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关键词： 组织金属蛋白酶抑制因子1   白细胞介素1β   增生性瘢痕  

摘要： 目的 探讨TIMP1和IL-1β在增生性瘢痕组织中的表达情况及相关性。方法 选取6例正常瘢痕和6例增生性瘢痕组织，HE染色观察瘢痕组织病理学改变，免疫组织化学染色检测TIMP1在组织中的表达情况，Westernblot和ELISA方法检测TIMP1和IL-1β在组织中的表达情况，通过皮尔逊相关系数分析TIMP1和IL-1β在组织中的相关性。结果 增生性瘢痕组织中的胶原纤维密度增高，且排列紊乱，并且成纤维细胞和炎症细胞显著上调；TIMP1和IL-1β在增生性瘢痕组织中表达显著高于正常组织，且二者表达呈正相关。结论TIMP1和IL-1β在增生性瘢痕组织中表达上调，并且二者表达呈正相关。

关键词： FLIP   L   caspase-8   protease activity   TNF   necroptosis   CP: Molecular biology  

摘要： The pro-form of caspase-8 prevents necroptosis by functioning in a proteolytically active complex with its catalytic-dead homolog, FLICE (FADD [Fas-associated death domain]-like interleukin 1b-converting enzyme)-like inhibitory protein long-form (FLIPL). However, how FLIPL imparts caspase-8 the catalytic activity to suppress necroptosis remains elusive. Here, we show that the protease-like domain of FLIPL is essential for the activity of the caspase-8-FLIPL heterodimer in blocking necroptosis. While substitution of two amino acids whose difference may contribute to the pseudo-caspase property of FLIPL with the corresponding amino acids in caspase-8 does not restore the protease activity of FLIPL, one of the amino acid replacements, tyrosine (Y) 362 to cysteine (C), is sufficient to completely abolish the activity of the caspase-8-FLIPL heterodimer in cleaving receptor-interacting protein 1 (RIP1), thus releasing the necroptosis blockade. Unconstrained necroptosis is observed in embryonic day (E)10.5-E11.5 embryos of FLIPL-Y362C knockin mice. Collectively, these results reveal that the protease-like domain of FLIPL has a special structure that imparts the pro-caspase-8-FLIPL heterodimer a unique catalytic activity toward RIP1 to prevent necroptosis.

关键词： regulatory T cells   IL-2   CTLA-4   anergy   tolerance   CP: Immunology  

摘要： Interleukin-2 (IL-2) variants with increased CD25 dependence that selectively expand Foxp3+ regulatory T (TR) cells are in clinical trials for treating inflammatory diseases. Using an Fc-fused IL-2 mutein (***-2 mutein) we developed that prevents diabetes in non-obese diabetic (NOD) mice, we show that ***-2 mutein induced an activated TR population with elevated proliferation, a transcriptional program associated with Stat5- and T cell receptor-dependent gene modules, and high IL-10 and CTLA-4 expression. Increased IL- 10 signaling limited surface major histocompatibility complex class II upregulation during conventional dendritic cell (cDC) maturation, while increased CTLA-4-dependent transendocytosis led to the transfer of CD80 and CD86 co-stimulatory ligands from maturing cDCs to TR cells. In NOD mice, ***-2 mutein treatment promoted the suppression of cDCs in the inflamed pancreas and pancreatic lymph nodes, resulting in T cell anergy. Thus, IL-2 mutein-expanded TR cells have enhanced functional properties and restrict cDC function, offering promise for targeted immunotherapy use in autoimmune disease.

关键词： 无细胞百日咳疫苗   脱毒   效价   细胞免疫  

摘要： 目的：探索无细胞百日咳疫苗中FHA和PRN组分脱毒处理前后的理化及免疫效果差异。方法：采用差示扫描荧光法对脱毒前后的FHA/PRN进行热稳定性分析，评估脱毒步骤对蛋白Tm值的影响；采用小鼠脑腔攻击实验（MICA法）、百日咳血清学效力检测（PSPT）方法评估脱毒前后FHA/PRN制成单组分或与PT制成三组分疫苗的保护效力；采用ELISpot以及流式细胞术评估疫苗样品免疫小鼠脾脏中细胞因子及免疫细胞差异。结果：未脱毒FHA的Tm值约57℃，脱毒FHA具有两个Tm值，分别约58℃及81℃；脱毒前后PRN的Tm值均约84℃。脱毒步骤没有显著影响疫苗样品的保护效力及小鼠脾脏中IL-4、IFN-γ细胞因子分泌。此外，脱毒步骤对小鼠脾脏中CD4+T细胞、CD8+T细胞、CD44+CD62L+记忆性T细胞水平也无显著影响。结论：无细胞百日咳疫苗中FHA和PRN组分脱毒处理仅对FHA的Tm值有影响，没有影响疫苗的免疫保护效果。

关键词： cancer vaccine   dendritic cell   IL-2  

摘要： One driver of the high failure rates of clinical trials for therapeutic cancer vaccines is likely the inability to sufficiently engage conventional dendritic cells (cDCs), the antigen-presenting cell (APC) subset that is specialized in priming antitumor T cells. Here, we demonstrate that, relative to vaccination with an injectable mesoporous silica rod (MPS) vaccine alone (Vax), combining MPS vaccines with CD122-biased IL-2/anti-IL-2 antibody complexes (IL-2cx) drives ~3-fold expansion of cDCs at the vaccination sites, vaccine-draining lymph nodes, and spleens of treated mice. Furthermore, relative to Vax alone, Vax+IL-2cx led to a ~3-fold increase in the numbers of CD8+ T cells and ~15-fold increase in the numbers of NK cells at the vaccination site. Notably, with both the model protein antigen OVA as well as various peptide neoantigens, Vax+IL-2cx induced ~5 to 30-fold greater numbers of circulating antigen-specific CD8+ T cells relative to Vax alone. We further demonstrate that Vax+IL-2cx leads to significantly improved efficacy in the MC38 colon carcinoma model relative to either monotherapy alone, driving complete regressions in 50% of mice in a cDC-dependent manner. Relative to vaccine alone, Vax+IL-2cx led to comparable numbers of CD8+ T cells, but markedly greater numbers of NK cells and activated cDCs in the B16F10 melanoma tumor microenvironment post-therapy. Taken together, these findings suggest that the administration of factors that engage both the cDC-CD8+ T cell and cDC-NK cell axes can boost the potency of therapeutic cancer vaccines. Copyright © 2024 the Author(s).

关键词： Cancer immunotherapy   Hematology   Immunology   Leukemias   T cells  

摘要： Despite the revolutionary achievements of chimeric antigen receptor (CAR) T cell therapy in treating cancers, especially leukemia, several key challenges still limit its therapeutic efficacy. Of particular relevance is the relapse of cancer in large part, as a result of exhaustion and short persistence of CAR-T cells in vivo. IL-2-inducible T cell kinase (ITK) is a critical modulator of the strength of T-cell receptor (TCR) signaling, while its role in CAR signaling is unknown. By electroporation of clustered regularly interspaced short palindromic repeats (CRISPR) associated protein 9 (Cas9) ribonucleoprotein (RNP) complex into CAR-T cells, we successfully deleted ITK in CD19-CAR-T cells with high efficiency. Bulk and single-cell RNA sequencing (scRNA-seq) analyses revealed down-regulation of exhaustion and up-regulation of memory gene signatures in ITK-deficient CD19-CAR-T cells. Our results further demonstrated a significant reduction of T cell exhaustion and enhancement of T cell memory, with significant improvement of CAR-T cell expansion and persistence both in vitro and in vivo. Moreover, ITK-deficient CD19-CAR-T cells showed better control of tumor relapse. Our work provides a promising strategy of targeting ITK to develop sustainable CAR-T products for clinical use.

关键词： HLA-immunized platelet transfusion refractoriness   HLA-matched PLT units   non-immune factors  

摘要： Immunized platelet transfusion refractoriness (IPTR) has a high incidence of death and complications. HLA-matched platelet unit can effectively treat HLA-immunized PTR. A cohort study was undertaken on nine patients with IPTR hematological disorders who underwent HLA-matched PLT units due to HLA immunization at the First Affiliated Hospital of Soochow University between April 2022 and April 2023. We calculated the 14-hour corrected count increments (14h-CCI) to evaluate the effect of PLT transfusions. A 14h- CCI > 5000 was considered to be a successful transfusion. A total of 113 PLT units were transfused to the nine patients with HLA-immunized PTR. Of the 113 PLT units, 50 were random, 34 were cross-matched, and 29 were HLA-matched. The median 14h-CCI values were 1683 for random PLT units, 5246 for cross-matched PLT units, and 5643 for HLA-matched PLT units (P = 0.02). Moreover, 10, 25, and 43.8% of transfusions were successful for random, cross-matched, and HLA-matched PLT units with non-immune factors (P = 0.013). Regarding non-immune factors, we confirmed that random PLT units, infection, splenomegaly, and bleeding affected platelet transfusion increments (P < 0.05).

关键词： H6亚型禽流感病毒   HA蛋白   单克隆抗体   无HI   中和活性   保护性  

摘要： 为研制抗H6禽流感病毒HA蛋白单克隆抗体，本研究利用小鼠适应毒株E-Teal/417（MA E-Teal/417）H6禽流感病毒免疫BALB/c小鼠，利用小鼠杂交瘤细胞技术，经间接免疫荧光试验（IFA）和血凝抑制（HI）试验等方法筛选和亚克隆，获得 4株没有HI效价的单克隆抗体，分别命名为1D4、4A7、4G2和5E9。IFA结果表明，4株单克隆抗体抗体均能识别转染pDP2002-HA（MA E-Teal/417）质粒的293T细胞中表达的HA蛋白。Western blot试验表明，单克隆抗体4G2和5E9能识别感染MA E-Teal/417病毒的MDCK细胞中表达的HA蛋白。中和试验结果发现，单克隆抗体1D4、4A7和4G2具有一定的体外中和活性，抑制MA E-Teal/417在MDCK细胞中感染和复制。小鼠保护性试验进一步表明，单克隆抗体4A7能提供良好的被动免疫抵抗致死性MA E-Teal/417感染。以上获得的4组无血凝抑制活性、但具有一定中和活性的抗HA单克隆抗体为H6禽流感病毒HA蛋白新型中和表位解析、疫苗设计及诊断技术开发等提供生物材料。

关键词： RSV   bronchiolitis   ALRI   asthma   microbiome   IL-6   trans   -signaling   neutrophils   maternal diet   liver   low grade systemic inflammation   CP: Immunology   CP: Metabolism  

摘要： A poor maternal diet during pregnancy predisposes the infant to severe lower respiratory tract infections (sLRIs), which, in turn, increases childhood asthma risk;however, the underlying mechanisms remain poorly understood. Here, we show that the offspring of high-fat diet (HFD)-fed mothers (HFD-reared pups) developed an sLRI following pneumovirus inoculation in early life and subsequent asthma in later life upon allergen exposure. Prior to infection, HFD-reared pups developed microbial dysbiosis and low-grade systemic inflammation (LGSI), characterized by hyperneutropoiesis in the liver and elevated inflammatory cytokine expression, most notably granulocyte-colony stimulating factor (G-CSF), interleukin-17A (IL-17A), IL-6 and soluble IL-6 receptor (sIL-6R) (indicative of IL-6 trans-signaling) in the circulation and multiple organs but most prominently the liver. Inhibition of IL-6 trans-signaling using sgp130Fc transgenic mice or via specific genetic deletion of IL-6Ra on neutrophils conferred protection against both diseases. Taken together, our findings suggest that a maternal HFD induces neonatal LGSI that predisposes to sLRI and subsequent asthma via neutrophil-mediated IL-6 trans-signaling.

关键词： Mathematical model   Tumor immunity   PD-1/PD-L1   Cytokines   Immunotherapy  

摘要： The tumor microenvironment constitutes a complex system shaped by the intricate interactions among tumor cells, immune cells, and cytokines. Within this environment, the interplay between immune cells and cytokines is crucial in influencing tumor growth and progression. Despite advancements in clinical tumor immunotherapy, there remains a gap in comprehensive simulations of tumor immune responses, particularly regarding cytokine-driven processes. This study aims to address this gap by investigating the regulatory interactions among tumor cells, immune cells, and cytokines to simulate the complexities of tumor immunotherapy. We develop a comprehensive modeling and computational framework incorporating PD-1 inhibitors and interleukin-10 (IL-10) antibodies. Through detailed mathematical analysis, we elucidate the impact of changes in the immune microenvironment on tumor cells number. Our findings highlight the significant therapeutic effect of anti-PD-1 and IL-10 inhibitors, with increased drug dosage correlating with a reduction in tumor burden. Furthermore, combination therapy demonstrates a marked extension of survival with reduced dosages compared to monotherapy. Based on model simulations, we proposed prognostic predictions by assessing the microenvironmental status before treatment. The findings indicate a promising method for enhancing treatment effectiveness and offering potential advantages to patients receiving tumor immunotherapy.