest stem cells can modify the expression of pro-inflammatory factor tumor necrosis factor α and anti-inflammatory factor interleukin-4 during facial nerve regeneration to promote the functional and morphological recovery of the facial nerve. METHODS: Epidermal neural crest stem cells were extracted from one 4-day-old SD rat, cultured and identified. Fifty-four adult SD rats were used to construct the bridge model of facial nerve trunk defect with the autologous venous catheter. Models were randomly and equally divided into the normal saline group, Dulbecco's modified eagle medium group and epidermal neural crest stem cell group. Tumor necrosis factor α and interleukin-4 expression levels were detected at postoperative 4 to 14 days using western blotting, immunohistochemistry, and immunofluorescence. The facial nerve function of rats was scored 12 weeks after the operation and the morphology of the facial nerve was observed by hematoxylin-eosin staining. RESULTS AND CONCLUSION: (1) Double positive expression of Nestin and p75NTR could be observed in epidermal neural crest stem cells and the cell purity was over 90%. (2) Compared with the other two groups, tumor necrosis factor α expression in the epidermal neural crest stem cell group turned weaker 7 days after surgery (P < 0.05), and interleukin-4 expression in the epidermal neural crest stem cell group was enhanced on postoperative 3, 7 and 14 days, respectively (P < 0.05). (3) The recovery score of facial nerve function in the epidermal neural crest stem cell group was significantly better than that in the other two groups (P < 0.05). (4) There were abundant nutrient vessels and regenerative axons in the graft segment. Compared with the other two groups, the arrangement of nerve fibers was more orderly and the wrapped myelin sheath was thicker in the graft and distal segment in the epidermal neural crest stem cell group. (5) The data inferred that epidermal neural crest stem cells regulate the local expression of tumor necrosis factor α and interleukin-4 at the early stage of facial nerve injury, inhibit the local inflammatory reaction, and improve facial nerve function and morphology of the regenerated nerve. [ABSTRACT FROM AUTHOR]

Cryptosporidiosis is an intestinal infection that is triggered by the protozoan parasite Cryptosporidium spp. Cryptosporidium oocysts can spread from one host to another either through direct contact with infected hosts' faeces or through indirect means (consumption of contaminated water or food). Significant numbers of oocysts are produced as a result of the rapid growth of the parasite within the infected hosts. For proper care of cryptosporidiosis, a laboratory diagnosis is necessary. Therefore, this study aimed to produce anti-Cryptosporidium parvum (C. parvum) oocyst immunoglobulin (Ig)G polyclonal antibodies (pAbs). The produced pAbs were used in the detection of C. parvum oocysts antigens in stool and serum samples of infected calves. Moreover, pAbs were tested in protection of balb-c male mice from cryptosporidiosis infection. C. parvum oocysts were used in the preparation of antigens to be used in the immunization of New Zealand white rabbits. pAb was purified by ammonium sulphate precipitation method, caprylic acid purification method and diethylaminoethyl (DEAE) anion exchange chromatographic method. Sandwich enzyme-linked immunosorbent assay (ELISA) (using prepared pAb) scored higher sensitivity (85% and 95% for serum and stool samples) than that (80%) of microscopic examination of stool samples. Moreover, pAb significantly reduced the oocysts shedding, decreased inflammatory cytokines and enhanced the loss in the body weight of protected animals. The prepared pAb succeeded in the diagnosis of cryptosporidiosis in calves with high sensitivity either in the serum or stool samples. Our results indicated the usefulness of using the prepared pAb in protection against cryptosporidiosis. Key points: Importance of pAb in the diagnosis of cryptosporidiosis in calves. Protection of balb-c mice by anti-Cryptosporidium oocysts pAb. Sandwich ELISA has a higher sensitivity than microscopy. [ABSTRACT FROM AUTHOR]

Summary: The structurally and functionally related interleukin‐4 (IL‐4) and IL‐13 cytokines play pivotal roles in shaping immune activity. The IL‐4/IL‐13 axis is best known for its critical role in T helper 2 (Th2) cell‐mediated Type 2 inflammation, which protects the host from large multicellular pathogens, such as parasitic helminth worms, and regulates immune responses to allergens. In addition, IL‐4 and IL‐13 stimulate a wide range of innate and adaptive immune cells, as well as non‐hematopoietic cells, to coordinate various functions, including immune regulation, antibody production, and fibrosis. Due to its importance for a broad spectrum of physiological activities, the IL‐4/IL‐13 network has been targeted through a variety of molecular engineering and synthetic biology approaches to modulate immune behavior and develop novel therapeutics. Here, we review ongoing efforts to manipulate the IL‐4/IL‐13 axis, including cytokine engineering strategies, formulation of fusion proteins, antagonist development, cell engineering approaches, and biosensor design. We discuss how these strategies have been employed to dissect IL‐4 and IL‐13 pathways, as well as to discover new immunotherapies targeting allergy, autoimmune diseases, and cancer. Looking ahead, emerging bioengineering tools promise to continue advancing fundamental understanding of IL‐4/IL‐13 biology and enabling researchers to exploit these insights to develop effective interventions. [ABSTRACT FROM AUTHOR]

Introduction: Interleukin-4 (IL-4) and interleukin-13 (IL-13) are two essential cytokines involved in the T helper 2 (Th2)-mediated inflammatory response to diseases, such as atopic dermatitis (AD). AK120 is a humanized immunoglobulin G subclass 4 (IgG4) monoclonal antibody (mAb) directed against the IL-4 receptor alpha (IL-4Rα) subunit shared by the IL-4 and IL-13 receptor complexes. This mAb inhibits the signaling of the IL-4 and IL-13 cytokines. Methods: The study consisted of two parts. Part 1 was a single ascending dose (SAD) study with five cohorts (receiving 15, 50, 150, 300 or 600 mg of AK120, respectively) of healthy subjects; part 2 was a multiple ascending dose (MAD) study with four cohorts (receiving AK120 at doses of 300 mg once every 2 weeks [Q2W], 300 mg once weekly [QW], 150 mg QW or 75 mg QW) of subjects with AD. A total of 81 subjects (40 in part 1, 41 in part 2) were enrolled in the study. Results: The compound was safe and well tolerated in both a SAD up to 600 mg in healthy subjects and in a MAD from 75 to 600 mg in subjects with AD. The exposure of AK120 increased in an approximately dose-dependent manner upon subcutaneous dosing. The levels of the biomarkers serum thymus and activation-regulated chemokine ligand 17 (TARC/CCL17) and immunoglobulin E decreased from baseline after AK120 administration, indicating the inhibition of the IL-4/IL-13 signaling pathways. AK120 showed improved Eczema Area and Severity Index (EASI) scores, and the proportion of subjects with Investigator Global Assessment (IGA) score 0/1 increased after AK120 treatment. Conclusions: AK120 exhibited an acceptable safety profile in healthy and AD subjects, and showed preliminary efficacy. These findings support the continued investigation of AK120 for treating AD. Clinical Trial Registration: ClinicalTrials.gov identification number: NCT04256174. [ABSTRACT FROM AUTHOR]

Background: Dupilumab, a monoclonal anti‐IL‐4Rα antibody, is approved for several type 2 mediated inflammatory diseases like asthma, atopic dermatitis, and diffuse type 2 chronic rhinosinusitis (CRS). Clinical studies had reported a transient increase in blood eosinophils during dupilumab therapy. This study aimed to assess the impact of elevated blood eosinophils on clinical outcome and to investigate the cause of high blood eosinophil levels under dupilumab therapy. Methods: Patients suffering from diffuse type 2 CRS treated with dupilumab were examined on days 0, 28, 90, and 180 after therapy start. Sino‐Nasal‐Outcome‐Test Score (SNOT‐22), Total Nasal Polyp Score (TNPS), and blood samples were collected. Cytokine measurements and proteomics analysis were conducted. Flow cytometry analysis measured receptor expression on eosinophils. Results: Sixty‐eighty patients were included. Baseline eosinophilia ≥0.3G/L was observed in 63.2% of patients, and in 30.9% of patients, eosinophils increased by ≥0.5G/L under dupilumab. Subjects with eosinophilia ≥0.3G/L at baseline had the best SNOT‐22 mean change compared to no eosinophilia. Eosinophil elevation during dupilumab therapy had no impact on clinical scores. The eosinophil adhesion molecule VCAM‐1 decreased significantly during therapy in all patients. The chemokine receptor CXCR4 was significantly down‐ and IL‐4 upregulated in subjects with eosinophil increase. Conclusion: Our findings suggest that increased eosinophils in type 2 CRS are associated with a good clinical response to dupilumab. Patients with elevated IL‐4 at baseline developed dupilumab‐induced transient eosinophilia. We identified the downregulation of VCAM‐1 and surface markers CD49d and CXCR4 on eosinophils as possible explanations of dupilumab‐induced eosinophilia. [ABSTRACT FROM AUTHOR]

The liver is a tolerogenic organ that exhibits hypo-responsiveness to antigens circulating in the portal vein. Antigens that are orally administered at high doses reach the liver. In our previous study, we demonstrated that administering ovalbumin (OVA) orally at high doses generates unique CD4+ T cells and tolerogenic dendritic cells, both of which can suppress T helper type 1 (Th1) responses, in the livers of two groups of mice: DO11.10 mice with transgenic CD4+ T cell receptors for OVA and BALB/c mice that received OVA-specific CD4+ T cells through adoptive transfer. This study aimed to investigate whether oral administration of OVA at high doses inhibits the development of hepatitis in the presence of OVA-specific CD4+ T cells. Oral administration of OVA at high doses inhibited the development of OVA-specific and concanavalin A (Con A)-induced hepatitis in DO11.10 mice, and these effects were associated with the downregulation of Th1 responses. Furthermore, the adoptive transfer of CD4+ T cells from the liver of OVA-fed DO11.10 mice inhibited the development of Con A-induced hepatitis in recipient BALB/c mice through the downregulation of Th1 responses. Finally, oral administration of OVA at high doses inhibited the development of Con A-induced hepatitis in BALB/c mice bearing naïve OVA-specific CD4+ T cells. These results suggest that the oral administration of antigens at high doses suppresses Th1-mediated hepatitis in an antigen-non-specific manner in the presence of antigen-specific CD4+ T cells. • Oral administration of OVA inhibits OVA-specific and Con A-induced hepatitis in mice. • Downregulation of Th1 responses is associated with suppression of OVA-specific and Con A-induced hepatitis. • Oral administration of OVA generates the hepatic immune environment suppressing Th1-mediated liver injury. [ABSTRACT FROM AUTHOR]

The purpose of this study was to examine the efficacy of ICG-mediated fluorescence molecular imaging (FMI) in debridement of necrotic tissue. 96 wound-infected rats were randomly divided into control group, ICG group, excitation light (EL)group and FMI group for debridement of necrotic tissue (n = 24). (I) Control group: only debridement; (II) ICG group: ICG injection before debridement; (III) EL group: Debridement under EL; (IV) FMI group: Debridement guided by ICG-mediated FMI. On the 3rd, 6th, and 9th days, the wound tissues of the rats in each group were collected for histological examination, and the levels of serum interleukin-4 (IL-4) and interferon-γ (INF-γ) were analyzed. The wound healing rate, wound score and body weight of the rats in each group were followed up until the wound healed. The results showed that the infected wounds of the rats in the FMI group had significant fluorescence development. The level of serum IL-4 in the FMI group was higher than that in the other three groups on the 6th day (p<0.01), while the level of INF-γ was lower than that in the other three groups on the 6th and 9th day (p<0.05). The results of dynamic wound tissue H&E staining indicated that the wound healing in the FMI group was better than the other three groups. The in vivo follow-up results showed that the wound healing rate and wound score of the FMI group were better than the other three groups, and the growth of rats had no difference with the other groups. ICG-mediated FMI can achieve accurate imaging of necrotic tissue for debridement, and so can accelerate wound healing, which has good clinical application prospects. [ABSTRACT FROM AUTHOR]

Objective: To explore the effect of Bairui granule combined with ceftezole sodium in the treatment of children with acute bronchitis. Methods: 102 children with acute bronchitis who were admitted to our hospital from April 2020 to January 2022 were divided into control group (ceftezole sodium treatment) and observation group (Bairui granule combined with ceftezole sodium treatment) according to random number table method, with 51 cases in each group. The curative effects, clinical indicators, blood routine indicators, serum inflammatory factor levels changes and the incidence of adverse reactions were compared in the two groups. Results: Compared with the control group, the total clinical effective rate in the observation group was significantly increased (P<0.05). Compared with the control group, sputum, asthma, pulmonary rales, cough and other symptoms disappeared time in the observation group were shorter (P<0.05). 7 d after treatment, the white blood cell count, neutrophil percentage and lymphocyte percentage in the two groups were decreased, and the observation group was lower than the control group in the same period (P<0.05). 7 d after treatment, C-reactive protein (CRP), interleukin-1β (IL-1β), interleukin-4 (IL-4), interleukin-6 (IL-6), interleukin-8 (IL-8) and interleukin-13 (IL-13) in the two groups were decreased, and the observation group was lower than the control group in the same period (P<0.05). There were no serious adverse reactions in the two groups. Conclusion: Bairui granule combined with ceftezole sodium in the treatment of children with acute bronchitis can effectively improve clinical symptoms, reduce inflammatory reaction and improve blood routine indicators. [ABSTRACT FROM AUTHOR]

The high incidence of children with recurrent episodes of acute obstructive bronchitis is a widespread problem. Correct identification of children at risk of developing bronchial asthma at school age may improve treatment and prevention approaches to this pathology, but the ability to identify these children remains limited. The purpose of the study was to determine the effectiveness of recombinant interferon alpha-2β in children with recurrent episodes of acute obstructive bronchitis in the course of treatment based on the assessment of cytokine profile. The study examined 59 children of the main group with recurrent episodes of acute obstructive bronchitis and 30 children of the comparison group who suffered from acute bronchitis, aged 2–8 years, who were in the hospital. The results of laboratory studies were compared with the data of 30 healthy children. In children with recurrent episodes of acute obstructive bronchitis, the content of serum interferon-γ and interleukin-4 was significantly reduced compared to healthy children, after treatment with recombinant human interferon alpha-2β, the content of interferon-γ and interleukin-4 in children significantly increased. The content of interleukin-1β in children with recurrent episodes of acute obstructive bronchitis was significantly higher than in healthy children, after immunomodulatory therapy with recombinant interferon alpha-2β, interleukin-4 normalized to its level in healthy children. It was found that children with recurrent episodes of acute obstructive bronchitis have an imbalance of cytokines, the effectiveness of recombinant human interferon alpha-2β therapy, which normalized the levels of the studied cytokines in the serum. The study examined 59 children of the main group with recurrent episodes of acute obstructive bronchitis and 30 children of the comparison group who suffered from acute bronchitis, aged 2 to 8 years. It was found that children with recurrent episodes of acute obstructive bronchitis have an imbalance of cytokines, the effectiveness of recombinant human interferon alpha-2β therapy, which normalized the levels of the studied cytokines in the serum. [ABSTRACT FROM AUTHOR]

• Developed an ensemble learning model, namely Meta-IL4, for accurate and robust prediction of IL4-inducing peptides. • RF, XGB, LGBM, & ETC were used as classification algorithms apart from the Meta-IL4 model. • Our Meta-IL4 model achieved the best results. The cytokine interleukin-4 (IL-4) plays an important role in our immune system. IL-4 leads the way in the differentiation of naïve T-helper 0 cells (Th0) to T-helper 2 cells (Th2). The Th2 responses are characterized by the release of IL-4. CD4+ T cells produce the cytokine IL-4 in response to exogenous parasites. IL-4 has a critical role in the growth of CD8+ cells, inflammation, and responses of T-cells. We propose an ensemble model for the prediction of IL-4 inducing peptides. Four feature encodings were extracted to build an efficient predictor: pseudo-amino acid composition, amphiphilic pseudo-amino acid composition, quasi-sequence-order, and Shannon entropy. We developed an ensemble learning model fusion of random forest, extreme gradient boost, light gradient boosting machine, and extra tree classifier in the first layer, and a Gaussian process classifier as a meta classifier in the second layer. The outcome of the benchmarking testing dataset, with a Matthews correlation coefficient of 0.793, showed that the meta-model (Meta-IL4) outperformed individual classifiers. The highest accuracy achieved by the Meta-IL4 model is 90.70%. These findings suggest that peptides that induce IL-4 can be predicted with reasonable accuracy. These models could aid in the development of peptides that trigger the appropriate Th2 response. [ABSTRACT FROM AUTHOR]

Respiratory virus infections are the main causes of pediatric diseases. Human metapneumovirus (hMPV) is an enveloped RNA virus similar to severe acute respiratory syndrome coronavirus type 2, both of which have emerged as important new respiratory viruses. Recent studies have found that interleukin-4 (IL-4) is involved in the replication of a variety of viruses, and its role differs in different viruses. The purpose of this study was to investigate the effect of IL-4 on hMPV and to elucidate its mechanism of action. We found that hMPV infection promoted the expression of IL-4 in human bronchial epithelial cells. The replication of the virus was reduced using small interfering RNA knockdown of IL-4 expression, while the addition of exogenous recombinant human IL-4 to IL-4 knockdown cells restored viral replication ability. These results demonstrate that the expression of IL-4 is closely related to the replication of hMPV; moreover, further experiments revealed that IL-4 promotes the replication of hMPV through a mechanism dependent on the Janus kinase/signal transductor and transcription activator 6 signaling pathway. Therefore, anti-IL-4 strategies may be a promising avenue for the treatment of hMPV infection, representing an important breakthrough for children at risk from hMPV infection. [ABSTRACT FROM AUTHOR]

Activation of the interleukin-4 (IL-4) pathway ameliorates secondary injury mechanisms after experimental traumatic brain injury (TBI); therefore, we assessed the effect of a therapeutic IL-4 administration on secondary brain damage after experimental TBI. We subjected 100 C57/Bl6 wildtype mice to controlled cortical impact (CCI) and administered IL-4 or a placebo control subcutaneously 15 min thereafter. Contusion volume (Nissl staining), neurological function (hole board, video open field, and CatWalkXT®), and the immune response (immunofluorescent staining) were analyzed up to 28 days post injury (dpi). Contusion volumes were significantly reduced after IL-4 treatment up to 14 dpi (e.g., 6.47 ± 0.41 mm3 vs. 3.80 ± 0.85 mm3, p = 0.011 3 dpi). Macrophage invasion and microglial response were significantly attenuated in the IL-4 group in the acute phase after CCI (e.g., 1.79 ± 0.15 Iba-1+/CD86+ cells/sROI vs. 1.06 ± 0.21 Iba-1/CD86+ cells/sROI, p = 0.030 in the penumbra 3 dpi), whereas we observed an increased neuroinflammation thereafter (e.g., mean GFAP intensity of 3296.04 ± 354.21 U vs. 6408.65 ± 999.54 U, p = 0.026 in the ipsilateral hippocampus 7 dpi). In terms of functional outcome, several gait parameters were improved in the acute phase following IL-4 treatment (e.g., a difference in max intensity of −7.58 ± 2.00 U vs. −2.71 ± 2.44 U, p = 0.041 3 dpi). In conclusion, the early single-dose administration of IL-4 significantly reduces secondary brain damage in the acute phase after experimental TBI in mice, which seems to be mediated by attenuation of macrophage and microglial invasion. [ABSTRACT FROM AUTHOR]

This study aimed to examine the impact of 12 weeks of aerobic interval, resistance and combined exercise on inflammatory markers of TNF-a, IFN-y, Metrnl and IL-4 in obese men. Forty-four individuals (35–55 years; body mass index (BMI): 30–40 kg/m2) voluntarily participated, and were randomly divided into four groups: aerobic interval exercise (n = 10) (5 × 3-min intervals at ∼40–80% peak heart rate), resistance exercise (n = 11) (2–3 sets, 8–20 repetitions at 40–80% one-repetition maximum), combined exercise (n = 12) (a combination of both aerobic and resistance exercises) and control (n = 11). The exercise protocol in all three groups was 12 weeks with a frequency of three sessions per week. IL-4 increased significantly in aerobic interval (P = 0.005), resistance (P = 0.005), and combined (P = 0.022) exercise groups. TNF-α decreased significantly only in the aerobic (P = 0.005) and combined (P = 0.012) groups, but it increased in the resistance group (P = 0.025). None of the exercise interventions resulted in a significant change in Meteorin-like protein (Metrnl) and IFN-γ levels (P > 0.05). The results suggest that aerobic and combined exercises may have more beneficial effects in improving the inflammatory status than resistance exercise alone in obese individuals. [ABSTRACT FROM AUTHOR]

Eczema is an inflammatory skin disease and may be the result of environmental factors, genetic factors, and immune factors. Itching, persistent scratching, skin redness, and dryness are the primary signs of the illness. Many Cytokines are essential to the pathophysiology of disease, including the role of interleukin-4 and interleukin-25 as major players in the immune pathogenesis of eczema. This study was conducted with the aim of evaluating some seromic criteria that clarify the effect of some immune and inflammatory factors on the pathological condition in eczema patients and comparing them with the healthy group. This can help determine whether there is a relationship between the level of interleukin-4 and the high level of interleukin-25 and the clinical symptoms of the disease. The current study was conducted in Marjan Medical City in Babylon Governorate. Samples of (50) patients with eczema were collected after being diagnosed by dermatologists, and samples of (50) ostensibly healthy people were collected as a control group. The enzyme-associated immuno-absorption method (Elisa) was used to measure the level of interleukin-4 and interleukin-25 in the serum of all patients and the control group Serum levels of interleukin-4 and interleukin-25 are significantly higher in eczema patients at the probability level (P <0.05), while they decrease in the control group for the same criteria rates According to the results of the current investigation, patients with eczema reported higher levels of interleukin-4 and interleukin-25 in the serum than the control group. [ABSTRACT FROM AUTHOR]

Backgrounds: Interleukin 4 (IL 4) is a cytokine associated with the cause of several allergic diseases such as asthma due to their function in the differentiation of T helper type 2 lymphocytes and induction of the IgE isotype switch. Object: The study aims to determine the association IL-4 level with total serum IgE levels and asthma severity. Methods: A cross-section study was performed. Eighty-seven subjects were recruited from Karbala Teaching Hospital for Children in the period extending from January 25 to May 24, 2022, including children with asthma, were subjected to measure IL 4 level using Elabscience ELISA kit and measured total IgE level using AccuBind IgE ELISA kit. Results: Eighty-seven subjects of asthmatic children in which the mean age was 7.833 ± 3.652. There are 65.52% and 34.48% of asthmatic children (male and female, respectively). Total serum IgE was 398.889 ± 227.156 IU/ml, while the IL-4 level was 5.18 ± 8.224. There was a significant difference in IL 4 levels depending on asthma severity (P= 0.037). IL-4 levels in severe persistent asthma were higher than IL-4 levels in mild and moderate persistent asthma. Further, there was a highly significant difference in IL 4 levels depending on asthma controlled (P= 0.004). IL-4 levels in not well-controlled asthma were higher than IL-4 levels in well and partial controlled asthma. In addition, there was no significant correlation between IL-4 levels and total serum IgE levels (P=0.436). Conclusion: IL 4 has an important role in predicting asthma severity and asthma control in children. These findings have important implications for the treatment of IgE-mediated asthma. Despite this, IL 4 levels have no significant correlation with total serum IgE levels. [ABSTRACT FROM AUTHOR]

Immunoassays, which use antigen–antibody reactions, are the primary techniques used to selectively quantify specific disease markers in blood. Conventional immunoassays, such as the microplate-based enzyme-linked immunosorbent assay (ELISA) and paper-based immunochromatography, are widely used, but they have advantages and disadvantages in terms of sensitivity and operating time. Therefore, in recent years, microfluidic-chip-based immunoassay devices with high sensitivity, rapidity and simplicity, which are compatible with whole blood assays and multiplex assays, have been actively investigated. In this study, we developed a microfluidic device using gelatin methacryloyl (GelMA) hydrogel to form a wall-like structure in a microfluidic channel and perform immunoassays inside the wall-like structure, which can be used for rapid and highly sensitive multiplex assays with extremely small sample amounts of ~1 μL. The characteristics of GelMA hydrogel, such as swelling rate, optical absorption and fluorescence spectra, and morphology, were carefully studied to adapt the iImmunowall device and immunoassay. Using this device, a quantitative analysis of interleukin-4 (IL-4), a biomarker of chronic inflammatory diseases, was performed and a limit of detection (LOD) of 0.98 ng/mL was achieved with only 1 μL sample and 25 min incubation time. The superior optical transparency over a wide range of wavelengths and lack of autofluorescence will help to expand the application field of the iImmunowall device, such as to a simultaneous multiple assay in a single microfluidic channel, and provide a fast and cost-effective immunoassay method. [ABSTRACT FROM AUTHOR]

Cytokines secreted by individual immune cells regulate tissue regeneration and allow communication between various cell types. Cytokines bind to cognate receptors and trigger the healing process. Determining the orchestration of cytokine interactions with their receptors on their cellular targets is essential to fully understanding the process of inflammation and tissue regeneration. To this end, we have investigated the interactions of Interleukin-4 cytokine (IL-4)/Interleukin-4 cytokine receptor (IL-4R) and Interleukin-10 cytokine (IL-10)/Interleukin-10 cytokine receptor (IL-10R) using in situ Proximity Ligation Assays in a regenerative model of skin, muscle and lung tissues in the mini-pig. The pattern of protein–protein interactions was distinct for the two cytokines. IL-4 bound predominantly to receptors on macrophages and endothelial cells around the blood vessels while the target cells of IL-10 were mainly receptors on muscle cells. Our results show that in situ studies of cytokine–receptor interactions can unravel the fine details of the mechanism of action of cytokines. [ABSTRACT FROM AUTHOR]

BACKGROUND: Ultrasound‐assisted glycation is a promising method for decreasing the allergenicity of α‐lactalbumin (ALA). However, there is a lack of in vivo studies on the allergenicity of ultrasound‐assisted glycated ALA. In this study, the effects of the ultrasound‐assisted glycation of ALA on the allergenicity and intestinal microflora were characterized using a BALB/c mouse model. RESULTS: Increased immunoglobulin ‐G/ immunoglobulin‐E (IgG/IgE) and interleukin‐4/6 (IL‐4/6) secretions, and reduced interferon‐γ (IFN‐γ) secretions were found in the serum of ALA sensitized and challenged, mice in comparison with a control group. However, there was no significant difference between the mice fed with ultrasound‐assisted glycated ALA and the control group. Mice that were sensitized and challenged with ALA showed disrupted intestinal microflora, manifesting in significantly decreased Firmicutes and significantly increased Proteobacteria. It was found that 100ALA‐gal could maintain the intestinal microflora of mice in a normal state. Pearson's rank correlation showed that Proteobacteria and Spirochaetota were correlated positively with the IL‐4/IL‐6 level and were correlated negatively with the expression of IFN‐γ. Proteobacteria were also significantly positively correlated with IL‐6 and negatively correlated with IFN‐γ (P < 0.05). CONCLUSION: These results suggested that ultrasound‐assisted glycation on ALA can maintain the intestinal microflora in a normal state thus balancing the proportion of Th1/Th2 to decrease allergic reaction. © 2022 Society of Chemical Industry. [ABSTRACT FROM AUTHOR]

Background: Atopic dermatitis (AD) is an inflammatory skin condition distinguished by an activated Th2 immune response. The local skin microbial dysbiosis is a contributing factor to the development of AD. The pathogenic coagulase-positive Staphylococcus aureus is the primary species responsible for the progression of AD. Even though Staphylococcus pseudintermedius is an animal-origin pathogen, it is increasingly becoming a source of concern in human diseases. As another coagulase-positive Staphylococci, it is crucial to pay more attention to S. pseudintermedius isolated from the lesion site. Results: In our investigation, we presented a case of cheilitis in a patient with atopic dermatitis (AD). We utilized culture and next-generation genomic sequencing (NGS) to identify the bacteria present on the skin swabs taken from the lip sites both prior to and following treatment. Our findings indicated that the predominant bacteria colonizing the lesion site of AD were S. pseudintermedius and S. aureus, both of which were eradicated after treatment. The Multi-locus sequence typing (MLST) of S. pseudintermedius and S. aureus demonstrated coordinated antibiotic susceptibility, with ST2384 and ST22 being the respective types. Although the skin abscess area resulting from S. pseudintermedius infection was significantly smaller than that caused by S. aureus in mice, the expression of cytokines interleukin-4 (IL-4) and interleukin-5 (IL-5) were significantly higher in the S. pseudintermedius-infected mice. Conclusions: The S. pseudintermedius strain isolated from the lesion site of the AD patient exhibited a higher expression of IL-4 and IL-5 when colonized on mouse skin, as compared to S. aureus. This observation confirms that S. pseudintermedius can effectively induce the Th2 response in vivo. Our findings suggest that animal-origin S. pseudintermedius may play a role in the development of AD when colonized on the skin, emphasizing the importance of taking preventive measures when in contact with animals. [ABSTRACT FROM AUTHOR]

Myoblast fusion is essential for skeletal muscle development, growth, and regeneration. However, the molecular mechanisms underlying myoblast fusion and differentiation are not fully understood. Previously, we reported that interleukin-4 (IL-4) promotes myoblast fusion; therefore, we hypothesized that IL-4 signaling might regulate the expression of the molecules involved in myoblast fusion. In this study, we showed that in addition to fusion, IL-4 promoted the differentiation of C2C12 myoblast cells by inducing myoblast determination protein 1 (MyoD) and myogenin, both of which regulate the expression of myomerger and myomaker, the membrane proteins essential for myoblast fusion. Unexpectedly, IL-4 treatment increased the expression of myomerger, but not myomaker, in C2C12 cells. Knockdown of IL-4 receptor alpha (IL-4Rα) in C2C12 cells by small interfering RNA impaired myoblast fusion and differentiation. We also demonstrated a reduction in the expression of MyoD, myogenin, and myomerger by knockdown of IL-4Rα in C2C12 cells, while the expression level of myomaker remained unchanged. Finally, cell mixing assays and the restoration of myomerger expression partially rescued the impaired fusion in the IL-4Rα-knockdown C2C12 cells. Collectively, these results suggest that the IL-4/IL-4Rα axis promotes myoblast fusion and differentiation via the induction of myogenic regulatory factors, MyoD and myogenin, and myomerger. [ABSTRACT FROM AUTHOR]

The rabbit VX2 bone tumor model is an ideal animal model for studying malignant bone tumors. Cytokines have been reported to play a role in tumor initiation and promotion, angiogenesis, and metastasis. However, few studies have investigated the relationship between cytokines and VX2 bone tumor development. This study investigated the effect of interferon-γ (IFN-γ) and interleukin-4 (IL-4) on proliferating cell nuclear antigen (PCNA) expression in tumor tissue. Thirty Japanese white rabbits were randomly divided into group A (n = 15) and group B (n = 15). The rabbit VX2 bone tumor model was constructed by implanting VX2 tumors on the medial side of the upper tibia. Group A was sacrificed in the first week of implantation, and group B in the second week of implantation. Peripheral venous blood, tumor tissue from the medullary cavity at the implantation site, and surrounding bone and soft tissue were harvested before implantation and execution in both experimental groups. IFN-γ and IL-4 expression levels in peripheral blood and PCNA levels in tumor tissues were measured by enzyme-linked immunosorbent assay (ELISA). The tumor tissue of the medullary cavity and surrounding bone and soft tissue was harvested for pathological examination. By the end of the experiment, 30 rabbits were included in the study. There was no significant difference in IFN-γ, IL-4 and PCNA expression levels in group A compared to group B before implantation (t = 1.187, p value = 0.255; t = 1.282, p value = 0.221; t = 0.499, p value = 0.626). IFN-γ and IL-4 expression levels before execution in group A were not significantly different from those before implantation (t = -1.280, p value = 0.213; t = 0.952, p value = 0.349), and PCNA expression levels were higher than those before implantation (t = 2.469, p value = 0.020). Group B had significantly lower IFN-γ expression levels before execution than before implantation (t = -3.741, p value = 0.001) and significantly higher IL-4 and PCNA expression levels before execution than before implantation (t = 6.279, p value < 0.01; t = 13.031, p value < 0.001). IFN-γ expression levels before execution in group B was significantly lower than those before execution in group A (t = 17.184, p value < 0.001), and IL-4 and PCNA expression before execution in group B was significantly higher than that before execution in group A (t = -26.235, p value < 0.001; t = -24.619, p value < 0.001). The correlation between IFN-γ and PCNA levels before execution in groups A and B was negative (r = -0.566, p value = 0.028; r = -0.604, p value = 0.017), and the correlation between IL-4 and PCNA levels was positive (r = 0.583, p value = 0.023; r = 0.884, p value < 0.001). In the rabbit VX2 bone tumor model, extending the period of time after tumor implantation resulted in a negative correlation between IFN-γ and PCNA levels and a positive correlation between IL-4 and PCNA levels. [ABSTRACT FROM AUTHOR]

1. Introduction. Multiple cytokines have been studied for their role in the propagation of the inflammatory process related to inflammatory bowel diseases (IBD), but the role of interleukin-4 remains controversial. The aim of this study was to evaluate the role of two IL-4 gene single nucleotide polymorphisms (SNPs) in disease susceptibility and phenotypic expression. 2. Materials and Methods. A group of 160 patients with IBD (86CD/74UC) and 160 healthy controls were genotyped for IL-4 rs2243250/−590C/T and rs2070874/−34C/T using real-time polymerase chain reaction with TaqMan assay. 3. Results. The analysis of IBD patients and controls revealed a significantly reduced frequency of the minor allele T of both SNPs in CD patients (p = 0.03, OR 0.55 and p = 0.02, OR 0.52) and for the entire IBD group (p = 0.01, OR 0.57 and p = 0.01, OR 0.55). Haplotype analysis identified the most frequent haplotype (rs2243250/rs2070874 CC) associated with a high risk for developing IBD (either UC or CD) (p = 0.003). IBD patients with extraintestinal manifestations had significantly increased frequency of the minor alleles T. We also found an association between the presence of allele C of rs2070874 and response to antiTNF treatment. 4. Conclusions. This is the first study to investigate the IL-4 gene's relation to IBD susceptibility conducted in Romania. Both SNPs were found to be associated with disease susceptibility and phenotypic features, such as extraintestinal manifestations and response to antiTNF agents. [ABSTRACT FROM AUTHOR]

Priming of macrophages with interferon-gamma (IFNγ) or interleukin-4 (IL-4) leads to polarisation into pro-inflammatory or anti-inflammatory subtypes, which produce key enzymes such as inducible nitric oxide synthase (iNOS) and arginase 1 (ARG1), respectively, and in this way determine host responses to infection. Importantly, L-arginine is the substrate for both enzymes. ARG1 upregulation is associated with increased pathogen load in different infection models. However, while differentiation of macrophages with IL-4 impairs host resistance to the intracellular bacterium Salmonella enterica serovar Typhimurium (S.tm), little is known on the effects of IL-4 on unpolarised macrophages during infection. Therefore, bone-marrow-derived macrophages (BMDM) from C57BL/6N, Tie2Cre+/−ARG1fl/fl (KO), Tie2Cre−/−ARG1fl/fl (WT) mice were infected with S.tm in the undifferentiated state and then stimulated with IL-4 or IFNγ. In addition, BMDM of C57BL/6N mice were first polarised upon stimulation with IL-4 or IFNγ and then infected with S.tm. Interestingly, in contrast to polarisation of BMDM with IL-4 prior to infection, treatment of non-polarised S.tm-infected BMDM with IL-4 resulted in improved infection control whereas stimulation with IFNγ led to an increase in intracellular bacterial numbers compared to unstimulated controls. This effect of IL-4 was paralleled by decreased ARG1 levels and increased iNOS expression. Furthermore, the L-arginine pathway metabolites ornithine and polyamines were enriched in unpolarised cells infected with S.tm and stimulated with IL-4. Depletion of L-arginine reversed the protective effect of IL-4 toward infection control. Our data show that stimulation of S.tm-infected macrophages with IL-4 reduced bacterial multiplication via metabolic re-programming of L-arginine-dependent pathways. [ABSTRACT FROM AUTHOR]

Physical and psychological stress modulates the hypothalamic pituitary adrenal (HPA) axis, and the redox and inflammatory systems. Impairments in these systems have been extensively reported in major depression (MD) patients. Therefore, our study aimed to investigate the effects of the intranasal administration of interleukin-4 (IL-4) in mice with depressive-like behavior induced by chronic unpredictable mild stress (CUMS) for 28 days. On the 28th day, mice received IL-4 intranasally (1 ng/mouse) or vehicle (sterile saline), and after 30 min, they were submitted to behavioral tests or euthanasia for blood collection and removal of the adrenal glands, axillary lymph nodes, spleen, thymus, prefrontal cortices (PFC), and hippocampi (HC). A single administration of IL-4 reversed CUMS-induced depression-like behavior in the tail suspension test and splash test, without evoking locomotor changes. IL-4 administration reduced the plasma levels of corticosterone and the increased weight of suprarenal glands in stressed mice. Moreover, IL-4 restored the expression of nuclear factor erythroid 2-related factor 2 (NRF2), nuclear factor kappa B (NF-kB), interleukin 1 beta (IL-1β), IL-4, brain derived neurotrophic factor (BDNF), and indoleamine 2,3-dioxygenase (IDO) in the PFC and HC and modulated oxidative stress markers in these brain structures in stressed mice. Our results showed for the first time the antidepressant-like effect of IL-4 through the modulation of neuroinflammation and oxidative stress. The potential effect of IL-4 administered intranasally arises as an innovative strategy for MD treatment. [ABSTRACT FROM AUTHOR]

Background: Insight into the pathomechanism of atopic diseases demonstrated a pivotal role of the cytokines interleukin‐4 (IL‐4) and IL‐13, which has spurred the development of tailored therapeutics targeting their common IL‐4 receptor (IL‐4R). However, several aspects of the IL‐4R system remain ill‐defined in humans. Methods: We used multicolor spectral flow cytometry to characterize IL‐4R subunit expression in 28 human immune cell subsets on protein and mRNA levels and assessed their subcellular distribution by applying a specifically adapted protocol that avoided influence of fixation and permeabilization on fluorochrome and antibody performance. In patients, we investigated possible changes in IL‐4Rα distribution before and during treatment with dupilumab, a monoclonal antibody‐targeting IL‐4Rα. Results: Whereas all immune cell subsets investigated expressed IL‐4Rα and common γ chain protein and mRNA, expression of IL‐13Rα1 was restricted to myeloid and B cells. Interestingly, some cells contained considerably more intracellular IL‐4R protein than on their surface. Naive B cells were found to carry the highest levels of IL‐4Rα distributed evenly between surface and intracellular space, whereas IL‐4Rα was found predominantly in intracellular pools in neutrophils. In patients with atopic diseases treated with dupilumab, we observed that engagement of IL‐4Rα by dupilumab resulted in internalization of the antibody and decreased total IL‐4Rα expression. Notably, even after months of treatment not all intracellular IL‐4Rα molecules were occupied by dupilumab, indicating the presence of a "dormant" intracellular IL‐4Rα pool that could be mobilized upon certain extrinsic or intrinsic cues. Conclusion: Collectively, our findings suggest that distinct human immune cell subsets contain surface and intracellular IL‐4R pools, which are differently affected by targeted biologic treatment. [ABSTRACT FROM AUTHOR]