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Genomics and proteomics of immune modulatory effects of a butanol fraction of echinacea purpurea in human dendritic cells.

Wang CY, Staniforth V, Chiao MT, Hou CC, Wu HM, Yeh KC, Chen CH, Hwang PI, Wen TN, Shyur LF, Yang NS - BMC Genomics (2008)

Bottom Line: Bioinformatics analysis of genes expressed in [BF/S+L/Ep]-treated DCs revealed a key-signaling network involving a number of immune-modulatory molecules leading to the activation of a downstream molecule, adenylate cyclase 8.Proteomic analysis showed increased expression of antioxidant and cytoskeletal proteins after treatment with [BF/S+L/Ep] and cichoric acid.This study provides information on candidate target molecules and molecular signaling mechanisms for future systematic research into the immune-modulatory activities of an important traditional medicinal herb and its derived phytocompounds.

View Article: PubMed Central - HTML - PubMed

Affiliation: Agricultural Biotechnology Research Center, Academia Sinica, Taipei 115, Taiwan. sally@mail.biowell.com.tw

ABSTRACT

Background: Echinacea spp. extracts and the derived phytocompounds have been shown to induce specific immune cell activities and are popularly used as food supplements or nutraceuticals for immuno-modulatory functions. Dendritic cells (DCs), the most potent antigen presenting cells, play an important role in both innate and adaptive immunities. In this study, we investigated the specific and differential gene expression in human immature DCs (iDCs) in response to treatment with a butanol fraction containing defined bioactive phytocompounds extracted from stems and leaves of Echinacea purpurea, that we denoted [BF/S+L/Ep].

Results: Affymetrix DNA microarray results showed significant up regulation of specific genes for cytokines (IL-8, IL-1beta, and IL-18) and chemokines (CXCL 2, CCL 5, and CCL 2) within 4 h after [BF/S+L/Ep] treatment of iDCs. Bioinformatics analysis of genes expressed in [BF/S+L/Ep]-treated DCs revealed a key-signaling network involving a number of immune-modulatory molecules leading to the activation of a downstream molecule, adenylate cyclase 8. Proteomic analysis showed increased expression of antioxidant and cytoskeletal proteins after treatment with [BF/S+L/Ep] and cichoric acid.

Conclusion: This study provides information on candidate target molecules and molecular signaling mechanisms for future systematic research into the immune-modulatory activities of an important traditional medicinal herb and its derived phytocompounds.

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Specific bioactivities of three subfractions of stem and leaf (S+L) extracts of E. purpurea in human immature dendritic cells (iDC). iDCs were treated for 24 h with the S+L tissue extracts and the derived ethyl acetate (EA), butanol (BuOH), or water fractions. Test cells were subsequently analyzed for cell-surface marker CD83 expression by flow cytometry.
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Figure 1: Specific bioactivities of three subfractions of stem and leaf (S+L) extracts of E. purpurea in human immature dendritic cells (iDC). iDCs were treated for 24 h with the S+L tissue extracts and the derived ethyl acetate (EA), butanol (BuOH), or water fractions. Test cells were subsequently analyzed for cell-surface marker CD83 expression by flow cytometry.

Mentions: Flow cytometry revealed that [BF/S+L/Ep] enhanced the expression of CD83, a key marker for DC maturation in iDCs, as compared with vehicle (0.1% DMSO) treatment (Figure 1) and the percentage of CD83+ expressing cells increased, from 20% to 45% with 10, 50 and 100 μg/ml [BF/S+L/Ep]. However, treatment with the ethyl acetate (EA) fraction of the S+L extract at 1, 10, or 50 μg/ml significantly decreased the percentage of CD83+ expressing cells from 20% to 0% (Figure 1). The observed effects were not caused by cellular mechanisms related to cytotoxicity, since MTT assay results indicated that treatment with EA (1 to 50 μg/ml) or [BF/S+L/Ep] (1 to 100 μg/ml) showed 98% to 125% cell viability as compared to vehicle treatment (data not shown). Thus, the [BF/S+L/Ep] fraction but not the EA fraction could enhance the maturation of iDCs. To reach this conclusion, it is essential for us to rule out the possibility that lipopolysaccharide (LPS), as a bacterial endotoxin contamination of the [BF/S+L/Ep] extract preparations, might have contributed to the observed results in the present CD83 assay and the subsequent functional genomics and proteomics studies. By using a LAL assay (see Methods), we have obtained a firm negative results (<0.125 EU/ml) on the presence of a significant level of LPS in our [BF/S+L/Ep] extract fraction. EchinaforceTM, a standardized commercial product from Swiss registered E. purpurea (L.) Moench fresh plant tincture, was reported to contain an endotoxin level of <0.5 EU/ml, and this tincture was used in a randomized double-blind clinical study [27]. In comparison, our plant extract fraction, [BF/S+L/Ep], was detected to contain a considerably lower level of possible endotoxin contamination (<0.125 EU/ml). We therefore are confident that a LPS effect on various bioactivities can virtually be ruled out from our present studies.


Genomics and proteomics of immune modulatory effects of a butanol fraction of echinacea purpurea in human dendritic cells.

Wang CY, Staniforth V, Chiao MT, Hou CC, Wu HM, Yeh KC, Chen CH, Hwang PI, Wen TN, Shyur LF, Yang NS - BMC Genomics (2008)

Specific bioactivities of three subfractions of stem and leaf (S+L) extracts of E. purpurea in human immature dendritic cells (iDC). iDCs were treated for 24 h with the S+L tissue extracts and the derived ethyl acetate (EA), butanol (BuOH), or water fractions. Test cells were subsequently analyzed for cell-surface marker CD83 expression by flow cytometry.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2571112&req=5

Figure 1: Specific bioactivities of three subfractions of stem and leaf (S+L) extracts of E. purpurea in human immature dendritic cells (iDC). iDCs were treated for 24 h with the S+L tissue extracts and the derived ethyl acetate (EA), butanol (BuOH), or water fractions. Test cells were subsequently analyzed for cell-surface marker CD83 expression by flow cytometry.
Mentions: Flow cytometry revealed that [BF/S+L/Ep] enhanced the expression of CD83, a key marker for DC maturation in iDCs, as compared with vehicle (0.1% DMSO) treatment (Figure 1) and the percentage of CD83+ expressing cells increased, from 20% to 45% with 10, 50 and 100 μg/ml [BF/S+L/Ep]. However, treatment with the ethyl acetate (EA) fraction of the S+L extract at 1, 10, or 50 μg/ml significantly decreased the percentage of CD83+ expressing cells from 20% to 0% (Figure 1). The observed effects were not caused by cellular mechanisms related to cytotoxicity, since MTT assay results indicated that treatment with EA (1 to 50 μg/ml) or [BF/S+L/Ep] (1 to 100 μg/ml) showed 98% to 125% cell viability as compared to vehicle treatment (data not shown). Thus, the [BF/S+L/Ep] fraction but not the EA fraction could enhance the maturation of iDCs. To reach this conclusion, it is essential for us to rule out the possibility that lipopolysaccharide (LPS), as a bacterial endotoxin contamination of the [BF/S+L/Ep] extract preparations, might have contributed to the observed results in the present CD83 assay and the subsequent functional genomics and proteomics studies. By using a LAL assay (see Methods), we have obtained a firm negative results (<0.125 EU/ml) on the presence of a significant level of LPS in our [BF/S+L/Ep] extract fraction. EchinaforceTM, a standardized commercial product from Swiss registered E. purpurea (L.) Moench fresh plant tincture, was reported to contain an endotoxin level of <0.5 EU/ml, and this tincture was used in a randomized double-blind clinical study [27]. In comparison, our plant extract fraction, [BF/S+L/Ep], was detected to contain a considerably lower level of possible endotoxin contamination (<0.125 EU/ml). We therefore are confident that a LPS effect on various bioactivities can virtually be ruled out from our present studies.

Bottom Line: Bioinformatics analysis of genes expressed in [BF/S+L/Ep]-treated DCs revealed a key-signaling network involving a number of immune-modulatory molecules leading to the activation of a downstream molecule, adenylate cyclase 8.Proteomic analysis showed increased expression of antioxidant and cytoskeletal proteins after treatment with [BF/S+L/Ep] and cichoric acid.This study provides information on candidate target molecules and molecular signaling mechanisms for future systematic research into the immune-modulatory activities of an important traditional medicinal herb and its derived phytocompounds.

View Article: PubMed Central - HTML - PubMed

Affiliation: Agricultural Biotechnology Research Center, Academia Sinica, Taipei 115, Taiwan. sally@mail.biowell.com.tw

ABSTRACT

Background: Echinacea spp. extracts and the derived phytocompounds have been shown to induce specific immune cell activities and are popularly used as food supplements or nutraceuticals for immuno-modulatory functions. Dendritic cells (DCs), the most potent antigen presenting cells, play an important role in both innate and adaptive immunities. In this study, we investigated the specific and differential gene expression in human immature DCs (iDCs) in response to treatment with a butanol fraction containing defined bioactive phytocompounds extracted from stems and leaves of Echinacea purpurea, that we denoted [BF/S+L/Ep].

Results: Affymetrix DNA microarray results showed significant up regulation of specific genes for cytokines (IL-8, IL-1beta, and IL-18) and chemokines (CXCL 2, CCL 5, and CCL 2) within 4 h after [BF/S+L/Ep] treatment of iDCs. Bioinformatics analysis of genes expressed in [BF/S+L/Ep]-treated DCs revealed a key-signaling network involving a number of immune-modulatory molecules leading to the activation of a downstream molecule, adenylate cyclase 8. Proteomic analysis showed increased expression of antioxidant and cytoskeletal proteins after treatment with [BF/S+L/Ep] and cichoric acid.

Conclusion: This study provides information on candidate target molecules and molecular signaling mechanisms for future systematic research into the immune-modulatory activities of an important traditional medicinal herb and its derived phytocompounds.

Show MeSH
Related in: MedlinePlus