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Blood, 15 February 2007, Vol. 109, No. 4, pp. 1611-1619.
Prepublished online as a Blood First Edition Paper on October 10, 2006; DOI 10.1182/blood-2006-03-008441.
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IMMUNOBIOLOGY
CpG DNA activation and plasma-cell differentiation of CD27 naive human B cells
Jennifer Huggins1,
Tina Pellegrin2,
Raymond E. Felgar3,
Chungwen Wei1,
Miguel Brown2,
Bo Zheng1,
Eric C. B. Milner1,
Steven H. Bernstein4,
Ignacio Sanz1, and
Martin S. Zand2
1 Division of Allergy, Immunology and Rheumatology,
2 Division of Nephrology,
3 Department of Pathology,
4 James P. Wilmot Cancer Center, University of Rochester Medical Center, NY
Unmethylated CpG DNA activation of naive CD27 B cells has been reported to require B-cellreceptor (BCR) cross-linking. We describe a culture system using CpG DNA with sequential steps for T-cellindependent activation of naive CD19+CD27 human peripheral blood B cells that induces efficient CD138+ plasma-cell differentiation. CD27+ and CD27 B cells were cultured in a 3-step system: (1) days 0 to 4: CpG, IL-2/10/15; (2) days 4 to 7: IL-2/6/10/15 and anti-CD40L; (3) days 7 to 10: IL-6/15, IFN- , hepatocyte growth factor, and hyaluronic acid. Both CD27+ and CD27 B cells up-regulated intracytoplasmic TLR-9 following CpG DNA activation. CD27 B-cell activation required cell-cell contact. Both naive and memory B cells progressed to a plasma-cell phenotype: CD19lowCD20lowCD27+CD38+HLA-DRlow. Seventy percent of the CD27-derived CD138+ cells demonstrated productive V chain rearrangements without somatic mutations, confirming their origin from naive precursors. Plasma cells derived from CD27+ B cells were primarily IgG+, while those from CD27 B cells were IgM+. Our results indicate that under certain conditions, naive B cells increase TLR-9 expression and proliferate to CpG DNA stimulation without BCR signaling. In addition to its immunologic significance, this system should be a valuable method to interrogate the antigenic specificity of naive B cells.

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