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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 induces IL-6 and IL-10 production in mice B cells.A. MACS-purified CD19+ B cells were treated with the indicated dosages of YZP for 72 h, and the culture supernatant was then collected for IgM measurements. B. MACS-purified CD19+ B cells were treated with YZP (10 μg/mL) for 72 h, and subsequently the culture supernatant was collected for IgM and IgG measurements. C. and D. MACS-purified CD19+ B cells were treated with the indicated dosages of YZP for 72 h, and subsequently the culture supernatant was collected for cytokine measurements.
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pone-0072422-g004: YZP induces IL-6 and IL-10 production in mice B cells.A. MACS-purified CD19+ B cells were treated with the indicated dosages of YZP for 72 h, and the culture supernatant was then collected for IgM measurements. B. MACS-purified CD19+ B cells were treated with YZP (10 μg/mL) for 72 h, and subsequently the culture supernatant was collected for IgM and IgG measurements. C. and D. MACS-purified CD19+ B cells were treated with the indicated dosages of YZP for 72 h, and subsequently the culture supernatant was collected for cytokine measurements.

Mentions: We examined the immunoglobulin (Ig) production ability of YZP-stimulated B cells and discovered a dose-dependent secretion of IgM (Figure 4A), whereas IgG production was unaltered (Figure 4B). In addition to Ab-secretion, B lymphocytes also modulate immune responses through the production of cytokines [26]. We screened for the presence of cytokines in the culture supernatant of B cells stimulated with YZP (2.5–20 μg/mL) and observed significant productions of IL-6 (Figure 4C) and IL-10 (Figure 4D). Intracellular staining experiments also confirm the capability of YZP to activate IL-10 expression by splenocytes and purified B cells (Figure 5C and 5D).


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 induces IL-6 and IL-10 production in mice B cells.A. MACS-purified CD19+ B cells were treated with the indicated dosages of YZP for 72 h, and the culture supernatant was then collected for IgM measurements. B. MACS-purified CD19+ B cells were treated with YZP (10 μg/mL) for 72 h, and subsequently the culture supernatant was collected for IgM and IgG measurements. C. and D. MACS-purified CD19+ B cells were treated with the indicated dosages of YZP for 72 h, and subsequently the culture supernatant was collected for cytokine measurements.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3760908&req=5

pone-0072422-g004: YZP induces IL-6 and IL-10 production in mice B cells.A. MACS-purified CD19+ B cells were treated with the indicated dosages of YZP for 72 h, and the culture supernatant was then collected for IgM measurements. B. MACS-purified CD19+ B cells were treated with YZP (10 μg/mL) for 72 h, and subsequently the culture supernatant was collected for IgM and IgG measurements. C. and D. MACS-purified CD19+ B cells were treated with the indicated dosages of YZP for 72 h, and subsequently the culture supernatant was collected for cytokine measurements.
Mentions: We examined the immunoglobulin (Ig) production ability of YZP-stimulated B cells and discovered a dose-dependent secretion of IgM (Figure 4A), whereas IgG production was unaltered (Figure 4B). In addition to Ab-secretion, B lymphocytes also modulate immune responses through the production of cytokines [26]. We screened for the presence of cytokines in the culture supernatant of B cells stimulated with YZP (2.5–20 μg/mL) and observed significant productions of IL-6 (Figure 4C) and IL-10 (Figure 4D). Intracellular staining experiments also confirm the capability of YZP to activate IL-10 expression by splenocytes and purified B cells (Figure 5C and 5D).

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