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1.
Mob DNA ; 13(1): 22, 2022 Sep 06.
Article in English | MEDLINE | ID: mdl-36068622

ABSTRACT

BACKGROUND: Bats are a major reservoir of zoonotic viruses, and there has been growing interest in characterizing bat-specific features of innate immunity and inflammation. Recent studies have revealed bat-specific adaptations affecting interferon (IFN) signaling and IFN-stimulated genes (ISGs), but we still have a limited understanding of the genetic mechanisms that have shaped the evolution of bat immunity. Here we investigated the transcriptional and epigenetic dynamics of transposable elements (TEs) during the type I IFN response in little brown bat (Myotis lucifugus) primary embryonic fibroblast cells, using RNA-seq and CUT&RUN. RESULTS: We found multiple bat-specific TEs that undergo both locus-specific and family-level transcriptional induction in response to IFN. Our transcriptome reassembly identified multiple ISGs that have acquired novel exons from bat-specific TEs, including NLRC5, SLNF5 and a previously unannotated isoform of the IFITM2 gene. We also identified examples of TE-derived regulatory elements, but did not find strong evidence supporting genome-wide epigenetic activation of TEs in response to IFN. CONCLUSION: Collectively, our study uncovers numerous TE-derived transcripts, proteins, and alternative isoforms that are induced by IFN in Myotis lucifugus cells, highlighting candidate loci that may contribute to bat-specific immune function.

2.
Methods Mol Biol ; 1634: 203-210, 2017.
Article in English | MEDLINE | ID: mdl-28819853

ABSTRACT

Enumerating circulating tumor cells (CTCs) in blood and disseminated tumor cells (DTCs) in bone marrow has shown to be clinically useful, as elevated numbers of these cells predict poor clinical outcomes. Accurate detection and quantification is, however, difficult and technically challenging because CTCs and DTCs are extremely rare. We have developed a novel quantitative detection method for enumeration of CTCs and DTCs. Our approach consists of two steps: (1) EPCAM-based immunomagnetic enrichment followed by (2) flow cytometry (IE/FC). The assay takes approximately 2 h to complete. In addition to tumor cell enumeration, IE/FC offers opportunities for direct isolation of highly pure tumor cells for downstream molecular characterization.


Subject(s)
Flow Cytometry/methods , Immunomagnetic Separation/methods , Magnetite Nanoparticles/chemistry , Neoplasms/diagnosis , Neoplastic Cells, Circulating/pathology , Antibodies, Monoclonal/chemistry , Antibodies, Monoclonal/metabolism , Biomarkers, Tumor/genetics , Biomarkers, Tumor/immunology , Biomarkers, Tumor/metabolism , Bone Marrow Cells/immunology , Bone Marrow Cells/metabolism , Bone Marrow Cells/pathology , Cell Count/instrumentation , Cell Count/methods , Cell Line, Tumor , Epithelial Cell Adhesion Molecule/genetics , Epithelial Cell Adhesion Molecule/immunology , Epithelial Cell Adhesion Molecule/metabolism , Equipment Design , Flow Cytometry/instrumentation , Fluorescent Dyes/chemistry , Humans , Immunoconjugates/chemistry , Immunomagnetic Separation/instrumentation , Leukocyte Common Antigens/genetics , Leukocyte Common Antigens/immunology , Leukocyte Common Antigens/metabolism , Neoplasms/blood , Neoplasms/immunology , Neoplasms/pathology , Neoplastic Cells, Circulating/immunology , Neoplastic Cells, Circulating/metabolism , Protein Binding
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