Project description:This experiment investigates the transcriptional effect of Interferon Alpha or Interferon Lambda upon Induced Hepatocyte-Like Cells (iHLCs). Peginterferon lambda-1a (Lambda) is a type III interferon that acts through a unique receptor complex expressed primarily on hepatocytes. In Phase 2b clinical studies, a combined regimen of Lambda with ribavirin (RBV) was associated with more rapid declines in viral load compared to a peginterferon alfa (PEGASYS; alfa) plus RBV regimen. This correlated with improved virologic response rates at Weeks 4 and 12 of treatment. To gain insight into the potential molecular mechanisms of these early robust responses with Lambda, we investigated the effects of alfa and Lambda on transcription in iHLCs obtained from Cellular Dynamics.. These samples are a subset of a larger experiment to be published at a later date.
Project description:Whereas most body cells respond to stimulation with type I (alpha and beta) interferon, type III (lambda) interferon was shown to specifically stimulate epithelial cells in the gastrointestinal and respiratory tract. Although both cytokines activate different receptor molecules (IFNAR versus IL-28R/IL-10R, respectively) the downstream signalling pathways and the ultimate response have been reported to be very similar. Here we examined the response of polarized gut intestinal epithelial cells to type I versus type III interferon in order to study potential differences in the transcriptional response.
Project description:RATIONALE: Drugs used in chemotherapy use different ways to stop tumor cells from dividing so they stop growing or die. Interferon alfa may interfere with the growth of cancer cells. Colony-stimulating factors such as filgrastim may increase the number of immune cells found in bone marrow or peripheral blood and may help a person recover from the side effects of chemotherapy. Combining chemotherapy with interferon alfa may kill more tumor cells.
PURPOSE: Phase II trial to study the effectiveness of combining chemotherapy and interferon alfa followed by filgrastim in treating patients who have gastrointestinal tract cancer.
Project description:Respiratory tract epithelium infection is considered crucial for airborne transmission of Nipah virus (NiV). Knowledge about infection dynamics and host responses to NiV infection in respiratory tract epithelia is scarce. Various studies in non-differentiated primary respiratory tract cells or cell lines indicate limitations in interferon responses. However, comprehensive studies determining complex reaction patterns in differentiated respiratory tract epithelia to understand efficient NiV replication and spread in swine populations are lacking. Here we characterized infection and spread of NiV in differentiated primary porcine bronchial epithelial cells (PBEC) cultivated at the air-liquid-interface (ALI). After initial infection of only a few apical cells, lateral spread for 12 days with disruption of the epithelium was observed without releasing substantial amounts of infectious virus from the apical or basal sides. Deep time course proteomics revealed pronounced upregulation of genes related to type I/II interferon (IFN), immunoproteasomal subunits, TAP-mediated peptide transport and MHC I antigen presentation, whereas spliceosomal factors were downregulated. We deduce a model in which NiV replication in pig bronchial epithelia is slowed by a potent and broad type I/II interferon host response with concurrent conversion from 26S proteasomal to immunoproteasomal antigen processing and improved MHC I presentation as a crucial step in adaptive immunity priming. Moreover, strong cytopathic effects observed in infected areas could reflect focal release of cell-associated NiV. Cell-associated NiV may translate into the source of efficient airborne viral spread in vivo with a high local infectious dose leading to high and fast spread of NiV throughout pig herds.
Project description:We designed a long-term culture system for porcine intestinal organoids from intestinal crypt or single Lgr5+ stem cells by combining previously defined insights in the growth requirements of intestinal epithelium of human and mouse. We showed that long-term cultured swine intestinal organoids were expanded in vitro more than six months at least and maintained the potential to differentiate into different types of cells. These organoids were successfully infected with porcine enteric coronavirus including porcine epidemic diarrhea virus (PEDV) and transmissible gastroenteritis virus (TGEV). RNA-seq analysis showed that robust induction of transcripts associated with antiviral signaling in response to enteric coronavrius infection, including a number of interferon-stimulated genes and cytokines. Moreover, gene set enrichment analysis indicated that PEDV infection could suppress immune response in organoids. This 3D intestinal organoid model offers a long-term, renewable resource for investigating porcine intestinal infections with a variety of pathogens.
Project description:Interferons (IFNs) induced early after SARS-CoV-2 infection are crucial for shaping immunity and preventing severe COVID-19. We previously demonstrated that injection of pegylated interferon-lambda1 (PEG-IFN-λ) accelerated viral clearance in COVID-19 patients. To determine if the viral decline was mediated by enhanced immunity, we assessed in vivo responses to PEG-IFN-λ by single cell RNA sequencing and measured SARS-CoV-2-specific T cell and antibody responses between placebo and PEG-IFN-λ-treated patients. PEG-IFN-λ treatment induced interferon stimulated genes in peripheral immune cells expressing IFNLR1, including plasmacytoid dendritic cells and B cells. PEG-IFN-λ did not affect SARS-CoV-2-specific antibody levels or the magnitude of virus-specific T cells. However, we identified delayed T cell responses in older adults, suggesting that PEG-IFN-λ can overcome delays in adaptive immunity to accelerate viral clearance in high-risk patients. Altogether, PEG-IFN-λ offers an early COVID-19 treatment option for outpatients to boost innate antiviral defenses without dampening peripheral adaptive immunity.
Project description:RNAseq of intestinal epithelial cell (IEC) organoids derived from biopsy of ileum or colon from healthy subjects and treated with type I interferon (IFN beta), type II interferon (IFN gamma), or type III interferon (IFN lambda 2).
Project description:RNAseq of intestinal epithelial cell (IEC) organoids and treated with PBS, interferon beta (IFNB), or interferon lambda (IFNL). RNAseq of IECs sorted from neonatal mouse intestines following treatment with PBS, IFNB, or IFNL.
Project description:RATIONALE: Interferon alfa may interfere with the growth of the cancer cells and slow the growth of non-Hodgkin’s lymphoma.
PURPOSE: Phase II trial to study the effectiveness of interferon alfa-2b in treating patients who have advanced low-grade non-Hodgkin’s lymphoma.
Project description:The objectives of this study is to evaluate the chromatin accesibility changes after deoxyshikonin stimulation in porcine intestinal epithelial cells.