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摘要： Intratumoral immunotherapy (ITIT) strives to generate effective antitumor immunity by directly stimulating the immune system in tumors to reverse local tumor-mediated immune suppression. In vivo expression of Interleukin-12 (IL-12) using in vivo plasmid transfection as an intratumoral cancer immunotherapy entered Phase II clinical trials for metastatic melanoma but to limited clinical success. We sought to improve the efficacy of in vivo IL-12 electroporation by the addition of a CD154 (CD40 ligand)- expressing plasmid to the IL-12 encoding plasmid treatment and assessing efficacy against solid tumors. Mice with intradermal B16F10 melanoma or MC38 murine colon carcinoma tumors received 2 weekly intratumoral (IT) injections of plasmids encoding IL-12 and CD154, followed by in vivo electroporation. The addition of CD154 to IL-12 was superior to IL-12 alone and resulted in frequent tumor clearance of treated tumors, marked by an increase in CD8 T cells and a drastic reduction in T regulatory cells in the tumor microenvironment. Tumor treatment responses were abrogated in mice which lack conventional DC1 cells (BatF3 KO) or lack CD8 T cells. These findings highlight the potential of adding CD154 to IL-12 plasmid electroporation as a cancer immunotherapy and suggest that other combinations would be therapeutically valuable.

关键词： autoimmune disease   high endothelial venules   interleuikin-17A   pemphigus vegetans   rituximab   tissue-resident memory T cell  

摘要： A 49-year-old man with pemphigus vegetans (PVeg) relapsed several times due to cessation of therapy. At the age of 66, vegetative plaques developed at the left oral commissure following mechanical plucking of approximately 500 beard hairs. Treatment with prednisolone (20 mg/day) was continued but yielded no clinical improvement. Subsequently, rituximab (1000 mg twice, 2 weeks apart) led to a rapid decline of anti-desmoglein 3 (Dsg3) autoantibody and clinical improvement. The lesion resolved after 18 months, and prednisolone was tapered without recurrence. Based on the site-specific recurrence and chronicity, we hypothesized that tissue-resident memory T (TRM) cells and high endothelial venules (HEV)-like vessels contributed to disease persistence. Immunohistochemical analysis revealed a substantial presence of IL-17A-producing CD69 ++ CD103 ++ TRM cells in the dermis. In addition, PNAd++ HEV-like vessels were increased, accompanied by infiltrates of B cells and plasma cells. Notably, Dsg3-specific B cells were also detected, indicating that rituximab-mediated B cell depletion likely contributed to lesion resolution in this case.

关键词： Macrophages  

摘要： The in vitro expanded seed cells and the supraphysiological doses of exogenous growth factors both pose huge safety risks in bone regeneration. In this study, a novel IL-8/tDM/PLGA composite scaffold was developed, where chemokine interleukin-8 (IL-8) and transferable decellularized matrix (tDM) were uniformly overlaid on exterior and interior surfaces of poly(lactic-co-glycolic acid) (PLGA) porous substrates. The in vitro experiments confirmed that the synergy of tDM and IL-8 achieved the obvious promotion of osteogenesis and angiogenesis via enhancing chemotaxis, adhesion, spreading, osteogenic differentiation and mineralization of bone marrow mesenchymal stem cells (BMSCs). IL-8 mediated the in situ recruitment of BMSCs and macrophages via binding with C-X-C motif chemokine receptor 2 (CXCR2), while maintaining cellular viability without inducing macrophage polarization. Moreover, tDM improved BMSC adhesion and spreading via the recognition and binding of the affinitive ligand existing in tDM by the cell adhesion molecules (CAMs) on the BMSC cytomembrane. Furthermore, tDM promoted the osteogenic differentiation and mineralization of BMSCs, benefiting from the retained growth factors. In a rat femoral defect model, the IL-8/tDM/PLGA scaffold significantly accelerated new bone mineralization and maturation through synergistic regulations of cell recruitment, matrix adhesion, and osteogenic signaling pathways. After 8 weeks post-implantation of the IL-8/tDM/PLGA scaffolds, the bone volume fraction of the newly formed bone, trabecular number, and trabecular separation at the defect site were 47%, 1.21 mm-2, and 0.50 mm, respectively, which presented significantly better bone repair effects than those in other groups. These results demonstrated that the innovative bone regeneration strategy combining chemokine-driven recruitment and endogenous tDM regulation offered a potential solution for clinical repair of large-sized bone defects.

关键词： EIS   heparan sulfate   interleukin 8   metal ions   X-ray photoelectron spectroscopy  

摘要： Heparan sulfate (HS) interactions with interleukin 8 (IL-8) are crucial for immune system response. The structural features of the HS and the environmental entities, such as metal ions, can regulate these interactions. However, it is challenging to evaluate the effect of each parameter on the interactions because of low accessibility to well-defined saccharides and the lack of characteristic features to be determined by analytical tools. We evaluated the effect of the HS structural features on IL-8 binding affinity utilizing electrochemical impedance spectroscopy (EIS) and X-ray photoelectron spectroscopy (XPS). We showed that the metal ions Ca2+ and Mg2+ dissimilarly mediate the interactions of HS and IL-8 in structure-dependent manner of the HS. We showed that in all glycans, a positive synergistic effect on IL-8 binding was observed. For several glycans, the presence of ions resulted in a dramatic increase in the affinity to IL-8, while for other glycans, a milder effect was observed. This demonstrated that both structural motifs and environmental features are crucial for maintaining the interactions between the HS and IL-8.

关键词： LRRK2   Crohn's Disease   Parkinson's Disease   B Cells   Interleukin-4  

摘要： Mutations in Leucine-Rich Repeat protein Kinase 2 (LRRK2) are associated with Parkinson's disease (PD) and Crohn's disease (CD), but the regulation of LRRK2 during inflammation remains relatively unexplored. Here we describe the development of a flow cytometry-based assay to assess LRRK2 activity in individual cells and the generation of an EGFP-Lrrk2 knock-in reporter mouse to analyse cell-specific LRRK2 expression. Using these tools, we measured LRRK2 levels and activity in murine splenic and intestinal immune cells and in human blood. Anti-CD3 induced inflammation increases LRRK2 expression and activity in B cells and monocytes, while in mature neutrophils, inflammation stimulates activity but reduces LRRK2 expression. A kinase-activating PD-associated LRRK2-R1441C mutation exacerbates inflammation-induced activation of LRRK2 specifically in monocytes and macrophages. We identify IL-4 as a novel T-cell-derived factor that upregulates LRRK2 expression and activity in B cells, replicating inflammatory effects observed in vivo. Our findings provide valuable new insights into the regulation of the LRRK2 pathway in immune cells, crucial for understanding LRRK2 and its therapeutic potential in inflammatory diseases such as CD.

关键词： 黄曲霉   细胞外囊泡   巨噬细胞   细胞因子   极化   大蜡螟  

摘要： 目的 探讨黄曲霉细胞外囊泡（EVs）对黄曲霉感染的治疗效果。方法 使用超速离心法提取黄曲霉的EVs，通过纳米颗粒跟踪分析（NTA）检测和鉴定EVs；qRT-PCR和酶联免疫吸附试剂盒检测黄曲霉EVs对骨髓源巨噬细胞（BMDMs）极化的影响和几种细胞因子的表达水平，包括肿瘤坏死因子α（TNF-α）、干扰素-γ（IFN-γ）、白介素-6（IL-6）和白介素-10（IL-10）；构建大蜡螟感染模型，设置黄曲霉EVs的3个浓度组（0.2、2、20μg/条幼虫）作为实验组，PBS作为感染模型对照组，同时设置3种形式的阴性对照组，包括PBS组、穿刺组，空白对照组，通过大蜡螟感染模型的存活率评价黄曲霉EVs对黄曲霉感染的治疗效果。结果 黄曲霉EVs粒径在20～550nm之间；黄曲霉EVs能够使BMDMs向M1表型和M2表型极化，并产生细胞因子，包括TNF-α、IFN-γ、IL-6和IL-10；大蜡螟感染模型结果显示黄曲霉EVs可提高黄曲霉感染后大蜡螟的存活率。结论 黄曲霉产生的EVs可促进BMDMs的促炎细胞因子和抗炎细胞因子表达，促进BMDMs向M1表型和M2表型极化，同时增加大蜡螟感染黄曲霉后的存活率。

关键词： Allergy   Asthma   Cholesterol   Immunology   Pulmonology  

摘要： Group 2 innate lymphoid cells (ILC2s) play a crucial role in inducing type 2 inflammation in the lungs in response to allergens. Our study investigated the regulatory mechanism of IL-10 production by ILC2s and its impact on airway hyperreactivity (AHR), focusing on the role of ICOS. We found that inhibiting ICOS in pulmonary ILC2s significantly enhances IL-10 production. The absence of ICOS reprograms ILC2 steroid metabolism, leading to increased cholesterol and cortisol biosynthesis, and subsequent Glucocorticoid receptor (GR) activation. This reprogramming regulates MAF and NFIL3 activation, promoting IL-10 production. Notably, in vivo GR inhibition or ILC2-specific GR deficiency exacerbated AHR development in multiple mouse models. We extended these findings to human ILC2s, demonstrating concordant results between murine models and human cells. Our results indicate that ICOS negatively regulates IL-10 production in ILC2s by controlling cholesterol and cortisol biosynthesis. This mechanism provides new insights into the complex interplay between ILC2s, ICOS, and glucocorticoid signaling in the context of allergic airway inflammation.

关键词： Anti-obesity drugs   Bariatric surgery   Chirurgie bariatrique   Chirurgie métabolique   Metabolic surgery   Obesity   Obésité   Traitements médicamenteux de l’obésité  

摘要： Les patients qui souffrent d’une insuffisance respiratoire chronique peuvent être en situation d’obésité. L’obésité peut être à l’origine de l’insuffisance respiratoire chronique, notamment dans le cadre du syndrome d’obésité hypoventilation (SOH) ou de certaines formes d’asthme. Elle peut également majorer certains symptômes comme la dyspnée. Dans ce contexte, il est souvent demandé au patient de s’engager dans une modification des habitudes de vie, voire dans un projet de chirurgie bariatrique, pour permettre une perte de poids. Cette perte de poids est-elle toujours souhaitable ? Nous discuterons dans cet article les possibilités actuelles pour obtenir une perte de poids significative et durable, les arguments en faveur d’une stratégie de perte de poids chez les patients ayant une insuffisance respiratoire chronique et ceux qui incitent à la prudence. Patients suffering from chronic respiratory failure may present with obesity, which can cause respiratory failure, particularly in cases of Obesity Hypoventilation Syndrome (OHS) or certain forms of asthma. It can also aggravate symptoms such as dyspnea. In this context, patients are often asked to change their lifestyle, or even to undergo bariatric surgery, the common objective being to help them lose weight. That said, a question may be raised: is weight loss always desirable? In the present review of relevant articles, we report on: (a) the current possibilities for achieving significant and lasting weight loss, (b) the arguments in favor of a weight loss strategy for patients with respiratory failure, and (c) the arguments suggesting a need for caution.

关键词： cytotoxicity   hyperthermia   lung cancer   major histocompatibility complex-I   programmed death ligand-1  

摘要： Hyperthermia has recently been applied to treat human non-small cell lung cancer (NSCLC). However, the mechanisms underlying cytotoxic sensitivity of NSCLC cells to hyperthermia are not fully understood. In this study, five NSCLC cell lines with different epidermal growth factor receptor (EGFR), Kirsten rat sarcoma and tumor protein p53 mutation profiles (A549, H292, H1299, PC9 and H1975) were used to evaluate effects of hyperthermia. All tested cell lines except H1975 were sensitive to hyperthermia-induced cytotoxicity. Annexin V-propidium iodide double staining, Poly(ADP-ribose) polymerase (PARP) cleavage and scanning electron microscopy revealed that apoptosis and necrosis were induced by hyperthermia in different lines. Tetramethylrhodamine, ethyl ester analysis further revealed that hyperthermia affected mitochondrial function in the four hyperthermia-sensitive lines. Transmission electron microscopic analysis revealed degeneration of cristae and ruptured mitochondria upon exposure to hyperthermia. Hyperthermia also caused elevation of reactive oxygen species in sensitive cells. In addition to these effects, hyperthermia impacted cell survival-related signalling proteins (EGFR, FAK and Akt). In particular, hyperthermia increased phosphorylated EGFR but suppressed total EGFR, phosphorylated Akt and total Akt in sensitive cells. Moreover, hyperthermia could modulate immunomodulatory molecules. Major histocompatibility complex-I (MHC-I) and surface programmed death ligand-1 (PD-L1) were both elevated by hyperthermia in all tested NSCLC cell lines except PC9. Taken together, our findings provide insights into the potential influence of different somatic mutations in NSCLC cells on hyperthermia-induced cytotoxicity and regulation of key immunomodulatory molecules.

摘要： La malnutrizione, intesa come nutrizione deficitaria e/o inappropriata, ha svolto storicamente un ruolo chiave per diagnosi cliniche relative al dismetabolismo glicolipidico, quale elemento necessario e sufficiente per un ragionamento diagnostico parsimonioso. Caso emblematico è la diagnosi di diabete mellito da malnutrizione, un disordine glicolipidico noto dagli inizi del XX secolo, divenuto attuale in Italia in relazione all’immigrazione dai paesi dove era stato originariamente identificato (Africa, Asia). Caratterizzato a metà del 1900 come diabete J (da Jamaica), si presentava come una forma “ibrida” tra diabete mellito Tipo 1 e 2 in un fenotipo sottopeso giovane che, di recente, sappiamo avere similitudini con quello normopeso metabolicamente insano. Questo diabete conduce di rado alla chetosi, richiede dosaggi insulinici elevati, non si associa a steatosi epatica, mantiene una riserva insulinica, potrebbe rispondere favorevolmente a nutrizione iperproteica e terapia combinata con insulina e ipoglicemizzanti orali e, quest’anno, è stato classificato dalla International Diabetes Federation come diabete mellito Tipo 5.