title: Apoptotic effect of rituximab on peripheral blood B cells in rheumatoid arthritis.
abstract: Rituximab (RTX) has proven efficacious in the treatment of rheumatoid arthritis (RA). Herein, we assessed the apoptosis-inducing capability of RTX in vitro on RA peripheral blood B-cell subsets and also compared the effects of RTX on B cells from rheumatoid factor-positive (RF+) and RF- patients. The likely relevance of B cells in disease was assessed by measuring B-cell-modulating serum cytokines. Peripheral blood B cells were isolated and cultured with the presence or absence of RTX. The levels of apoptosis within the naïve, memory and IgD+CD27+ B-cell subpopulations were determined by cytofluorometric analysis and caspase 3/7 assays. Levels of serum cytokines were measured with a multiplex cytokine array system. RTX induced significant apoptosis in all B-cell subsets in both RA and controls. In naïve and memory B cells from RA patients, RTX induced significantly higher levels of apoptosis than in controls. RTX induced apoptosis of B cells in RF+ and RF- patients. Serum levels of interleukin-1beta (IL-1beta), IL-4, IL-10 and IL-13 were profoundly increased in RF+ patients compared to RF- patients and controls. Although our cohort was small (10 RA patients), the data suggest that RTX induces apoptosis in all investigated subsets of B cells from RA patients. Interestingly, memory B cells from RA patients were more sensitive to RTX than memory cells from normal controls, suggesting that the delay in treatment response to RTX observed in clinical trials may be due in part to memory cell depletion. The apoptotic effects of RTX were similar in RF+ and RF- patients, but serum levels of B-cell-activating cytokine levels were only elevated in RF+ but not RF- patients. These data suggest that RTX is less effective in RF- RA because B cells play a less significant role in RA pathogenesis in RF- patients.
authors.affiliation: Broegelmann Research Laboratory, The Gade Institute, University of Bergen, Armauer Hansen Building, N-5021 Bergen, Norway. peter.szodoray@gades.uib.no
authors.forename: P
authors.initials: P
authors.lastname: Szodoray
authors.forename: P
authors.initials: P
authors.lastname: Alex
authors.forename: V
authors.initials: V
authors.lastname: Dandapani
authors.forename: B
authors.initials: B
authors.lastname: Nakken
authors.forename: J
authors.initials: J
authors.lastname: Pesina
authors.forename: X
authors.initials: X
authors.lastname: Kim
authors.forename: G L
authors.initials: GL
authors.lastname: Wallis
authors.forename: P C
authors.initials: PC
authors.lastname: Wilson
authors.forename: R
authors.initials: R
authors.lastname: Jonsson
authors.forename: M
authors.initials: M
authors.lastname: Centola
mesh_terms: D000911:Antibodies, Monoclonal / Q000494:pharmacology*; D058846:Antibodies, Monoclonal, Murine-Derived; D018501:Antirheumatic Agents / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects* / Q000276:immunology; D001172:Arthritis, Rheumatoid / Q000097:blood* / Q000188:drug therapy / Q000276:immunology; D016175:B-Lymphocyte Subsets / Q000187:drug effects / Q000276:immunology; D001402:B-Lymphocytes / Q000187:drug effects* / Q000276:immunology; D053148:Caspase 3; D020169:Caspases / Q000276:immunology; D016207:Cytokines / Q000276:immunology; D005260:Female; D005434:Flow Cytometry; D006801:Humans; D007118:Immunoassay; D008297:Male; D012217:Rheumatoid Factor / Q000276:immunology; D000069283:Rituximab; D018709:Statistics, Nonparametric
chemical_list: D000911:Antibodies, Monoclonal; D058846:Antibodies, Monoclonal, Murine-Derived; D018501:Antirheumatic Agents; D016207:Cytokines; D000069283:Rituximab; D012217:Rheumatoid Factor; C505407:CASP3 protein, human; D053148:Caspase 3; D020169:Caspases