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摘要： Lysosomal damage is an endogenous danger signal to the cell, but its significance for innate immunity and how specific signaling pathways are engaged by this stressor remain unclear. Here, we uncover an immune-inducible pathway that connects lysosomal damage to mitochondrial DNA (mtDNA) efflux and type I IFN production. Lysosomal damage elicits mitochondrial outer membrane permeabilization (MOMP) via BAK/BAX macropores; however, the inner mitochondrial membrane (IMM) prevents wholesale mtDNA release in resting cells. Priming with type II IFN (IFN-γ) induced the antibacterial effector apolipoprotein L-3 (APOL3), which upon transient lysosomal damage, targets mitochondria undergoing MOMP and selectively permeabilizes the IMM to enhance mtDNA release and activate cGAS/STING signaling. Biochemical and cellular reconstitution revealed that analogous to its bactericidal detergent-like mechanism, APOL3 solubilizes cardiolipin to permeabilize the IMM. Our findings illustrate how cells use an antibacterial protein to expedite the breakdown of endosymbiosis and facilitate a heightened response to injury and infection.

关键词： IFN-gamma inhibitory molecule   EmSAG   T cell subsets   immune evasion   immunomodulation  

摘要： Eimeria maxima (E. maxima) infection inhibits the expression of IFN-gamma, a cytokine that is essential for the Th1 immune response and plays a key role in combating this parasite. In our preliminary investigations, we identified the E. maxima surface antigen (EmSAG) as an inhibitory molecule of IFN-gamma. EmSAG was screened and characterised from an E. maxima sporozoite cDNA expression library. The present study aimed to evaluate the immunomodulatory effects of EmSAG on chicken peripheral blood mononuclear cells (PBMCs) and various T cell subsets. We analysed cell proliferation, nitric oxide (NO) release, and cytokine transcription. The results revealed that EmSAG boosts PBMC proliferation and promotes differentiation of CD4+/CD8+ T cells. Additionally, stimulation with EmSAG significantly inhibited NO release and IFN-gamma transcription while enhancing the transcription of IL-4, IL-10, and TGF-beta 1 in chicken PBMCs. The sorting purity of T cell subsets was as follows: CD8+ (96.90%), CD4+ (86.25%), CD4+CD25- (89.14%), and CD4+CD25+ regulatory T cells (Tregs;92.16%). These purified subsets were co-incubated with EmSAG to analyse the transcription of hallmark cytokines associated with Th1, Th2, and Treg responses. EmSAG significantly inhibited the transcription of IFN-gamma and IL-2 in both CD4+ and CD8+ T cells, while promoting the expression of IL-10, TGF-beta 1, and CTLA-4 in Tregs. Moreover, depletion of CD25+ cells reversed the EmSAG-induced suppression of IL-2 transcription and reduced its stimulating effects on IL-4 and IL-10 transcription in CD4+CD25- T cells. These findings highlight the role of EmSAG as an inhibitor of IFN-gamma, facilitating immune evasion by attenuating the Th1 immune response and modulating Treg cell function. This study provides critical insights into the immune evasion mechanisms utilised by chicken coccidia.

摘要： CAR T cell efficacy in solid tumors is limited due in part to the immunosuppressive TME. To improve anti-tumor responses, we hypothesized that enabling CAR T cells to secrete bifunctional fusion proteins consisting of a cytokine modifier (e.g., TGFβtrap, IL15, or IL12) combined with an immune checkpoint inhibitor (e.g., αPDL1) will provide tumor localized immunomodulation to improve CAR T cell functionality. To that end, we engineered CAR T cells to secrete TGFβtrap, IL15, or IL12 molecules fused to αPDL1 scFv, and assessed in vitro functionality and in vivo safety and efficacy in prostate and ovarian cancer models. CAR T cells engineered with αPDL1-IL12 were superior in safety and efficacy compared to CAR T cells alone and to those engineered with αPDL1 fused with TGFβtrap or IL15. Further, αPDL1-IL12 engineered CAR T cells improved T cell trafficking and tumor infiltration, localized IFNγ production, TME modulation, and anti-tumor responses, with reduced systemic inflammation-associated toxicities. We believe our αPDL1-IL12 engineering strategy presents an opportunity to improve CAR T cell clinical efficacy and safety across multiple solid tumor types.

关键词： kujigamberol   NFAT   T cells   interferon gamma   interleukin-2   interleukin-4   Fas ligand   eomesodermin  

摘要： Kujigamberol, a dinorlabdane compound isolated from Kuji amber, exerts multiple biological effects, including anti-allergic and anti-inflammatory activities. The present study demonstrated that kujigamberol inhibited cytokine production by T cells. In response to a phorbol 12-myristate 13-acetate (PMA) and ionomycin (IM) stimulation, kujigamberol suppressed interferon-gamma (IFN-gamma) and interleukin-2 (IL-2) mRNA expression in murine T-cell lymphoma BW5147 cells stably transfected with the T-box transcription factor eomesodermin. IL-4 and Fas ligand mRNA expression was also inhibited by kujigamberol. In the murine cytotoxic T-cell line CTLL-2, kujigamberol more strongly decreased IFN-gamma mRNA expression induced by IM alone than that induced by the combination of PMA and IM. A luciferase reporter assay showed that kujigamberol preferentially reduced nuclear factor of activated T cell (NFAT)-dependent transcription in human embryonic kidney 293T cells. Unlike the calcineurin inhibitor FK506, kujigamberol did not markedly affect NFATc2 protein levels in BW5147 cells but interfered with the binding of NFATc2 to the IFN-gamma and IL-2 promoters. These results indicate that kujigamberol inhibited IFN-gamma and IL-2 mRNA expression by preventing the binding of NFATc2 to their promoters;therefore, it has potential as an immunosuppressive agent.

关键词： eosinophilic esophagitis   interferon-gamma   interleukin-10   regulatory B cells  

摘要： Background: Eosinophilic Esophagitis (EoE) is a chronic food allergy that causes esophageal inflammation and fibrosis and manifests with symptoms of reflux, chest pain, swallowing difficulty, and food impactions. Though the prevalence of EoE is increasing by similar to 15% each year, our understanding of EoE immunopathology is limited. A noted feature of EoE is the presence of food-specific IgG4 antibodies in the circulation and esophageal tissue. Production of IgG4 is confined to IL-10+ B cells (Bregs) in other allergic diseases, suggesting Bregs may be present in EoE. Methods: We examined circulating Bregs in patients with EoE milk allergy. In parallel, we performed mechanistic investigations of the role of Bregs in a murine model of food-antigen-dependent EoE. Flow cytometry and histologic analyses were used to assess esophageal and draining lymph node immune cells, and in vitro assays were used to evaluate Breg functional capacity. Results: Breg frequency was reduced in both EoE milk allergic subjects and an EoE disease model. Murine Breg suppressive capacity was impaired during EoE-like inflammation. Inducible deletion of Breg-derived IL-10 worsened EoE-like inflammation, while adoptive transfer of IL-10 sufficient Bregs suppressed DC activation and improved esophageal eosinophilia. IFN gamma was sufficient to suppress Breg expansion and IL-10 production in vitro and contributed to Breg dysfunction and esophageal inflammation in vivo. Conclusion: Bregs play an immunoregulatory role during EoE by controlling esophageal eosinophilia but are functionally impaired due to IFN gamma-mediated signaling. These findings have important implications for understanding EoE's etiology and implementing future therapies that target IFN gamma.

关键词： kidney   mismatch   non-HLA   transplantation  

摘要： HLA-matching between donor (D) and recipient (R) is routinely performed in kidney allocation to optimize allograft survival but explains only a moderate variability of these outcomes. Recent findings suggest that donor-to-recipient mismatches outside the HLA region contribute to alloimmunity and graft loss, but the extent varies in different publications. We therefore conducted a systematic review of publications on this subject using a broad search string in our literature review in accordance with current guidelines for systematic reviews. The effect sizes were analyzed by a meta-analysis. A total of 1890 publications from 2019-25 within three different repositories (465 Medline, 1408 Embase, 17 Central) were systematically screened using the PICOTS (Population, Intervention, Comparator, Outcome, Time and Setting) system, which resulted in 12 eligible papers that met the inclusion criteria. Cohort studies that investigated the association of D-R non-HLA SNP mismatch and graft rejection/loss in renal transplant patients were included. We found that overall SNP mismatch between D-R pairs outside the HLA region was independently numerically associated with rejection hazard ratio (HR) 1.26 [95% confidence interval (CI) 0.97-1.65] and graft loss HR 1.35 (95% CI 0.86-2.12). Furthermore, loss of function mutation of the gene LIMS1 in the recipient who received a transplant organ with at least one functioning copy (collision genotype) was numerically associated with rejection HR 1.23 (95% CI 0.68-2.23) and graft loss HR 1.43 (95% CI 0.61-3.36). The exact quantification of the effect size of these mismatches varied by publication and needs further investigation. Based on these data, the strength of immunosuppression may be guided by the load of D-R mismatches in the future. 10.1093/ndt/gfaf073 Video Abstract Watch the video abstract of this contribution at https://***/ndt/pages/author_videos gfaf073Media1 6373014838112

关键词： Chemoresistance   Colorectal cancer   Inflammation   Nanocrystals   Xanthatin  

摘要： Poor water-solubility of emerging new chemotherapeutic drugs lead to low absorption and tissue bioavailability. Improved drug delivery has therefore recently been achieved through the versatile physico-chemical properties of nanocrystals (NCs) in targeted cancer therapies. Here, nanocrystalization was used with xanthatin, a not highly water-soluble natural sesquiterpene lactone compound that possesses anti-tumour properties and which was recently investigated for potential use in the treatment of cancer and autoimmune diseases. Given that tumour-promoting inflammation is a hallmark of colorectal cancer (CRC), and that epidemiological studies associated inflammatory biomarkers to CRC poor prognosis and therapy resistance, the anti-inflammatory properties of xanthatin NCs were assessed in 2D monolayers and in 3D spheroids of a human HT29 CRC cell model. The 3D spheroids being a model recapitulating a cancer stem cells and chemoresistant phenotype. HT29 2D monolayer cell response was first tested against four pro-inflammatory inducers including phorbol-12-myristate-13-acetate, tumour necrosis factor alpha (TNF alpha), transforming growth factor beta, and Concanavalin A. Of these inducers, HT29 cell response to TNF alpha resulted in the most elevated expression of cyclooxygenase (COX)-2 which was prevented by commercial xanthatin along with the phosphorylation of the extracellular signal-regulated kinase 1/2 and of IkappaB (I kappa B). Alteration of 3D spheroids formation and of the inflammatory/immunity transcriptomic signature was also found better altered by xanthatin NCs in comparison to commercial xanthatin and the isolated molecule. Collectively, our data indicate that xanthatin nanocrystallization did not alter the potential in vitro anti-inflammatory and anticancer properties of xanthatin against a 3D CRC chemoresistance cellular model. These properties make NCs a significant advancement in the field of cancer theranostics to improve patient outcomes.

摘要： OBJECTIVE:To investigate the role of senescent epidermal basal cells in cutaneous lupus erythematosus (CLE) pathogenesis using skin samples from patients with CLE and a mouse model of systemic lupus erythematosus (SLE). METHODS:Cellular senescence profiling utilized datasets from the NCBI Gene Expression Omnibus database and Accelerating Medicines Partnership® (AMP®) Phase 1-Metro. Gene array data from GSE184989 (CLE: n = 68, control: n = 4), single-cell RNA sequencing data from GSE186476 (CLE: n = 7, control: n = 14), and AMP® Phase 1-Metro (SLE: n = 17) were utilized. In vitro experiments further examined the expression of p21, type I interferon (IFN), human leukocyte antigen class I (HLA-I), and epidermal growth factor receptor (EGFR) signalling. Pharmacological clearance of senescent cells was performed using the senolytic drug fisetin in the SLE mouse model. RESULTS:p21-high senescent epidermal basal cells in patients with CLE exhibit high type I IFN expression. These cells enhance IFN signalling in surrounding normal epidermal basal cells, leading to increased HLA-I expression. In contrast, senescent epidermal cells upregulate EGFR signalling, which downregulates HLA-I expression, allowing them to evade immune surveillance. This heterogeneity of HLA-I expression promotes CD8-positive T-mediated toxicity against normal epidermal basal cells, resulting in their apoptosis. Pharmacological clearance of senescent epidermal basal cells improved SLE-like skin lesions. CONCLUSION:Senescent cells create a microenvironment that directs cytotoxic T-cell-mediated responses against normal epidermis in CLE, contributing to disease pathology. Targeting senescent cells and their signalling pathways may offer novel therapeutic strategies for CLE and SLE skin lesions.

关键词： 布鲁杆菌病   CD8 +CD28 -T淋巴细胞   免疫检查点   免疫失调  

摘要： 目的:探讨布鲁杆菌病（Brucellosis，简称布病）患者外周血CD8 +T淋巴细胞亚群与不同免疫检查点的表达水平、相关性及诊断效能。 方法:病例对照研究。选取2021年2月1日至2023年12月31就诊于新疆维吾尔自治区人民医院的32例急性期，男27例，女5例，年龄36（30，43）岁;38例慢性期，男30例，女8例，年龄40（32，48）岁;布病患者及同期体检的30名健康对照者，男24名，女6名，年龄39（32，46）岁。受试者均于清晨空腹采血1次，应用流式细胞术检测其外周血淋巴细胞亚群比例及CD8 +T淋巴细胞表面程序性细胞死亡受体-1（PD-1）、T淋巴细胞免疫球蛋白和ITIM结构蛋白（TIGIT）、免疫球蛋白和黏蛋白结构域-3（TIM-3）、穿孔素和颗粒酶B的表达水平，观察以上指标在疾病急、慢性期的变化，并利用Spearman方法进行相关性分析，采用绘制受试者工作特征曲线（ROC）分析具有差异性的免疫学指标评估急慢性布病的诊断价值。 结果:CD3 +T淋巴细胞慢性组（70.71%±8.78%），健康对照组（74.65%±7.31%），两组比较差异有统计学意义（ P<0.05），CD4 +T淋巴细胞急性期（39.52%±5.85%），健康对照组（45.10%±5.18%），两组比较差异有统计学意义（ P<0.01），CD8 +T淋巴细胞急性组（31.73%±5.87%），慢性期（26.75%±4.71%），两组比较差异有统计学意义（ P<0.001），CD8 +CD28 -T淋巴细胞急性组69.85%（58.62%，78.55%），慢性组86.46%（73.30%，92.52%），健康对照组25.39%（20.60%，32.90%），三组间两两比较差异有统计学意义（ P<0.001）。免疫检查点PD-1、TIGIT、TIM-3在CD8 +T淋巴细胞表面的表达水平在急、慢性组均高于健康对照组（ P<0.001）。CD8 +T淋巴细胞分泌穿孔素在急、慢性组中的表达量低于健康对照组（ P<0.05），颗粒酶B在急、慢性组的表达量高于健康对照组（ P<0.01）。布病患者CD8 +CD28 -T淋巴细胞比例与TIGIT、TIM-3的表达水平呈正相关（ r=0.624、0.406， P均<0.001）。ROC曲线联合CD8 +CD28 -T淋巴细胞比例和CD8+T淋巴细胞表面TIGIT比例鉴别急慢性布病，曲线下面积（AUC）为0.973，对于临床区分急慢性病患者具有一定提示意义。 结论:布病患者的CD8 +T淋巴细胞亚群呈现动态变化，并伴随相关免疫检查点分子的变化，调节布病患者的免疫状态的激活与抑制。CD8 +CD28 -T细胞与TIGIT/TIM-3的协同作用可能是驱动慢性化的关键机制，二者联合诊断可作为临床分期标志。

关键词： 银屑病   固本祛湿化瘀方   树突状细胞   白细胞介素-17   免疫应答反应   中药复方  

摘要： 目的 探讨固本祛湿化瘀方（GQHF）是否通过调控树突状细胞（DCs）为核心的免疫应答机制治疗银屑病。方法 体内实验：将18只Balb/c小鼠随机分组，分别为正常组（N）、模型组（M）、固本祛湿化瘀方治疗组（GQHF），每组各6只。N组不做任何处理。其余组通过5%咪喹莫特乳膏62.5 mg构建银屑病疾病模型，同时从造模后第1天开始灌胃。其中GQHF组以9.1 g/kg的GQHF灌胃，M组予等体积的饮用水灌服，连续7天。期间记录和评估小鼠银屑病样皮损红斑、鳞屑、浸润程度及严重程度指数（PASI）评分，通过苏木精—伊红染色（HE）法检测表皮层厚度、酶联免疫吸附试验（ELISA）法检测炎症因子白细胞介素（IL）-17的表达水平，流式细胞术（FCM）检测腋下及腹股沟淋巴结DCs成熟标志物表面分子（CD80、CD86）的表达情况。体外实验：将从4只Balb/c小鼠股骨中培养诱导获得的DCs分为空白对照组（BK）、空白血清对照组（NC）、15%GQHF含药血清干预组（15%GQHF）及20%GQHF含药血清干预组（20%GQHF）。除BK组外，其余组通过脂多糖（LPS）刺激DCs成熟，模拟银屑病的炎症状态。通过细胞增殖及细胞毒性检测试剂（MTT）法分别检测在24、48、72 h时GQHF含药血清对DCs细胞增殖的影响。另外根据MTT结果，将DCs分为BK组、NC组及20%GQHF组。各组共培养48 h后通过FCM检测表达CD80、CD86的DCs百分比变化。结果 体内实验：与N组比较，M组小鼠银屑病样皮损PASI评分升高（P<0.01），表皮层厚度增厚（P<0.01），淋巴结中表达CD80、CD86的DCs百分比增加（P<0.05），血清的炎症因子IL-17表达水平升高（P<0.05）。与M组比较，GQHF组小鼠银屑病样皮损PASI评分降低（P<0.05），表皮层厚度变薄（P<0.01），淋巴结中表达CD80、CD86的DCs百分比减少，其中CD11c+CD80+的百分比减少有统计学意义（P<0.01）；血清IL-17表达水平降低（P<0.05）。体外实验：GQHF含药血清抑制DCs细胞增殖的生物学活性随浓度的提高而增加，且随着共培养时间的延长而增加，呈浓度和时间依赖性。与NC组比较，20%浓度的GQHF含药血清与DCs共培养48 h时对DCs的增殖抑制作用（P<0.05）。此外，GQHF含药血清抑制DCs的成熟表达。与BK组比较，NC组经LPS刺激的DCs成熟标志物表面分子CD80、CD86的表达水平升高（P<0.01）。与NC组比较，GQHF组下调DCs表达CD80、CD86的DCs细胞百分比（P<0.05）。结论 GQHF可能以调控DCs为核心的免疫应答机制，降低下游关键炎症因子IL-17表达水平而改善银屑病的皮肤损害。