Analysis of MZ and FO B cells following BM transplantation.

<p>(A) and (B) Two months following BM transplantation, the four groups of mice were sacrificed and spleen cells were stained using anti-CD19, anti-CD21 and anti-CD23 (A), or anti-CD19, anti-CD21 and anti-CD24 (B) and then analyzed using FCM. The cells were initially gated using anti-CD19. CD24^hiCD21^hi cells represent MZ/T2 B cells, and CD21^medCD24^hi cells represent T1 cells. The numbers denote the percentages of cells in the indicated squares. Data are representative of five independent experiments. (C) Absolute numbers of MZ and FO B cells. Numbers were calculated by multiplying the fraction of each FCM B cell subset analyzed in (A) by the total number of CD19⁺ cells. Data are mean±S.D. obtained from five independent experiments. (D) The ratio of MZ to FO cell number was calculated. (E) Immunofluorescent staining of splenic cryosections with Cy3-anti-IgM and FITC-anti–MOMA-1. MOMA-1⁺ metalophilic macrophages define the border between the MZ B and FO B cells. Data are representative of five independent experiments. Scale bar, 100 µm. (F) Southern blot analysis of the DNA from the BM and spleen of the recipient mice. 4 Kb fragment represents wild-type, and 3 Kb fragment represent deletion.