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Epigenetic Regulation of Antibody Responses by the Histone H2A Deubiquitinase MYSM1.

Jiang XX, Chou Y, Jones L, Wang T, Sanchez S, Huang XF, Zhang L, Wang C, Chen SY - Sci Rep (2015)

Bottom Line: In this study, we found that mice deficient in the histone H2A deubiquitinase MYSM1, despite their severe defect in B cell development, exhibit an enhanced antibody response against both T cell-dependent and independent antigens.Mechanistic studies demonstrated that MYSM1 is a transcriptional activator of Pax5, the repressors of plasma cell differentiation, by facilitating key transcriptional factor recruitment and coordinating histone modifications at the Pax5 loci.Importantly, this study further provides a new target and strategy to modulate antibody production and responses with profound therapeutic implications.

View Article: PubMed Central - PubMed

Affiliation: Department of Molecular Microbiology and Immunology, Norris Comprehensive Cancer Center Keck School of Medicine, University of Southern California, Los Angeles, California, 90033, USA.

ABSTRACT
B cell-mediated antibody response plays critical roles in protective immunity, as well as in the pathogenesis of allergic and autoimmune diseases. Epigenetic histone and DNA modifications regulate gene transcription and immunity; however, so far, little is known about the role of epigenetic regulation in antibody responses. In this study, we found that mice deficient in the histone H2A deubiquitinase MYSM1, despite their severe defect in B cell development, exhibit an enhanced antibody response against both T cell-dependent and independent antigens. We revealed that MYSM1 intrinsically represses plasma cell differentiation and antibody production. Mechanistic studies demonstrated that MYSM1 is a transcriptional activator of Pax5, the repressors of plasma cell differentiation, by facilitating key transcriptional factor recruitment and coordinating histone modifications at the Pax5 loci. Hence, this study uncovers a critical role for MYSM1 in epigenetically repressing plasma cell differentiation and antibody production, in addition to its opposing, active role in B cell development. Importantly, this study further provides a new target and strategy to modulate antibody production and responses with profound therapeutic implications.

No MeSH data available.


Related in: MedlinePlus

MYSM1 intrinsically represses Ig production by plasma cells.(a) Plasma cells (CD138+) from naïve WT and Mysm1−/− mice were cultured at 10,000 cells/well in triplicate. Supernatants were collected at 36 h and were assayed for total IgM, IgG3, and IgG1 secretion by ELISA. (b–d) Supernatant ELISA analysis of NP-specific (b), MUC1-specific (c), or OVA-specific (d) IgG1, IgG3, and IgG2b secretion in sorted CD138+ cells from WT and Mysm1−/− mice that were immunized with the indicated antigens in alum. CD138+ cells were seeded at 10,000 cells/well in triplicate and the supernatants were harvested after 36 h of culture. (e) Quantitative RT-PCR analysis of mRNA levels of indicated genes in sorted CD138+ cells from WT and Mysm1−/− mice. Data were normalized to Hprt and is presented as relative to that of WT sample, set as 1, from one of two independent experiments. **p < 0.01, Mysm1−/−vs. WT.
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f4: MYSM1 intrinsically represses Ig production by plasma cells.(a) Plasma cells (CD138+) from naïve WT and Mysm1−/− mice were cultured at 10,000 cells/well in triplicate. Supernatants were collected at 36 h and were assayed for total IgM, IgG3, and IgG1 secretion by ELISA. (b–d) Supernatant ELISA analysis of NP-specific (b), MUC1-specific (c), or OVA-specific (d) IgG1, IgG3, and IgG2b secretion in sorted CD138+ cells from WT and Mysm1−/− mice that were immunized with the indicated antigens in alum. CD138+ cells were seeded at 10,000 cells/well in triplicate and the supernatants were harvested after 36 h of culture. (e) Quantitative RT-PCR analysis of mRNA levels of indicated genes in sorted CD138+ cells from WT and Mysm1−/− mice. Data were normalized to Hprt and is presented as relative to that of WT sample, set as 1, from one of two independent experiments. **p < 0.01, Mysm1−/−vs. WT.

Mentions: Even with enhanced plasma cell differentiation, the total numbers of plasma cells in Mysm1−/− mice were still significantly lower than those in WT littermates (Fig. 3b). Accordingly, we investigated whether MYSM1 controls Ig production by plasma cells, in addition to its role in repressing plasma cell differentiation. We isolated CD138+B220− plasma cells from naïve WT and Mysm1−/− mice and seeded an equal number of plasma cells on 96-well plates. The supernatants were harvested for ELISA assays. Figure 4a shows a drastic increase in the production of total IgM, IgG1, and IgG3 production by Mysm1−/− CD138+ plasma cells, indicating an enhanced ability of Mysm1−/− plasma cells to produce Ig. To further test this possibility, we isolated CD138+ plasma cells from WT and Mysm1−/− mice that were immunized with NP-KLH and seeded an equal number of isolated plasma cells onto plates. In agreement, the production of IgG antibodies against NP, including IgG1, IgG2b, and IgG3 isotypes, by plasma cells from immunized Mysm1−/− mice was also drastically enhanced (Fig. 4b). To further examine the repressive role of MYSM1 in antibody production, Mysm1−/− and WT mice were immunized with OVA, or 20-mer MUC1 peptides emulsified in IFA2526. The production of anti-MUC1 IgG1, IgG3, and IgG2b was significantly enhanced by sorted CD138+ Mysm1−/− plasma cells from the immunized mice (Fig. 4c). Enhanced levels of anti-OVA IgG1, IgG3, and IgG2b were also produced by CD138+ Mysm1−/− plasma cells from the immunized mice (Fig. 4d). In addition, the expression of the representative genes that are important for Ig production was all significantly enhanced in CD138+ Mysm1−/− plasma cells (Fig. 4e). Thus, these data demonstrate that MYSM1 represses plasma cell differentiation, as well as Ig production by plasma cells.


Epigenetic Regulation of Antibody Responses by the Histone H2A Deubiquitinase MYSM1.

Jiang XX, Chou Y, Jones L, Wang T, Sanchez S, Huang XF, Zhang L, Wang C, Chen SY - Sci Rep (2015)

MYSM1 intrinsically represses Ig production by plasma cells.(a) Plasma cells (CD138+) from naïve WT and Mysm1−/− mice were cultured at 10,000 cells/well in triplicate. Supernatants were collected at 36 h and were assayed for total IgM, IgG3, and IgG1 secretion by ELISA. (b–d) Supernatant ELISA analysis of NP-specific (b), MUC1-specific (c), or OVA-specific (d) IgG1, IgG3, and IgG2b secretion in sorted CD138+ cells from WT and Mysm1−/− mice that were immunized with the indicated antigens in alum. CD138+ cells were seeded at 10,000 cells/well in triplicate and the supernatants were harvested after 36 h of culture. (e) Quantitative RT-PCR analysis of mRNA levels of indicated genes in sorted CD138+ cells from WT and Mysm1−/− mice. Data were normalized to Hprt and is presented as relative to that of WT sample, set as 1, from one of two independent experiments. **p < 0.01, Mysm1−/−vs. WT.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4562257&req=5

f4: MYSM1 intrinsically represses Ig production by plasma cells.(a) Plasma cells (CD138+) from naïve WT and Mysm1−/− mice were cultured at 10,000 cells/well in triplicate. Supernatants were collected at 36 h and were assayed for total IgM, IgG3, and IgG1 secretion by ELISA. (b–d) Supernatant ELISA analysis of NP-specific (b), MUC1-specific (c), or OVA-specific (d) IgG1, IgG3, and IgG2b secretion in sorted CD138+ cells from WT and Mysm1−/− mice that were immunized with the indicated antigens in alum. CD138+ cells were seeded at 10,000 cells/well in triplicate and the supernatants were harvested after 36 h of culture. (e) Quantitative RT-PCR analysis of mRNA levels of indicated genes in sorted CD138+ cells from WT and Mysm1−/− mice. Data were normalized to Hprt and is presented as relative to that of WT sample, set as 1, from one of two independent experiments. **p < 0.01, Mysm1−/−vs. WT.
Mentions: Even with enhanced plasma cell differentiation, the total numbers of plasma cells in Mysm1−/− mice were still significantly lower than those in WT littermates (Fig. 3b). Accordingly, we investigated whether MYSM1 controls Ig production by plasma cells, in addition to its role in repressing plasma cell differentiation. We isolated CD138+B220− plasma cells from naïve WT and Mysm1−/− mice and seeded an equal number of plasma cells on 96-well plates. The supernatants were harvested for ELISA assays. Figure 4a shows a drastic increase in the production of total IgM, IgG1, and IgG3 production by Mysm1−/− CD138+ plasma cells, indicating an enhanced ability of Mysm1−/− plasma cells to produce Ig. To further test this possibility, we isolated CD138+ plasma cells from WT and Mysm1−/− mice that were immunized with NP-KLH and seeded an equal number of isolated plasma cells onto plates. In agreement, the production of IgG antibodies against NP, including IgG1, IgG2b, and IgG3 isotypes, by plasma cells from immunized Mysm1−/− mice was also drastically enhanced (Fig. 4b). To further examine the repressive role of MYSM1 in antibody production, Mysm1−/− and WT mice were immunized with OVA, or 20-mer MUC1 peptides emulsified in IFA2526. The production of anti-MUC1 IgG1, IgG3, and IgG2b was significantly enhanced by sorted CD138+ Mysm1−/− plasma cells from the immunized mice (Fig. 4c). Enhanced levels of anti-OVA IgG1, IgG3, and IgG2b were also produced by CD138+ Mysm1−/− plasma cells from the immunized mice (Fig. 4d). In addition, the expression of the representative genes that are important for Ig production was all significantly enhanced in CD138+ Mysm1−/− plasma cells (Fig. 4e). Thus, these data demonstrate that MYSM1 represses plasma cell differentiation, as well as Ig production by plasma cells.

Bottom Line: In this study, we found that mice deficient in the histone H2A deubiquitinase MYSM1, despite their severe defect in B cell development, exhibit an enhanced antibody response against both T cell-dependent and independent antigens.Mechanistic studies demonstrated that MYSM1 is a transcriptional activator of Pax5, the repressors of plasma cell differentiation, by facilitating key transcriptional factor recruitment and coordinating histone modifications at the Pax5 loci.Importantly, this study further provides a new target and strategy to modulate antibody production and responses with profound therapeutic implications.

View Article: PubMed Central - PubMed

Affiliation: Department of Molecular Microbiology and Immunology, Norris Comprehensive Cancer Center Keck School of Medicine, University of Southern California, Los Angeles, California, 90033, USA.

ABSTRACT
B cell-mediated antibody response plays critical roles in protective immunity, as well as in the pathogenesis of allergic and autoimmune diseases. Epigenetic histone and DNA modifications regulate gene transcription and immunity; however, so far, little is known about the role of epigenetic regulation in antibody responses. In this study, we found that mice deficient in the histone H2A deubiquitinase MYSM1, despite their severe defect in B cell development, exhibit an enhanced antibody response against both T cell-dependent and independent antigens. We revealed that MYSM1 intrinsically represses plasma cell differentiation and antibody production. Mechanistic studies demonstrated that MYSM1 is a transcriptional activator of Pax5, the repressors of plasma cell differentiation, by facilitating key transcriptional factor recruitment and coordinating histone modifications at the Pax5 loci. Hence, this study uncovers a critical role for MYSM1 in epigenetically repressing plasma cell differentiation and antibody production, in addition to its opposing, active role in B cell development. Importantly, this study further provides a new target and strategy to modulate antibody production and responses with profound therapeutic implications.

No MeSH data available.


Related in: MedlinePlus