The Journal of Experimental Medicine
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Published online April 23, 2007
doi:10.1084/jem.20061952
The Journal of Experimental Medicine, Vol. 204, No. 5, 1145-1156
The Rockefeller University Press, 0022-1007 $30.00
© 2007 Crouch et al.
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ARTICLE

 Regulation of AID expression in the immune response

Elizabeth E. Crouch1, Zhiyu Li1, Makiko Takizawa1, Stefan Fichtner-Feigl2, Polyxeni Gourzi4, Carolina Montaño1, Lionel Feigenbaum5, Patrick Wilson6, Siegfried Janz3, F. Nina Papavasiliou4, and Rafael Casellas1

1 Genomic Integrity and Immunity, National Institute of Arthritis and Musculoskeletal and Skin Diseases (NIAMS), 2 Mucosal Immunity Section, National Institute of Allergy and Infectious Diseases (NIAID), and 3 Laboratory of Genetics, National Cancer Institute (NCI), National Institutes of Health (NIH), Bethesda, MD 20892
4 Laboratory of Lymphocyte Biology, The Rockefeller University, New York, NY 10021
5 Laboratory Animal Science Program, Science Applications International Corporation (SAIC), NCI, NIH, Frederick, MD 21702
6 Molecular Immunogenetics, Oklahoma Medical Research Foundation, Oklahoma City, OK 73104

CORRESPONDENCE R. Casellas: casellar{at}mail.nih.gov

The B cell–specific enzyme activation-induced cytidine deaminase (AID) has been shown to be essential for isotype switching and affinity maturation of antibody genes during the immune response. Conversely, AID activity has also been linked to autoimmunity and tumorigenesis. Determining how AID expression is regulated in vivo is therefore central to understanding its role in health and disease. Here we use phylogenetic footprinting and high-resolution histone acetylation mapping to accurately demarcate AID gene regulatory boundaries. Based on this strategy, we identify a novel, positive regulatory element required for AID transcription. Furthermore, we generate two AID indicator mouse strains using bacterial artificial chromosomes that faithfully recapitulate endogenous AID expression. The first strain uses a green fluorescent protein reporter to identify B cells that actively express AID during the immune response. In the second strain, AID transcription affects the permanent expression of a yellow fluorescent protein reporter in post–germinal center and terminally differentiated lymphocytes. We demonstrate the usefulness of these novel strains by resolving recent contradictory observations on AID expression during B cell ontogeny.

Abbreviations used: Ab-MLV, Abelson murine leukemia virus; AID, activation-induced cytidine deaminase; BAC, bacterial artificial chromosome; CNS, conserved noncoding sequence; GC, germinal center; ILF, isolated lymphoid follicle; NP, nitrophenol; PP, Peyer's patch; QM, quasimonoclonal; SC-RT-PCR, single cell RT-PCR strategy; YFP, yellow fluorescent protein.

E.E. Crouch and Z. Li contributed equally to this work.


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