Contributions
Abstract: P458
Type: Poster
Abstract Category: Pathology and pathogenesis of MS - 15 Immunology
Objective: Determine stability across time of clonally related cerebrospinal fluid (CSF) B cells in patients with multiple sclerosis (MS).
Background: Anti-CD20 therapy efficacy highlights the need to better understand the role B cells play in MS pathophysiology. We investigated whether related
B cell clonotypes persist in the CSF over time.
Design and methods: IgM and IgG B cell receptor (BCR) heavy chain variable region immune repertoires were generated on an Ion Torrent machine using RNA extracted from CSF and peripheral blood B cells of ten MS patients at an untreated time point (a) and at a later time point (b) (1.18 years later (+/- 0.28)). A custom bioinformatics pipeline based on MiXCR1 identified CDR3 sequences. Clonally related BCRs were identified by comparing CDR3 sequences using a distance metric approach.
Results: In five of the ten patients, related CSF B cell clonotypes were found at both time points. Persistent clonally related B cells were more often plasmablast/plasma cell (15/31) and switched memory (14/31) phenotype rather than unswitched memory (1/31) double negative (1/31) or naïve (0/31). Persistent CSF B cell relatives' CDR3s were unique to each patient.
Conclusions: To our knowledge, this is the first longitudinal B cell immune repertoire study in the CSF compartment of MS patients. Persistent related CSF B cells are more likely to be found in patients with a recent relapse, suggesting that recurring recruitment or intrathecal persistence of disease-associated
B cells may be a marker of active relapsing disease. These persistent B cell clonal relatives will also serve as a population to further investigate at a single cell level.
1. Bolotin, D. A. et al. MiXCR: software for comprehensive adaptive immunity profiling. Nature Methods 12, 380-381 (2015).
Disclosure:
S. L. Hauser currently serves on the SAB of Symbiotix, Annexon, Bionure, and Molecular Stethoscope and on the BOT of Neurona.
Dr. Hauser also has received travel reimbursement and writing assistance from F. Hoffman-La Roche Ltd. for CD20-related meetings and presentations.
Bruce Cree has received personal compensation for consulting from Abbvie, Biogen, EMD Serono and Novartis.
At the time of submission, Dr. von Büdingen is an employee of F. Hoffman-La Roche, Basel, Switzerland and holds an adjunct faculty position at UCSF. This work does not represent the opinion of Roche. Dr. von Büdingen has received compensation for consulting activities from Roche, Novartis, and Genzyme, and research funding from Roche, Genentech, and Pfizer.
Dr. Wilson has received research funding from Roche-Genentech.
Dr. Greenfield has nothing to disclose.
Erica Eggers has nothing to disclose.
Hao Wu has nothing to disclose.
Dr. Laurent has nothing to disclose.
Dr. Michel has nothing to disclose.
William Harkin has nothing to disclose.
Natalie Pierson has nothing to disclose.
Abstract: P458
Type: Poster
Abstract Category: Pathology and pathogenesis of MS - 15 Immunology
Objective: Determine stability across time of clonally related cerebrospinal fluid (CSF) B cells in patients with multiple sclerosis (MS).
Background: Anti-CD20 therapy efficacy highlights the need to better understand the role B cells play in MS pathophysiology. We investigated whether related
B cell clonotypes persist in the CSF over time.
Design and methods: IgM and IgG B cell receptor (BCR) heavy chain variable region immune repertoires were generated on an Ion Torrent machine using RNA extracted from CSF and peripheral blood B cells of ten MS patients at an untreated time point (a) and at a later time point (b) (1.18 years later (+/- 0.28)). A custom bioinformatics pipeline based on MiXCR1 identified CDR3 sequences. Clonally related BCRs were identified by comparing CDR3 sequences using a distance metric approach.
Results: In five of the ten patients, related CSF B cell clonotypes were found at both time points. Persistent clonally related B cells were more often plasmablast/plasma cell (15/31) and switched memory (14/31) phenotype rather than unswitched memory (1/31) double negative (1/31) or naïve (0/31). Persistent CSF B cell relatives' CDR3s were unique to each patient.
Conclusions: To our knowledge, this is the first longitudinal B cell immune repertoire study in the CSF compartment of MS patients. Persistent related CSF B cells are more likely to be found in patients with a recent relapse, suggesting that recurring recruitment or intrathecal persistence of disease-associated
B cells may be a marker of active relapsing disease. These persistent B cell clonal relatives will also serve as a population to further investigate at a single cell level.
1. Bolotin, D. A. et al. MiXCR: software for comprehensive adaptive immunity profiling. Nature Methods 12, 380-381 (2015).
Disclosure:
S. L. Hauser currently serves on the SAB of Symbiotix, Annexon, Bionure, and Molecular Stethoscope and on the BOT of Neurona.
Dr. Hauser also has received travel reimbursement and writing assistance from F. Hoffman-La Roche Ltd. for CD20-related meetings and presentations.
Bruce Cree has received personal compensation for consulting from Abbvie, Biogen, EMD Serono and Novartis.
At the time of submission, Dr. von Büdingen is an employee of F. Hoffman-La Roche, Basel, Switzerland and holds an adjunct faculty position at UCSF. This work does not represent the opinion of Roche. Dr. von Büdingen has received compensation for consulting activities from Roche, Novartis, and Genzyme, and research funding from Roche, Genentech, and Pfizer.
Dr. Wilson has received research funding from Roche-Genentech.
Dr. Greenfield has nothing to disclose.
Erica Eggers has nothing to disclose.
Hao Wu has nothing to disclose.
Dr. Laurent has nothing to disclose.
Dr. Michel has nothing to disclose.
William Harkin has nothing to disclose.
Natalie Pierson has nothing to disclose.