Phenotyping and auto-antibody production by liver-infiltrating B cells in primary sclerosing cholangitis and primary biliary cholangitis

Brian K Chung; Bardia T Guevel; Gary M Reynolds; D B R K Gupta Udatha; Eva Kristine Klemsdal Henriksen; Zania Stamataki; Gideon M Hirschfield; Tom Hemming Karlsen; Evaggelia Liaskou

doi:10.1016/j.jaut.2016.10.003

Phenotyping and auto-antibody production by liver-infiltrating B cells in primary sclerosing cholangitis and primary biliary cholangitis

J Autoimmun. 2017 Feb:77:45-54. doi: 10.1016/j.jaut.2016.10.003. Epub 2016 Oct 24.

Authors

Brian K Chung¹, Bardia T Guevel², Gary M Reynolds², D B R K Gupta Udatha³, Eva Kristine Klemsdal Henriksen⁴, Zania Stamataki², Gideon M Hirschfield⁵, Tom Hemming Karlsen⁶, Evaggelia Liaskou²

Affiliations

¹ Centre for Liver Research and National Institute for Health Research (NIHR) Birmingham Liver Biomedical Research Unit, Institute of Immunology and Immunotherapy, College of Medical and Dental Sciences, University of Birmingham, Birmingham, UK; Birmingham Health Partners, Institute of Translational Medicine, University Hospitals Birmingham, Birmingham, UK; Norwegian PSC Research Center, Department of Transplantation Medicine, Division of Surgery, Inflammatory Medicine and Transplantation, Oslo University Hospital Rikshospitalet, Oslo, Norway; Institute of Clinical Medicine, Faculty of Medicine, University of Oslo, Oslo, Norway.
² Centre for Liver Research and National Institute for Health Research (NIHR) Birmingham Liver Biomedical Research Unit, Institute of Immunology and Immunotherapy, College of Medical and Dental Sciences, University of Birmingham, Birmingham, UK; Birmingham Health Partners, Institute of Translational Medicine, University Hospitals Birmingham, Birmingham, UK.
³ Norwegian PSC Research Center, Department of Transplantation Medicine, Division of Surgery, Inflammatory Medicine and Transplantation, Oslo University Hospital Rikshospitalet, Oslo, Norway; Research Institute of Internal Medicine, Division of Surgery, Inflammatory Medicine and Transplantation, Oslo University Hospital Rikshospitalet, Oslo, Norway; K.G. Jebsen Inflammation Research Centre, Institute of Clinical Medicine, University of Oslo, Oslo, Norway.
⁴ Norwegian PSC Research Center, Department of Transplantation Medicine, Division of Surgery, Inflammatory Medicine and Transplantation, Oslo University Hospital Rikshospitalet, Oslo, Norway; Institute of Clinical Medicine, Faculty of Medicine, University of Oslo, Oslo, Norway; Research Institute of Internal Medicine, Division of Surgery, Inflammatory Medicine and Transplantation, Oslo University Hospital Rikshospitalet, Oslo, Norway; K.G. Jebsen Inflammation Research Centre, Institute of Clinical Medicine, University of Oslo, Oslo, Norway.
⁵ Centre for Liver Research and National Institute for Health Research (NIHR) Birmingham Liver Biomedical Research Unit, Institute of Immunology and Immunotherapy, College of Medical and Dental Sciences, University of Birmingham, Birmingham, UK; Birmingham Health Partners, Institute of Translational Medicine, University Hospitals Birmingham, Birmingham, UK. Electronic address: g.hirschfield@bham.ac.uk.
⁶ Norwegian PSC Research Center, Department of Transplantation Medicine, Division of Surgery, Inflammatory Medicine and Transplantation, Oslo University Hospital Rikshospitalet, Oslo, Norway; Institute of Clinical Medicine, Faculty of Medicine, University of Oslo, Oslo, Norway; Research Institute of Internal Medicine, Division of Surgery, Inflammatory Medicine and Transplantation, Oslo University Hospital Rikshospitalet, Oslo, Norway; K.G. Jebsen Inflammation Research Centre, Institute of Clinical Medicine, University of Oslo, Oslo, Norway. Electronic address: t.h.karlsen@medisin.uio.no.

PMID: 27784538
DOI: 10.1016/j.jaut.2016.10.003

Abstract

Primary biliary cholangitis (PBC) and primary sclerosing cholangitis (PSC) are immune-mediated biliary diseases that demonstrate prominent and restricted genetic association with human leukocyte antigen (HLA) alleles. In PBC, anti-mitochondrial antibodies (AMA) are specific and used as diagnostic biomarkers. PSC-relevant auto-antibodies remain controversial despite a distinct HLA association that mirrors archetypical auto-antigen driven disorders. Herein, we compared antibody-secreting B cells (ASCs) in PSC and PBC liver explants to determine if liver-infiltrating ASCs represent an opportune and novel source of disease-relevant auto-antibodies. Using enzymatic digestion and mechanical disruption, liver mononuclear cells (LIMCs) were isolated from fresh PSC and PBC explants and plasmablast (CD19+CD27+CD38^hiCD138-) and plasma cell (CD19+CD27+CD38^hiCD138+) ASCs were enumerated by flow cytometry. We observed 45-fold fewer plasma cells in PSC explants (n = 9) compared to PBC samples (n = 5, p < 0.01) and 10-fold fewer IgA-, IgG- and IgM-positive ASCs (p < 0.05). Liver-infiltrating ASCs from PSC and PBC explants were functional and produced similar concentrations of IgA, IgG and IgM following 2 weeks of culture. Antibody production by PBC ASCs (n = 3) was disease-specific as AMA to pyruvate dehydrogenase complex E2 subunit (PDC-E2) was detected by immunostaining, immunoblotting and ELISA. Antibody profiling of PSC supernatants (n = 9) using full-length recombinant human protein arrays (Cambridge Protein Arrays) revealed reactivities to nucleolar protein 3 (5/9) and hematopoietic cell-specific Lyn substrate 1 (3/9). Array analysis of PBC supernatants (n = 3) detected reactivities to PDC-E2 and hexokinase 1 (3/3). In conclusion, we detected unique frequencies of liver-infiltrating ASCs in PSC and PBC and in so doing, highlight a feasible approach for understanding disease-relevant antibodies in PSC.

Keywords: Antibody-secreting B cells; Auto-antibodies; Autoimmunity; Biomarkers; Primary biliary cholangitis; Primary sclerosing cholangitis; Protein arrays.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Adolescent
Adult
Aged
Antibody Formation / immunology
Antigens, CD20 / metabolism
Autoantibodies / blood
Autoantibodies / immunology*
Autoimmunity
B-Lymphocytes / immunology*
B-Lymphocytes / pathology
Biomarkers
Cholangitis, Sclerosing / diagnosis*
Cholangitis, Sclerosing / immunology*
Cholangitis, Sclerosing / metabolism
Enzyme-Linked Immunosorbent Assay
Female
Fluorescent Antibody Technique
Humans
Immunophenotyping
Liver / immunology
Liver / metabolism
Liver / pathology
Liver Cirrhosis, Biliary / diagnosis*
Liver Cirrhosis, Biliary / immunology*
Liver Cirrhosis, Biliary / metabolism
Lymphocyte Count
Male
Middle Aged
Phenotype*
Plasma Cells / immunology
Plasma Cells / metabolism
Young Adult

Substances

Antigens, CD20
Autoantibodies
Biomarkers

Grants and funding

MC_PC_13057/MRC_/Medical Research Council/United Kingdom