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Identification of regulatory CD8+ T cells that suppress pathological immune responses is an importunate pursuit. In a recent issue of Science , Li et al. demonstrated that human KIR+CD8+ T cells suppress autoimmunity by eliminating pathogenic CD4+ T cells. [ABSTRACT FROM AUTHOR]

Background: Cytochrome P450 (CYP) isoenzymes are an important phase I enzyme system. In the present study, we investigated the effects of Oenanthe javanica (Blume) DC on CYP1A1 and CYP1A2. Findings: Whole plants were completely dried and then divided into leaves, stems, and roots for extraction. The human liver hepatocellular carcinoma cell line HepG2 was treated with ethanol extracts of these organs for 72 h and mRNA and protein expression levels were assessed. The root extract of O. javanica significantly elevated the expression of both CYP1A1 and CYP1A2 mRNAs (by 68 and 102 %, respectively). Similarly, the CYP1A1 and CYP1A2 protein levels were increased by the root extract (by 112 and 157 %, respectively). The effects of the root extract were much more pronounced than those of leaf and stem extracts. Subsequent GC-MS analysis revealed that the levels of major coumarin derivatives, xanthotoxin, bergapten, and isopimpinellin, were significantly higher in O. javanica root extracts than in leaf or stem extracts. Of note, 5 μM xanthotoxin (the most abundant furanocoumarin in O. javanica) induced the expression of CYP1A1 mRNA as well as CYP1A2 mRNA and protein, albeit the CYP1A1 protein level was elevated only at 10 μM xanthotoxin. Conclusions: Although it is difficult to extrapolate such effects to metabolic outcomes because of the inherent limitations of in vitro experiments, it is important to note that dietary exposure to O. javanica may modulate phase I enzymes and thereby affect various xenobiotic metabolism. [ABSTRACT FROM AUTHOR]

Few studies have investigated Seomae mugwort (a Korean native mugwort variety of Artemisia argyi H. Lév. & Vaniot), exclusively cultivated in the southern Korean peninsula, and the possibility of its use as a food resource. In the present study, we compared the nutritional and chemical properties as well as sensory attributes of Seomae mugwort and the commonly consumed species Artemisia princeps Pamp. In comparison with A. princeps, Seomae mugwort had higher contents of polyunsaturated fatty acids, total phenolic compounds, vitamin C, and essential amino acids. In addition, Seomae mugwort had better radical scavenging activity and more diverse volatile compounds than A. princeps as well as favorable sensory attributes when consumed as tea. Given that scant information is available regarding the Seomae mugwort and its biological, chemical, and sensory characteristics, the results herein may provide important characterization data for further industrial and research applications of this mugwort variety. [ABSTRACT FROM AUTHOR]

Here, monophosphoryl lipid A (MPLA) and aluminum salt (Alum) were introduced into a hyaluronic acid (HA)-based combination vaccine adjuvant for hepatitis B vaccine (HBV). Although Alum is a well-known hepatitis B vaccine adjuvant that induces an enhanced humoral immune response, it cannot induce the cellular immune responses. On the other hand, MPLA has been generally reported to promote IFN-γ production via antigen-specific CD4 + T cells, but it is not water soluble as a result of its long hydrophobic alkyl chains. To this end, water insoluble MPLA could be solubilized in an aqueous solution with the help of HA, which contains many carboxyl and hydroxyl groups that can be used to attach to the hydroxyl head groups of MPLA via hydrogen bonds. Three groups of mice were treated with either hepatitis B surface antigen (HBsAg) alone, HBsAg_Alum complex, or HBsAg_Alum_MPLA/HA complex. The group immunized with the HBsAg_Alum_MPLA/HA complex exhibited a high increase in cellular immune response as well as in humoral immune response relative to the other two groups. The antibody, cytokine and T cell levels were most elevated in the group of mice immunized with HBsAg_Alum_MPLA/HA complex, even at a 1 μg/mice dose, and the magnitude was still maintained even after 8 weeks. Specifically, the antibody value was 120 times larger in mice vaccinated with HBsAg_Alum_MPLA/HA complex than in mice vaccinated with HBsAg_Alum complex designed similar to commercially available hepatitis B vaccine, Engerix B. The cytokine and T cell proliferation levels were 2 times and 6 times larger in mice adjuvanted with HBsAg_Alum_MPLA/HA complex than in those vaccinated with HBsAg_Alum. The results therefore indicate that incorporating MPLA and Alum with HA can be a potent strategy to increase both the magnitude and the persistence of HBsAg-specific immune responses to protect hosts against hepatitis B virus infection. [ABSTRACT FROM AUTHOR]

Biological polyunsaturated fatty acids (PUFAs) are important precursors of secondary messengers that modulate inflammatory responses, cellular growth, and cholesterol metabolism. The optimal n-6/n-3 ratio is extremely important for maintaining normal homeostasis because n-3 and n-6 PUFAs are competitively metabolized. To date, a widely accepted analytical method to determine the biological n-6/n-3 ratio is gas chromatography-mass spectrometry (GC-MS) on dried whole blood samples. However, this technique has several drawbacks, including the intrusive nature of collecting blood samples, high expenses involved, and length of time required to use the GC/MS instrument. To overcome these limitations, we introduced Raman spectroscopy (RS) to distinguish PUFAs present in the epididymal adipose tissue (EAT) isolated from experimental rats that were fed three different high-fat diets (HFDs) with multivariate analysis, including principal component analysis (PCA) and linear discriminant analysis (LDA). The diets comprised HFD, HFD + perilla oil (HFD + PO [n-3 rich oil]), and HFD + corn oil (HFD + CO [n-6 rich oil]). This method allows for quantitative, label-free, noninvasive, and rapid monitoring of biochemical changes in the EAT with high sensitivity. In RS, the Raman bands of the EAT from three different diet groups (HFD, HFD + PO, and HFD + CO) detected and distinguished peaks at 1079 (C–C stretching vibration), 1300 (CH2 deformation), 1439 (CH2 deformation), 1654 (amide I), 1746 (C = O stretching vibration), and 2879 cm−1 (–C–H stretching vibration). The PCA-LDA analysis results showed that PUFAs in the EAT of animals receiving the three different dietary interventions can be determined according to the three groups (HFD, HFD + PO, and HFD + CO). In conclusion, we investigated the possibility of determining PUFA profiles in specimens using RS. [ABSTRACT FROM AUTHOR]

In this study, the protective effects of Brussels sprouts extract and its major constituents against oxidative stress-induced damages were investigated in rat pheochromocytoma cells and Institute of Cancer Research mice. The major constituents of Brussels sprouts (3,4′,5,7-tetrahydroxyflavone (kaempferol), indole-3-carbinol, and phenethyl isothiocyanate) were selectively tested. Of these, the flavonoid compound, kaempferol exhibited the highest potency in radical scavenging activity (1,1-diphenyl-2-picryl hydrazyl assay and oxygen radical absorbance capacity assay) and was most protective against oxidative stress in neuronal cell assays (measurement of intracellular oxidative stress levels and cell viability). In mice, after 4 weeks of kaempferol administration, significant protection against amyloid beta (A β) peptide-induced neurotoxicity was also observed, as assessed through the passive avoidance test. Taken together, the results suggest that Brussels sprouts could be protective against A β-induced neurotoxicity, possibly due to the antioxidative capacity of its major constituent, kaempferol. [ABSTRACT FROM AUTHOR]

The surface presentation of peptides by major histocompatibility complex (MHC) class I molecules is critical to all CD8+ T-cell adaptive immune responses, including those against tumors. The generation of peptides and their loading on MHC class I molecules is a multistep process involving multiple molecular species that constitute the so-called antigen processing and presenting machinery (APM). The majority of class I peptides begin as proteasome degradation products of cytosolic proteins. Once transported into the endoplasmic reticulum by TAP (transporter associated with antigen processing), peptides are not bound randomly by class I molecules but are chosen by length and sequence, with peptidases editing the raw peptide pool. Aberrations in APM genes and proteins have frequently been observed in human tumors and found to correlate with relevant clinical variables, including tumor grade, tumor stage, disease recurrence, and survival. These findings support the idea that APM defects are immune escape mechanisms that disrupt the tumor cells’ ability to be recognized and killed by tumor antigen–specific cytotoxic CD8+ T cells. Detailed knowledge of APM is crucial for the optimization of T cell–based immunotherapy protocols. [ABSTRACT FROM PUBLISHER]

Abstract: The fatty acid profiles of commercially-grown Runner-type peanut cultivars (i.e., 10 cultivars, n =151) collected over two production years (2005 and 2006) were determined by gas–liquid chromatography. Eight major fatty acids were identified in the sample set including palmitic (C16:0), stearic (C18:0), oleic (C18:1, ω9), linoleic (C18:2, ω6), arachidic (C20:0), gondoic (C20:1, ω9), behenic (C22:0), and lignoceric (C24:0) acids. Based on the oleic to linoleic acid (O/L) ratio, these cultivars were denoted as normal, mid-, and high-oleic peanut types. Correlation coefficients (r) between the eight major fatty acids identified were generated and revealed an inverse association between oleic and linoleic acids (r =–0.997, P <0.001), while oleic acid and linoleic acid were positively correlated to gondoic acid (r =0.818, P <0.001) and palmitic acid (r =0.967, P <0.001), respectively. Principal component analysis (PCA) of the fatty acid data yielded three significant PCs (i.e., eigenvalues⩾1), which together account for 87.18% of the total variance in the data set; with PC1 contributing 60.45% of the total. Eigen analysis of the correlation matrix loadings of the three significant PCs revealed that PC1 was mainly contributed to by palmitic, oleic, linoleic, and gondoic acids; PC2, by behenic acid; and PC3, by lignoceric acid. The score plot generated between PC1 and PC2 successfully segregated normal, mid- and high-oleic peanut cultivars, while the PC1–PC3 plot segregated normal and the combination of mid- and high-oleic acid cultivars. [Copyright &y& Elsevier]

IFN-gamma is known as the initial and primary inducer of immunoproteasomes during viral infections. We now report that type I IFN induced the transcription and translation of immunoproteasome subunits, their incorporation into the proteasome complex, and the generation of an immunoproteasome-dependent CD8 T cell epitope in vitro and provide in vivo evidence that this mechanism occurs prior to IFN-gamma responses at the site of viral infection. Type I IFN-mediated generation of immunoproteasomes was initiated by either poly(I:C) or HCV RNA in human hepatoma cells and was inhibited by neutralization of type I IFN. In serial liver biopsies of chimpanzees with acute HCV infection, increases in immunoproteasome subunit mRNA preceded intrahepatic IFN-gamma responses by several weeks, instead coinciding with intrahepatic type I IFN responses. Thus, viral RNA-induced innate immune responses regulate the antigen-processing machinery, which occurs prior to the detection of IFN-gamma at the site of infection. This mechanism may contribute to the high effectiveness (95%) of type I IFN-based therapies if administered early during HCV infection. [ABSTRACT FROM AUTHOR]

alpha-Synuclein, a presynaptic protein of the central nervous system, has been implicated in the synaptic events such as neuronal plasticity during development and learning, and neuronal degeneration under pathological conditions. As an effort to understand the biological function of alpha-synuclein, we examined the expression patterns of alpha-synuclein in various human hematopoietic cells, and in Drosophila at different developmental stages. The alpha-synuclein was ubiquitously expressed in all the tested hematopoietic cells including T cells, B cells, NK cells, and monocytes, as well as in the lymphoma cell lines, Jurkat and K562. A potential alpha-synuclein homologue was also expressed in Drosophila, and its expression appeared to be temporally and spatially regulated during development. Our data suggest that alpha-synuclein may function in invertebrates as well as in vertebrates and its function may not be restricted to the neuron. [ABSTRACT FROM AUTHOR]

The ligation-mediated PCR was used to map DNA alkylation sites induced by altromycin B at nucleotide resolution in genomic DNA purified from cultured human colon carcinoma. Altromycin B, one of the pluramycin group of antitumor antibiotics, is characterized as intercalator with the added ability to alkylate N7 guanine. DNA adducts formed in genomic DNA were cleaved into DNA strand breaks by hot piperidine treatment, and fragments containing ligatable breaks were then amplified in a single-sided, ligation-mediated PCR. The alkylation sites observed in exon 9 of the p53 gene revealed that the most high reactivity sites for altromycin B were found to be N7 of guanine in a 5'-AG[sup *] sequence. Determination of the DNA alkylation sites in naked radiolabeled plasmid DNA also showed that altromycin B preferred N7 of guanine in a 5'-AG[sup *] sequence. Thus, it can be concluded that the sequence selective DNA adduct formation induced by the intercalating alkylator, altromycin B, in genomic DNA is similar to that observed in naked plasmid DNA. [ABSTRACT FROM AUTHOR]

This study investigated changes in nutrients (fatty acids, amino acids, and minerals), ginsenosides, and volatile flavors, and antioxidant activities during food processing of mountain-cultivated ginseng (MCG) with the cocktail lactic acid bacteria. Fatty acid content increased, but the free amino acid content decreased, and minerals were practically unaffected during processing. Total phenolic and flavonoid contents and maillard reaction products increased markedly according to processing stage. The total ginsenosides levels increased from 31.25 mg/g (DMCG) to 32.36 mg/g (red MCG, RMCG) and then decreased (27.27 mg/g, at fermented RMCG) during processing. Particularly, the contents of F2 (0.31 → 1.02 → 2.27 mg/g), Rg3 (0.36 → 0.77 → 1.93 mg/g), and compound K (0.5 → 1.68 → 4.13 mg/g) of ginsenosides and β-panasinsene (17.28 → 22.69 → 31.61%), biocycloelemene (0.11 → 0.84 → 0.92%), δ-cadinene (0.39 → 0.5 → 0.94%), and alloaromadendrene (1.64 → 1.39 → 2.6%) of volatile flavor compounds increased during processing, along with to the antioxidant effects (such as DPPH, ABTS, and hydroxyl radical scavenging activities, and FRAP). This study may provide several choices for the use of ginseng in functional foods and functional cosmetics. [ABSTRACT FROM AUTHOR]

Direct-acting antiviral agents (DAAs) that allow for rapid clearance of hepatitis C virus (HCV) may evoke immunological changes. Some cases of rapid de novo hepatocellular carcinoma (HCC) development or early recurrence of HCC after DAA treatment have been reported. During chronic HCV infection, natural killer (NK) cells exhibited a deviant functional phenotype with decreased production of antiviral cytokines and increased cytotoxicity; however, DAA treatment rapidly decreased their cytotoxic function. Effective DAA therapy also suppressed the intrahepatic activation of macrophages/monocytes. This was followed by a decrease in mucosal-associated invariant T (MAIT) cell cytotoxicity without normalization of cytokine production. Rapid changes in the phenotypes of NK and MAIT cells after DAA treatment may attenuate the cytotoxicity of these cells against cancer cells. Moreover, DAA treatment did not normalize the increased frequencies of regulatory T cells even after clearance of HCV infection. Thus, the persistently increased frequency of regulatory T cells may contribute to a local immunosuppressive milieu and hamper the clearance of cancer cells. This review will focus on recent studies describing the changes in innate and adaptive immune responses after DAA treatment in patients with chronic HCV infection in the context of de novo occurrence or recurrence of HCC. [ABSTRACT FROM AUTHOR]

Achieving a functional cure for chronic hepatitis B virus (HBV) infection or complete elimination of HBV covalently closed circular DNA (cccDNA) has been challenging in the treatment of patients with chronic HBV infection. Although novel antivirals are being investigated, improving HBV-specific adaptive immune responses is also important for durable viral clearance. Tissue-resident memory CD8+ T (TRM) cells were recently reported as a T-cell population that resides in peripheral tissues and does not recirculate. TRM cells have been studied in the livers of mice and humans. Liver TRM cells have distinct characteristics compared to T cells in peripheral blood or other tissues, which may be associated with the unique microenvironment of the liver. In this review, we describe the characteristics of liver TRM cells and their implications in chronic HBV infection. We emphasize that liver TRM cells can be an immunotherapeutic target for the treatment of chronic HBV infection. [ABSTRACT FROM AUTHOR]

When interferons (IFNs) bind to their receptors, they upregulate numerous IFN-stimulated genes (ISGs) with antiviral and immune regulatory activities. Hepatitis C virus (HCV) is a single-stranded, positive-sense RNA virus that affects over 71 million people in the global population. Hepatocytes infected with HCV produce types I and III IFNs. These endogenous IFNs upregulate a set of ISGs that negatively impact the outcome of pegylated IFN-α and ribavirin treatments, which were previously used to treat HCV. In addition, the IFNL4 genotype was the primary polymorphism responsible for a suboptimal treatment response to pegylated IFN-α and ribavirin. However, recently developed direct-acting antivirals have demonstrated a high rate of sustained virological response without pegylated IFN-α. Herein, we review recent studies on types I and III IFN responses in HCV-infected hepatocytes. In particular, we focused on open issues related to IFN responses in the direct-acting antiviral era. [ABSTRACT FROM AUTHOR]

Hepatitis C virus (HCV) infects ~71 million people worldwide, and 399,000 people die annually due to HCV-related liver cirrhosis and hepatocellular carcinoma. The use of direct-acting antivirals results in a sustained virologic response in >95% of patients with chronic HCV infection. However, several issues remain to be solved to eradicate HCV. At the 26th International Symposium on Hepatitis C Virus and Related Viruses (HCV2019) held in Seoul, South Korea, October 5–8, 2019, virologists, immunologists, and clinical scientists discussed these remaining issues and how we can achieve the elimination of HCV. [ABSTRACT FROM AUTHOR]

Purpose: Patients with STAT1 gain-of-function (GOF) mutations often exhibit autoimmune features. The JAK1/2 inhibitor ruxolitinib can be administered to alleviate autoimmune symptoms; however, it is unclear how immune cells are molecularly changed by ruxolitinib treatment. Then, we aimed to investigate the trnscriptional and epigenetic status of immune cells before and after ruxolitinib treatment in a patient with STAT1 GOF. Methods: A patient with a heterozygous STAT1 GOF variant (p.Ala267Val), exhibiting autoimmune features, was treated with ruxolitinib, and peripheral blood mononuclear cells (PBMCs) were longitudinally collected. PBMCs were transcriptionally analyzed by single-cell cellular indexing of the transcriptomes and epitopes by sequencing (CITE-seq), and epigenetically analyzed by assay of transposase-accessible chromatin sequencing (ATAC-seq). Results: CITE-seq analysis revealed that before treatment, the patient's PBMCs exhibited aberrantly activated inflammatory features, especially IFN-related features. In particular, monocytes showed high expression levels of a subset of IFN-stimulated genes (ISGs). Ruxolitinib treatment substantially downregulated aberrantly overexpressed ISGs, and improved autoimmune features. However, epigenetic analysis demonstrated that genetic regions of ISGs—e.g., STAT1, IRF1, MX1, and OAS1—were highly accessible even after ruxolitinib treatment. When ruxolitinib was temporarily discontinued, the patient's autoimmune features were aggravated, which is in line with sustained epigenetic abnormality. Conclusions: In a patient with STAT1 GOF, ruxolitinib treatment improved autoimmune features and downregulated aberrantly overexpressed ISGs, but did not correct epigenetic abnormality of ISGs. [ABSTRACT FROM AUTHOR]

Preclinical studies have shown that radiation therapy modulates antitumor immune responses. However, circulating T-cell responses after radiation therapy in patients with cancer have been poorly characterized. This study aims to explore the changes in circulating T cells after stereotactic body radiation therapy (SBRT). Peripheral blood samples of 30 patients with breast cancer who underwent SBRT for bone metastasis were analyzed using multicolor flow cytometry. Phenotypes of PD-1+ CD8+ T cells and regulatory T (T REG) cells were examined. Additionally, plasma protein levels were analyzed using a bead-based immunoassay. Circulating PD-1+ CD8+ T cells, which are enriched for tumor-specific clonotypes, were activated at 1 week after SBRT. However, circulating T REG cells were also activated after SBRT; this pattern was also evident among effector Foxp3hiCD45RA− T REG cells. We observed no difference in T-cell responses according to the fraction size and number. Notably, activation of T REG cells was more prominent in patients who experienced greater activation of PD-1+ CD8+ T cells. Plasma level changes in TGF-β1, soluble CTLA-4, and soluble 4-1BB at 1 week after SBRT were associated with PD-1+ CD8+ T-cell responses. Activation of T REG cells at 1 week after SBRT was associated with worse progression-free survival. Clinical factors including molecular subtype were not associated with the T-cell responses. SBRT induced activation of both potentially tumor-specific CD8+ T cells and T REG cells, which were tightly associated with each other. These results may support the use of T REG cell-modulating strategies with SBRT to improve the antitumor immune response. [ABSTRACT FROM AUTHOR]

Background: Recent studies using single-cell transcriptomic analysis have reported several distinct clusters of neoplastic epithelial cells and cancer-associated fibroblasts in the pancreatic cancer tumor microenvironment. However, their molecular characteristics and biological significance have not been clearly elucidated due to intra- and inter-tumoral heterogeneity. Methods: We performed single-cell RNA sequencing using enriched non-immune cell populations from 17 pancreatic tumor tissues (16 pancreatic cancer and one high-grade dysplasia) and generated paired spatial transcriptomic data from seven patient samples. Results: We identified five distinct functional subclusters of pancreatic cancer cells and six distinct cancer-associated fibroblast subclusters. We deeply profiled their characteristics, and we found that these subclusters successfully deconvoluted most of the features suggested in bulk transcriptome analysis of pancreatic cancer. Among those subclusters, we identified a novel cancer cell subcluster, Ep_VGLL1, showing intermediate characteristics between the extremities of basal-like and classical dichotomy, despite its prognostic value. Molecular features of Ep_VGLL1 suggest its transitional properties between basal-like and classical subtypes, which is supported by spatial transcriptomic data. Conclusions: This integrative analysis not only provides a comprehensive landscape of pancreatic cancer and fibroblast population, but also suggests a novel insight to the dynamic states of pancreatic cancer cells and unveils potential therapeutic targets. [ABSTRACT FROM AUTHOR]

The article presents a study on killer cells in during chronic viral infections. It was found that lymphocytes are losing its ability to grow and reproduce during infection. It is aimed to boost the actions of CD8 T cells in response to the effects of viral infection to suppress the retardation of the cell proliferation and develop the effector functions.

Pet owners think of their animals as part of their family, which further promotes the growth of the pet food market, encouraging pet owners to select nutritious, palatable, and high-quality foods for pets. Therefore, the evaluation of taste and volatile compounds in pet foods is essential to improve palatability. In this study, the sensory characteristics of taste and odor compounds in 10 commercially available dry dog foods were investigated using electronic tongue (E-tongue), electronic nose (E-nose), gas chromatography–mass spectrometry (GC-MS), and gas chromatography–olfactometry (GC-O). Dry dog foods were separated based on the sensory properties of taste and volatile compounds through the multivariate analysis of integrated results of the E-tongue and E-nose. A total of 67 odor active compounds were detected through GC-MS and GC-O, and octanal, nonanal, 2-pentyl furan, heptanal, and benzaldehyde were identified as key odor compounds which may have positive effects on food intake. The multivariate analysis was used to classify samples based on key odor compounds. Volatile compounds responsible for aroma properties of samples were evaluated using GC-O and multivariate analysis in this present study for the first time. These results are expected to provide fundamental data for sensory evaluation in producing new dog foods with improved palatability. [ABSTRACT FROM AUTHOR]

The maintenance of bone is dependent on both osteoclasts, which break down bone, and osteoblasts, which build new bone. Various bone-related disorders, including osteoporosis, can occur as a result of an imbalance between these two cell types. Prolonged use of currently available bone resorption inhibitors may show side effects. Therefore, developing a novel preventive material which effectively inhibits osteoclast differentiation could be beneficial. This study planned to investigate the inhibitory effect of wheat sprout ethanolic extracts (Saegeumgang [SGG] and Arriheuk [ARH]) on the differentiation of osteoclasts induced by RANKL, as well as the mechanisms why fundamental to these effects. The effects of SGG and ARH on bone resorption and osteoclast differentiation were evaluated using RAW 264.7 cells and assessed through TRAP cell count, pit formation, and activity. The expressions of mRNA and protein were accomplished using western blotting, and reverse transcription quantitative polymerase chain reaction analyses were conducted. SGG and ARH were found to suppress osteoclast differentiation in RANKL-stimulated RAW264.7 cells without causing cytotoxic effects. In addition, treatment with SGG and ARH led to a reduction in the number of cells with positive staining for TRAP and TRAP activity. SGG and ARH treatment dose-dependently decreased the pit area in pit formation assays, showing a notable reduction compared to the pit area created by mature osteoclasts. SGG and ARH inhibited osteoclast activity by 84.9% and 95.7% at 200 μg/mL, respectively. In addition, SGG and ARH suppressed the transcriptional activation of various osteoclast-related genes, such as RANK, NFATc1, cathepsin K, c-Fos, TRAP, matrix metallopeptidase-9, dendritic cell-specific transmembrane protein, ATPase H+ transporting v0 subunit d2, and osteoclast-associated receptor in RAW264.7 cells treated with RANKL. SGG and ARH extracts were found to affect the expression of NFATc1 and genes that are specific to osteoclasts during osteoclast differentiation, suggesting their potential use as functional foods or as therapeutic interventions targeting bone health. [ABSTRACT FROM AUTHOR]

Summary: Roasting processing was commonly applied to enhance flavour characteristics, and thus this study was to investigate the taste and aroma compounds between broccoli floret and stem during oven‐roasting processes. In electronic tongue analysis, sourness, saltiness, umami, and sweetness in florets showed an increasing tendency during the roasting, however, decreasing bitterness. Besides, In electronic nose analysis, methanethiol was reduced after the roasting processes and other sulfur compounds were generated in oven‐roasted florets and stems. Furthermore, furans were generated after the roasting. Pyridine and pyrazines were only identified in oven‐roasted florets, however, pyrrole was only identified in oven‐roasted stems. Principal component analysis and cluster analysis were conducted based on combined taste and aroma profiles. Broccoli floret roasted at 15 min was affected by numerous variables compared with the others, and broccoli stems roasted at 15 and 20 min were affected by numerous variables. Additionally, the floret roasted at 15 min was the highest dissimilarity amongst all florets, and the stems roasted at 15 and 20 min were the highest dissimilarity amongst all stems. Accordingly, oven‐roasting induced higher changes in taste and aroma characteristics with increasing times. In addition, this research may provide a basis for enhancing the flavour characteristics of broccoli. [ABSTRACT FROM AUTHOR]

Buah Merah oil (BMO) is unrefined edible oil containing a high level of free fatty acids (FFA; ∼30% w/w). This study was aimed at preparing deacidified BMO from BMO via lipase‐catalyzed esterification of FFA in BMO with added glycerol, using Duolite A568‐immobilized Eversa Transform 2.0 (Thermomyces lanuginosus lipase) as biocatalyst. BMO containing 2.4% w/w FFA and 94.6% w/w triacylglycerol was obtained under optimal reaction conditions (temperature, 70°C; FFA‐to‐glycerol molar ratio, 3:1; enzyme loading based on the protein quantity, 3.75 mg/g BMO, and reaction time, 48 h). No significant difference was found in the contents of β‐carotene, tocopherols, and phytosterols between raw and deacidified BMO. The induction period of oxidation was significantly longer in deacidified BMO (16.37 h) than in raw BMO (0.03 h). These results suggest that deacidified BMO could be enzymatically prepared without the loss of health‐beneficial minor components while enhancing the oxidative stability. Practical Application: Although BMO has recently received much attention for its potential biological activities, the commercial use of BMO as a healthy oil has been limited due to its high FFA content. Unlike conventional alkali and steam refining, enzymatic deacidification of BMO employed in this study might help the commercialization of BMO, because this procedure enables the improvement of oil yield and the retaining of health‐beneficial minor components. [ABSTRACT FROM AUTHOR]

Comparative analyses of SARS-CoV-2-specific immune responses elicited by diverse prime-boost regimens are required to establish efficient regimens for the control of COVID-19. In this prospective observational cohort study, spike-specific immunoglobulin G (IgG) and neutralizing antibodies (nAbs) alongside spike-specific T -cell responses in age-matched groups of homologous BNT162b2/BNT162b2 or AZD1222/AZD1222 vaccination, heterologous AZD1222/BNT162b2 vaccination, and prior wild-type SARS-CoV-2 infection/vaccination were evaluated. Peak immune responses were achieved after the second vaccine dose in the naïve vaccinated groups and after the first dose in the prior infection/vaccination group. Peak titers of anti-spike IgG and nAb were significantly higher in the AZD1222/BNT162b2 vaccination and prior infection/vaccination groups than in the BNT162b2/BNT162b2 or AZD1222/AZD1222 groups. However, the frequency of interferon-γ-producing CD4+ T cells was highest in the BNT162b2/BNT162b2 vaccination group. Similar results were observed in the analysis of polyfunctional T cells. When nAb and CD4+ T -cell responses against the Delta variant were analyzed, the prior infection/vaccination group exhibited higher responses than the groups of other homologous or heterologous vaccination regimens. nAbs are efficiently elicited by heterologous AZD1222/BNT162b2 vaccination, as well as prior infection/vaccination, whereas spike-specific CD4+ T -cell responses are efficiently elicited by homologous BNT162b2 vaccination. Variant-recognizing immunity is more efficiently generated by prior infection/vaccination than the other homologous or heterologous vaccination regimens. [ABSTRACT FROM AUTHOR]

This study investigated the effects of four brewing methods (cold and hot brew, espresso, and cezve) on the chemical sensory properties of green and roasted coffee beans (cv. Yellow Bourbon) extract. The caffeine and chlorogenic acid contents of the coffee were analyzed using HPLC. The taste and volatile aromatic compounds of coffee were analyzed using an electronic tongue and nose, respectively, and the results were analyzed using principal component analysis. For the taste components analyzed using the electronic tongue, the degree of separation was relatively large depending on the extraction method, and the degree of separation was larger depending on roasting for the volatile compounds analyzed using the electronic nose. Our findings provide basic data for the coffee industry. Practical Application: The use of an electronic sensor will provide flavor characteristics for four different types of coffee extracted from green beans and roasted beans. In this study, it was confirmed that the extraction method had a greater effect on the taste of coffee, and in the case of the volatile aromatic compounds of coffee, there was a large difference depending on the green beans and roasted beans. Therefore, our findings will provide data based on the sensory properties of coffee. [ABSTRACT FROM AUTHOR]

Associations between immunosenescence and radiation pneumonitis (RP) are largely unknown. We aimed to identify a peripheral blood T cell senescence biomarker to predict RP in patients with non-small cell lung cancer (NSCLC). Patients with locally advanced NSCLC who received definitive concurrent chemoradiotherapy (dCRT) were prospectively registered (cohort 1, n=23; cohort 2, n=31). Peripheral blood was collected at baseline, during dCRT, and at 1 month post-dCRT. Patients were dichotomized to grade ≥2 (G2+) RP and grade 0-1 (G0-1) RP. Flow cytometry was performed to assess phenotypes and functional properties of T cell subsets. RP incidence was estimated via competing risk analysis. Five and six patients exhibited G2+ RP following dCRT in cohorts 1 and 2, respectively. Patients with G2+ RP exhibited a more aged T cell pool and higher frequencies of senescent CD57+CD28−CD8+ T cells than patients with G0-1 RP at baseline, during dCRT, and at 1 month post-dCRT. These senescent cells exhibited increased granzyme B, IFN-γ, and TNF-α production. Higher baseline frequency of CD57+CD28−CD8+ T cells was an independent predictor of G2+ RP (hazard ratio, 8.42; 95% confidence interval, 2.58–27.45; P<0.001). Recursive partitioning analysis revealed three distinct risk groups stratified by baseline CD57+CD28−CD8+ T cell frequency and lung V 20 Gy , with 1-year cumulative G2+ RP incidences of 50.0%, 16.7%, and 0% for high-, intermediate-, and low-risk groups, respectively (P=0.002). Higher baseline frequencies of CD57+CD28−CD8+ T cells correlated with increased G2+ RP risks. Our results suggest the need for further investigation of the role of T cell senescence on radiation-induced organ damage. [ABSTRACT FROM AUTHOR]

Immune checkpoint inhibitors are effective for advanced hepatocellular carcinoma (HCC), but there remains a need for peripheral blood biomarkers to predict the clinical response. Here, we analyzed the peripheral blood of 45 patients with advanced HCC who underwent nivolumab. During treatment, frequency of classical monocytes (CD14+CD16−) was increased on day 7, and the fold increase in the frequency on day 7 over day 0 (cMonocyteD7/D0) was significantly higher in patients with durable clinical benefit (DCB) than in patients with non-DCB (NDB). When we analyzed transcriptomes of classical monocytes, CD274, gene encoding PD-L1, was upregulated in NDB patients compared to DCB patients at day 7. Notably, gene signature of suppressive tumor-associated macrophages, or IL4l1+PD-L1+IDO1+ macrophages, was enriched after treatment in NDB patients, but not in DCB patients. Accordingly, the fold increase in the frequency of PD-L1+ classical monocytes at day 7 over day 0 (cMonocyte-PDL1D7/D0) was higher in NDB patients than DCB patients. The combined biomarker cMonocyteD7/D0/cMonocyte-PDL1D7/D0 was termed the "monocyte index", which was significantly higher in DCB patients than NDB patients. Moreover, the monocyte index was an independent prognostic factor for survival. Overall, our results suggest that early changes of circulating classical monocytes, represented as a monocyte index, could predict clinical outcomes of advanced HCC patients undergoing anti-PD-1 therapy. [ABSTRACT FROM AUTHOR]

Age-related changes in the immune system, commonly termed 'immunosenescence,' contribute to deterioration of the immune response and fundamentally impact the health and survival of elderly individuals. Immunosenescence affects both the innate and adaptive immune systems; however, the most notable changes are in T cell immunity and include thymic involution, the collapse of T cell receptor (TCR) diversity, an imbalance in T cell populations, and the clonal expansion of senescent T cells. Senescent T cells have the ability to produce large quantities of proinflammatory cytokines and cytotoxic mediators; thus, they have been implicated in the pathogenesis of many chronic inflammatory diseases. Recently, an increasing body of evidence has suggested that senescent T cells also have pathogenic potential in cardiovascular diseases, such as hypertension, atherosclerosis, and myocardial infarction, underscoring the detrimental roles of these cells in various chronic inflammatory responses. Given that cardiovascular disease is the number one cause of death worldwide, there is great interest in understanding the contribution of age-related immunological changes to its pathogenesis. In this review, we discuss general features of age-related alterations in T cell immunity and the possible roles of senescent T cells in the pathogenesis of cardiovascular disease. [ABSTRACT FROM AUTHOR]

Fatigue is a common complaint among people under stress, causing an array of negative effects on physical function. In this study, we investigated the antifatigue and anti-inflammatory effects of Cervus elaphus L., Angelica gigas Nakai, and Astragalus membranaceus Bunge complex extracts (CAA) using a treadmill stress test in animal models. The mice were administered various doses of CAA (50–200 mg/kg bw per day) once daily for 21 days. After exhaustive treadmill exercise, the running time of CAA-treated mice increased 1.5 times; fatigue-related biochemical parameters, including lactate dehydrogenase (∼30%), creatine kinase (∼20%), and proinflammatory cytokines interleukin (IL)-1β (∼10%), and IL-6 (∼10%) in the serum and muscle tissue were downregulated compared with those in exercised control mice. This study provides strong evidence for the prevention of CAA-induced inflammatory incidences mediated by the blockade of nuclear factor-κB activation. Collectively, our results indicate that CAA can alleviate symptoms of fatigue in mice as an effective anti-inflammatory agent. [ABSTRACT FROM AUTHOR]

Chronic exposure to airborne particulate matter (PM) causes respiratory damage in humans owing to oxidative stress and inflammation. Chrysanthemum zawadskii Herbich var. latilobum (Maxim.) Kitamura (CZL) has been used in traditional medicine to treat several inflammatory diseases; however, studies on inflammatory pulmonary diseases are scarce. This study investigated the protective effects of CZL extract against PM10-induced lung injury in BALB/c mice. Cell type specific signaling pathways were explored using A549 and RAW264.7 cell lines. CZL extract noticeably attenuated PM10-induced lung injury and inflammatory cell infiltration in a mouse model. Protein markers, such as p-AKT, p-ERK, and p-NF-κB for PM10 induced lung inflammation were effectively reduced in CZL extract-treated mice and cells. Furthermore, CZL extracts considerably reduced the generation of reactive oxygen species and nitric oxide in cells. Collectively, CZL extract effectively reduced PM10-induced lung injury by suppressing pulmonary inflammation, potentially due to its anti-inflammatory and antioxidant properties. [ABSTRACT FROM AUTHOR]

Summary [ABSTRACT FROM AUTHOR]

Asthma is one of the most common inflammatory diseases of the lung worldwide. There has been considerable progress in recent studies to treat and prevent allergic asthma, however, various side effects are still observed in clinical practice. Six-week-old male BALB/c mice were orally administered with either sword bean pod extracts (SBP; 100 or 300 mg/kg) or dexamethasone (DEX; 5 mg/kg) once daily over 3 weeks, followed by ovalbumin sensitization (OVA/Alum.; intraperitoneal administration, 50 μg/2 mg/per mouse). Scoring of lung inflammation was performed to observe pathological changes in response to SBP treatment compared to OVA/Alum.-induced lung injury. Additionally, inflammatory cytokines were quantified in serum, bronchoalveolar lavage fluid (BALF), and lung tissue using ELISA and Western blot analyses. SBP treatment significantly reduced the infiltration of inflammatory cells, and release of histamine, immunoglobulin E, and leukotriene in serum and BALF. Moreover, the therapeutic effect of SBP was also assessed to analyze the inflammatory changes in the lung tissues. SBP markedly suppressed the activation of the MAPK signaling pathway and the expression of key inflammatory proteins (e.g., TNF-α) and Th2 type cytokines (IL-5 and IL-13). SBP was effective in ameliorating the allergic inflammation against OVA/Alum.-induced asthma by suppressing pulmonary inflammation. [ABSTRACT FROM AUTHOR]

The liver provides a unique niche of lymphocytes enriched with a large proportion of innate-like T cells. However, the heterogeneity and functional characteristics of the hepatic T-cell population remain to be fully elucidated. We obtained liver sinusoidal mononuclear cells from the liver perfusate of healthy donors and recipients with HBV-associated chronic liver disease (CLD) during liver transplantation. We performed a CITE-seq analysis of liver sinusoidal CD45+ cells in combination with T cell receptor (TCR)-seq and flow cytometry to examine the phenotypes and functions of liver sinusoidal CD8+ T cells. We identified a distinct CD56hiCD161-CD8+ T-cell population characterized by natural killer (NK)-related gene expression and a uniquely restricted TCR repertoire. The frequency of these cells among the liver sinusoidal CD8+ T-cell population was significantly increased in patients with HBV-associated CLD. Although CD56hiCD161-CD8+ T cells exhibit weak responsiveness to TCR stimulation, CD56hiCD161-CD8+ T cells highly expressed various NK receptors, including CD94, killer immunoglobulin-like receptors, and NKG2C, and exerted NKG2C-mediated NK-like effector functions even in the absence of TCR stimulation. In addition, CD56hiCD161-CD8+ T cells highly respond to innate cytokines, such as IL-12/18 and IL-15, in the absence of TCR stimulation. We validated the results from liver sinusoidal CD8+ T cells using intrahepatic CD8+ T cells obtained from liver tissues. In summary, the current study found a distinct CD56hiCD161-CD8+ T-cell population characterized by NK-like activation via TCR-independent NKG2C ligation. Further studies are required to elucidate the roles of liver sinusoidal CD56hiCD161-CD8+ T cells in immune responses to microbial pathogens or liver immunopathology. The role of different immune cell populations in the liver is becoming an area of increasing interest. Herein, we identified a distinct T-cell population that had features similar to those of natural killer (NK) cells – a type of innate immune cell. This distinct population was expanded in the livers of patients with chronic liver disease and could thus have pathogenic relevance. [Display omitted] • Single-cell analysis revealed heterogeneity among liver sinusoidal CD8+ T cells. • The CD56hiCD161-CD8+ T-cell population expands in HBV-associated chronic liver disease. • CD56hiCD161-CD8+ T cells have NK-like transcriptomes and unique TCR repertoire. • CD56hiCD161-CD8+ T cells exert TCR-independent, NKG2C-mediated effector functions. • CD56hiCD161-CD8+ T cells exhibit hyper-responsiveness to IL-12/IL-18 and IL-15. [ABSTRACT FROM AUTHOR]

• Excessive oven-roasting of pomegranate seeds breaks down the innate carotenoid pigments and reduces the a* (redness) value. • Oven roasting increases the total phenolic and flavonoid content of pomegranate seeds, which improves their antioxidant capacities. • Proper roasting conditions increase antioxidant properties and positive flavors such as furan derivatives, pyrazines. • Higher temperatures and a long roasting time increase the amounts of off-flavor such as sulfur compounds and amines. This study investigated the effects of oven-roasting temperature (160, 180, and 200 ℃) and time (5, 10, 15, and 20 min) on pomegranate seeds. Physicochemical properties, such as color (L*, a*, and b* values), browning index (BI), total phenolic and flavonoid contents, 2,2-diphenyl-1-picrylhydrazyl radical scavenging capacity, and chemosensory properties, including taste and volatile compounds, were analyzed. The L* and a* values, and level of sourness, umami, sweetness, and terpenes decreased, whereas the b* value, BI, and level of saltiness, bitterness, furan derivatives, pyrazines, and sulfur-containing compounds, increased with roasting time. The findings of this study showed that the positive roasting conditions for pomegranate seeds were 10–20 min at 160 ℃ and, 5–10 min at 180 ℃. This study is expected to be used as a primary reference for selecting the optimal oven-roasting conditions in which positive effects appear and for developing products utilizing pomegranate seeds. [ABSTRACT FROM AUTHOR]

Purpose: Chemoradiotherapy (CRT) followed by consolidation immune checkpoint inhibitors significantly improves survival in unresectable locally advanced non-small cell lung cancer. However, the optimal sequence for CRT and immune checkpoint inhibitors has not yet been established. We investigated the dynamics of peripheral blood immune cells during CRT to determine the best sequence for treatment.Methods and Materials: Peripheral blood samples were prospectively collected pretreatment, weekly during CRT for 6 weeks, and 1 month posttreatment in 24 patients with locally advanced non-small cell lung cancer who received definitive CRT. Immune cell analysis was performed by flow cytometry. Ex vivo PD-1 blockade assays were performed by IFN-γ intracellular cytokine staining.Results: Lymphopenia was prominently observed during CRT and mostly recovered 1 month post-CRT. Robust proliferation of CD8+ T cells was induced, peaking in the last week during CRT and decreasing post-CRT. The robust proliferation of CD8+ T cells led to an increase in the frequency of CD28-CD57+ replicative senescent and terminally differentiated cells post-CRT. Tumor-reactive CD8+ T cells increased during CRT and peaked in the last week. One month post-CRT, the frequency of tumor-reactive CD8+ T cells decreased and TOXhiTCF1lo terminally exhausted CD8+ T cells significantly increased. Anti-PD-1-induced functional restoration of PD-1+CD8+ T cells was maximized in the last week of CRT and significantly decreased post-CRT.Conclusions: The findings suggest that earlier administration of PD-1 blockade may be associated with superior efficacy compared with delayed administration after completion of CRT. These findings provide an immunologic rationale for optimal timing of combining immune checkpoint inhibitors with CRT in clinical trials. [ABSTRACT FROM AUTHOR]

This study investigated the antiobesogenic effects of the inhalation of volatile compounds derived from basil essential oil (BEO) in high fat diet‐induced obese rats. A total of 47 volatile compounds were identified in BEO using gas chromatography‐mass spectrometry. Major volatile compounds identified by olfactory testing include linalool oxide, linalool, 1‐menthene, and carvone. White adipose tissue significantly decreased in the rats that inhaled 0.3% BEO (more than +10%) compared to the control. Plasma marker analysis showed increased high‐density lipoprotein‐cholesterol (ca. double fold) and decreased low‐density lipoprotein‐cholesterol (more than −30%) levels in inhaled 1% BEO group compared to the control. Leptin significantly decreased in the 0.3 and 1% BEO groups (more than −70 and −85%, respectively). Last, systolic blood pressure at week 12 was significantly lower in inhaled 1% BEO group (more than −15%) compared to the control. The results of this study suggest that BEO inhalation may be effective in managing plasma lipid markers (cholesterols and leptin) and possibly metabolic disorders such as obesity. Practical Application: Changes in metabolic health markers, which are effected by inhalation of volatiles in basil (Ocimum basilicum) essential oil, will provide physiological variations in vivo to the public. In this study, the opposite effects were identified between 0.3% and 1% inhalation, respectively. Therefore, our findings will provide optimized and useful guidance for inhalation of basil essential oil. [ABSTRACT FROM AUTHOR]

This study aims to investigate the taste and odor characteristics of broccoli by different thermal processes. For electronic tongue analysis, sourness and umami tastes increased after thermal processing, however, saltiness, sweetness, and bitterness decreased after thermal processing. Among all broccolis, air‐fried broccoli was the highest sourness (6.9) and umami (7.8) tastes, however, saltiness (8.7), sweetness (6.2), and bitterness (10.0) were the highest in the raw broccoli. For electronic nose analysis, a total of 28 volatiles were identified. Among sulfur‐containing compounds, methanethiol and 2,4,5‐tirmethylthiazole were identified as major volatiles in the raw broccoli, however, these volatiles were decreased upon thermal processes. In addition, dimethyl disulfide, 2‐methyl‐3‐furanthiol, and 2‐acetylthiazoline may be generated by thermal processes. In multivariate analysis, the flavor dissimilarity between oven‐heated and microwaved broccolis showed the lowest dissimilarity among all broccolis. Raw broccoli showed the highest dissimilarity among all broccolis. This study is believed to be the first study demonstrating flavors in broccoli by different thermal processing. Novelty impact statement: The present work further investigates the effects of various heating processes, including blanching, oven‐heating, microwave heating, and air‐frying. Thus, this study will be utilized by researchers and food industry that are interested in consuming cooked broccoli. The changes in tastes and odors in thermally processed broccolis were rarely reported. Therefore, this study will be considered worthy that contributes to basic data for further study to characterize flavor and taste in broccoli. [ABSTRACT FROM AUTHOR]

Hepatitis C virus (HCV) is endemic in many countries due to its high propensity for establishing persistence. The presence of HCV-specific T cells in subjects repeatedly exposed to HCV who test negative for HCV RNA and antibodies and who do not have any history of HCV infection has been interpreted as T cell-mediated protection. Here, we show in nonhuman primates that repeated exposure to human plasma with trace amounts of HCV induced HCV-specific T cells without seroconversion and systemic viremia but did not protect upon subsequent HCV challenge. Rather, HCV-specific recall and de novo T cell responses, as well as intrahepatic T cell recruitment and interferon-γ (IFN-γ) production, were suppressed upon HCV challenge, concomitant with quantitative and qualitative changes in regulatory T cells (Treg cells) that occurred after subinfectious HCV exposure and increased after HCV challenge. In vitro Treg cell depletion restored HCV-specific T cell responses. Thus, T cells primed by trace amounts of HCV do not generate effective recall responses upon subsequent HCV infection. Subinfectious HCV exposure predisposes to Treg cell expansion, which suppresses effector T cells during subsequent infection. Strategies to reverse this exposure-induced immune suppression should be examined to aid in the development of T cell−based vaccines against HCV and other endemic pathogens. [ABSTRACT FROM AUTHOR]

Peripheral blood mononuclear cell (PBMC) separation is routinely performed by centrifugation with a density gradient medium, and is essential for immunological research and clinical applications. PBMC separation highly relies on laborious, time‐consuming blood layering and PBMC collection by careful pipetting. Current methods to address this problem use a physical barrier formed between the separation medium and a blood sample to avoid the need for delicate pipetting, but still pose a risk of red blood cell (RBC) contamination. Here, a blood divider (B‐DV) is proposed as a centrifuge tube insert that allows blood cells to pass through a separation medium in one direction, while blocking fluid passage in the opposite direction and thus preventing RBC contamination during PBMC harvest. The B‐DV is a 3D‐printed mesh in which a fluid can divide into two, above and below the mesh face as soon as inverting the centrifuge tube. Such complete fluid division enables minimizing the volume of the separation medium to the extent of RBC volume to be packed, thus reducing the sedimentation distance for RBCs, and facilitating rapid PBMC isolation by simple tube inverting. This technology enables rapid, easy PBMC separation without user‐to‐user variability on PBMC separation efficiencies. [ABSTRACT FROM AUTHOR]

There is insufficient data on the longevity of immunity acquired following severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection. We aimed to evaluate the duration of SARS-CoV-2-specific humoral and cellular immunity according to the clinical severity of coronavirus disease 2019 (COVID-19). The study population comprised asymptomatic (n=14), symptomatic/non-pneumonic (n=42), and pneumonic (n=41) patients. The anti-SARS-CoV-2 IgG and neutralizing antibody (NAb) titers lasted until six months after diagnosis, with positivity rates of 66.7% and 86.9%, respectively. Older age, prolonged viral shedding and accompanying pneumonia were more frequently found in patients with sustained humoral immunity. SARS-CoV-2 specific T-cell response was strongly observed in pneumonic patients and prominent in individuals with sustained humoral immunity. In conclusion, most (> 85%) patients carries NAb until six months after diagnosis of SARS-CoV-2 infection, providing insights for establishing vaccination strategies against COVID-19. [ABSTRACT FROM AUTHOR]

Objectives: Simultaneous transplantation of a solid organ and bone marrow from the same donor is a possible means of achieving transplant tolerance. Here, we attempted to identify biomarkers that indicate transplant tolerance for discontinuation of immunosuppressants in combined kidney and bone marrow transplantation (CKBMT). Methods: Conventional kidney transplant (KT) recipients (n = 20) and CKBMT recipients (n = 6) were included in this study. We examined various immunological parameters by flow cytometry using peripheral blood mononuclear cells (PBMCs), including the frequency and phenotype of regulatory T (Treg) cell subpopulations. We also examined the suppressive activity of the Treg cell population in the setting of mixed lymphocyte reaction (MLR) with or without Treg cell depletion. Results: Among six CKBMT recipients, three successfully discontinued immunosuppressants (tolerant group) and three could not (non‐tolerant group). The CD45RA−FOXP3++ Treg cell subpopulation was expanded in CKBMT recipients compared to conventional kidney transplant patients, and this was more obvious in the tolerant group than the non‐tolerant group. In addition, high suppressive activity of the Treg cell population was observed in the tolerant group. The ratio of CD45RA−FOXP3++ Treg cells to CD45RA−FOXP3+ cells indicated good discrimination between the tolerant and non‐tolerant groups. Conclusion: Thus, our findings propose a biomarker that can distinguish CKBMT patients who achieve transplant tolerance and are eligible for discontinuation of immunosuppressants and may provide insight into tolerance mechanisms in CKBMT. [ABSTRACT FROM AUTHOR]

Background/Aims: The hepatitis B virus X protein (HBx), a major viral transactivator, is implicated in hepatic inflammation, since it induces many pro-inflammatory cytokines at transcriptional level. The aim of this study was to investigate role of HBx in expression of interleukin 18 (IL-18), a newly identified cytokine that up-regulates Fas ligand (FasL) expression.Methods: Chang X-34 that expressing HBx under the control of a doxycycline-inducible promoter, and hepatitis B virus (HBV)-integrated hepatoma cell lines were examined for IL-18 expression by Northern and Western blotting analysis. To test the role of IL-18 produced by hepatoma cells, FasL expression was examined by flow cytometry after treatment with neutralizing anti-IL-18 antibodies. Further, IL-18 expression was examined in the liver tissues of HBx-transgenic mice.Results: Induction of IL-18 following HBx expression in Chang X-34 and the pattern of IL-18 expression in HBV-integrated cell lines, implicated that HBx transcriptionally induces IL-18 expression. Neutralizing anti-IL-18 antibodies blocked the expression of FasL, suggesting that IL-18 plays a critical role in FasL expression. Further, IL-18 expression in the HBx-transgenic liver, was correlated with the degree of hepatitis.Conclusions: Our results demonstrated that HBx induces IL-18 expression in liver, which may be associated with hepatic injury by amplifying FasL expression during HBV infection. [Copyright &y& Elsevier]

Purpose: To understand the tumor immune microenvironment precisely, it is important to secure the quantified data of tumor-infiltrating immune cells, since the immune cells are true working unit. We analyzed unit immune cell number per unit volume of core tumor tissue of high-grade gliomas (HGG) to correlate their immune microenvironment characteristics with clinical prognosis and radiomic signatures. Methods: The number of tumor-infiltrating immune cells from 64 HGG core tissue were analyzed using flow cytometry and standardized. After sorting out patient groups according to diverse immune characteristics, the groups were tested if they have any clinical prognostic relevance and specific radiomic signature relationships. Sparse partial least square with discriminant analysis using multimodal magnetic resonance images was employed for all radiomic classifications. Results: The median number of CD45 + cells per one gram of HGG core tissue counted 865,770 cells which was equivalent to 8.0% of total cells including tumor cells. There was heterogeneity in the distribution of immune cell subpopulations among patients. Overall survival was significantly better in T cell-deficient group than T cell-enriched group (p = 0.019), and T8 dominant group than T4 dominant group (p = 0.023). The number of tumor-associated macrophages (TAM) and M2-TAM was significantly decreased in isocitrate dehydrogenase mutated HGG. Radiomic signature classification showed good performance in predicting immune phenotypes especially with features extracted from apparent diffusion coefficient maps. Conclusions: Absolute quantification of tumor-infiltrating immune cells confirmed the heterogeneity of immune microenvironment in HGG which harbors prognostic impact. This immune microenvironment could be predicted by radiomic signatures non-invasively. [ABSTRACT FROM AUTHOR]

Objectives: In patients with severe combined immunodeficiency (SCID), the immune system often fails to eradicate maternal cells that enter the foetus via the placenta, resulting in transplacental maternal engraftment (TME) syndrome. However, the clinical significance of TME has not been comprehensively elucidated. Methods: Here, we describe a patient with SCID with a novel frameshift IL2RG mutation associated with maternal engrafted CD8+ T cells that had been expanded by viral infection. To evaluate the origin of the expanded T cells, we HLA‐typed the myeloid and T cells of the patient and analysed the immunological characteristics of the expanded CD8+ T cells using T‐cell receptor (TCR) repertoire and flow cytometry analysis. Results: In our patient, the maternal engrafted CD8+ T cells expanded and exerted in vitro antiviral function against human cytomegalovirus (CMV) infection before and after haematopoietic cell transplantation (HCT). After haploidentical HCT from the maternal donor, maternal engrafted CMV‐specific CD8+ T cells were maintained, successfully proliferated and activated against CMV. We found no evidence of acute graft‐versus‐host disease or infectious complications other than recurrent episodes of CMV viraemia, which were well controlled by ganciclovir and, possibly by, the maternal engrafted CMV‐specific CD8+ T cells. Conclusion: Our findings elucidate a possible functional role of TME in controlling CMV infection in patient with SCID and suggest an optimal strategy for donor selection in patients with SCID with TME. [ABSTRACT FROM AUTHOR]

Background: The heterogeneous immune landscapes of intrahepatic cholangiocarcinoma (ICC) remain largely unknown. Here we aimed to investigate the implications of tissue‐resident memory (TRM)‐related features of tumour‐infiltrating CD8+ T cells (CD8+ TILs) from ICC patients. Methods: From ICC patients, we obtained blood samples and ICC surgical specimens (n = 33). We performed multicolour flow cytometry, multiplexed immunohistochemistry and RNA sequencing. Results: When compared to peripheral CD8+ T cells, the CD8+ TILs included significantly higher proportions of the CD69+CD103− and CD69+CD103+ TRM‐like subsets (P <.001 for both). Relative to CD69− and CD69+CD103− cells, the CD69+CD103+ CD8+ TILs harboured higher levels of T‐cell markers representing tumour specificity (ie CD39), proliferation (ie Ki‐67) and T‐cell activation (ie HLA‐DR and CD38) (all P <.001). Moreover, compared to the stroma, the tumour margin and core density each had a significantly higher density of CD103+ CD8+ TILs (P <.001 for both). ICCs with high proportions of CD69+CD103+ cells displayed higher levels of parameters associated with response to immune checkpoint inhibitors (ICIs)—including number of CD8+ TIL infiltrates (P =.019), PD‐L1 expression in the tumour (P =.046) and expression of the T cell‐inflamed gene signature (P <.001). ICCs with lower proportions of CD69+CD103+ CD8+ TILs exhibited significant enrichment of genes related to the Wnt/β‐catenin (P <.001) and TGF‐β pathways (P =.002). Conclusion: CD69+CD103+ TRM‐like CD8+ TILs represent prominent tumour‐specific immune responses and hold promise as a potential therapeutic target in ICC patients. Differential TRM‐related features of ICCs may help develop future immunotherapeutic strategies such as maximizing TRM responses or inhibiting pathways contributing to immune evasion. [ABSTRACT FROM AUTHOR]

Exhausted T cells in the tumor microenvironment are major targets of immunotherapies. However, the exhaustion status of CD8+ tumor-infiltrating lymphocytes (TILs) in bladder cancer has not been comprehensively evaluated. Herein, we examined distinct exhaustion status of CD8+ TILs based on the level of programmed cell death-1 (PD-1) and thymocyte selection-associated high mobility group box protein (TOX) expression in urothelial bladder cancer. We also evaluated the reinvigoration of exhausted CD8+ TILs upon ex vivo treatment with inhibitory checkpoint blockers. TOX-expressing PD-1highCD8+ TILs had the highest expression of immune checkpoint receptors (ICRs), the most terminally exhausted features, and the highest tumor antigen reactivity among PD-1+CD8+ TILs. Bladder cancer patients with a high percentage of PD-1highTOX+CD8+ TILs had more progressed T-cell exhaustion features and higher programmed death-ligand 1 expression in tumor tissues. TIGIT was the most frequent co-expressed ICR on PD-1+CD8+ TILs, and TIGIT blockade enhanced the PD-1 blockade-mediated cytokine production by CD8+ TILs from bladder cancer patients. Our findings provide an improved understanding of the heterogeneous exhaustion status of CD8+ TILs and additional immunotherapy strategies to improve outcomes of bladder cancer patients. [ABSTRACT FROM AUTHOR]

Programmed cell death-1 (PD-1) inhibitor treatment can cause hyperprogressive disease (HPD), but the incidence, outcome, and predictive factors of HPD are unknown in patients with hepatocellular carcinoma (HCC). Herein, we assessed the existence and factors predictive of HPD in patients with advanced HCC treated with nivolumab. We enrolled 189 patients with advanced HCC treated with nivolumab. Occurrence of HPD was investigated using tumour growth dynamics based on tumour growth kinetics (TGK) and tumour growth rate (TGR) before and after treatment, or time to treatment failure. We additionally analysed patients treated with regorafenib (n = 95) or best supportive care (BSC)/placebo (n = 103) after progression on sorafenib to compare tumour growth dynamics. Flare-up of tumour growth was observed in a fraction of patients upon PD-1 blockade, indicating the occurrence of HPD. Based on distinct patterns of disease progression exclusively observed in the nivolumab-treated cohort, but not in the regorafenib- or BSC/placebo-treated cohorts, 4-fold increases in TGK and TGR ratios as well as a 40% increase in TGR were the cut-off values used to define HPD; 12.7% of the patients (24/189) treated with nivolumab met all these criteria. Patients with HPD had worse progression-free survival (hazard ratio [HR] 2.194; 95% CI 1.214–3.964) and overall survival (HR 2.238; 95% CI 1.233–4.062) compared to patients with progressive disease without HPD. More than 90% of patients with HPD missed the opportunity for subsequent treatment because of rapid clinical deterioration. An elevated neutrophil-to-lymphocyte ratio (>4.125) was associated with HPD and an inferior survival rate. HPD occurs in a fraction of patients with HCC who receive PD-1 inhibitor treatment. Analyses of the baseline immune profile and on-treatment tumour growth dynamics could enable optimal patient selection and earlier identification of HPD. Hyperprogressive disease is an unexpected response pattern observed in patients treated with an immune checkpoint inhibitor. This study revealed that hyperprogressive disease occurs in a fraction of patients with advanced hepatocellular carcinoma treated with an anti-PD-1 antibody, providing evidence to encourage careful monitoring of patients to prevent clinical deterioration induced by PD-1 blockade. • HPD occurs in a fraction of patients with HCC treated with PD-1 inhibitors. • HPD is associated with worse PFS and OS, depriving patients of the chance to receive subsequent treatments. • Elevated neutrophil-to-lymphocyte ratio predicts the occurrence of HPD and inferior survival rate after PD-1 blockade. [ABSTRACT FROM AUTHOR]

The predictive value of immune monitoring with circulating CD8+ T lymphocytes for treatment response to programmed cell death protein 1 (PD-1) inhibitors has not been explored in non-small-cell lung cancer (NSCLC), prompting us to investigate whether dynamic changes in PD-1+CD8+ T lymphocytes have predictive value for durable clinical benefit (DCB) and survival after PD-1 blockade. Patients with recurrent and/or metastatic NSCLC treated with PD-1 inhibitors were enrolled (discovery cohort; n = 94). Peripheral blood was obtained immediately before and after one cycle of treatment with PD-1 blockade. Phenotyping of circulating CD8+ T lymphocytes was conducted using multi-colour flow cytometry. Predictive values of dynamic changes in circulating PD-1+CD8+ T lymphocytes during the first cycle were validated in an independent cohort (validation cohort; n = 54) of a prospective trial with a PD-1 inhibitor (NCT03486119). Circulating PD-1+CD8+ T lymphocytes were enriched with effector/memory populations with elevated expression of activation- and exhaustion-related markers. Reduction in the frequency of PD-1+ cells among CD8+ T lymphocytes after one cycle of treatment was associated with a higher probability of DCB and superior survival outcomes in the discovery cohort. Similar results were obtained in the analysis of tumour antigen NY-ESO-1-specific CD8+ T lymphocytes and the validation cohort. Mechanistically, PD-1 molecule expression on CD8+ T lymphocytes suppresses the effector functions of tumour antigen-specific CD8+ T lymphocytes. Dynamic changes in circulating PD-1+CD8+ T lymphocytes predict clinical, and survival benefit from PD-1 blockade treatment in NSCLC, providing a useful tool to identify patient subgroups who will optimally benefit from PD-1 inhibitors. • Current biomarkers for PD-1 blockade are suboptimal and require invasive procedures. • Reduction of PD-1+ cells in CD8+ T lymphocytes predicts favourable treatment outcomes. • Suppressive function of circulating PD-1+CD8+ T lymphocytes was uncovered. [ABSTRACT FROM AUTHOR]