Characterization of orange-spotted grouper (Epinephelus coioides) interferon regulatory factor 4 regulated by heat shock factor 1 during heat stress in response to antiviral immunity.

Interferon regulatory factor 4 (IRF4), in conjunction with thermogenic regulation, is a negative regulator of immune responses. Therefore, we examined whether temperature changes regulated the antiviral response of IRF4 in nervous necrosis virus (NNV)-infected orange-spotted groupers. We found that osgIRF4 mRNA expression was responsive to poly I:C stimulation and NNV infection. In vitro overexpression of osgIRF4 caused a marked decrease in the promoter activity of the antiviral protein Mx1, and magnified NNV replication. Notably, we showed that the IAD domain of osgIRF4 exerted a dominant inhibitory effect on the Mx1 promoter. Furthermore, on exposure to high temperatures, the action of osgIRF4 was dependent on heat shock factor 1 (HSF1) expression. Additionally, small interfering RNA knockdown of HSF1 abrogated high temperature-mediated osgIRF4 activity. These findings suggest that osgIRF4 is an essential negative regulator of innate antiviral immunity and enhances viral replication during heat stress in the orange-spotted grouper.

Investigation of Antigen-Specific T-Cell Receptor Clusters in Human Cancers.

PURPOSE: Cancer antigen-specific T cells are key components in antitumor immune response, yet their identification in the tumor microenvironment remains challenging, as most cancer antigens are unknown. Recent advance in immunology suggests that similar T-cell receptor (TCR) sequences can be clustered to infer shared antigen specificity. This study aims to identify antigen-specific TCRs from the tumor genomics sequencing data. EXPERIMENTAL DESIGN: We used the TRUST (Tcr Repertoire Utilities for Solid Tissue) algorithm to assemble the TCR hypervariable CDR3 regions from 9,700 bulk tumor RNA-sequencing (RNA-seq) samples, and developed a computational method, iSMART, to group similar TCRs into antigen-specific clusters. Integrative analysis on the TCR clusters with multi-omics datasets was performed to profile cancer-associated T cells and to uncover novel cancer antigens. RESULTS: Clustered TCRs are associated with signatures of T-cell activation after antigen encounter. We further elucidated the phenotypes of clustered T cells using single-cell RNA-seq data, which revealed a novel subset of tissue-resident memory T-cell population with elevated metabolic status. An exciting application of the TCR clusters is to identify novel cancer antigens, exemplified by our identification of a candidate cancer/testis gene, HSFX1, through integrated analysis of HLA alleles and genomics data. The target was further validated using vaccination of humanized HLA-A*02:01 mice and ELISpot assay. Finally, we showed that clustered tumor-infiltrating TCRs can differentiate patients with early-stage cancer from healthy donors, using blood TCR repertoire sequencing data, suggesting potential applications in noninvasive cancer detection. CONCLUSIONS: Our analysis on the antigen-specific TCR clusters provides a unique resource for alternative antigen discovery and biomarker identification for cancer immunotherapies.

Anti-inflammatory effect on genes expression after four days of Qigong training in peripheral mononuclear blood cells in healthy women.

INTRODUCTION: Some studies have shown the influence of Qigong on gene expression in different cells, but there is little data associated with the influence of this kind of therapy on genes expression in pheripheral monocellucar blood cells. OBJECTIVE: The aim of this study was to evaluate changes in the expression of genes associated with cellular stress response in peripheral mononuclear blood cells (PMBC) in healthy women. MATERIAL AND METHODS: The experiment took place at the Japanese Martial Arts Centre "DOJO" in Stara Wies, Poland, conducted over the course of a 4-day qigong training session. To evaluate the genes effect of this training, blood samples were taken before and after the training period. This experiment involved 20 healthy women (aged 56.2±9.01, body height 164.8±6.5 and mass 65.5±8.2). To determine the expression of HSF-1, HSPA1A, NF-kB, IL10 and CCL2 mRNA, 3 ml of venous blood was collected. The blood samples were placed in tubes allowing for separation (BD Vacutainer CPT TM) before and after the 4 days of qigong training. Isolated PMBC were used to determine gene expression using real-time qRT-PCR (quantitative reverse transcription polymerase chain reaction). RESULTS: Significant decreases in NF-kB and CCL2 mRNA and increases in IL10, HSF1 and HSPA1A m-RNA were detected after 4 days of qigong training. The obtained findings suggest that qigong caused a reduction in the inflammatory and intensified anti-inflammatory gene expression, as well as a higher expression of HSF-1 and HSPA1A. CONCLUSIONS: The adaptive response to qigong training was similar to the adaptive response to physical activity and was detected through gene expression in PMBC. Furthermore, this kind of training is especially indicated for women because of their higher susceptibility to psychosocial stress when compared to men.

NF­κB regulates HSF1 and c­Jun activation in heat stress­induced intestinal epithelial cell apoptosis.

Heat stress may induce intestinal epithelial cell apoptosis; however, the molecular mechanisms have not yet been identified. The present study used IEC­6 rat small intestinal epithelial cells to investigate heat stress­induced production of reactive oxygen species (ROS), which may be involved in nuclear factor (NF)­κB activation during heat stress. IEC­6 cells were transfected with NF­κB p65­specific small interfering RNA (siRNA), and observed a significant increase in cell apoptosis and caspase­3 cleavage; however, in cells transfected with adenovirus that constitutively overexpressed p65, the opposite results were obtained. Furthermore, p65 knockdown increased the heat stress­induced expression and activity of heat shock transcription factor 1 (HSF1); conversely, p65 overexpression slightly decreased HSF1 activity. The levels of heat stress­induced c­Jun phosphorylation were also examined: Knockdown of p65 resulted in a reduction of c­Jun phosphorylation, whereas p65 overexpression resulted in increased phosphorylation. Furthermore, siRNA­mediated knockdown of HSF1 in IEC­6 cells significantly increased heat stress­induced apoptosis. Cells pretreated with c­Jun peptide, an inhibitor of c­Jun activation, exhibited a significant reduction in apoptosis. These findings indicated that heat stress stimulation in IEC­6 cells induced the pro­apoptotic role of NF­κB by regulating HSF1 and c­Jun activation.

Autoantibodies against HSF1 and CCDC155 as Biomarkers of Early-Stage, High-Grade Serous Ovarian Cancer.

Background: Tumor-directed circulating autoantibodies (AAb) are a well-established feature of many solid tumor types, and are often observed prior to clinical disease manifestation. As such, they may provide a good indicator of early disease development. We have conducted a pilot study to identify novel AAbs as markers of early-stage HGSOCs.Methods: A rare cohort of patients with early (FIGO stage Ia-c) HGSOCs for IgG, IgA, and IgM-mediated AAb reactivity using high-content protein arrays (containing 9,184 individual proteins). AAb reactivity against selected antigens was validated by ELISA in a second, independent cohort of individual patients.Results: A total of 184 antigens were differentially detected in early-stage HGSOC patients compared with all other patient groups assessed. Among the six most highly detected "early-stage" antigens, anti-IgA AAbs against HSF1 and anti-IgG AAbs CCDC155 (KASH5; nesprin 5) were significantly elevated in patients with early-stage malignancy. Receiver operating characteristic (ROC) analysis suggested that AAbs against HSF1 provided better detection of early-stage malignancy than CA125 alone. Combined measurement of anti-HSF1, anti-CCDC155, and CA125 also improved efficacy at higher sensitivity.Conclusions: The combined measurement of anti-HSF1, anti-CCDC155, and CA125 may be useful for early-stage HGSOC detection.Impact: This is the first study to specifically identify AAbs associated with early-stage HGSOC. The presence and high frequency of specific AAbs in early-stage cancer patients warrants a larger scale examination to define their value for early disease detection at primary diagnosis and/or recurrence. Cancer Epidemiol Biomarkers Prev; 27(2); 183-92. ©2017 AACR.