This product comprises a CRISPR/Cas9-edited polyclonal KLRB1 knockout cell population derived from the Ca Ski cell line, offering a reliable loss-of-function model for investigating CD161-mediated immune regulatory mechanisms. The targeted disruption of KLRB1 in these cells enables functional interrogation of the inhibitory receptor CD161 in a human cervical carcinoma background without clonal isolation, preserving genetic heterogeneity and facilitating robust comparative studies.
Ca Ski cells represent a well-established adherent epithelial model of human cervical squamous cell carcinoma, maintaining stable integration of human papillomavirus type 16 (HPV16). This line is widely utilized to study HPV-driven carcinogenesis, including the roles of viral oncoproteins E6 and E7, and provides a physiologically relevant host for examining tumor-intrinsic and immune-modulatory pathways in the cervical microenvironment.
KLRB1 encodes CD161, an inhibitory C-type lectin receptor predominantly expressed on natural killer (NK) cells and subsets of T cells. Engagement by its ligand LLT1 (CLEC2D) triggers phosphorylation of the intracellular immunoreceptor tyrosine-based inhibitory motif (ITIM), which recruits SHP-1 and SHP-2 phosphatases. These phosphatases dephosphorylate key proximal signaling molecules, including Vav1, LAT, and PLC??1, thereby attenuating activation signals. This cascade suppresses NK cell degranulation and reduces secretion of pro-inflammatory cytokines such as IFN-?? and TNF-??. Expression of KLRB1 is regulated by cytokines including IL-12, IL-15, and IL-18, along with transcription factors Eomes and T-bet, while interacting partners include Lck and LLT1.
In the cervical cancer context, where HPV16 oncoproteins contribute to an immunosuppressive milieu, CD161?CLLT1 interactions may promote immune evasion by dampening anti-tumor effector functions. The Ca Ski knockout model allows dissection of CD161’s contribution to immune checkpoint-like signaling within the HPV16-positive epithelial background, providing insights into how tumor cells might indirectly influence NK and T cell responses via this receptor axis.
Practical applications include co-culture assays with primary NK cells or T cells to evaluate alterations in cytotoxicity (e.g., LDH release or CD107a degranulation), cytokine profiles (ELISA or intracellular flow cytometry), and phospho-signaling events (phospho-ITIM, phospho-SHP-1). The cells are also suitable for assessing LLT1-dependent interactions, gene expression studies by RT-qPCR, protein confirmation by Western blot, and immunofluorescence-based localization. For further technical details, please contact Ascent Research.
