Gene expression profiling of human tissue-resident immune cells: Comparing blood and liver
Autor: | Andre Boonstra, Jun Hou, Simon P. Fletcher, Anuj Gaggar, Robert J. de Knegt, Marieke van der Heide-Mulder, Lauke L. Boeijen, Li Li, Gertine W. van Oord |
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Přispěvatelé: | Gastroenterology & Hepatology |
Rok vydání: | 2019 |
Předmět: |
Adult
Male 0301 basic medicine Immunology System Biology & Immunogenetics Cells: Natural Killer Brief Conclusive Report Tissue/System: Digestive Inflammation Biology Virus 03 medical and health sciences 0302 clinical medicine Immune system Cells: T Lymphocytes Gene expression medicine Humans Immunology and Allergy Lymphocytes RNA Messenger Gene Process: Lymphoid Cell Mediated Immunity Techniques: Multiparameter FACS Gene Expression Profiling Process: Gene Regulation Cell Biology Middle Aged Cell sorting Hepatitis B medicine.disease Gene expression profiling Tissue/System: Lymphoid 030104 developmental biology Liver Process: Host‐Pathogen Interactions 030220 oncology & carcinogenesis Female medicine.symptom Viral hepatitis Signal Transduction |
Zdroj: | Journal of Leukocyte Biology Journal of Leukocyte Biology, 105(3), 603-608. John Wiley & Sons Inc. |
ISSN: | 1938-3673 0741-5400 |
Popis: | In this study, we describe a method to reliably characterize intrahepatic leukocyte populations using flow cytometry and next‐generation RNA sequencing on fresh human liver biopsies. Over the last decades, immune responses of viral hepatitis patients, and of other liver diseases, have been incompletely characterized. Most studies include peripheral blood samples only, foregoing the possibility to investigate the site of inflammation directly. Here, we show that with an optimized protocol that combines cell sorting and RNA sequencing, we can perform a side by side comparison of both intrahepatic and peripheral immune cells. Using core liver biopsies from chronic hepatitis B virus patients, we show that the expression levels of IFN‐stimulated genes and leukocyte‐specific genes are markedly different in the liver compartment as compared to the peripheral blood. These observations emphasize the need to sample the liver directly. The variation of gene expression profiles in these chronic hepatitis B patients was considerable, despite the uniform treatment with nucleotide analogs and absence of liver inflammation in these patients. Finally, we show that this method can provide a detailed characterization of previously undetected liver‐specific effects of novel candidate therapeutic compounds. The use of fluorescence‐activated cell sorting and next generation RNA sequencing to maximize the information yield of studies investigating tissue‐resident leukocytes in human liver. |
Databáze: | OpenAIRE |
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