Perez, Ricardo E.Saleiro, DianaIlut, LilianaSchiltz, Gary E....
9页
查看更多>>摘要：The novel coronavirus, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has been shown to hijack angiotensin converting enzyme 2 (ACE2) for entry into mammalian cells. A short isoform of ACE2, termed deltaACE2 (dACE2), has recently been identified. In contrast to ACE2, the short dACE2 isoform lacks the ability to bind the spike protein of SARS-CoV-2. Several studies have proposed that expression of ACE2 and/or dACE2 is induced by interferons (IFNs). Here, we report that drug-targeted inhibition or silencing of Unc51-like kinase 1 (ULK1) results in repression of type I IFN-induced expression of the dACE2 isoform. Notably, dACE2 is expressed in various squamous tumors. In efforts to identify pharmacological agents that target this pathway, we found that fisetin, a natural flavonoid, is an ULK1 inhibitor that decreases type I IFN-induced dACE2 expression. Taken together, our results establish a requirement for ULK1 in the regulation of type I IFN-induced transcription of dACE2 and raise the possibility of clinical translational applications of fisetin as a novel ULK1 inhibitor.
查看更多>>摘要：Bovine endometritis is a serious hazard to husbandry, so it is necessary to know the mechanism of endometritis. In past research, we found microRNAs (miRNAs) might be regulators in inflammation, including miR-196b, but the mechanism of miR-196b in bovine endometritis was unknown. Therefore, we tended to find out what role miR-196b would play in bovine endometritis. As a result, we found miR-196b up-regulated in the endometritis tissue and the high concentration lipopolysaccharide (LPS)-stimulated bovine endometrial epithelial (BEND) cell line, but down-regulated in the low concentration. And, over-expression of miR-196b inhibited the expressions of some inflammatory factors, such as tumor necrosis factor-alpha (TNF-alpha), interleukin-1 beta (IL-1 beta), interleukin-6 (IL6), and neuroblastoma RAS (NRAS)/nuclear factor (NF)-kappa B pathway proteins. Furthermore, the dual-luciferase reporter assay and NRAS knockdown confirmed that miR-196b inhibited activation of the downstream pathway by directly targeting NRAS. In conclusion, we provide evidence that miR-196b alleviates LPS-induced inflammatory injury by targeting NRAS.
查看更多>>摘要：IL-7 promotes the development of thymic double negative (DN) T cells during beta-selection, which might contribute to the remission of aging-associated thymic involution. Methylation levels of CpG sites is correlated with aging and modulates the development. To determine the involvement of DNA methylation/demethylation instructed by IL-7 signaling during the expansion of double negative (DN) T cells, the aged mice were treated with recombinant Adeno-Associated Virus 2-mediated IL-7 (rAAV2-IL-7) and the DNA methylation/demethylation modifications in this process were analyzed. The results showed that rAAV2-IL-7 increased the number of thymocytes, especially the DN3 thymocytes during beta-selection in aged mice. With aging, the methylation levels of Bcl2 and c-Myc promoter regions were increased in DN3 cells. Following rAAV2-IL-7 treatment, DNA methyl-transferase Dnmt3a and Dnmt3b decreased, DNA demethylation factors TET2 and TET3 increased, and the methylation levels of Bcl2 and c-Myc in DN3 cells were reduced during DN3 stage in aged mice, consequently, resulting in the upregulation of Bcl2 and c-Myc and the larger increase of DN3 cells in thymus. In conclusion, these findings showed that Bcl2 and c-Myc genes of DN3 cells had an increased DNA methylation levels in aged mice compared to the young, and the hypermethylation in aged mice could be restored following rAAV2-IL-7 treatment.
查看更多>>摘要：Tumors can induce the generation and accumulation of immunosuppressive cells in -the tumor microenvironment (TME). Among them, tumor-educated dendritic cells (TEDCs) involved in tolerance induction contribute greatly to the progression of tumors. However, the mechanisms governing the immunosuppressive function of dendritic cells in the TME are unclear. In this study, we found that the expression of transcription factor EB (TFEB) was significantly increased in TEDCs induced by cancer cell supernatant. TFEB knockdown significantly promoted the differentiation and maturation of TEDCs, with upregulated expression of CD11c and costimulatory molecules (CD86 and MHC-II) but reduced expression of the inhibitory molecule PD-L1, and enhanced their ability to induce Th1 proliferation and differentiation. Moreover, TEDCs with TFEB knockdown significantly reduced tumor growth with increased infiltration of CD11c(+)MHC-II+ dendritic cells and effector T cells in tumor masses, thus leading to a delay in tumor progression. These findings demonstrate a critical role of TFEB in regulating the immunosuppression of TEDCs, with potential implications as an antitumor immune therapeutic approach.
查看更多>>摘要：Background: The tumour microenvironment reshapes the specific gene expression of regulatory T cells (Tregs). A better definition of Treg subpopulations in the non-small-cell lung cancer (NSCLC) milieu is expected to clarify the identity and functional mode of Tregs and lead to the identification of better therapeutic targets.Methods: A total of 53 peripheral blood (PB) samples from 36 NSCLC patients and 17 control subjects and 42 matched bronchoalveolar lavage fluid (BALF) samples from 31 NSCLC patients and 11 control subjects were obtained to examine the frequencies of Treg subgroups through flow cytometry. Fifteen PB samples from healthy individuals were collected to explore the differential functions of Treg subsets. The PB samples of 5 patients after chemotherapy were obtained to evaluate the effect of chemotherapy on Treg subsets. Serum CYFRA 21-1 levels in NSCLC patients were determined using an electrochemiluminescence immunoassay.Results: The proportions of CD4(+)CD25(+)FoxP3(+) Tregs in both PB and BALF were increased in NSCLC patients compared to controls. In BALF, the TIGIT(+), Helios(+), and TIGIT(+)Helios(+) Treg subset levels were significantly elevated; the levels of the last two subsets were associated with NSCLC development, while the level of TIGIT-Helios- Tregs was decreased. The proportions of overall Tregs and TIGIT(+), Helios(+), and Helios(+)TIGIT(+) Tregs were positively correlated with the serum CYFRA 21-1 levels in all patients. Functional differences were observed between Helios(+)TIGIT(+) and Helios-TIGIT- Tregs. After chemotherapy, regardless of the reduction in serum CYFRA 21-1 levels, the proportions of Tregs and Treg subsets did not change. Conclusions: Elevated TIGIT(+)Helios(+) and Helios(+) Treg levels may play a role in NSCLC tumour progression, and targeting TIGIT and Helios on Tregs may be an effective treatment for NSCLC.
查看更多>>摘要：Purpose: To explore the therapeutic effect of chondroitin sulfate (CS) on Aspergillus fumigatus (A. fumigatus) keratitis. Methods: The nontoxic concentration of CS was determined using the Draize eye test and cell counting kit-8. Cell scratch test and cell proliferation test were evaluated the impact of CS on the proliferation and migration of human corneal epithelial cells (HCECs). Adherence assay and plate counting were used to detect fungal load in vivo and in vitro, respectively. Clinical score and hematoxylin-eosin (HE) staining were applied to assess the therapeutic effects of CS in an A. fumigatus keratitis mice model. The neutrophil infiltration and activity were detected by flow cytometry (FCM), immunofluorescence staining, and myeloperoxidase (MPO) assay. Toll-like receptor 4 (TLR-4) expression in RAW 264.7 cells and mouse cornea was detected by immunofluorescence staining. Real-time PCR (RT-PCR), western blot, and enzyme-linked immunosorbent assay (ELISA) were applied to examine the expression of inflammatory mediators. Results: CS at 400 mu g/mL (non-cytotoxic) significantly promoted proliferation and migration of HCECs. In an A. fumigatus keratitis mice model, CS treatment alleviated fungal keratitis (FK) severity by decreasing corneal fungal load and inhibiting neutrophil infiltration. In RAW 264.7 cells, the mRNA and protein levels of TLR-4, phosphorylated nuclear factor (NF)-kappa B (p-NF-kappa B), interleukin-1 beta (IL-1 beta), interleukin-6 (IL-6), tumor necrosis factor-a (TNF-alpha), cyclooxygenase 2 (COX-2), and macrophage inflammatory protein-2 (MIP-2) were remarkably lower in the siTLR-4 treated group, while higher in rTLR-4 treated group than in the corresponding control group. CS treatment suppressed rTLR-4 induced the mRNA and protein expression of TLR-4, p-NF-kappa B, IL-1 beta, IL-6, COX-2, TNF-alpha, and MIP-2 in RAW cells. Conclusion: CS may ameliorate the prognosis of A. fumigatus keratitis by promoting corneal epithelial proliferation, inhibiting the recruitment and activity of neutrophils, and inhibiting the inflammatory response by downregulation of the TLR-4/NF-kappa B signaling.
查看更多>>摘要：T-cell immunoglobulin and ITIM domain (TIGIT) is a novel type of immune checkpoint. Importantly, immune checkpoint molecules promote cancer progression by various antitumor suppressive mechanisms. TIGIT is an inhibitory receptor expressed on T cells, natural killer cells, and regulatory T cells that was recently attracted attention as a major emerging target for cancer immunotherapy. Regulatory T cells (Tregs) play crucial roles in immune homeostasis. Specifically, tumor-infiltrating Tregs promote cancer progression by restricting antitumor immunity and supporting tumor immune escape. In this review, we summarized the current understanding on TIGIT and tumor-infiltrating Tregs. Here, we reviewed the latest advances in the understanding of mechanisms causing tumor-infiltrating Tregs abundance to optimize Tregs targeted therapy. Collectively, anti-TIGIT targeting Tregs hold great promise for potent cancer target therapy.
查看更多>>摘要：Background: Angiogenesis associates with chondrocytes differentiation in inflammatory arthritis. Interleukin (IL)- 1 beta stimulated SW1353 cells have a phenotype similar to this kind of chondrocytes. IL-17A, a target in T helper 17 (Th17)/IL-17 signaling pathways, was expressed by SW1353 cells. The study aimed to explore the role of IL-35 on angiogenesis in IL-1 beta stimulated SW1353 cells and its related signaling pathways. Methods: Microarray dataset was downloaded from the Gene Expression Omnibus database of arthritis cartilage. The protein-protein interaction (PPI) was analyzed for IL-35, pro-angiogenic factors and the differentially expressed genes (DEGs). We studied the effects of IL-35 on proliferation and apoptosis in IL-1 beta stimulated SW1353 cells using cell counting kit-8 (CCK-8) assay and flow cytometry. The expression of pro-angiogenic factors and IL-17A were assessed by western blot and real-time PCR. Added plumbagin (inhibitor of IL-17A) to repeat the above experiment. The secretion of IL-17A was assessed by ELISA. Results: IL-35, pro-angiogenic factors interacted with DEGs to affect the function of arthritis chondrocytes. IL-35 promoted IL-1 beta-stimulated SW1353 cells proliferation, inhibited apoptosis, and decreased pro-angiogenic molecules and IL-17A expression in a concentration dependent manner. IL-35 inhibited IL-17A secretion in the supernatants of these cells. Blocking the Th17/IL-17 related pathways with plumbagin abolished the effects of IL-35 on IL-1 beta-stimulated SW1353 cells. Conclusion: These results suggested that IL-35 regulated differentiation and pro-angiogenic molecules expression in IL-1 beta stimulated SW1353 cells via Th17/IL-17 related signaling pathways. Our findings may reveal the mechanisms of novel angiogenesis molecules in inflammatory chondrocyte lesion.
Asmani, FarzanehKhavari-Nejad, Ramazan AliSalmanian, Ali HatefAmani, Jafar...
9页
查看更多>>摘要：Enterotoxigenic Escherichia coli is the most important bacterial agent causing traveller's diarrhea in developing countries. Enterotoxins (LT & ST) and colonization factors (CFs) are two important factors in the ETEC pathogenesis. In the present study, a recombinant four-part fusion protein containing CFAB*ST, CFAE, and LTB (CCL) was expressed in hairy roots of Nicotiana tabacum. The synthetic gene sequence and gene order were designed based on bioinformatics analysis that predicted the best arrangement for antigenicity and stimulation of the immune response. Codon usage was optimized for expression in tobacco plant, under the control of promoter CaMV35S in plasmid pBI121. CCL was efficiently expressed in tobacco hairy roots to yield 1.11% soluble protein as determined by quantitative ELISA and Western blot. In this study, mice were immunized with purified CCL via the oral and subcutaneous route. Humoral immunity especially mucosal immunity with antigen specific IgG and IgA detected in serum and feces. The ability of CCL to elicit neutralizing antibodies was evaluated in the rabbit ileal loop model, using anti-CCL antibodies derived from immunized mice, and co-incubated with ETEC strains. A decrease in fluid accumulation in the intestinal lumen of rabbit ileal loops challenged with ETEC LT and ST positive strains, correlated with the presence of anti-CCL antibodies capable of toxin neutralization. The ability of these antibodies to neutralize toxin confirmed the recognition of epitopes, either linear or conformational, displayed by the recombinant chimeric protein expressed in transgenic tobacco hairy roots. Transgenic plants containing multivalent immunogenic vaccine candidates have the potential to be used for immunization with protection against gastrointestinal pathogens like ETEC.
查看更多>>摘要：Background: Primary Sjo spacing diaeresis gren's syndrome (pSS) is a common chronic autoimmune disease characterized by lymphocytic infiltration of salivary and lacrimal glands. The current study was performed to investigate the roles of follicular helper T (Tfh) and follicular regulatory T (Tfr) subsets in patients with pSS, and to evaluate the effects of sirolimus on these cells. Methods: Levels of circulating Tfh and Tfr subsets in 58 pSS patients and 26 healthy controls (HC) were deter-mined by flow cytometry. These T cell subsets were also analyzed in 12 patients before and after treatment with sirolimus. Clinical features and correlations with follicular T cells were analyzed systematically. The discrimi-native ability of the cells and ratios was evaluated based on the area under the receiver operating characteristic curves. Results: Patients with pSS had higher percentage and absolute number of PD-1+ICOS+Tfh cells, while lower percentage and absolute number of Tfr, activated regulatory T (aTreg) cells, and CD45RA-Foxp3(high) activated Tfr cells. Furthermore, increased number of PD-1+ICOS+Tfh cells was associated with B cells, while decreased numbers of Tfr and their subsets was strongly associated with aTreg cells in pSS patients. Also, the higher proportion of PD-1+ICOS+Tfh cells was positively correlated with higher level of autoantibodies, ESR, IgG, cytokines (IL-2, IL-4, IL-10, IL-17, IFN-gamma, TNF-alpha, IL-21 and sIL-2 alpha R), and disease activity. Unexpectedly, the elevated PD-1+ICOS+Tfh:CD45RA-Foxp3(high) activated Tfr ratio had the greatest ability to discriminate between pSS and HC, and sirolimus therapy restored the PD-1+ICOS+Tfh cells:CD45RA-Foxp3(high) activated Tfr ratio, and controlled disease activity. Conclusion: The novel ratio of PD-1+ICOS+Tfh to CD45RA-Foxp3(high) activated Tfr cells can effectively discriminate the pSS patients from controls, and Tfr cell subsets may resemble Treg cell lineages. Furthermore, the PD-1+ICOS+Tfh cells can be used as a biomarker of disease activity and to verify the therapeutic effects of sirolimus in pSS.
NSTL
Elsevier