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Specific medicinal plant polysaccharides effectively enhance the potency of a DC-based vaccine against mouse mammary tumor metastasis.

Chang WT, Lai TH, Chyan YJ, Yin SY, Chen YH, Wei WC, Yang NS - PLoS ONE (2015)

Bottom Line: Treatments with Am, Cp and [Am+Cp] also resulted in augmented expression of CD40, CD80 and CD86 markers in test DCs.Biochemical analysis revealed that Am and Cp are composed mainly of polysaccharides containing a high level (70-95%) glucose residues, but few or no (< 1%) mannose residues.In summary, our findings suggest that the specific plant polysaccharides Am and Cp extracted from traditional Chinese medicines can be effectively used instead of bacterial LPS as a potent adjuvant in the formulation of a DC-based vaccine for cancer immunotherapies.

View Article: PubMed Central - PubMed

Affiliation: Department of Food and Nutrition, Providence University, Taichung, Taiwan, R. O. C; Institute of Agricultural Biotechnology Research Center, Academia Sinica, Taipei, Taiwan, R. O. C.

ABSTRACT
Dendritic cell (DC) vaccines are a newly emerging immunotherapeutic approach for the treatment and prevention of cancer, but major challenges still remain particularly with respect to clinical efficacy. Engineering and optimization of adjuvant formulations for DC-based vaccines is one strategy through which more efficacious treatments may be obtained. In this study, we developed a new ex vivo approach for DC vaccine preparation. We evaluated two highly purified mixed polysaccharide fractions from the root of Astragalus membranaceus and Codonopsis pilosulae, named Am and Cp, for their use in enhancing the efficiency of a DC-based cancer vaccine against metastasis of 4T1 mammary carcinoma in mice. Mixed lymphocyte reaction showed all Am-, Cp- and [Am+Cp]-treated DCs enhanced mouse CD4+ and CD8+ T-cell proliferation. [Am+Cp]-treated DCs exhibited the strongest anti-4T1 metastasis activity in test mice. Treatments with Am, Cp and [Am+Cp] also resulted in augmented expression of CD40, CD80 and CD86 markers in test DCs. Bioinformatics analysis of the cytokine array data from treated DCs identified that [Am+Cp] is efficacious in activation of specific immune functions via mediating the expression of cytokines/chemokines involved in the recruitment and differentiation of defined immune cells. Biochemical analysis revealed that Am and Cp are composed mainly of polysaccharides containing a high level (70-95%) glucose residues, but few or no (< 1%) mannose residues. In summary, our findings suggest that the specific plant polysaccharides Am and Cp extracted from traditional Chinese medicines can be effectively used instead of bacterial LPS as a potent adjuvant in the formulation of a DC-based vaccine for cancer immunotherapies.

No MeSH data available.


Related in: MedlinePlus

Effect of candidate adjuvants and tumor cell lysate on cell viability and T-cell proliferation.(A) Effect of candidate adjuvants (i.e., Cp, Am and [Am+Cp]), and (B) tumor cell lysate on viability of dendritic cells. DCs (2 × 105) were treated with Cp, Am or [Am+Cp] at a dose between 1 to 1000 μg/ml or treated with TCL at concentrations between 50 and 500 μg/ml for 24 h. Cell viability was performed by MTT assay. Data represent the mean ± SD of three replicates. Optimal dosage/concentration of TCL, Cp, Am and [Am+Cp] phytoextracts for stimulating DC-mediated activation of splenocyte or T-cell proliferation. (C) DCs as stimulator cells were pulsed with TCLs at 50–500 μg/ml in medium supplemented with LPS at 1 μg/ml. (D) and (E), TCL-loaded DCs as stimulator cells were treated with Cp, Am or [Am+Cp] phytoextracts at 100 or 200 μg/ml, or with 1 μg/ml LPS (positive control). The (C), (D) and (E) sets of DC stimulator cells were then co-cultured with splenocytes (C), CD8+ T cells (D) or CD4+ T cells (E), as responder cells, for 4 days. Cell proliferation activities of (C) splenocytes, (D) CD8+ cell and (E) CD4+ cells are represented as the fold change over control (i.e., splenocytes or T cells only). Data represent the mean ± SE obtained from three independent experiments. A P value of less than 0.05 was considered significant (*, P < 0.05; **, P < 0.01; ***, P < 0.001; n.s, no significance).
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pone.0122374.g001: Effect of candidate adjuvants and tumor cell lysate on cell viability and T-cell proliferation.(A) Effect of candidate adjuvants (i.e., Cp, Am and [Am+Cp]), and (B) tumor cell lysate on viability of dendritic cells. DCs (2 × 105) were treated with Cp, Am or [Am+Cp] at a dose between 1 to 1000 μg/ml or treated with TCL at concentrations between 50 and 500 μg/ml for 24 h. Cell viability was performed by MTT assay. Data represent the mean ± SD of three replicates. Optimal dosage/concentration of TCL, Cp, Am and [Am+Cp] phytoextracts for stimulating DC-mediated activation of splenocyte or T-cell proliferation. (C) DCs as stimulator cells were pulsed with TCLs at 50–500 μg/ml in medium supplemented with LPS at 1 μg/ml. (D) and (E), TCL-loaded DCs as stimulator cells were treated with Cp, Am or [Am+Cp] phytoextracts at 100 or 200 μg/ml, or with 1 μg/ml LPS (positive control). The (C), (D) and (E) sets of DC stimulator cells were then co-cultured with splenocytes (C), CD8+ T cells (D) or CD4+ T cells (E), as responder cells, for 4 days. Cell proliferation activities of (C) splenocytes, (D) CD8+ cell and (E) CD4+ cells are represented as the fold change over control (i.e., splenocytes or T cells only). Data represent the mean ± SE obtained from three independent experiments. A P value of less than 0.05 was considered significant (*, P < 0.05; **, P < 0.01; ***, P < 0.001; n.s, no significance).

Mentions: In order to characterize the applicability, and biosafety of the candidate adjuvants for a DC-based cancer vaccine, the cytotoxicity of Cp, Am and [Am+Cp] and 4T1 tumor cell lysate on test DCs was determined by MTT assay. Cp, Am and [Am+Cp] at a dose between 1 and 1000 μg/ml did not impair the viability of test DCs after treatment for 24 h (Fig 1A). In addition, TCL at a concentration between 50 and 500 μg/ml also did not exhibit a cytotoxic effect on test DCs (Fig 1B).


Specific medicinal plant polysaccharides effectively enhance the potency of a DC-based vaccine against mouse mammary tumor metastasis.

Chang WT, Lai TH, Chyan YJ, Yin SY, Chen YH, Wei WC, Yang NS - PLoS ONE (2015)

Effect of candidate adjuvants and tumor cell lysate on cell viability and T-cell proliferation.(A) Effect of candidate adjuvants (i.e., Cp, Am and [Am+Cp]), and (B) tumor cell lysate on viability of dendritic cells. DCs (2 × 105) were treated with Cp, Am or [Am+Cp] at a dose between 1 to 1000 μg/ml or treated with TCL at concentrations between 50 and 500 μg/ml for 24 h. Cell viability was performed by MTT assay. Data represent the mean ± SD of three replicates. Optimal dosage/concentration of TCL, Cp, Am and [Am+Cp] phytoextracts for stimulating DC-mediated activation of splenocyte or T-cell proliferation. (C) DCs as stimulator cells were pulsed with TCLs at 50–500 μg/ml in medium supplemented with LPS at 1 μg/ml. (D) and (E), TCL-loaded DCs as stimulator cells were treated with Cp, Am or [Am+Cp] phytoextracts at 100 or 200 μg/ml, or with 1 μg/ml LPS (positive control). The (C), (D) and (E) sets of DC stimulator cells were then co-cultured with splenocytes (C), CD8+ T cells (D) or CD4+ T cells (E), as responder cells, for 4 days. Cell proliferation activities of (C) splenocytes, (D) CD8+ cell and (E) CD4+ cells are represented as the fold change over control (i.e., splenocytes or T cells only). Data represent the mean ± SE obtained from three independent experiments. A P value of less than 0.05 was considered significant (*, P < 0.05; **, P < 0.01; ***, P < 0.001; n.s, no significance).
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4380423&req=5

pone.0122374.g001: Effect of candidate adjuvants and tumor cell lysate on cell viability and T-cell proliferation.(A) Effect of candidate adjuvants (i.e., Cp, Am and [Am+Cp]), and (B) tumor cell lysate on viability of dendritic cells. DCs (2 × 105) were treated with Cp, Am or [Am+Cp] at a dose between 1 to 1000 μg/ml or treated with TCL at concentrations between 50 and 500 μg/ml for 24 h. Cell viability was performed by MTT assay. Data represent the mean ± SD of three replicates. Optimal dosage/concentration of TCL, Cp, Am and [Am+Cp] phytoextracts for stimulating DC-mediated activation of splenocyte or T-cell proliferation. (C) DCs as stimulator cells were pulsed with TCLs at 50–500 μg/ml in medium supplemented with LPS at 1 μg/ml. (D) and (E), TCL-loaded DCs as stimulator cells were treated with Cp, Am or [Am+Cp] phytoextracts at 100 or 200 μg/ml, or with 1 μg/ml LPS (positive control). The (C), (D) and (E) sets of DC stimulator cells were then co-cultured with splenocytes (C), CD8+ T cells (D) or CD4+ T cells (E), as responder cells, for 4 days. Cell proliferation activities of (C) splenocytes, (D) CD8+ cell and (E) CD4+ cells are represented as the fold change over control (i.e., splenocytes or T cells only). Data represent the mean ± SE obtained from three independent experiments. A P value of less than 0.05 was considered significant (*, P < 0.05; **, P < 0.01; ***, P < 0.001; n.s, no significance).
Mentions: In order to characterize the applicability, and biosafety of the candidate adjuvants for a DC-based cancer vaccine, the cytotoxicity of Cp, Am and [Am+Cp] and 4T1 tumor cell lysate on test DCs was determined by MTT assay. Cp, Am and [Am+Cp] at a dose between 1 and 1000 μg/ml did not impair the viability of test DCs after treatment for 24 h (Fig 1A). In addition, TCL at a concentration between 50 and 500 μg/ml also did not exhibit a cytotoxic effect on test DCs (Fig 1B).

Bottom Line: Treatments with Am, Cp and [Am+Cp] also resulted in augmented expression of CD40, CD80 and CD86 markers in test DCs.Biochemical analysis revealed that Am and Cp are composed mainly of polysaccharides containing a high level (70-95%) glucose residues, but few or no (< 1%) mannose residues.In summary, our findings suggest that the specific plant polysaccharides Am and Cp extracted from traditional Chinese medicines can be effectively used instead of bacterial LPS as a potent adjuvant in the formulation of a DC-based vaccine for cancer immunotherapies.

View Article: PubMed Central - PubMed

Affiliation: Department of Food and Nutrition, Providence University, Taichung, Taiwan, R. O. C; Institute of Agricultural Biotechnology Research Center, Academia Sinica, Taipei, Taiwan, R. O. C.

ABSTRACT
Dendritic cell (DC) vaccines are a newly emerging immunotherapeutic approach for the treatment and prevention of cancer, but major challenges still remain particularly with respect to clinical efficacy. Engineering and optimization of adjuvant formulations for DC-based vaccines is one strategy through which more efficacious treatments may be obtained. In this study, we developed a new ex vivo approach for DC vaccine preparation. We evaluated two highly purified mixed polysaccharide fractions from the root of Astragalus membranaceus and Codonopsis pilosulae, named Am and Cp, for their use in enhancing the efficiency of a DC-based cancer vaccine against metastasis of 4T1 mammary carcinoma in mice. Mixed lymphocyte reaction showed all Am-, Cp- and [Am+Cp]-treated DCs enhanced mouse CD4+ and CD8+ T-cell proliferation. [Am+Cp]-treated DCs exhibited the strongest anti-4T1 metastasis activity in test mice. Treatments with Am, Cp and [Am+Cp] also resulted in augmented expression of CD40, CD80 and CD86 markers in test DCs. Bioinformatics analysis of the cytokine array data from treated DCs identified that [Am+Cp] is efficacious in activation of specific immune functions via mediating the expression of cytokines/chemokines involved in the recruitment and differentiation of defined immune cells. Biochemical analysis revealed that Am and Cp are composed mainly of polysaccharides containing a high level (70-95%) glucose residues, but few or no (< 1%) mannose residues. In summary, our findings suggest that the specific plant polysaccharides Am and Cp extracted from traditional Chinese medicines can be effectively used instead of bacterial LPS as a potent adjuvant in the formulation of a DC-based vaccine for cancer immunotherapies.

No MeSH data available.


Related in: MedlinePlus