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Scutellaria extract and wogonin inhibit tumor-mediated induction of Treg cells via inhibition of TGF-β1 activity

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Abstract

A number of studies have implicated tumor-induced Treg cell activity in the sub-optimal response to therapeutic vaccines. Development of neo-adjuvant strategies targeting Treg cells is therefore imperative. Scutellaria extracts or constituent flavonoids have shown encouraging efficacy against various tumors, including gliomas, in both pre-clinical and clinical studies. We report here, for the first time, that Scutellaria ocmulgee leaf extract (SocL) and flavonoid wogonin could inhibit TGF-β1-induced Treg activity in malignant gliomas. F344 rats, subcutaneously transplanted with F98 gliomas, were treated with SocL. There was a significant inhibition of intra-tumoral TGF-β1 and Treg cell frequency as well as peripheral blood TGF-β1 levels in SocL-treated animals compared to the controls. SocL extract and wogonin also inhibited glioma-induced, TGF-β1-mediated Treg activity in vitro. SocL extract and wogonin also inhibited the secretion of IL-10 in Treg culture; whereas the level of IL-2 was either unchanged or marginally enhanced. We also observed an inhibition of Smad-3, GSK-3β and ERK1/2 signaling by SocL and wogonin in Treg cells, while phosphorylation of P38 MAPK was considerably enhanced, indicating that SocL or wogonin could inhibit the T cells’ response to TGF-β1 via modulation of both Smad and non-Smad signaling pathways. Overall, this study suggests that Scutellaria can potentially reverse tumor-mediated immune suppression via inhibition of TGF-β1 secretion as well as via inhibition of T cells’ response to TGF-β1. This may provide an opportunity for develo** a novel adjuvant therapeutic strategy for malignant gliomas, combining Scutellaria with immunotherapy and chemo/radio-therapeutic regimen, which could potentially improve the disease outcome.

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Abbreviations

CM:

Conditioned medium

ERK:

Extracellular-regulated kinase

GSK:

Glycogen synthase kinase

nT:

Naïve T cells

IHC:

Immunohistochemistry

SocL:

Scutellaria ocmulgee leaf extract

Treg :

Regulatory T cells

TGF:

Transforming growth factor

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Acknowledgments

We acknowledge research support from the USDA-NIFA (GEOX-2008-02989) and Fund for Medical Research and Education (FMRE). The Microscopy, Imaging and Cytometry Resources Core is supported, in part, by NIH Center grant P30CA22453 to The Karmanos Cancer Institute, Wayne State University and the Perinatology Research Branch of the National Institutes of Child Health and Development, Wayne State University. We are grateful to Dr. Larry Tait for the invaluable help with fluorescent IHC. We are thankful to Dr. Indrajit Sinha for his critical evaluation of the manuscript and also for his help with the preparation of the Figures.

Conflict of interest

The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Department of Neurosurgery, Wayne State University, 550 E. Canfield, Lande Bldg. #460, Detroit, MI, 48201, USA

    Sagar Dandawate, Linford Williams, Sandeep Mittal & Prahlad Parajuli

  2. Agricultural Research Station, Fort Valley State University, Fort Valley, GA, USA

    Nirmal Joshee

  3. USDA-ARS, Natural Products Utilization Research Unit, University, MS, USA

    Agnes M. Rimando

  4. Department of Oncology, Medicine and Immunology and Microbiology, Karmanos Cancer Institute, Wayne State University, Detroit, MI, USA

    Archana Thakur & Lawrence G. Lum

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  1. Sagar Dandawate
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Correspondence to Prahlad Parajuli.

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262_2011_1130_MOESM1_ESM.ppt

Supplementary Figure S1. SocL preferentially inhibits intra-tumoral FOXP3 + T reg cell activity without affecting total T-cell infiltration. F344 rats were subcutaneously transplanted with 1 × 106 F98 glioma cells on the right flank. SocL administration was performed as described. Rats were euthanized on day 29; the tumor was resected and processed for IHC, as described in the ‘Methods’ section, to determine the presence of FOXP3+ and CD3+ T cells. Result shown is from one representative paraffinized glioma specimen out of three studied. The micrographs were imaged at 200× magnifications. (PPT 12789 kb)

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Dandawate, S., Williams, L., Joshee, N. et al. Scutellaria extract and wogonin inhibit tumor-mediated induction of Treg cells via inhibition of TGF-β1 activity. Cancer Immunol Immunother 61, 701–711 (2012). https://doi.org/10.1007/s00262-011-1130-3

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