Dysfunction of Cytotoxic T Lymphocyte Induced by Hepatoma Cells through the Gln-GLS2-Endoplasmic Reticulum Stress Pathway

Wei Wang; Mengnan Guo; Zekun Bai; Wen Bai; Weixi Chen; Yali Su; Jinghua Wu

doi:10.31083/j.fbl2708243

Frontiers in Bioscience-Landmark (Aug 2022)

Dysfunction of Cytotoxic T Lymphocyte Induced by Hepatoma Cells through the Gln-GLS2-Endoplasmic Reticulum Stress Pathway

Wei Wang,
Mengnan Guo,
Zekun Bai,
Wen Bai,
Weixi Chen,
Yali Su,
Jinghua Wu

Affiliations

Wei Wang: Laboratory Department, Tangshan Maternal and Child Health Care Hospital Affiliated to North China University of Science and Technology, 063000 Tangshan, Hebei, China
Mengnan Guo: Laboratory Department, Tangshan Maternal and Child Health Care Hospital Affiliated to North China University of Science and Technology, 063000 Tangshan, Hebei, China
Zekun Bai: Laboratory Department, Tangshan Maternal and Child Health Care Hospital Affiliated to North China University of Science and Technology, 063000 Tangshan, Hebei, China
Wen Bai: Laboratory Department, Tangshan Maternal and Child Health Care Hospital Affiliated to North China University of Science and Technology, 063000 Tangshan, Hebei, China
Weixi Chen: Laboratory Department, Tangshan Maternal and Child Health Care Hospital Affiliated to North China University of Science and Technology, 063000 Tangshan, Hebei, China
Yali Su: Laboratory Department, Tangshan Maternal and Child Health Care Hospital Affiliated to North China University of Science and Technology, 063000 Tangshan, Hebei, China
Jinghua Wu: Laboratory Department, Tangshan Maternal and Child Health Care Hospital Affiliated to North China University of Science and Technology, 063000 Tangshan, Hebei, China

DOI: https://doi.org/10.31083/j.fbl2708243
Journal volume & issue: Vol. 27, no. 8
p. 243

Abstract

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Background: Metabolic activities of tumor cells lead to a depletion of nutrients within the tumor microenvironment, which results in the dysfunction of infiltrating T cells. Here, we explored how glutamine (gln) metabolism, which is essential for biosynthesis and cellular function, can affect the functions of cytotoxic T lymphocytes (CTLs). Methods: Activated CTLs were co-cultured with hepatoma cells. Western blot was used to analyze changes of proteins and ELISA was used to analyze changes of effector. RNA-sequencing was used to detect differentially expressed genes in CTLs. The status of the endoplasmic reticulum (ER) was investigated using transmission electron microscopy experiments. Results: Co-culturing CTLs and hepatoma cells revealed that CTLL-2 cells in the co-culture group expressed high levels of PD-1 (Programmed cell death protein 1), TIM-3 (T cell immunoglobulin and mucin domain-containing protein-3), GRP78 (Glucose regulated protein 78), and P-PERK (phosphorylated protein kinase RNA-activated-like endoplasmic reticulum kinase) and secreted low levels of Granzyme B and perforin. Additionally, the substructure of the ER was severely damaged. When CTLs were treated with an inhibitor of ER stress, their functions were restored. Next, complete medium without Gln was used to culture cells, causing CTLs to display dysfunction and ER stress. WB results revealed decreased expression levels of GLS2 and SLC1A5 (Solute carrier family 1 member 5) in CTLs in the co-culture group. Subsequently, glutaminase (GLS) inhibitors were added to the cultures. As expected, CTLs treated with a GLS2 inhibitor had increased protein content of PD-1 and TIM-3, decreased secretion of Granzyme B and perforin, and an enhanced ER stress response. Conclusions: In summary, CTLs are functionally downregulated induced by hepatoma cells through the Gln-GLS2-ERS pathway.

Published in Frontiers in Bioscience-Landmark

ISSN: 2768-6701 (Print); 2768-6698 (Online)
Publisher: IMR Press
Country of publisher: Singapore
LCC subjects: Science: Chemistry: Organic chemistry: Biochemistry; Science: Biology (General)
Website: https://www.imrpress.com/journal/FBL

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