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Rheumatology Advance Access originally published online on December 20, 2005
Rheumatology 2006 45(5):549-557; doi:10.1093/rheumatology/kei170
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© The Author 2005. Published by Oxford University Press on behalf of the British Society for Rheumatology. All rights reserved. For Permissions, please email: journals.permissions@oxfordjournals.org

Limited VH gene usage in B-cell clones established with nurse-like cells from patients with rheumatoid arthritis

S. Nakamura-Kikuoka1, K. Takahi2, H. Tsuboi2, T. Toyosaki-Maeda1, M. Maeda-Tanimura1, C. Wakasa1, N. Kikuchi1, S. Norioka3, M. Iwasaki1, T. Matsutani4, T. Itoh4, S. Yamane1, H. Takemoto1, Y. Tsuruta1, Y. Shimaoka2, M. Yukioka5, R. Suzuki1,6 and T. Ochi6

1 Discovery Research Laboratories, Shionogi & Co., Ltd, 2 Department of Orthopaedics, Osaka University Medical School, 3 Graduate School of Frontier Biosciences, Osaka University, 4 Department of Cell Biology, Tohoku University School of Medicine, Sendai, 5 Yukioka Hospital, Osaka and 6 Clinical Research Center for Rheumatology and Allergy, National Sagamihara Hospital, Kanagawa, Japan.

Correspondence to: T. Ochi, Clinical Research Center for Rheumatology and Allergy, National Sagamihara Hospital, 18-1 Sakuradai, Sagamihara, Kanagawa 228-8522, Japan. E-mail: t-ochi{at}sagamihara-hosp.gr.jp

Objectives. Nurse-like stromal cells (NLC) in synovia and bone marrow of patients with rheumatoid arthritis (RA) can support pseudoemperipolesis, protect from apoptosis and enhance immunoglobulin production of peripheral blood B cells isolated from healthy individuals, suggesting the profound contribution of hyperactivation of B cells in RA. In the course of establishing RA-NLC from RA patients, we observed the growth of B cells in the presence of RA-NLC.

Methods. We cloned B cells from the synovium or bone marrow of RA patients using the limiting dilution technique. For established clones, nucleotide sequences of immunoglobulin and surface antigens were investigated. To investigate the dependence of these clones on NLC, differences in the proliferation and the amount of immunoglobulin produced in the presence or absence of NLC were compared. Immunocytochemical staining of various cells was performed using the antibody these clones produced.

Results. Nine B-cell clones established from RA patients showed RA-NLC-dependent growth. These B-cell clones expressed CD19, CD20, CD38, CD39 and CD40, suggesting that the cloned cells were mature and activated. All clones secreted immunoglobulins in culture media, which were specific for intracellular components of various cell lines, including RA-NLC. Interestingly, we found limited usage of immunoglobulin heavy-chain variable regions (VH) among B-cell clones from RA patients. These repertoires were reported to be detected preferentially in fetal livers.

Conclusion. The present study provides a novel insight into the involvement of RA-NLC in the immunopathogenesis of RA via an autoreactive B cell development and/or activation mechanism.

KEY WORDS: Rheumatoid arthritis, B cell, Immunoglobulin VH gene, Autoantibody


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