Limits...
c-Myb is required for plasma cell migration to bone marrow after immunization or infection.

Good-Jacobson KL, O'Donnell K, Belz GT, Nutt SL, Tarlinton DM - J. Exp. Med. (2015)

Bottom Line: Here, we detail the critical role of the transcription factor c-Myb in determining plasma cell location.This was correlated with a dramatic reduction of plasma cells in peripheral blood, mislocalization in spleen, and an inability of c-Myb-deficient plasma cells to migrate along a CXCL12 gradient.Therefore, c-Myb plays an essential, novel role in establishing the long-lived plasma cell population in the BM via responsiveness to chemokine migration cues.

View Article: PubMed Central - HTML - PubMed

Affiliation: Walter and Eliza Hall Institute of Medical Research, Parkville, Victoria 3052, Australia Department of Medical Biology, University of Melbourne, Parkville, Victoria 3010, Australia jacobson@wehi.edu.au tarlinton@wehi.edu.au.

Show MeSH

Related in: MedlinePlus

Absence of Ag-specific ASCs in the BM during an immune response.Cd23Cre/+ (black bars or closed squares) and c-Mybfl/flCd23Cre/+ (white bars or open circles) mice were immunized with NP-KLH precipitated in alum. (A–E) ELISPOT analysis of NP+IgG1+ ASCs in the BM at day 14 (A and B), over time (C), kinetics in the spleen (D), and affinity of splenic ASCs (E). n ≥ 4 mice per experiment, results are combined from 1 (day 10, day 42), 2 (day 14), or 3 (day 7, day 28) independent experiments per time point. *, P < 0.05; **, P < 0.01; ***, P < 0.001. (Mann-Whitney nonparametric, two-tailed test; mean ± SEM). Bar, 3 mm. (F) c-Mybfl/flAicdaCre/+ and littermate controls were immunized with NP-KLH in alum and ELISPOT analysis of BM ASCs was performed 14 d after immunization (n ≥ 3 mice per genotype). (G–I) Number of NP+Fas+ GC B cells (G), frequency of NP+Fas+ GC B cells that are IgG1+ (H), and number (I) of IgG1+NP+Fas+ GC B cells assessed by flow cytometry 7 d (n ≥ 11 mice per genotype; combined from three independent experiments), 14 d (n ≥ 6 mice per genotype; combined from two independent experiments), and 28 d (n = 4 mice per genotype) after immunization.
© Copyright Policy - openaccess
Related In: Results  -  Collection

License 1 - License 2
getmorefigures.php?uid=PMC4493410&req=5

fig1: Absence of Ag-specific ASCs in the BM during an immune response.Cd23Cre/+ (black bars or closed squares) and c-Mybfl/flCd23Cre/+ (white bars or open circles) mice were immunized with NP-KLH precipitated in alum. (A–E) ELISPOT analysis of NP+IgG1+ ASCs in the BM at day 14 (A and B), over time (C), kinetics in the spleen (D), and affinity of splenic ASCs (E). n ≥ 4 mice per experiment, results are combined from 1 (day 10, day 42), 2 (day 14), or 3 (day 7, day 28) independent experiments per time point. *, P < 0.05; **, P < 0.01; ***, P < 0.001. (Mann-Whitney nonparametric, two-tailed test; mean ± SEM). Bar, 3 mm. (F) c-Mybfl/flAicdaCre/+ and littermate controls were immunized with NP-KLH in alum and ELISPOT analysis of BM ASCs was performed 14 d after immunization (n ≥ 3 mice per genotype). (G–I) Number of NP+Fas+ GC B cells (G), frequency of NP+Fas+ GC B cells that are IgG1+ (H), and number (I) of IgG1+NP+Fas+ GC B cells assessed by flow cytometry 7 d (n ≥ 11 mice per genotype; combined from three independent experiments), 14 d (n ≥ 6 mice per genotype; combined from two independent experiments), and 28 d (n = 4 mice per genotype) after immunization.

Mentions: To assess the contribution of c-Myb to a humoral response, we generated c-Mybfl/fl mice carrying a Cd23Cre/+ transgene that deletes c-Myb at the T2 stage of B cell development (Emambokus et al., 2003; Kwon et al., 2008). Strikingly, immunized c-Mybfl/flCd23Cre/+ had no IgG1+ Ag-specific ASCs in the BM (Fig. 1, A–C). Although the frequency of Ag-specific IgG1+ ASCs in the BM of Cd23Cre/+ control mice increased throughout the immune response (Fig. 1 C), there were few or no BM IgG1+ ASCs detected in the absence of c-Myb. This was not due to a lack of production, as assessment of NP+IgG1+ ASCs in the spleen demonstrated comparable frequencies over time (Fig. 1 D). Similarly, affinity maturation of splenic NP+IgG1+ ASCs appeared normal (Fig. 1 E). c-Mybfl/fl mice carrying an AicdaCre/+ allele, in which c-Myb was deleted after Ag activation of mature B cells (Kwon et al., 2008), also revealed a lack of NP+IgG1+ ASCs in the BM during an immune response (Fig. 1 F). This indicates a role for c-Myb during the processes of ASC differentiation and migration rather than in establishing a preexisting condition in naive B cells. NP+ GC B cells formed normally in the absence of c-Myb at day 7 after immunization, but by day 14 there was a twofold decrease, suggesting persistence of these cells was not optimal in the absence of c-Myb (Fig. 1 G). Within the c-Myb–deficient NP+ GC compartment, however, the frequency of IgG1+ cells was increased at day 14 and 28 after immunization compared with controls (Fig. 1 H). Thus, the number of IgG1+NP+ GC B cells, which are arguably the precursors to IgG1+ ASCs in the BM, was similar between knockout and controls at days 7 and 14, and significantly decreased at day 28 (Fig. 1 I). Thus, the lack of ASC in the BM of c-Myb–deficient mice was not caused by an absence of IgG1+NP+ GC B cells.


c-Myb is required for plasma cell migration to bone marrow after immunization or infection.

Good-Jacobson KL, O'Donnell K, Belz GT, Nutt SL, Tarlinton DM - J. Exp. Med. (2015)

Absence of Ag-specific ASCs in the BM during an immune response.Cd23Cre/+ (black bars or closed squares) and c-Mybfl/flCd23Cre/+ (white bars or open circles) mice were immunized with NP-KLH precipitated in alum. (A–E) ELISPOT analysis of NP+IgG1+ ASCs in the BM at day 14 (A and B), over time (C), kinetics in the spleen (D), and affinity of splenic ASCs (E). n ≥ 4 mice per experiment, results are combined from 1 (day 10, day 42), 2 (day 14), or 3 (day 7, day 28) independent experiments per time point. *, P < 0.05; **, P < 0.01; ***, P < 0.001. (Mann-Whitney nonparametric, two-tailed test; mean ± SEM). Bar, 3 mm. (F) c-Mybfl/flAicdaCre/+ and littermate controls were immunized with NP-KLH in alum and ELISPOT analysis of BM ASCs was performed 14 d after immunization (n ≥ 3 mice per genotype). (G–I) Number of NP+Fas+ GC B cells (G), frequency of NP+Fas+ GC B cells that are IgG1+ (H), and number (I) of IgG1+NP+Fas+ GC B cells assessed by flow cytometry 7 d (n ≥ 11 mice per genotype; combined from three independent experiments), 14 d (n ≥ 6 mice per genotype; combined from two independent experiments), and 28 d (n = 4 mice per genotype) after immunization.
© Copyright Policy - openaccess
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4493410&req=5

fig1: Absence of Ag-specific ASCs in the BM during an immune response.Cd23Cre/+ (black bars or closed squares) and c-Mybfl/flCd23Cre/+ (white bars or open circles) mice were immunized with NP-KLH precipitated in alum. (A–E) ELISPOT analysis of NP+IgG1+ ASCs in the BM at day 14 (A and B), over time (C), kinetics in the spleen (D), and affinity of splenic ASCs (E). n ≥ 4 mice per experiment, results are combined from 1 (day 10, day 42), 2 (day 14), or 3 (day 7, day 28) independent experiments per time point. *, P < 0.05; **, P < 0.01; ***, P < 0.001. (Mann-Whitney nonparametric, two-tailed test; mean ± SEM). Bar, 3 mm. (F) c-Mybfl/flAicdaCre/+ and littermate controls were immunized with NP-KLH in alum and ELISPOT analysis of BM ASCs was performed 14 d after immunization (n ≥ 3 mice per genotype). (G–I) Number of NP+Fas+ GC B cells (G), frequency of NP+Fas+ GC B cells that are IgG1+ (H), and number (I) of IgG1+NP+Fas+ GC B cells assessed by flow cytometry 7 d (n ≥ 11 mice per genotype; combined from three independent experiments), 14 d (n ≥ 6 mice per genotype; combined from two independent experiments), and 28 d (n = 4 mice per genotype) after immunization.
Mentions: To assess the contribution of c-Myb to a humoral response, we generated c-Mybfl/fl mice carrying a Cd23Cre/+ transgene that deletes c-Myb at the T2 stage of B cell development (Emambokus et al., 2003; Kwon et al., 2008). Strikingly, immunized c-Mybfl/flCd23Cre/+ had no IgG1+ Ag-specific ASCs in the BM (Fig. 1, A–C). Although the frequency of Ag-specific IgG1+ ASCs in the BM of Cd23Cre/+ control mice increased throughout the immune response (Fig. 1 C), there were few or no BM IgG1+ ASCs detected in the absence of c-Myb. This was not due to a lack of production, as assessment of NP+IgG1+ ASCs in the spleen demonstrated comparable frequencies over time (Fig. 1 D). Similarly, affinity maturation of splenic NP+IgG1+ ASCs appeared normal (Fig. 1 E). c-Mybfl/fl mice carrying an AicdaCre/+ allele, in which c-Myb was deleted after Ag activation of mature B cells (Kwon et al., 2008), also revealed a lack of NP+IgG1+ ASCs in the BM during an immune response (Fig. 1 F). This indicates a role for c-Myb during the processes of ASC differentiation and migration rather than in establishing a preexisting condition in naive B cells. NP+ GC B cells formed normally in the absence of c-Myb at day 7 after immunization, but by day 14 there was a twofold decrease, suggesting persistence of these cells was not optimal in the absence of c-Myb (Fig. 1 G). Within the c-Myb–deficient NP+ GC compartment, however, the frequency of IgG1+ cells was increased at day 14 and 28 after immunization compared with controls (Fig. 1 H). Thus, the number of IgG1+NP+ GC B cells, which are arguably the precursors to IgG1+ ASCs in the BM, was similar between knockout and controls at days 7 and 14, and significantly decreased at day 28 (Fig. 1 I). Thus, the lack of ASC in the BM of c-Myb–deficient mice was not caused by an absence of IgG1+NP+ GC B cells.

Bottom Line: Here, we detail the critical role of the transcription factor c-Myb in determining plasma cell location.This was correlated with a dramatic reduction of plasma cells in peripheral blood, mislocalization in spleen, and an inability of c-Myb-deficient plasma cells to migrate along a CXCL12 gradient.Therefore, c-Myb plays an essential, novel role in establishing the long-lived plasma cell population in the BM via responsiveness to chemokine migration cues.

View Article: PubMed Central - HTML - PubMed

Affiliation: Walter and Eliza Hall Institute of Medical Research, Parkville, Victoria 3052, Australia Department of Medical Biology, University of Melbourne, Parkville, Victoria 3010, Australia jacobson@wehi.edu.au tarlinton@wehi.edu.au.

Show MeSH
Related in: MedlinePlus