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摘要： BACKGROUND The recurrence of IgA nephropathy (IgAN) after kidney transplantation (KT) varies between 9% and 51%. Our analysis aimed to identify risk and protective HLA alleles for the development of IgAN and its recurrence after KT. MATERIAL AND METHODS This retrospective single-center analysis included all patients after KT. Patients underwent HLA typing prior to being listed on the waiting list for KT. Comparisons were performed with a cohort of 10 000 healthy donors from the Eurotransplant registry. RESULTS A total of 470 patients were included. Biopsy-proven IgAN, as the underlying cause of renal failure, was found in 7.2% (n=48), of whom 77% were male (n=37). The DRB1*11 allele was present in 47.92% and was identified as a significant risk factor for IgAN (OR 2.09, P=0.0048). The DRB1*03 allele was detected in 4.17%, and we identified it as potentially protective (OR 0.2, P=0.5472). Recurrence of IgAN was detected in 20.8%, 100% of whom were male. The mean time to confirm recurrence was 56.1 months. We identified the DRB1*11 risk allele in 77.8% of patients with confirmed recurrence but did not identify it as an independent risk factor (HR 2.3967; P=0.3956). We found a significant correlation of IgAN recurrence with the development of DSA after KT (r 0.3980, P=0.0218). CONCLUSIONS Our study identified a 20.8% incidence of IgAN recurrence after KT; 77.8% of these patients had the HLA-DRB1*11 allele, which we also identified as a risk allele for the development of IgAN in native kidneys. De novo DSA after KT was correlated with IgAN recurrence.

关键词： autoimmune brain disorder   neuroimmunology   neuropsychiatric   transcriptomics  

摘要： The pathogenesis of Sydenham chorea remains unclear. We report a 10-year-old girl presenting with subacute chorea and mild carditis following Streptococcal throat infection. Single-cell RNA sequencing on 30,794 peripheral immune cells from the patient and two sex-matched controls revealed nine immune cell clusters. We found up-regulated antigen processing pathways in B cells, monocytes, and eosinophils, enriched by HLA genes, particularly HLA-DRB5 and HLA-DQB1, and immunoglobulin (IGH) genes in B cells. Up-regulated proteasomal pathways in plasma cells were enriched by PSM genes. These insights support the B cell-mediated autoantibody hypothesis in Sydenham chorea. We highlight the potential of scRNA-seq in rare diseases to inform targeted therapies.

关键词： 刚地弓形虫   四抗原融合蛋白   蛋白疫苗   免疫保护  

摘要： 目的 探讨刚地弓形虫棒状体蛋白18 （ROP18）、中心蛋白家族蛋白TGME49＿237490、卵囊壁特异性蛋白直系同源物TGME49＿268230和微线体蛋白13 （MIC13）四抗原融合蛋白（简称4×）的免疫保护作用。方法PCR扩增获得4×目的基因，构建重组质粒并进行鉴定。将构建成功的质粒转化至大肠埃希菌BL21感受态细胞，进行目的蛋白的诱导表达和亲和纯化，十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）与蛋白免疫印迹分析（Western blotting）鉴定目的蛋白表达情况。36只雌性BALB/c小鼠随机分成免疫组与对照组，各18只，免疫组注射4×蛋白加弗氏佐剂，对照组注射等量PBS加弗氏佐剂，共免疫3次，每次间隔2周。分别于免疫后第0、2、4、6周采集小鼠眼眶血，分离血清，ELISA检测血清中IgG抗体水平，第6周加测IgG抗体亚型水平。末次免疫后2周各取10只小鼠无菌取脾，制备脾细胞悬液，5只小鼠采用细胞增殖及毒性检测试剂盒检测细胞增殖水平，5只小鼠通过微球免疫法流式多因子检测试剂盒检测细胞因子水平。末次免疫后2周取5只小鼠外周血，流式细胞术检测小鼠T细胞亚群比例。末次后免疫2周各取8只小鼠通过腹腔注射100个弓形虫RH株速殖子，3只小鼠通过荧光定量PCR检测脏器弓形虫负荷量，5只小鼠记录存活时间，绘制生存曲线，采用GraphPad Prism 8软件进行统计学分析。结果 4×基因PCR扩增片段约为3 576 bp，与预期大小一致。重组质粒经PCR、酶切鉴定均获得预期大小的条带。目的蛋白被成功表达、纯化，SDS-PAGE与Western bloting均在相对分子质量（Mr） 110 000处可见明显条带。ELISA检测结果显示，免疫组IgG抗体水平在免疫后第0、2、4、6周的吸光度(A450值)分别为0.079±0.004、 0.759±0.179、1.670±0.243、2.461±0.056，对照组分别为0.080±0.006、0.067±0.009、0.080±0.014、0.076±0.011，免疫组除第0周（t=0.05,P> 0.05）外均高于对照组（t=9.88、23.33、34.66,P <0.05）；免疫组IgG1和IgG2a抗体亚型水平的A450值分别为3.202±0.401、3.725±0.066，均高于对照组的0.082±0.003、0.059±0.017 （t=13.73、96.83,P <0.05）。流式细胞术检测结果显示，免疫组细胞因子γ干扰素（INF-γ）、白细胞介素-2 （IL-2）、 IL-12、 IL-4、 IL-10水平分别为（4 998.52±2 131.24）、（5.76±1.02）、（1.38±0.86）、（28.83±1.64）、（3 376.57±218.48） pg/ml，对照组分别为（8.90±0.17）、（3.05±0.50）、（0.06±0.13）、（11.18±1.58）、（13.87±3.55） pg/ml，免疫组细胞因子水平均高于对照组（t=4.89、4.61、3.06、15.93、30.30,P <0.05）；免疫组小鼠的脾细胞增值水平（3.286±0.552）高于对照组（1.251±0.157）(t=9.15, P <0.05)。免疫组CD3+CD4+T细胞和CD3+CD8+T细胞的比例分别为（31.15±2.83）%、（18.06±2.82）%，对照组分别为（26.82±1.17）%、（9.33±1.22）%，两组差异有统计学意义（t=3.16、6.36,P <0.05）。qPCR结果显示，免疫组小鼠的肝、脾、肺虫荷量分别为（206.7±16.0）、（100.2±15.4）、（7.14±2.03） fg/mg，低于对照组的（20 086.0±1 310.0）、（26 254.0±9 658）、（5 300.0±741.2） fg/mg （t=26.29、4.69、12.37,P <0.05）。对照组小鼠在8 d内均死亡，免疫组小鼠在30 d的观察期内，存活1只，存活时间显著延长（X2=7.47,P <0.05）。结论 四抗原融合蛋白能在小鼠体内诱导产生有效的体液免疫和细胞免疫反应，具有显著的抗弓形虫感染保护效果。

摘要： Objectives Given the efficacy of B-cell depletion therapy in systemic lupus erythematosus (SLE) treatment and the capacity of engineered natural killer (NK) cells in B-cell elimination, we explored the potential of genetically modified NK cells to target CD19 for the treatment of SLE. Methods Peripheral blood-derived NK cells were engineered with ***19/interleukin (IL)-15 (XB-19.15) or ***19 alone. In vitro assays tested cytotoxicity, proliferation (Ki-67), and cytokine release (IL-15/interferon-gamma [IFN-γ]] against CD19+ cells (Raji, Nalm6, SLE B-cell). In vivo models included Nalm6-luc xenografts, humanised CD34+ mice, and SLE peripheral blood mononuclear cells xenografts to assess efficacy, persistence, and safety. A phase 1 trial enrolled 3 patients with refractory SLE receiving XB-19.15 as a proof-of-concept study. Results XB-19.15 showed superior cytotoxicity, sustained IL-15 secretion, and enhanced proliferation in vitro . In the mouse model, XB-19.15 showed significant dose-dependent tumour regression of Nalm-6 and B-cell depletion of humanised mice with long-term persistence in various lymphoid organs. The total levels of hIgG and anti-dsDNA antibodies were decreased in XB-19.15-treated groups with deep clearance of B-cell and plasma cells of bone marrow and spleen in the SLE xenograft model, indicating potential attenuation of SLE. Three patients received XB-19.15 achieved improvement in disease activity and reset of B-cell repertoires without severe adverse events. Conclusions XB-19.15 enables potent, durable B-cell depletion and immune resetting in SLE with off-the-shelf utility and favourable safety. Preclinical and early clinical data support further trials to optimise dosing and confirm long-term efficacy.

摘要： Human genetics supports a causal involvement of IL-6 signaling in atherosclerotic cardiovascular disease, prompting the clinical development of anti-IL-6 therapies. Genetic evidence has historically focused on IL6R missense variants, but emerging cardiovascular treatments target IL-6, not its receptor, questioning the translatability of genetic findings. Here we develop a genetic instrument for IL-6 signaling downregulation comprising IL6 locus variants that mimic the effects of the anti-IL-6 antibody ziltivekimab and use it to predict the effects of IL-6 inhibition on cardiometabolic and safety endpoints. Similar to IL6R, we found that genetically downregulated IL-6 signaling via IL6 perturbation is associated with lower lifetime risks of coronary artery disease, peripheral artery disease and ischemic atherosclerotic stroke in individuals of European and East Asian ancestry. Unlike IL6R missense variants linked to bacterial infections, the IL6 instrument was associated with lower risk of pneumonia hospitalization. Our data suggest that IL-6 inhibition can reduce cardiovascular risk without major unexpected safety concerns.

关键词： BALT   bronchus-associated lymphoid tissue   Tregs   regulatory T cells   AEC   alveolar epithelail cells   SeV   Sendai virus   IAV   Influenza A virus   PR8   Puerto Rico 8   EGFR   epidermal growth factor receptor   dpi   days post-infection   ISHLT   International Society of Heart and Lung Transplantation   UMAP   Uniform Manifold Approximation and Projection   PBS   phosphate-buffered saline   NP   nucleocapsid protein   AUC   area under the curve   Krt   keratin   IL   interleukin   H&E   hematoxylin and eosin   PAS   periodic acid-Schiff   PCR   polymerase chain reaction  

摘要： Tolerance after lung transplantation is associated with the induction of Foxp3 + regulatory T cell-enriched bronchus-associated lymphoid tissue, which suppresses local and systemic alloimmune responses. How this tolerogenic graft environment shapes responses to respiratory viral infections, a known contributor to adverse outcomes after lung transplantation, remains unknown. Using a mouse model of a seasonally circulating parainfluenza virus, we found that acute infection of tolerant lung allografts results in temporary reductions of both bronchus-associated lymphoid tissue size and abundance of graft-resident Foxp3 + cells, but doesn’t trigger rejection. At late time points, pathological chronic type-2 inflammatory responses characteristic of severe parainfluenza virus infection were decreased and Krt5 + and Krt8 + lesions were reduced in tolerant allografts when compared to infected native lungs or syngeneic grafts. This reduction in dysplastic alveolar regeneration in tolerant allografts was associated with an increase in amphiregulin-expressing Foxp3 + cells. Type II alveolar epithelial cells in lung allografts express genes related to injury when recipient Foxp3 + cells cannot produce amphiregulin. These findings shed new light on how immune pathways that are established in tolerant lung allografts, in addition to preventing rejection, protect against aberrant tissue repair after a clinically relevant respiratory viral infection.

关键词： TNF   IL-17   psoriasis   skin   inflammation   neutrophils   fibroblasts   chemokines   transcriptomics   anti-TNF   CP: Immunology  

摘要： Inflammation at epithelial barriers involves a complex cell communication network, with subsets of fibroblasts increasingly recognized as active players in immune cell recruitment. To investigate the role of fibroblasts in interleukin (IL)-17-mediated skin diseases, we analyzed single-cell transcriptomic data from human psoriatic skin and identified tumor necrosis factor (TNF) recognition by TNF receptor 1 (TNFR1) as the most upregulated pathway in immune-acting fibroblasts. Functionally, TNF induced TNFR1-dependent neutrophil chemokine expression in cultured fibroblasts. Transcriptomics of skin from patients treated with TNF inhibitors revealed that expression of fibroblast-derived neutrophil chemokines depends on TNF. To test the significance of TNF recognition by fibroblasts, we developed mice lacking TNFR1 specifically in fibroblasts that showed impaired neutrophil recruitment to the skin after intradermal IL-17A and TNF injection and topical imiquimod treatment. This study establishes TNF signaling in immune-acting fibroblasts as a key driver of type 17 skin inflammation, challenging the prevailing view that TNF acts predominantly through keratinocytes.

关键词： FADD   RIPK1   TNFR1   extrinsic apoptosis   filament  

摘要： Extrinsic apoptosis is initiated by signaling from death receptors, leading to the assembly of RIPK1, FADD, and caspase-8 complex. Subsequently, caspase-8 forms a filamentous structure through the oligomerization of its tandem death effector domain (tDED), resulting in caspase activation and cell death. Although the DED of FADD (DED) is homologous to the tDEDs of caspase-8 (tDED) and both oligomerize to function, the functional form of DED oligomer in extrinsic apoptosis remains unclear. Here, using cryogenic-electron microscopy, we elucidate the structure of DED filaments comprising three helical chains assembled through three types of iterative interactions. Mutations disrupting DED filament formation impair the recruitment of RIPK1 and caspase-8, and abrogate the cell death response, suggesting that DED filamentation represents an important mechanistic step in the initiation of TNF-induced extrinsic apoptosis. Contrary to the belief that the homotypic death domains of RIPK1 and FADD are solely responsible for their interaction, we here show this interaction requires DED filamentation. Furthermore, cFLIP can disrupt DED filaments, uncovering an additional antiapoptotic mechanism of cFLIP beyond its disruption of caspase-8 filament. Molecular dynamics simulations reveal that DED filament thermodynamically favors tDED monomer over DED monomer, thus explaining the hierarchy and stoichiometry of FADD/caspase-8 complex assembly. These findings highlight the hitherto unappreciated roles of DED filament formation in extrinsic apoptosis.

关键词： ER stress   HIV-1 peptide pool   MHC-I   THP-1 monocytes   TPEN   zinc transporters  

摘要： Monocytes, upon activation, are known to produce substantial levels of cytokines and increased expression of MHC-I. The status of zinc and the nature of the stimulating agent significantly influence the release of various cytokines as well as the extent of MHC-I expression. However, research exploring the combined effects of inflammation and zinc deficiency remains limited. One of the primary challenges in investigating the influence of zinc status on cytokine production is the specificity of the cell types utilized in experimental settings. This study investigates zinc depletion and inflammation using THP-1 monocytes as an model, with HIV-1 viral peptide pools and TPEN (,,' ,'-Tetrakis (2-pyridylmethyl) ethylenediamine), an intracellular zinc chelator. Zinc depletion was confirmed through the application of zinquin ethyl ester fluorescence microscopy. Additionally, we assessed the expression of zinc transporters, Bcl2 family proteins, and caspase-3 using quantitative RT-PCR and Western blot analyses. The production of TNF-α was evaluated through Golgi-Stop experiments, while MHC-I levels were measured following an 8 h incubation with HIV-1 viral peptides. The impact of PHA on monocytes concerning cytokine production and MHC-I levels was also examined. Our results revealed a rapid increase in TNF-α production and elevated MHC-I levels within 8 h post-stimulation. Furthermore, we observed an enhancement of endoplasmic reticulum (ER) stress markers, including CHOP, ATF6, and PERK, as well as BCl2 family-related genes, following treatment with the peptide pool. These findings highlight the hyperactivation of monocytes in response to HIV-1 viral peptides, which may contribute to immune system impairment through caspase-mediated apoptosis.