IL18 Receptor Signaling Inhibits Intratumoral CD8 + T-Cell Migration in a Murine Pancreatic Cancer Model

• Published in: Cells (Basel, Switzerland) Switzerland), 2023-01, Vol.12 (3), p.456
• Main Authors: Nasiri, Elena, Student, Malte, Roth, Katrin, Siti Utami, Nadya, Huber, Magdalena, Buchholz, Malte, Gress, Thomas M, Bauer, Christian
• Format: Article
• Language: English
• Subjects: Adenocarcinoma; Adoptive transfer; Analysis; Animals; Antigens; Carcinoma, Pancreatic Ductal; Care and treatment; CD8 antigen; CD8-Positive T-Lymphocytes; Cell adhesion & migration; Cell culture; Cell migration; Cell Movement; Cell-mediated cytotoxicity; Cytokines; Cytotoxicity; Diagnosis; Experiments; fluorophore; Fluorophores; Health aspects; Immunology; Immunotherapy; Inflammation; Interleukin 1 receptors; Interleukin 18; Interleukin 18 receptors; intravital two-photon microscopy animal models for intravital imaging; live cell imaging; Lymphatic system; Lymphocytes; Lymphocytes T; Metastases; Mice; Motility; Ovalbumin; Pancreatic cancer; Pancreatic Neoplasms; pausing phases/arrest coefficient; Spheroids; T cells; T-cell migration; Testing; Tumor cells; tumor spheroid coculture model; Tumors
• ISSN: 2073-4409; 2073-4409
• DOI: 10.3390/cells12030456

In pancreatic ductal adenocarcinoma (PDAC), the infiltration of CD8 cytotoxic T cells (CTLs) is an important factor in determining prognosis. The migration pattern and interaction behavior of intratumoral CTLs are pivotal to tumor rejection. NLRP3-dependent proinflammatory cytokines IL-1β and IL-18 play a prominent role for CTL induction and differentiation. Here, we investigate the effects of T-cellular IL-1R and IL-18R signaling for intratumoral T-cell motility. Murine adenocarcinoma cell line Panc02 was stably transfected with ovalbumin (OVA) and fluorophore H2B-Cerulean to generate PancOVA H2B-Cerulean tumor cells. Dorsal skinfold chambers (DSFC) were installed on wild-type mice, and PancOVA H2B-Cerulean tumor cells were implanted into the chambers. PancOVA spheroids were formed using the Corning Matrigel -based 3D cell culture technique. CTLs were generated from OT-1 mice, OT-1 mice, or OT-1 mice and were marked with fluorophores. This was followed by the adoptive transfer of CTLs into tumor-bearing mice or the application into tumor spheroids. After visualization with multiphoton microscopy (MPM), Imaris software was used to perform T-cell tracking. Imaris analysis indicates a significantly higher accumulation of CTLs in PancOVA tumors and a significant reduction in tumor volume compared to wild-type CTLs. CTLs covered a longer distance (track displacement length) in comparison to wild-type (WT) CTLs, and had a higher average speed (mean track speed). The analysis of instantaneous velocity suggests a higher percentage of arrested tracks (arrests: