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

Elena Nasiri; Malte Student; Katrin Roth; Nadya Siti Utami; Magdalena Huber; Malte Buchholz; Thomas M Gress; Christian Bauer

doi:10.3390/cells12030456

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

Cells. 2023 Jan 31;12(3):456. doi: 10.3390/cells12030456.

Authors

Elena Nasiri¹, Malte Student^{1

2}, Katrin Roth³, Nadya Siti Utami¹, Magdalena Huber⁴, Malte Buchholz¹, Thomas M Gress¹, Christian Bauer¹

Affiliations

¹ Department of Gastroenterology, Endocrinology, Infectious Diseases and Metabolism, University Hospital Marburg, Philipps University Marburg, 35043 Marburg, Germany.
² Department of Internal Medicine I, University Hospital Ulm, 89081 Ulm, Germany.
³ Core Facility Cellular Imaging, Center for Tumor Biology and Immunology, Philipps University Marburg, 35043 Marburg, Germany.
⁴ Institute for Medical Microbiology and Hospital Hygiene, Philipps University Marburg, 35043 Marburg, Germany.

Abstract

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, Il1r^-/- OT-1 mice, or Il18r^-/- 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 Il18r^-/- CTLs in PancOVA tumors and a significant reduction in tumor volume compared to wild-type CTLs. Il18r^-/- 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: <4 μm/min) for Il18r^-/- CTLs. Our data indicate the contribution of IL-18R signaling to T-cell effector strength, warranting further investigation on phenomena such as intratumoral T-cell exhaustion.

Keywords: T-cell migration; T-cell plasticity; cytokines; fluorophore; intravital two-photon microscopy animal models for intravital imaging; live cell imaging; pancreatic carcinoma; pausing phases/arrest coefficient; tumor spheroid coculture model.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
CD8-Positive T-Lymphocytes
Carcinoma, Pancreatic Ductal*
Cell Movement
Interleukin-18
Mice
Pancreatic Neoplasms*

Substances

Interleukin-18
Il18r1 protein, mouse

Grants and funding

This research was funded by DFG grants KFO325 BA 3824/3-1, BA 3824/3-2, RO 5599/1-1, and RO 5599/1-2, and the Behring-Röntgen-Foundation (project number 63-0010).