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A Role for the Transcription Factor Arid3a in Mouse B2 Lymphocyte Expansion and Peritoneal B1a Generation

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A Role for the Transcription Factor Arid3a in Mouse B2 Lymphocyte Expansion and Peritoneal B1a Generation

Katrin Habir et al. Front Immunol.

Abstract

The initiation, commitment, and terminal differentiation of the B cell lineage is stringently controlled by the coordinated action of various transcription factors. Among these, Arid3a has previously been implicated in regulating early B lymphopoiesis, humoral immune responses to phosphocholine, and furthermore to promote the B1 over the B2 cell lineage. We have now interrogated the function of Arid3a in the adult mouse using conditional mutagenesis. We demonstrate that loss of Arid3a does not affect early B cell development or lineage commitment but rather loss of this transcription factor results in a broad expansion of bone marrow B lymphopoiesis in a manner that reflects its developmental expression pattern. Furthermore, loss of Arid3a resulted in expanded splenic B cell numbers with the exception of the B1 lineage that was maintained at normal numbers. However, B1a lymphoyctes were reduced in the peritoneal cavity. In addition, antibody responses to phosphocholine were attenuated in the absence of Arid3a. Hence, functional Arid3a is required in mature B cells for specific immune responses and for generating normal numbers of B cells in a subset dependent manner.

Keywords: Arid3a; B cell development; antibody formation; bone marrow; spleen.

Figures

Figure 1
Figure 1
Conditional loss of Arid3a and B cell development. (A) Relative expression of Arid3a in B cell subsets in the bone marrow (common lymphoid progenitor and fractions A to E), spleen (transitional and mature stages), and peritoneal cavity using data from the ImmGen database. (B,C) Flow cytometry of bone marrow B cell development in Arid3afl/fl, Arid3afl/+ Cd79aCre/+, and Arid3afl/fl Cd79aCre/+ mice. Flow cytometry plots are shown on the left. Absolute cell numbers for B1 (CD19+B220lo), B2 (CD19+B220+), immature B (CD19+B220+IgM+IgD), recirculating B (CD19+B220+IgMIgD+), pro-B (CD19+B220+IgMIgD c-Kit+CD25), and pre-B (CD19+B220+IgMIgDc-KitCD25+) cell populations are shown on the right. Error bars represent SEM and n = 17–24 for each group. p-Values were determined by Student’s t-test and fold changes are indicated. (D) Deletion of the conditional Arid3a allele determined by RT-polymerase chain reaction in sorted pro-B cells from the bone marrow of Arid3afl/+ Cd79aCre/+ and Arid3afl/fl Cd79aCre/+ aged 4–6 weeks of age. The full length wild-type and floxed allele (WT/Fl) and exon 4 deleted allele (ΔE4) are indicated. HPRT was used as a loading control.
Figure 2
Figure 2
The in vivo function of Arid3a in splenic B cell development. (A) Flow cytometry of splenic B1 cells (CD19+B220lo) and B2 cells (CD19+B220+) in the left panel with quantification shown on the right for the indicated genotypes from mice aged 10–12 weeks. (B) Analysis of Transitional 1 (CD19+B220+IgM+CD23) and Transitional 2 (CD19+B220+IgM+CD23+) B cells. (C) Follicular B (CD19+B220+CD21loCD23+) and marginal zone B (CD19+B220+CD21hi CD23lo) cell populations in the spleen. Error bars represent SEM and n = 9–32 for each group. p-Values were determined by Student’s t-test and fold changes are indicated. (D) Expression of IgM on marginal zone and follicular B cells.
Figure 3
Figure 3
Arid3a is required for normal peritoneal cavity B1a cell development. (A) Flow cytometry of B1 cells (CD19+B220lo) and B2 cells (CD19+B220+) taken from peritoneal lavages of mice aged 10–12 weeks old. B1 cells were further gated for CD5 to determine B1a (CD19+B220loCD5+) and B1b (CD19+B220loCD5) populations shown in the bottom panel. Error bars represent SEM and n = 3–8 for each group. p-Values were determined by Student’s t-test and fold changes are indicated. (B) Cell surface phenotype of B1a (CD19+B220loCD5+) and B2 (CD19+B220+) cells. (C) Deletion of the conditional Arid3a allele was shown by RT-polymerase chain reaction in B1 and B2 cells sorted from peritoneal cavity in Arid3afl/+ Cd79aCre/+ and Arid3afl/fl Cd79aCre/+ mice, 10–12 weeks of age. The full length wild-type and floxed allele (WT/Fl) and exon 4 deleted allele (ΔE4) are indicated. HPRT was used as a loading control.
Figure 4
Figure 4
A requirement for Arid3a in humoral immunity. (A) Plasma was collected from naïve mice of the indicated genotypes aged 10–12 weeks old and analyzed for total IgM, IgA, and IgG levels with Enzyme-Linked ImmunoSorbent Assay. (B) IgG1, IgG2b, and IgG3 antibody titers. n = 15–26 for each group and p-values were determined by Student’s t-test.
Figure 5
Figure 5
Normal antibody production against phosphocholine requires Arid3a. (A) Enzyme-Linked ImmunoSorbent Assay measurements of anti-phosphocholine IgM in naïve mice from the indicated genotypes n = 15–28 for each group. (B) Anti-phosphocholine IgM levels 14 days after PC-KLH immunization n = 4–5 for each group. (C) Absolute number of cells with a germinal center phenotype (CD19+B220+IgDGL7+CD95+) in spleen, Peyer’s patches, and mesenteric lymph nodes after immunization with PC-KLH/Alum. n = 4–6 for each group. p-Values determined by Student’s t-test.

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References

    1. Kantor A. A new nomenclature for B cells. Immunol Today (1991) 12:388.10.1016/0167-5699(91)90135-G - DOI - PubMed
    1. Yuan J, Nguyen CK, Liu X, Kanellopoulou C, Muljo SA. Lin28b reprograms adult bone marrow hematopoietic progenitors to mediate fetal-like lymphopoiesis. Science (2012) 335:1195.10.1126/science.1216557 - DOI - PMC - PubMed
    1. Das C, Eaton S, Calame K, Tucker PW. Novel protein-DNA interactions associated with increased immunoglobulin transcription in response to antigen plus interleukin-5. Mol Cell Biol (1991) 11:5197.10.1128/MCB.11.10.5197 - DOI - PMC - PubMed
    1. Herrscher RF, Kaplan MH, Lelsz DL, Das C, Scheuermann R, Tucker PW. The immunoglobulin heavy-chain matrix-associating regions are bound by Bright: a B cell-specific trans-activator that describes a new DNA-binding protein family. Genes Dev (1995) 9:3067.10.1101/gad.9.24.3067 - DOI - PubMed
    1. Zhou Y, Li YS, Bandi SR, Tang L, Shinton SA, Hayakawa K, et al. Lin28b promotes fetal B lymphopoiesis through the transcription factor Arid3a. J Exp Med (2015) 212:569–80.10.1084/jem.20141510 - DOI - PMC - PubMed

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