The KLRB1 Knockout SK-HEP-1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human SK-HEP-1 liver endothelial-like cell line. These cells feature targeted disruption of the KLRB1 gene encoding CD161, an inhibitory C-type lectin receptor. The polyclonal format provides a heterogeneous pool of edited cells, serving as a robust loss-of-function model for studying KLRB1-mediated immune regulation in a liver sinusoidal endothelial context.
The parental SK-HEP-1 line originates from hepatic adenocarcinoma ascites and displays an endothelial phenotype positive for CD31 and von Willebrand factor. It is a well-established model for liver sinusoidal endothelium, exhibiting angiogenic behavior and adhesion molecule expression. This endothelial background makes the cells particularly suitable for investigating endothelial?Cimmune cell crosstalk within the liver microenvironment, including interactions with natural killer cells and T lymphocytes.
KLRB1 encodes CD161, which binds its ligand LLT1 (CLEC2D) on target cells. Upon ligation, CD161 recruits SHP-1 phosphatase via its ITIM, dephosphorylating activating kinases such as ZAP70 and Syk, thereby attenuating downstream PI3K/AKT and NFAT signaling. This cascade inhibits NK cell cytotoxicity and reduces production of IFN-?? and IL-17. KLRB1 expression is modulated by cytokines including IL-12, IL-18, IL-15, and TGF-??, and transcription factors ROR??t, T-bet, and Eomes, linking it to Th17 and natural killer cell pathways.
Disruption of KLRB1 in SK-HEP-1 cells removes CD161-mediated inhibition, allowing enhanced activation of co-cultured immune cells. This model is valuable for dissecting the KLRB1-LLT1 immune checkpoint axis in hepatocellular carcinoma, where liver sinusoidal endothelial cells may modulate anti-tumor immunity. It also facilitates examination of endothelial CD161 in lymphocyte adhesion, transmigration, and cytokine secretion under inflammatory conditions.
These cells support Western blotting for CD161 and phospho-ZAP70/Syk, flow cytometry, and co-culture cytotoxicity assays with LLT1+ targets. Applications include ELISA for IFN-?? and IL-17, RT-qPCR, and Sanger sequencing for indel confirmation. They enable investigation of CD161 as a checkpoint target in liver cancer, functional study of the KLRB1-LLT1 axis in endothelial-immune interactions, and drug target validation. For additional technical information, contact Ascent Research.
