The Journal of Experimental Medicine
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Published online
doi:10.1084/jem.20062571
The Journal of Experimental Medicine, Vol. 204, No. 9, 2103-2114
The Rockefeller University Press, 0022-1007 $30.00
© Anderson et al.
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Article

 New markers for murine memory B cells that define mutated and unmutated subsets

Shannon M. Anderson1, Mary M. Tomayko3, Anupama Ahuja2, Ann M. Haberman1, and Mark J. Shlomchik1,2

1 Section of Immunobiology, 2 Department of Laboratory Medicine, and 3 Department of Dermatology, Yale University School of Medicine, New Haven, CT 06510

CORRESPONDENCE Mark Shlomchik: mark.shlomchik{at}yale.edu

The study of murine memory B cells has been limited by small cell numbers and the lack of a definitive marker. We have addressed some of these difficulties with hapten-specific transgenic (Tg) mouse models that yield relatively large numbers of antigen-specific memory B cells upon immunization. Using these models, along with a 5-bromo-2'-deoxyuridine (BrdU) pulse-label strategy, we compared memory cells to their naive precursors in a comprehensive flow cytometric survey, thus revealing several new murine memory B cell markers. Most interestingly, memory cells were phenotypically heterogeneous. Particularly surprising was the finding of an unmutated memory B cell subset identified by the expression of CD80 and CD35. We confirmed these findings in an analogous V region knock-in mouse and/or in non-Tg mice. There also was anatomic heterogeneity, with BrdU+ memory cells residing not just in the marginal zone, as had been thought, but also in splenic follicles. These studies impact the current understanding of murine memory B cells by identifying new phenotypes and by challenging assumptions about the location and V region mutation status of memory cells. The apparent heterogeneity in the memory compartment implies either different origins and/or different functions, which we discuss.

Abbreviations used: Ab, antibody; AFC, antibody-forming cell; Ag, antigen; AP, alkaline phosphatase; APC, allophycocyanin; BCR, B cell receptor; CDR, complementarity-determining region; FW, framework region; GC, germinal center; MFI, mean fluorescence intensity; MZ, marginal zone; NIP, 4-hydroxy-3-nitroiodophenyl; NP, 4-hydroxy-3-nitrophenyl; Tg, transgenic.

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