Citation

  • Authors: Hu, X., Zhong, Y., Lambers, T. T., Jiang, W.
  • Year: 2019
  • Journal: Inflamm Res 68 715-722
  • Applications: in vitro / Protein/Peptide/Antibody / PULSin
  • Cell type: HCT-8
    Description: Human ileocecal colorectal adenocarcinoma cells

Method

GrB serine protease

Abstract

OBJECTIVE: Nutritional factors such as extensively hydrolyzed casein (eHC) have been proposed to exert anti-inflammatory activity and affect clinical outcomes such as tolerance development in cow's milk allergy. Granzyme B (GrB) induces apoptosis in target cells and also controls the inflammatory response. Whether eHC could affect the activity of granzyme B and play a role in GrB-mediated inflammatory responses in vitro was unknown. METHODS: The activity of GrB was measured using the substrate Ac-IEPD-pNA. Inflammatory responses were induced with GrB in HCT-8 and THP-1 cells, and pro-inflammatory cytokines were determined at the transcriptional and protein level. RESULTS: GrB could induce the expression of IL-1beta in HCT-8 cells, and IL-8 and MCP-1 in THP-1 cells, respectively. Interestingly, GrB acted synergistically on LPS-induced inflammation in HCT-8 cells and eHC reduced pro-inflammatory responses in both GrB and LPS-mediated inflammation. Further analyses revealed that eHC could inhibit the biological activities and cytotoxic activities of GrB and then could reduce GrB-mediated inflammatory response. CONCLUSION: The results from the current study suggest that anti-inflammatory activity of extensively hydrolyzed casein is, to a certain extent, mediated through modulation of granzyme B activity and responses.

Pubmed