Transfer of small resting B cells into immunodeficient hosts results in the selection of a self-renewing activated B cell population - PubMed (original) (raw)
Transfer of small resting B cells into immunodeficient hosts results in the selection of a self-renewing activated B cell population
F Agenès et al. J Exp Med. 1999.
Abstract
We studied the role of bone marrow B cell production in the renewal of peripheral B cells and the feedback mechanisms that control the entry of newly formed B cells into the peripheral B cell pools. When resting lymph node B cells are injected into B cell-deficient hosts, a fraction of the transferred cells expands and constitutes a highly selected population that survives for prolonged periods of time by continuous cell renewal at the periphery. Although the number of donor B cells recovered is low, a significant fraction shows an activated phenotype, and the serum immunoglobulin (Ig)M levels are as in normal mice. This population of activated B cells is resistant to replacement by a new cohort of B cells and is able to feedback regulate both the entry of newly formed B cells into the peripheral pool and terminal differentiation. These findings suggest that peripheral B cell selection follows the first come, first served rule and that IgM-secreting cells are generated from a pool of stable activated B cells with an independent homeostasis.
Figures
Figure 1
(A) Fate of LN B cells transferred into immunodeficient Rag2−/− mice. Each host mouse was injected with 107 cells (4 × 106 B cells). Each point represents the number of donor LN-derived B cells (B220+ μ+) recovered in the SPL of three to six host mice as a function of time. The curve joins the mean of the values obtained at each time point. (B) Phenotype of in vivo activated B cells. Rag2−/− mice were injected with 4 × 107 LN cells (1.8 × 107 B cells). The hosts were killed 2 mo later and the SPL cells analyzed by flow cytometry. Histograms are gated for the B220+ B cell population. Bold line, phenotypic analysis of LN-derived B cells. Thin line, phenotypic analysis of splenic B cells from C57Bl/6 control mice. (C) Contour plot showing the sorted B2 cells from CD3ε−/− donors before and after in vivo activation in Rag2−/− hosts. Similar observations were made when LN cells from the same donors were transferred. (D) Cytokine profile of in vivo activated B cells. RT-PCR was performed on sorted B cells (>99% pure). Lanes from the left to the right: splenic cells from Rag2−/− mice injected 1 mo before with LN cells from CD3ε−/− donors (pool of 10 mice); mature B cells (IgMlowIgDhigh) from the SPL of CD3ε−/− mice (pool of 5 mice); activated B cells (IgMhighIgDlow) from the SPL of CD3ε−/− mice (pool of 5 mice); B cells from the LN of CD3ε−/− mice (pool of 5 mice). (E) Number of IgM- secreting cells in the SPL of immunodeficient hosts injected with 107 cells (4 × 106 B cells) as a function of time. Each point represents one individual mouse and the curve joins the mean values obtained in each time point.
Figure 1
(A) Fate of LN B cells transferred into immunodeficient Rag2−/− mice. Each host mouse was injected with 107 cells (4 × 106 B cells). Each point represents the number of donor LN-derived B cells (B220+ μ+) recovered in the SPL of three to six host mice as a function of time. The curve joins the mean of the values obtained at each time point. (B) Phenotype of in vivo activated B cells. Rag2−/− mice were injected with 4 × 107 LN cells (1.8 × 107 B cells). The hosts were killed 2 mo later and the SPL cells analyzed by flow cytometry. Histograms are gated for the B220+ B cell population. Bold line, phenotypic analysis of LN-derived B cells. Thin line, phenotypic analysis of splenic B cells from C57Bl/6 control mice. (C) Contour plot showing the sorted B2 cells from CD3ε−/− donors before and after in vivo activation in Rag2−/− hosts. Similar observations were made when LN cells from the same donors were transferred. (D) Cytokine profile of in vivo activated B cells. RT-PCR was performed on sorted B cells (>99% pure). Lanes from the left to the right: splenic cells from Rag2−/− mice injected 1 mo before with LN cells from CD3ε−/− donors (pool of 10 mice); mature B cells (IgMlowIgDhigh) from the SPL of CD3ε−/− mice (pool of 5 mice); activated B cells (IgMhighIgDlow) from the SPL of CD3ε−/− mice (pool of 5 mice); B cells from the LN of CD3ε−/− mice (pool of 5 mice). (E) Number of IgM- secreting cells in the SPL of immunodeficient hosts injected with 107 cells (4 × 106 B cells) as a function of time. Each point represents one individual mouse and the curve joins the mean values obtained in each time point.
Figure 1
(A) Fate of LN B cells transferred into immunodeficient Rag2−/− mice. Each host mouse was injected with 107 cells (4 × 106 B cells). Each point represents the number of donor LN-derived B cells (B220+ μ+) recovered in the SPL of three to six host mice as a function of time. The curve joins the mean of the values obtained at each time point. (B) Phenotype of in vivo activated B cells. Rag2−/− mice were injected with 4 × 107 LN cells (1.8 × 107 B cells). The hosts were killed 2 mo later and the SPL cells analyzed by flow cytometry. Histograms are gated for the B220+ B cell population. Bold line, phenotypic analysis of LN-derived B cells. Thin line, phenotypic analysis of splenic B cells from C57Bl/6 control mice. (C) Contour plot showing the sorted B2 cells from CD3ε−/− donors before and after in vivo activation in Rag2−/− hosts. Similar observations were made when LN cells from the same donors were transferred. (D) Cytokine profile of in vivo activated B cells. RT-PCR was performed on sorted B cells (>99% pure). Lanes from the left to the right: splenic cells from Rag2−/− mice injected 1 mo before with LN cells from CD3ε−/− donors (pool of 10 mice); mature B cells (IgMlowIgDhigh) from the SPL of CD3ε−/− mice (pool of 5 mice); activated B cells (IgMhighIgDlow) from the SPL of CD3ε−/− mice (pool of 5 mice); B cells from the LN of CD3ε−/− mice (pool of 5 mice). (E) Number of IgM- secreting cells in the SPL of immunodeficient hosts injected with 107 cells (4 × 106 B cells) as a function of time. Each point represents one individual mouse and the curve joins the mean values obtained in each time point.
Figure 1
(A) Fate of LN B cells transferred into immunodeficient Rag2−/− mice. Each host mouse was injected with 107 cells (4 × 106 B cells). Each point represents the number of donor LN-derived B cells (B220+ μ+) recovered in the SPL of three to six host mice as a function of time. The curve joins the mean of the values obtained at each time point. (B) Phenotype of in vivo activated B cells. Rag2−/− mice were injected with 4 × 107 LN cells (1.8 × 107 B cells). The hosts were killed 2 mo later and the SPL cells analyzed by flow cytometry. Histograms are gated for the B220+ B cell population. Bold line, phenotypic analysis of LN-derived B cells. Thin line, phenotypic analysis of splenic B cells from C57Bl/6 control mice. (C) Contour plot showing the sorted B2 cells from CD3ε−/− donors before and after in vivo activation in Rag2−/− hosts. Similar observations were made when LN cells from the same donors were transferred. (D) Cytokine profile of in vivo activated B cells. RT-PCR was performed on sorted B cells (>99% pure). Lanes from the left to the right: splenic cells from Rag2−/− mice injected 1 mo before with LN cells from CD3ε−/− donors (pool of 10 mice); mature B cells (IgMlowIgDhigh) from the SPL of CD3ε−/− mice (pool of 5 mice); activated B cells (IgMhighIgDlow) from the SPL of CD3ε−/− mice (pool of 5 mice); B cells from the LN of CD3ε−/− mice (pool of 5 mice). (E) Number of IgM- secreting cells in the SPL of immunodeficient hosts injected with 107 cells (4 × 106 B cells) as a function of time. Each point represents one individual mouse and the curve joins the mean values obtained in each time point.
Figure 1
(A) Fate of LN B cells transferred into immunodeficient Rag2−/− mice. Each host mouse was injected with 107 cells (4 × 106 B cells). Each point represents the number of donor LN-derived B cells (B220+ μ+) recovered in the SPL of three to six host mice as a function of time. The curve joins the mean of the values obtained at each time point. (B) Phenotype of in vivo activated B cells. Rag2−/− mice were injected with 4 × 107 LN cells (1.8 × 107 B cells). The hosts were killed 2 mo later and the SPL cells analyzed by flow cytometry. Histograms are gated for the B220+ B cell population. Bold line, phenotypic analysis of LN-derived B cells. Thin line, phenotypic analysis of splenic B cells from C57Bl/6 control mice. (C) Contour plot showing the sorted B2 cells from CD3ε−/− donors before and after in vivo activation in Rag2−/− hosts. Similar observations were made when LN cells from the same donors were transferred. (D) Cytokine profile of in vivo activated B cells. RT-PCR was performed on sorted B cells (>99% pure). Lanes from the left to the right: splenic cells from Rag2−/− mice injected 1 mo before with LN cells from CD3ε−/− donors (pool of 10 mice); mature B cells (IgMlowIgDhigh) from the SPL of CD3ε−/− mice (pool of 5 mice); activated B cells (IgMhighIgDlow) from the SPL of CD3ε−/− mice (pool of 5 mice); B cells from the LN of CD3ε−/− mice (pool of 5 mice). (E) Number of IgM- secreting cells in the SPL of immunodeficient hosts injected with 107 cells (4 × 106 B cells) as a function of time. Each point represents one individual mouse and the curve joins the mean values obtained in each time point.
Figure 2
(A) Number of B cells recovered in the SPL of immunodeficient Rag2−/− mice as a function of time. Each curve corresponds to a group of mice (three to six individuals) injected with a particular number of cells: □, 106 LN cells (0.3 × 106 B); ⋄, 5 × 106 LN cells (1.5 × 106 B cells); ○, 10 × 106 LN cells (3 × 106 B cells); and ▵, 30 × 106 LN cells (9 × 106 B cells). (B) Number of IgM-secreting cells recovered in the SPL of immunodeficient mice as a function of time. Each curve corresponds to a group of mice (three to six individuals) injected with a particular number of cells: □, 106 LN cells (0.3 × 106 B cells); ⋄, 5 × 106 LN cells (1.5 × 106 B cells); ○, 10 × 106 LN cells (3 × 106 B cells); ▵, 30 × 106 LN cells (9 × 106 B cells). (C) Number of B cells recovered in the SPL (○), the LN (▵), and the BM (⋄) as a function of the number of LN B cells injected into immunodeficient mice two mo after transfer (mean ± SD of three to six mice). (D) Phenotype of B cells recovered in the BM, SPL, and LN of the same immunodeficient mouse reconstituted with 4 × 107 LN B cells. In the SPL the majority of B cells show an activated phenotype (IgMhighIgDlow), whereas resting B cells (IgMlowIgDhigh) accumulate in the SPL, LN, and BM.
Figure 2
(A) Number of B cells recovered in the SPL of immunodeficient Rag2−/− mice as a function of time. Each curve corresponds to a group of mice (three to six individuals) injected with a particular number of cells: □, 106 LN cells (0.3 × 106 B); ⋄, 5 × 106 LN cells (1.5 × 106 B cells); ○, 10 × 106 LN cells (3 × 106 B cells); and ▵, 30 × 106 LN cells (9 × 106 B cells). (B) Number of IgM-secreting cells recovered in the SPL of immunodeficient mice as a function of time. Each curve corresponds to a group of mice (three to six individuals) injected with a particular number of cells: □, 106 LN cells (0.3 × 106 B cells); ⋄, 5 × 106 LN cells (1.5 × 106 B cells); ○, 10 × 106 LN cells (3 × 106 B cells); ▵, 30 × 106 LN cells (9 × 106 B cells). (C) Number of B cells recovered in the SPL (○), the LN (▵), and the BM (⋄) as a function of the number of LN B cells injected into immunodeficient mice two mo after transfer (mean ± SD of three to six mice). (D) Phenotype of B cells recovered in the BM, SPL, and LN of the same immunodeficient mouse reconstituted with 4 × 107 LN B cells. In the SPL the majority of B cells show an activated phenotype (IgMhighIgDlow), whereas resting B cells (IgMlowIgDhigh) accumulate in the SPL, LN, and BM.
Figure 2
(A) Number of B cells recovered in the SPL of immunodeficient Rag2−/− mice as a function of time. Each curve corresponds to a group of mice (three to six individuals) injected with a particular number of cells: □, 106 LN cells (0.3 × 106 B); ⋄, 5 × 106 LN cells (1.5 × 106 B cells); ○, 10 × 106 LN cells (3 × 106 B cells); and ▵, 30 × 106 LN cells (9 × 106 B cells). (B) Number of IgM-secreting cells recovered in the SPL of immunodeficient mice as a function of time. Each curve corresponds to a group of mice (three to six individuals) injected with a particular number of cells: □, 106 LN cells (0.3 × 106 B cells); ⋄, 5 × 106 LN cells (1.5 × 106 B cells); ○, 10 × 106 LN cells (3 × 106 B cells); ▵, 30 × 106 LN cells (9 × 106 B cells). (C) Number of B cells recovered in the SPL (○), the LN (▵), and the BM (⋄) as a function of the number of LN B cells injected into immunodeficient mice two mo after transfer (mean ± SD of three to six mice). (D) Phenotype of B cells recovered in the BM, SPL, and LN of the same immunodeficient mouse reconstituted with 4 × 107 LN B cells. In the SPL the majority of B cells show an activated phenotype (IgMhighIgDlow), whereas resting B cells (IgMlowIgDhigh) accumulate in the SPL, LN, and BM.
Figure 2
(A) Number of B cells recovered in the SPL of immunodeficient Rag2−/− mice as a function of time. Each curve corresponds to a group of mice (three to six individuals) injected with a particular number of cells: □, 106 LN cells (0.3 × 106 B); ⋄, 5 × 106 LN cells (1.5 × 106 B cells); ○, 10 × 106 LN cells (3 × 106 B cells); and ▵, 30 × 106 LN cells (9 × 106 B cells). (B) Number of IgM-secreting cells recovered in the SPL of immunodeficient mice as a function of time. Each curve corresponds to a group of mice (three to six individuals) injected with a particular number of cells: □, 106 LN cells (0.3 × 106 B cells); ⋄, 5 × 106 LN cells (1.5 × 106 B cells); ○, 10 × 106 LN cells (3 × 106 B cells); ▵, 30 × 106 LN cells (9 × 106 B cells). (C) Number of B cells recovered in the SPL (○), the LN (▵), and the BM (⋄) as a function of the number of LN B cells injected into immunodeficient mice two mo after transfer (mean ± SD of three to six mice). (D) Phenotype of B cells recovered in the BM, SPL, and LN of the same immunodeficient mouse reconstituted with 4 × 107 LN B cells. In the SPL the majority of B cells show an activated phenotype (IgMhighIgDlow), whereas resting B cells (IgMlowIgDhigh) accumulate in the SPL, LN, and BM.
Figure 3
(A) IgM/IgD staining on SPL B cells recovered in a Rag2−/− mouse injected with two LN populations sequentially. (Top) Relative proportion of Ly51 cells in IgMlowIgDhigh subpopulation (mature B cells). (Bottom) Relative proportion of Ly51 cells in IgMhighIgDlow subpopulation (activated B cells). Note that activated B cells (IgMhighIgDlow) are primarily Ly51−, thus derived from the first population injected. (B) IgMb allotype concentrations in mice injected with B6Ly51 LN population (Ly51, IgHb) either alone (○) or that also received B6IgHa LN population (Ly52, IgHa) 1 mo before (□). Mice were killed 1 mo after the transfer of the B6Ly51 LN population.
Figure 3
(A) IgM/IgD staining on SPL B cells recovered in a Rag2−/− mouse injected with two LN populations sequentially. (Top) Relative proportion of Ly51 cells in IgMlowIgDhigh subpopulation (mature B cells). (Bottom) Relative proportion of Ly51 cells in IgMhighIgDlow subpopulation (activated B cells). Note that activated B cells (IgMhighIgDlow) are primarily Ly51−, thus derived from the first population injected. (B) IgMb allotype concentrations in mice injected with B6Ly51 LN population (Ly51, IgHb) either alone (○) or that also received B6IgHa LN population (Ly52, IgHa) 1 mo before (□). Mice were killed 1 mo after the transfer of the B6Ly51 LN population.
Figure 4
(A) Fate of LN-derived and BM-derived B cells in immunodeficient mice. Each host mouse was injected simultaneously with 4 × 106 LN B cells and 106 BM cells. Each point represents the number of donor LN-derived B cells (○) and BM-derived B cells (□) recovered in the SPL of three to six host mice as a function of time. The curves join the mean of the values obtained at each time point for each B cell population.
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