Hepatic Arterial Bland Embolization Increases Th17 Cell Infiltration in a Syngeneic Rat Model of Hepatocellular Carcinoma
- 268 Downloads
To determine the tumor immune cell landscape after transcatheter arterial bland embolization (TAE) in a clinically relevant rat hepatocellular carcinoma (HCC) model.
Materials and Methods
Buffalo rats (n = 21) bearing syngeneic McArdle RH-7777 rat hepatoma cells implanted into the left hepatic lobe underwent TAE using 70–150 µm beads (n = 9) or hepatic artery saline infusion (n = 12). HCC nodules, peritumoral margin, adjacent non-cancerous liver, and splenic parenchyma were collected and disaggregated to generate single-cell suspensions for immunological characterization 14 d after treatment. Changes in tumor-infiltrating immune subsets including CD4 T cells (Th17 and Treg), CD8 cytotoxic T cells (IFNγ), and neutrophils were evaluated by multiparameter flow cytometry. Migration and colony formation assays were performed to examine the effect of IL-17, a signature cytokine of Th17 cells, on McArdle RH-7777 hepatoma cells under conditions simulating post-embolization environment (i.e., hypoxia and nutrient privation). Statistical significance was determined by the Student unpaired t test or one-way ANOVA.
TAE induces increased infiltration of Th17 cells in liver tumors when compared with controls 14 d after treatment (0.29 ± 0.01 vs. 0.19 ± 0.02; p = 0.02). A similar pattern was observed in the spleen (1.41 ± 0.13 vs. 0.57 ± 0.08; p < 0.001), indicating both local and systemic effect. No significant differences in the percentage of FoxP3 + Tregs, IFNγ-producing CD4 T cells, and CD8 T cells were observed between groups (p > 0.05). In vitro post-embolization assays demonstrated that IL-17 reduces McA-RH7777 cell migration at 24–48 h (p = 0.003 and p = 0.002, respectively).
Transcatheter hepatic arterial bland embolization induces local and systemic increased infiltration of Th17 cells and expression of their signature cytokine IL-17. In a simulated post-embolization environment, IL-17 significantly reduced McA-RH7777 cell migration.
KeywordsTh17 cells IL-17 Transcatheter hepatic arterial bland embolization Tumor-infiltrating lymphocytes HCC immune landscape
This research was supported by BTG and SIO Immuno-Oncology-Interventional Oncology grant program, Biocompatibles UK Ltd (BTG), Surrey, UK, Grant# BTG-SP-08.004-F01, Sister Institution Network Fund (SINF) Grant# SINF-600801-80-115693-21, Department of Defense W81XWH-16-1-0100, and The University of Texas MD Anderson Cancer Center, and Center for Inflammation and Cancer Support Grant. The funders had no role in study design, data collection, analysis, interpretation of the data, decision to submit results, the decision to publish, or preparation of the manuscript.
Compliance with Ethical Standard
Conflict of interest
All authors declare that they have no conflict of interest.
All applicable international, national, and/or institutional guidelines for the care and use of animals were followed. All procedures performed in studies involving animals were in accordance with the ethical standards of the institution at which the studies were conducted. This article does not contain any studies with human participants performed by any of the authors.
- 3.Brown KT, Do RK, Gonen M, Covey AM, Getrajdman GI, Sofocleous CT, et al. Randomized trial of hepatic artery embolization for hepatocellular carcinoma using doxorubicin-eluting microspheres compared with embolization with microspheres alone. J Clin Oncol. 2016;34(17):2046–53. https://doi.org/10.1200/jco.2015.64.0821.CrossRefPubMedPubMedCentralGoogle Scholar
- 8.Duan XH, Li TF, Zhou GF, Han XW, Zheng CS, Chen PF, et al. Transcatheter arterial embolization combined with radiofrequency ablation activates CD8(+) T-cell infiltration surrounding residual tumors in the rabbit VX2 liver tumors. Onco Targets Ther. 2016;9:2835–44. https://doi.org/10.2147/ott.S95973.CrossRefPubMedPubMedCentralGoogle Scholar
- 17.Li Z, Li N, Li F, Zhou Z, Sang J, Chen Y, et al. Immune checkpoint proteins PD-1 and TIM-3 are both highly expressed in liver tissues and correlate with their gene polymorphisms in patients with HBV-related hepatocellular carcinoma. Medicine (Baltimore). 2016;95(52):e5749. https://doi.org/10.1097/md.0000000000005749.CrossRefGoogle Scholar
- 21.Munoz NM, Minhaj AA, Maldonado KL, Kingsley CV, Cortes AC, Taghavi H, et al. Comparison of dynamic contrast-enhanced magnetic resonance imaging and contrast-enhanced ultrasound for evaluation of the effects of sorafenib in a rat model of hepatocellular carcinoma. Magn Reson Imaging. 2019;57:156–64. https://doi.org/10.1016/j.mri.2018.11.012.CrossRefPubMedGoogle Scholar
- 24.Hendry S, Salgado R, Gevaert T, Russell PA, John T, Thapa B, et al. assessing tumor-infiltrating lymphocytes in solid tumors: a practical review for pathologists and proposal for a standardized method from the international immunooncology biomarkers working group: part 1: assessing the host immune response, TILs in invasive breast carcinoma and ductal carcinoma in situ, metastatic tumor deposits and areas for further research. Adv Anat Pathol. 2017;24(5):235–51. https://doi.org/10.1097/pap.0000000000000162.CrossRefPubMedPubMedCentralGoogle Scholar
- 25.Rhee TK, Larson AC, Prasad PV, Santos E, Sato KT, Salem R, et al. Feasibility of blood oxygenation level-dependent MR imaging to monitor hepatic transcatheter arterial embolization in rabbits. J Vasc Interv Radiol. 2005;16(11):1523–8. https://doi.org/10.1097/01.Rvi.0000182179.87340.D7.CrossRefPubMedGoogle Scholar
- 34.Liao Y, Wang B, Huang ZL, Shi M, Yu XJ, Zheng L, et al. Increased circulating Th17 cells after transarterial chemoembolization correlate with improved survival in stage III hepatocellular carcinoma: a prospective study. PLoS ONE. 2013;8(4):e60444. https://doi.org/10.1371/journal.pone.0060444.CrossRefPubMedPubMedCentralGoogle Scholar
- 36.Zhou Y, Xu X, Ding J, Jing X, Wang F, Wang Y, et al. Dynamic changes of T-cell subsets and their relation with tumor recurrence after microwave ablation in patients with hepatocellular carcinoma. J Cancer Res Ther. 2018;14(1):40–5. https://doi.org/10.4103/jcrt.JCRT_775_17.CrossRefPubMedGoogle Scholar
- 42.Erinjeri JP, Thomas CT, Samoilia A, Fleisher M, Gonen M, Sofocleous CT, et al. Image-guided thermal ablation of tumors increases the plasma level of interleukin-6 and interleukin-10. J Vasc Interv Radiol. 2013;24(8):1105–12. https://doi.org/10.1016/j.jvir.2013.02.015.CrossRefPubMedPubMedCentralGoogle Scholar
- 43.Rozenblum N, Zeira E, Bulvik B, Gourevitch S, Yotvat H, Galun E, et al. Radiofrequency ablation: inflammatory changes in the periablative zone can induce global organ effects, including liver regeneration. Radiology. 2015;276(2):416–25. https://doi.org/10.1148/radiol.15141918.CrossRefPubMedGoogle Scholar