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关键词： antibody   donors   female   HLA   low titer group O whole blood   parous   testing   transfusion-associated acute lung injury  

摘要： BackgroundAs low titer group O whole blood (LTOWB) increases in popularity, blood centers are finding innovative ways of maintaining the supply. One potential way is collecting LTOWB from parous female donors without testing for HLA antibodies. This in silico simulation predicted the risk of an LTOWB unit containing an HLA antibody and the subsequent risk for an HLA-incompatible *** LTOWB blood bank with 1 million units was simulated consisting of male, nulliparous, and parous female donors. The proportion of each donor type was modeled after the sex distribution at US blood centers. The parity of female donors was calculated based on the average number of live births per female depending on her age. HLA-alloimmunization risk was determined by her parity status. The HLA haplotypes of the simulated recipients were derived from the 100 most common HLA haplotypes in the US National Marrow Registry Program database. The proportion of different race/ethnic groups in the US was used to simulate 100,000 LTOWB recipients to whom between 1 and 10 units were ***, the risk of an LTOWB unit containing at least one HLA antibody was 12.2% and the rate of receiving an HLA-incompatible unit was 21.3%. The risk of receiving an HLA-incompatible unit rose from 4.8% after receipt of one unit to 36.5% after 10 *** collectors and hospitals should evaluate the potential TRALI risk against the benefit of a potentially expanded inventory of LTOWB before collecting plasma-containing products from non-HLA-tested parous female donors.

关键词： GBM   Immunotherapy   IL12   INCR1   PD-L1  

摘要： PurposeThe potent antitumor effects of interleukin 12 (IL12) gene therapy in glioblastoma (GBM) are significantly attenuated by the highly immunosuppressive microenvironment and the upregulation of the PD-1/PD-L1 immune checkpoint. However, combining IL12 gene therapy with PD-1/PD-L1 inhibitors failed to improve efficacy. This study aims to assess the effects of silencing the immunosuppressive long noncoding RNA INCR1 when combined with IL12 *** in situ hybridization was performed to analyze INCR1 and PD-L1 expression in tumor tissues from GBM patients pre- and post-IL12 gene therapy. Quantitative PCR was used to analyze immunosuppressive gene expression in patient-derived GBM cells co-cultured with immune cells stimulated with IL12. The effects of INCR1 and PD-L1 silencing on the expression of immunosuppressive genes were evaluated by RNA sequencing. 3D-cytotoxicity assays were performed to assess the activity of immune cells against GBM tumor ***1 and PD-L1 expression was upregulated in tumor tissue from GBM patients treated with IL12 gene therapy compared to the tumor tissue of the same patients before the IL12 treatment. Co-culture of patient-derived GBM cells with IL12-stimulated immune cells increased the expression of several immunosuppressive genes. Knocking down INCR1 was more effective than silencing PD-L1 in reducing the expression of multiple immunosuppressive genes. INCR1 silencing improved IL12-mediated immune cell antitumor activity compared to monoclonal antibodies targeting the PD-1/PD-L1 immune checkpoint ***1 silencing affects more immune evasive pathways than PD-L1. Targeting INCR1 may represent a valid approach to improve the efficacy of IL12 therapy in GBM.

关键词： HLA   HLA-DRB1*07:60:02   new allele   next generation sequencing  

摘要： HLA-DRB1*07:60:02 differs from HLA-DRB1*07:01:01:01 by one nucleotide substitution at codon 44 in exon 2.

关键词： Coxsackievirus A16   hSCARB2 mice   Nerve injury   TLR2/MYD88/TNF-alpha/Cxcl1 signaling pathway  

摘要： Coxsackievirus A16 (CVA16), a major pathogen responsible for hand-foot-and-mouth disease (HFMD) in children, has frequently replaced Enterovirus A71 as the predominant causative agent in China and other Asia-Pacific regions. The lack effective drugs and vaccines against this virus exacerbates the concerns on its outbreaks. Clinical reports and laboratory studies indicate that CVA16 infection may lead to neurological injury, but the precise mechanisms remain elusive. In this study, we meticulously established a CVA16 murine disease model using 3-week-old hSCARB2 knock-in mice through intracranial inoculation. Within 4-7 days post-infection, the infected mice exhibited severe neurological symptoms featured as limb paralysis, hind limb weakness and ataxia. Furthermore, high viral loads were detected in the brain, spleen, skeletal muscle tissues, indicating a systemic infection. A robust cytokine response was observed, characterized with the elevation of TNF-alpha, IL-12 (p40), IL-10 and MIP-1 beta. Histological and immunofluorescence staining revealed extensive inflammation, marked by the concentrated infiltration of astrocytes cells, as well as severe neurological injury, which included hypertrophic and extended pseudopodia microglia, increased astrocytes with long and stretched protuberances, markedly decreased neuronal cell bodies and nerve fibers in brain. No visible pathological changes were observed in spinal cord tissues. RNA sequencing and immunofluorescence staining of brain tissue verification assays indicated that the neurological injury may engage in TLR2/MYD88/TNF-alpha/CXCL1 signal pathway. Over all, this work addressed the gap in the availability of CVA16 disease rodent model for vaccine development and provided novel insights into the mechanisms underlying neurological injury caused by enteroviruses and other neurotropic viruses.

关键词： Inflammatory bowel disease   Interleukin-1 beta   Inflammasome   Calcium-sensing receptor   Estrogen receptor beta  

摘要： Introduction: Although several estrogen receptor (3 (ER(3) agonists have been reported to alleviate IBD, the pivotal mechanism remains obscure. Objectives: To examine the effects and mechanisms of ER(3 activation on cytokine/chemokine networks in colitis mice. Methods: Dextran sulfate sodium salt (DSS) and trinitro-benzene-sulfonic acid (TNBS) were used to induce mouse colitis model. Multiple molecular biological methods were employed to evaluate the severity of mouse colitis and the level of cytokine and/or chemokine. Results: Bioinformatics analysis, ELISA and immunofluorescence results showed that the targeted cytokines and/or chemokines associated with ER(3 expression and activation is IL-1(3, and the anti-colitis effect of ER(3 activation was significantly attenuated by the overexpression of AAV9-IL-1(3. Immunofluorescence analysis indicated that ER(3 activation led to most evident downregulation of IL-1(3 expression in colonic macrophages as compared to monocytes and neutrophils. Given the pivotal roles of NLRP3, NLRC4, and AIM2 inflammasome activation in the production of IL-1(3, we examined the influence of ER(3 activation on inflammasome activity. ELISA and WB results showed that ER(3 activation selectively blocked the NLRP3 inflammasome assembly-mediated IL-1(3 secretion. The calcium-sensing receptor (CaSR) and calcium signaling play crucial roles in the assembly of the NLRP3 inflammasome. WB and immunofluorescence results showed that ER(3 activation reduced intracellular CaSR expression and calcium signaling in colonic macrophages. Combination with CaSR overexpression plasmid reversed the suppressive effect of ER(3 activation on NLRP3 inflammasome assembly, and counteracting the downregulation of IL-1(3 secretion. Conclusion: Our research uncovers that the anti-colitis effect of ER(3 activation is accomplished through the reduction of IL-1(3 levels in colonic tissue, achieved by specifically decreasing CaSR expression in macrophages to lower intracellular

关键词： multiomics   proteomics   septic cardiomyopathy   transcriptomics  

摘要： Background: Septic cardiomyopathy (SCM) is a potentially fatal complication of sepsis. In this study, transcriptomic and proteomic analyzes of serum samples from sepsis patients were conducted to uncover the underlying pathological mechanisms and identify potential therapeutic targets for SCM. Methods: This retrospective, dual-center study investigated the progression of sepsis to SCM in patients admitted to intensive care units. A total of 50 patients were enrolled and divided into two groups: sepsis with cardiomyopathy (25 cases) and sepsis without cardiomyopathy (25 cases). Co-expression network analysis was employed to elucidate the biological significance of differentially expressed proteins. By integrating proteomic and transcriptomic data, molecular networks were constructed to visualize interactions among key molecules, aiming to enhance data interpretation and support the study's findings. Results: Proteomic analysis identified 216 differentially expressed proteins (Fold change > 1.5, p-value < 0.05) between the two groups. Transcriptomic analysis revealed two proteins, including Interleukin-27 subunit beta (IL-27B) and carbonic anhydrase, co-downregulated in patients with septic cardiomyopathy. IL-27B was associated with the immune response, and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis indicated its involvement in the cytokine-cytokine receptor interaction signaling pathway. Conclusion: Comprehensive integrated transcriptomic and proteomic analyzes identified significant changes in protein expression associated with SCM, primarily associated with inflammation-related pathways and amino acid metabolism. These findings provide new insights into the pathological mechanisms of SCM and highlight potential therapeutic targets for its treatment.

关键词： MAVS   Interferon   Intronic enhancer   IRFs   Large yellow croaker (Larimichthys crocea)  

摘要： The mitochondrial antiviral signaling protein (MAVS) relays signals from RIG-I-like receptors (RLRs) to induce type I interferon (IFN) production. In teleost fish, MAVS expression is significantly upregulated in response to viral infections or synthetic double-stranded RNA (dsRNA), whereas mammalian MAVS does not exhibit a similar response. However, the mechanisms regulating MAVS expression in teleosts remain unclear. In this study, we demonstrate that the viral mimic poly(I:C)-induced upregulation of Larimichthys crocea (Lc) MAVS occurs via the type I IFN signaling pathway. Inhibition of the JAK-STAT pathway significantly suppressed both poly(I:C)- and LcIFNi-induced LcMAVS expression. Further analysis revealed that an enhancer in the 5 '- untranslated region (UTR) intron of LcMAVS contains two functional interferon-stimulated response elements (ISREs), which are crucial for its activation. The enhancer activity of LcMAVS is regulated by interferon regulatory factors (IRFs), including IRF1, IRF3, IRF7, IRF9, and IRF11. These IRFs form several heterodimeric complexes, such as IRF1/3, IRF1/7, IRF3/7, and IRF3/11, to mediate LcMAVS enhancer activation. Structural analysis indicates that the ISRE motifs in the intronic enhancer can accommodate two or three DNA-binding domains (DBDs) from IRFs. These findings provide a potential explanation for the differential regulation of MAVS in response to stimuli in teleosts and mammals. Furthermore, our study demonstrates that MAVS is an interferon-stimulated gene (ISG) in a marine fish, providing insights into the evolutionary divergence of the vertebrate RLR signaling pathway.

关键词： Target Cells   Cytotoxic Assays   PHA Blasts  

摘要： Background & AimPeptide-pulsed PHA blasts (T cells activated with mitogen phytohemagglutinin) are often used as target cells in assays evaluating the function of antigen-specific cytotoxic T lymphocytes (CTLs). While CD8+ CTLs can recognize antigen in the context of HLA Class I, which is constitutively expressed on all T cells, CD4+ CTLs recognize antigen presented on HLA Class II, which is not expressed on the quiescent T cells. Activation of T cells causes transient upregulation of HLA Class II (HLA-DR), and the extent of this upregulation may affect the effectiveness of PHA blasts as targets for CD4+ CTLs. We explored the dynamics of HLA-DR expression on PHA blasts over time to determine the optimum time for using PHA blasts in CTL manufacturing/testing workflow. MethodologyPeripheral Blood Mononuclear Cell (PBMC) obtained from 4 healthy volunteers were stimulated with phytohemagglutinin (5 ug/mL) in the presence of IL-2 (50 IU/mL) over the course of 2 weeks. The trends in HLA-DR expression were assessed by flow cytometry. The effect of HLA-DR expression on antigen presentation by the PHA was assessed by exposing CTLs to peptide-loaded PHA with high and low HLA-DR expression, and comparing antigen reactivity (measured as % of IFN-g releasing cells). ResultsThe expression of HLA-DR was 3.6% (SD 1.3) at baseline, upregulated to the maximum (54.8%, SD 12.2) by day 7-10, and fell to 21.8 (SD 10.94) by day 11-14 (fig. 1A). Restimulation at Day 10 helped maintain, on average, 19.7% higher level of HLA expression (SD 4.9) for up to 7 additional days (fig. 1B). HLA-DR expressing T cells were predominantly CD8 (mean CD4:CD8 ratio 0.23, SD 0.03, fig. 2). Freezing and thawing of PHA blasts did not have a significant effect on the level of HLA-DR expression (49.0 vs 53.7%). PHA expressing high level HLA-DR were found to elicit higher CTL reactivity than PHA with low HLA-DR expression (0.22% for PHA with 37% HLA-DR vs 0.10% for PHA with 12% HLA-DR). ConclusionThe upregulation

关键词： Children   Complex febrile seizures   Neutrophils/lymphocyte ratio   Interleukin   Viral epidemic  

摘要： This report aimed to analyze the potential effects of cytokines and neutrophils-to-lymphocytes ratio (NLR) on the occurrence of febrile seizures (FS) in children during the epidemic of novel coronavirus and influenza virus. Between July 2022 and April 2023, clinical data of 422 children with FS hospitalized in the Pediatrics, Shaoxing People's Hospital and the First Affiliated Hospital of Shaoxing University, during the epidemic and non-epidemic periods of novel coronavirus and influenza virus were analyzed. By comparing the various clinical characteristics of children with FS at different periods, comparison of variations in peripheral blood cell count, neutrophils, lymphocytes, hypersensitive C-creation protein, NLR, and inflammatory cytokines. Among 422 cases, 130 cases were classified as complex FS (CFS), while 292 cases of simple FS (SFS) were identified. During the time of the viral epidemic, 315 FS cases were observed, among which the incidence of CFS was 33.65%. In the non-viral epidemic period, there were 105 cases of FS. The incidence of CFS accounted for 22.86%. In addition, the levels of IL-4, IL-10, and tumor necrosis factor alpha (TNF-alpha) in children in the viral epidemic period were significantly higher than those in the non-epidemic period (P < 0.05). ROC curve analysis revealed that during the epidemic period, NLR (AUC = 0.724), IL-4 (AUC = 0.826), and IL-10 (AUC = 0.688) exhibited high predictive value for CFS children, and the indexes were statistically significant (P < 0.05). This study suggests that IL-4, IL-10, and NLR may play an important role in the pathogenesis of children with FS, especially those with CFS.