TY - JOUR
T1 - Phenotyping and auto-antibody production by liver-infiltrating B cells in primary sclerosing cholangitis and primary biliary cholangitis
AU - Chung, Brian
AU - Guevel, Bardia T.
AU - Reynolds, Gary
AU - Gupta Udatha, D.B.R.K.
AU - Henriksen, Eva Kristine Klemsdal
AU - Stamataki, Zania
AU - Hirschfield, Gideon
AU - Karlsen, Tom Hemming
AU - Liaskou, Evaggelia
PY - 2016/10/24
Y1 - 2016/10/24
N2 - 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 (CD19þCD27þCD38hiCD138) and plasma cell (CD19þCD27þCD38hiCD138þ) ASCs were enumerated by flow cytometry. We observed 45-fold fewer plasma cells in PSC explants (n ¼ 9) compared to PBC
samples (n ¼ 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 ¼ 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 ¼ 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 ¼ 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.
AB - 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 (CD19þCD27þCD38hiCD138) and plasma cell (CD19þCD27þCD38hiCD138þ) ASCs were enumerated by flow cytometry. We observed 45-fold fewer plasma cells in PSC explants (n ¼ 9) compared to PBC
samples (n ¼ 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 ¼ 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 ¼ 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 ¼ 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.
KW - Primary biliary cholangitis
KW - Primary sclerosing cholangitis
KW - Antibody-secretaing B cells
KW - Autoimmunity
KW - Auto-antibodies
KW - Protein arrays
KW - Biomarkers
U2 - 10.1016/j.jaut.2016.10.003
DO - 10.1016/j.jaut.2016.10.003
M3 - Article
SN - 0896-8411
JO - Journal of Autoimmunity
JF - Journal of Autoimmunity
ER -