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. 2007 May 14;204(5):1145–1156. doi: 10.1084/jem.20061952

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Copyright © 2007, The Rockefeller University Press

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Figure 4. — Dynamics of AID expression in T cell–dependent and –independent immune responses. (A) AID-GFP mice were immunized with NP-CGG, and CD4⁻CD8⁻IgD⁻PI (or DAPI)⁻ lymphocytes recognizing NP were subdivided by B220 expression (left) and assessed for CD138 and AID-GFP expression 7–10 d after immunization (middle and right). Antibodies were IgD-biotin/Streptavidin-PE-Cy5, CD4-PE-Cy5, CD8-PE-Cy5, NP-APC, CD95-PE-Cy7, B220-APC-Cy7, and CD138-PE. (B) Left: NP staining of blood lymphocytes (IgM⁻IgD⁻Gr-1⁻F4/80⁻) recovered from unimmunized (day 0) and NP-CGG–immunized (day 7) wild-type mice. Staining was To-Pro-3, F4/80-Alexa 647, GR1-Alexa 647, IgM-APC, IgD-Alexa 647 (Dump channel) and NP-PE, IgG1-Biotin-Streptavidin-PercPCy5.5, and B220-FITC. Right: Analysis of AID transcription by SC-RT-PCR. NP⁺ cells were recovered by cell sorting from blood (72 cells analyzed) or lymph node GCs (72 cells) isolated from wild-type or AID^−/− mice. (C) Left: AID-GFP animals carrying the pre-recombined B1-8^hi heavy chain gene were immunized with NP-haptenated Ficoll and splenic B cells analyzed 5 d after immunization. AID-GFP expression in resting (B220⁺CD138⁻), T cell–independent GC (B220⁺CD138^low), plasmablasts (B220⁺CD138^high), and plasma cells (B220⁻CD138^high) is indicated with histograms. Bottom histograms show CD95 expression in AID-GFP⁺ cells from population 3 or population 4. Right: Analysis of AID transcription by SC-RT-PCR in the indicated populations isolated from B1-8^hi mice.