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Trametes versicolor protein YZP activates regulatory B lymphocytes - gene identification through de novo assembly and function analysis in a murine acute colitis model.

Kuan YC, Wu YJ, Hung CL, Sheu F - PLoS ONE (2013)

Bottom Line: YZP induced a greater than 60-fold increase in IL-10 secretion in mice B lymphocytes; moreover, YZP specifically triggered the differentiation of CD1d(+) B cells into IL-10-producing regulatory B cells (Bregs) and enhanced the expression of CD1d.Furthermore, YZP activated Breg function via interaction with TLR2 and TLR4 and up-regulation of the TLR-mediated signaling pathway.We purified a novel Breg-stimulating protein, YZP, from T. versicolor and developed an advanced approach combining RNA-seq and de novo assembly technologies.to clone its gene.

View Article: PubMed Central - PubMed

Affiliation: Center for Biotechnology, National Taiwan University, Taipei, Taiwan ; Department of Horticulture, National Taiwan University, Taipei, Taiwan.

ABSTRACT

Background: Trametes versicolor (Yun-Zhi) is a medicinal fungus used as a chemotherapy co-treatment to enhance anti-tumor immunity. Although the efficacies of T. versicolor extracts have been documented, the active ingredients and mechanisms underlying the actions of these extracts remain uncharacterized.

Results: We purified a new protein, YZP, from the fruiting bodies of T. versicolor and identified the gene encoding YZP using RNA-seq and de novo assembly technologies. YZP is a 12-kDa non-glycosylated protein comprising 139 amino acids, including an 18-amino acids signal peptide. YZP induced a greater than 60-fold increase in IL-10 secretion in mice B lymphocytes; moreover, YZP specifically triggered the differentiation of CD1d(+) B cells into IL-10-producing regulatory B cells (Bregs) and enhanced the expression of CD1d. YZP-induced B cells suppressed approximately 40% of the LPS-activated macrophage production of inflammatory cytokines in a mixed leukocyte reaction and significantly alleviated the disease activity and colonic inflammation in a DSS-induced acute colitis murine model. Furthermore, YZP activated Breg function via interaction with TLR2 and TLR4 and up-regulation of the TLR-mediated signaling pathway.

Conclusions: We purified a novel Breg-stimulating protein, YZP, from T. versicolor and developed an advanced approach combining RNA-seq and de novo assembly technologies.to clone its gene. We demonstrated that YZP activated CD1d(+) Breg differentiation through TLR2/4-mediated signaling pathway, and the YZP-stimulated B cells exhibited anti-inflammatory efficacies in vitro and in murine acute colitis models.

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YZP-induced B cells inhibit inflammatory cytokine release from LPS-activated macrophages.A. Mice peritoneal macrophages (2×106) were stimulated with or without LPS (1 μg/mL) for 12 h and subsequently co-cultured with or without an equal number (2×106) of YZP-induced B cells or control B cells for 12 h. The culture supernatant was subsequently collected for TNF-α and IL-1β measurements. B. Mice peritoneal macrophages (2×106) were stimulated with LPS (1 μg/mL) for 12 h and were subsequently co-cultured with the indicated numbers of YZP-induced B cells for 12 h. The culture supernatant was collected for TNF-α and IL-1β measurements. C. and D. Mice peritoneal macrophages (1×106) were stimulated with or without LPS (1 μg/mL) for 12 h and subsequently co-cultured with or without 2×106 YZP-induced B cells or control B cells for 12 h. The culture supernatant was subsequently collected for TNF-α and IL-1β measurements. The data is presented as the mean ± SD, where the asterisks indicate the statistical differences (*, p<0.05; **, p<0.01; N.S., non-significant) among groups.
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pone-0072422-g006: YZP-induced B cells inhibit inflammatory cytokine release from LPS-activated macrophages.A. Mice peritoneal macrophages (2×106) were stimulated with or without LPS (1 μg/mL) for 12 h and subsequently co-cultured with or without an equal number (2×106) of YZP-induced B cells or control B cells for 12 h. The culture supernatant was subsequently collected for TNF-α and IL-1β measurements. B. Mice peritoneal macrophages (2×106) were stimulated with LPS (1 μg/mL) for 12 h and were subsequently co-cultured with the indicated numbers of YZP-induced B cells for 12 h. The culture supernatant was collected for TNF-α and IL-1β measurements. C. and D. Mice peritoneal macrophages (1×106) were stimulated with or without LPS (1 μg/mL) for 12 h and subsequently co-cultured with or without 2×106 YZP-induced B cells or control B cells for 12 h. The culture supernatant was subsequently collected for TNF-α and IL-1β measurements. The data is presented as the mean ± SD, where the asterisks indicate the statistical differences (*, p<0.05; **, p<0.01; N.S., non-significant) among groups.

Mentions: YZP-induced B cells (B-YZP) comprised approximately 16% of IL-10+ Bregs (Figure 5B). To determine if B-YZP has immune suppressive activity, LPS-activated macrophages (MΦ-LPS) were co-cultured with B-YZP or untreated B cells (B-CTR) for 12 h, and the inflammatory cytokines present in the culture supernatant were quantified. Compared with untreated cells (MΦ-CTR), MΦ-LPS produced 2.6- and 33-fold higher amounts of TNF-α and IL-1β, respectively, in macrophage cultures. Co-culturing MΦ-LPS with B-YZP resulted in a significant reduced of TNF-α and IL-1β production by 46.9% and 37.0%, respectively (Figure 6A). Moreover, B cell-dependent reductions in TNF-α and IL-1β production were observed when MΦ-LPS were co-cultured with different amounts of B-YZP (Figure 6B). Collectively, these results confirmed the regulatory activity of B-YZP.


Trametes versicolor protein YZP activates regulatory B lymphocytes - gene identification through de novo assembly and function analysis in a murine acute colitis model.

Kuan YC, Wu YJ, Hung CL, Sheu F - PLoS ONE (2013)

YZP-induced B cells inhibit inflammatory cytokine release from LPS-activated macrophages.A. Mice peritoneal macrophages (2×106) were stimulated with or without LPS (1 μg/mL) for 12 h and subsequently co-cultured with or without an equal number (2×106) of YZP-induced B cells or control B cells for 12 h. The culture supernatant was subsequently collected for TNF-α and IL-1β measurements. B. Mice peritoneal macrophages (2×106) were stimulated with LPS (1 μg/mL) for 12 h and were subsequently co-cultured with the indicated numbers of YZP-induced B cells for 12 h. The culture supernatant was collected for TNF-α and IL-1β measurements. C. and D. Mice peritoneal macrophages (1×106) were stimulated with or without LPS (1 μg/mL) for 12 h and subsequently co-cultured with or without 2×106 YZP-induced B cells or control B cells for 12 h. The culture supernatant was subsequently collected for TNF-α and IL-1β measurements. The data is presented as the mean ± SD, where the asterisks indicate the statistical differences (*, p<0.05; **, p<0.01; N.S., non-significant) among groups.
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Related In: Results  -  Collection

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pone-0072422-g006: YZP-induced B cells inhibit inflammatory cytokine release from LPS-activated macrophages.A. Mice peritoneal macrophages (2×106) were stimulated with or without LPS (1 μg/mL) for 12 h and subsequently co-cultured with or without an equal number (2×106) of YZP-induced B cells or control B cells for 12 h. The culture supernatant was subsequently collected for TNF-α and IL-1β measurements. B. Mice peritoneal macrophages (2×106) were stimulated with LPS (1 μg/mL) for 12 h and were subsequently co-cultured with the indicated numbers of YZP-induced B cells for 12 h. The culture supernatant was collected for TNF-α and IL-1β measurements. C. and D. Mice peritoneal macrophages (1×106) were stimulated with or without LPS (1 μg/mL) for 12 h and subsequently co-cultured with or without 2×106 YZP-induced B cells or control B cells for 12 h. The culture supernatant was subsequently collected for TNF-α and IL-1β measurements. The data is presented as the mean ± SD, where the asterisks indicate the statistical differences (*, p<0.05; **, p<0.01; N.S., non-significant) among groups.
Mentions: YZP-induced B cells (B-YZP) comprised approximately 16% of IL-10+ Bregs (Figure 5B). To determine if B-YZP has immune suppressive activity, LPS-activated macrophages (MΦ-LPS) were co-cultured with B-YZP or untreated B cells (B-CTR) for 12 h, and the inflammatory cytokines present in the culture supernatant were quantified. Compared with untreated cells (MΦ-CTR), MΦ-LPS produced 2.6- and 33-fold higher amounts of TNF-α and IL-1β, respectively, in macrophage cultures. Co-culturing MΦ-LPS with B-YZP resulted in a significant reduced of TNF-α and IL-1β production by 46.9% and 37.0%, respectively (Figure 6A). Moreover, B cell-dependent reductions in TNF-α and IL-1β production were observed when MΦ-LPS were co-cultured with different amounts of B-YZP (Figure 6B). Collectively, these results confirmed the regulatory activity of B-YZP.

Bottom Line: YZP induced a greater than 60-fold increase in IL-10 secretion in mice B lymphocytes; moreover, YZP specifically triggered the differentiation of CD1d(+) B cells into IL-10-producing regulatory B cells (Bregs) and enhanced the expression of CD1d.Furthermore, YZP activated Breg function via interaction with TLR2 and TLR4 and up-regulation of the TLR-mediated signaling pathway.We purified a novel Breg-stimulating protein, YZP, from T. versicolor and developed an advanced approach combining RNA-seq and de novo assembly technologies.to clone its gene.

View Article: PubMed Central - PubMed

Affiliation: Center for Biotechnology, National Taiwan University, Taipei, Taiwan ; Department of Horticulture, National Taiwan University, Taipei, Taiwan.

ABSTRACT

Background: Trametes versicolor (Yun-Zhi) is a medicinal fungus used as a chemotherapy co-treatment to enhance anti-tumor immunity. Although the efficacies of T. versicolor extracts have been documented, the active ingredients and mechanisms underlying the actions of these extracts remain uncharacterized.

Results: We purified a new protein, YZP, from the fruiting bodies of T. versicolor and identified the gene encoding YZP using RNA-seq and de novo assembly technologies. YZP is a 12-kDa non-glycosylated protein comprising 139 amino acids, including an 18-amino acids signal peptide. YZP induced a greater than 60-fold increase in IL-10 secretion in mice B lymphocytes; moreover, YZP specifically triggered the differentiation of CD1d(+) B cells into IL-10-producing regulatory B cells (Bregs) and enhanced the expression of CD1d. YZP-induced B cells suppressed approximately 40% of the LPS-activated macrophage production of inflammatory cytokines in a mixed leukocyte reaction and significantly alleviated the disease activity and colonic inflammation in a DSS-induced acute colitis murine model. Furthermore, YZP activated Breg function via interaction with TLR2 and TLR4 and up-regulation of the TLR-mediated signaling pathway.

Conclusions: We purified a novel Breg-stimulating protein, YZP, from T. versicolor and developed an advanced approach combining RNA-seq and de novo assembly technologies.to clone its gene. We demonstrated that YZP activated CD1d(+) Breg differentiation through TLR2/4-mediated signaling pathway, and the YZP-stimulated B cells exhibited anti-inflammatory efficacies in vitro and in murine acute colitis models.

Show MeSH
Related in: MedlinePlus