Volume 10, Issue 5, Pages (May 1999)

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Volume 10, Issue 5, Pages 619-628 (May 1999) Mutations Affecting Either Generation or Survival of Cells Influence the Pool Size of Mature B Cells Antonius G Rolink, Thomas Brocker, Horst Bluethmann, Marie H Kosco-Vilbois, Jan Andersson, Fritz Melchers Immunity Volume 10, Issue 5, Pages 619-628 (May 1999) DOI: 10.1016/S1074-7613(00)80061-8

Figure 1 Defective B Cell Maturation in I-Aα−/− Mice (A) B220/pB130-140 expression on bone marrow and spleen of adult B6, B6 I-Aβ−/−, and B6 I-Aα−/− mice. Bone marrow and spleen cells were labeled with mAb 493 and anti-B220 mAb and were analyzed by flow cytometry. Numbers in quadrants represent percentage of total gated lymphoid cells. The absolute numbers of spleen cells were 6.6 ± 1.6 × 107 (B6), 5.9 ± 1.2 × 107 (B6 I-Aβ−/−), and 3.4 ± 0.9 × 107 (B6 I-Aα−/− mice). (B) BrdU-labeled B220+ spleen cells. Adult B6, B6 I-Aβ−/−, and B6 I-Aα−/− mice were given BrdU for 4 days in their drinking water. Splenic cells were analyzed for BrdU incorporation. Histograms represent BrdU labeling of B220+ gated spleen cells. Numbers represent percentage of BrdU-positive B220+ spleen cells. Immunity 1999 10, 619-628DOI: (10.1016/S1074-7613(00)80061-8)

Figure 2 Complementation of the B Cell Defect in I-Aα−/− Mice by an I-Eα Transgene (A) B220/pB130-140 expression on bone marrow and spleen cells of adult B6 I-Aα−/− mice complemented with the EAD-107 or the EAD-J transgene. Spleen and bone marrow cells were labeled with mAb 493 (anti-pB130-140) and an anti-B220 mAb and were analyzed by flow cytometry. (B) I-E expression on bone marrow and spleen cells of B6 I-Aα−/− mice complemented with the EAD-107 or the EAD-J transgene. Spleen and bone marrow cells were labeled with the I-E-specific mAb 14-4-4S (Pharmingen) and an anti-B220 mAb and then analyzed by flow cytometry. Numbers in quadrants indicate percentage of total gated lymphoid cells. Total number of spleen cells were 3.3 ± 0.6 × 107 (EAD-107) and 6.0 ± 0.7 × 107 (EAD-J). Immunity 1999 10, 619-628DOI: (10.1016/S1074-7613(00)80061-8)

Figure 3 Severe B Cell Maturation Defect in B6 I-Aα−/−/CBA/N Double Mutant Mice B220/pB130-140 expression on spleen cells from adult CBA/N and B6 I-Aα−/−/CBA/N double mutant mice. Spleen cells were labeled with mAb 493 and an anti-B220 mAb and were analyzed by flow cytometry. Numbers in quadrants indicate percentage of total gated lymphoid cells. Immunity 1999 10, 619-628DOI: (10.1016/S1074-7613(00)80061-8)

Figure 4 B6 I-Aα−/−/CBA/N Double Mutant Mice Lack B Cell Follicles Cryosections from (A) B6 I-Aα−/−, (B) CBA/N, and (C). B6 I-Aα−/−/CBA/N mice were incubated with Texas red–conjugated goat anti-mouse IgM (red) and rat anti-Thy 1 (T24) followed by mouse anti-rat IgG conjugated to FITC (green). While the single mutant mice show development of B cell zones (red) around the T cell containing PALS (green), these structures are absent or drastically reduced in the double mutant (C). Immunity 1999 10, 619-628DOI: (10.1016/S1074-7613(00)80061-8)