CAL-1 as Cellular Model System to Study CCR7-Guided Human Dendritic Cell Migration

Abstract

Dendritic cells (DCs) are potent and versatile professional antigen-presenting cells and central for the induction of adaptive immunity. The ability to migrate and transport peripherally acquired antigens to draining lymph nodes for subsequent cognate T cell priming is a key feature of DCs. Consequently, DC-based immunotherapies are used to elicit tumor-antigen specific T cell responses in cancer patients. Understanding chemokine-guided DC migration is critical to explore DCs as cellular vaccines for immunotherapeutic approaches. Currently, research is hampered by the lack of appropriate human cellular model systems to effectively study spatio-temporal signaling and CCR7-driven migration of human DCs. Here, we report that the previously established human neoplastic cell line CAL-1 expresses the human DC surface antigens CD11c and HLA-DR together with co-stimulatory molecules. Importantly, if exposed for three days to GM-CSF, CAL-1 cells induce the endogenous expression of the chemokine receptor CCR7 upon encountering the clinically approved TLR7/8 agonist Resiquimod R848 and readily migrate along chemokine gradients. Further, we demonstrate that CAL-1 cells can be genetically modified to express fluorescent (GFP)-tagged reporter proteins to study and visualize signaling or can be gene-edited using CRISPR/Cas9. Hence, we herein present the human CAL-1 cell line as versatile and valuable cellular model system to effectively study human DC migration and signaling.

Loss of β2-integrin function results in metabolic reprogramming of dendritic cells, leading to increased dendritic cell functionality and anti-tumor responses

“…# 10131035) selection at 37°C in a humidified atmosphere of 5% CO 2 . The human DC cell line CAL-1 39 was obtained from Leibniz-Institut Deutsche Sammlung von Mikroorganismen und Zellkulturen (DSMZ, #ACC923). Cells were cultured in a complete growth medium (RPMI 1640 supplemented with 10% FCS, 100 U/ml penicillin–streptomycin (penicillin, Orion, cat.…”

Exocrine gland–resident memory CD8 + T cells use mechanosensing for tissue surveillance

“…T RM migration through a confined 3D collagen type I matrix was performed in μ-Slide Chemotaxis ibiTreat chambers (ibidi, Switzerland) as described ( 95 ). Briefly, sorted LifeAct-GFP × mT/mG T RM were resuspended at 5 × 10 5 cells/ml in RPMI 1640 medium supplemented with 10% heat-inactivated fetal bovine serum.…”

Research progress of nanovaccine in anti-tumor immunotherapy

“…Lymph nodes (LNs) are small, bean-shaped organs located throughout the body that are an essential part of the immune system. Within the lymph nodes, immune cells such as DCs, macrophages, B cells, and T cells interact and collaborate to initiate and coordinate adaptive immune responses against foreign invaders like viruses, bacteria, and cancer cells (121). DCs residing in peripheral tissues capture antigens and then migrate through afferent lymphatic vessels toward draining lymph nodes under the action of the chemokine receptor CCR7 (122).…”