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International Immunology Advance Access published online on November 29, 2006
International Immunology, doi:10.1093/intimm/dxl122
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© The Japanese Society for Immunology. 2006. All rights reserved. For permissions, please e-mail: journals.permissions@oxfordjournals.org
Received June 22, 2006
Accepted October 23, 2006

Article

Extreme skewing of annexin II and S100A6 expression identified by proteomic analysis of peritoneal B-1 cells

Rubén Francés 1, Joseph R. Tumang 2, and Thomas L. Rothstein 3 *

1 Department of Medicine, Boston University School of Medicine, Boston University Medical Center, Boston, MA 02118, USA; Immunobiology Unit, Evans Memorial Department of Clinical Research, Boston University Medical Center, Boston, MA 02118, USA; Present address: Immunology Section, Hospital General Universitario, Pintor Baeza 12, 03010 Alicante, Spain
2 Department of Medicine, Boston University School of Medicine, Boston University Medical Center, Boston, MA 02118, USA; Immunobiology Unit, Evans Memorial Department of Clinical Research, Boston University Medical Center, Boston, MA 02118, USA; Present address: Center for Oncology and Cell Biology, The Feinstein Institute for Medical Research, 350 Community Drive, Manhasset, NY 11030, USA
3 Department of Medicine, Boston University School of Medicine, Boston University Medical Center, Boston, MA 02118, USA; Immunobiology Unit, Evans Memorial Department of Clinical Research, Boston University Medical Center, Boston, MA 02118, USA; Department of Microbiology, Boston University School of Medicine, Boston University Medical Center, Boston, MA 02118, USA; Present address: Center for Oncology and Cell Biology, The Feinstein Institute for Medical Research, 350 Community Drive, Manhasset, NY 11030, USA

* To whom correspondence should be addressed.
Thomas L. Rothstein, E-mail: tr{at}nshs.edu


  Abstract

B-1 cells differ phenotypically, biochemically and functionally from conventional B-2 cells. The origin of these differences remains uncertain. We hypothesized that unbiased analysis of differences in protein expression between B-1 and B-2 cells might provide information not otherwise available, and thus undertook 1-dimensional (1D) gel analysis combined with mass spectrometry. We identified annexin II and S100A6 in peritoneal B-1 cells (B-1P) but not in splenic B-2 cells (B-2S); these results were confirmed by western blot analysis and reflected in mRNA expression. Further analysis of mRNA indicated that elevated expression levels of annexin II and S100A6 were unique to B-1P among several naive lymphoid populations. However, expression of annexin II and S100A6 protein was up-regulated in mitogenically stimulated B-2S. In both naive B-1 cells and stimulated B-2 cells, annexin II and S100A6 formed Ca++-sensitive complexes. These results confirm that the emerging field of proteomics detects differentially expressed molecules independently of RNA screening methods. These results identify two proteins (annexin II and S100A6) that are unexpectedly differentially expressed in B-1 cells and, although members of larger families, may fulfill unique, subset-specific functions. These results also validate 1D GE/LC-MS/MS as a reliable screening tool in identifying final protein product expression differences between B-1P and B-2S.

Keywords: annexin; B cells; calcium; proteomics; rodent.
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