IDO1 expression in 171 PTs and 40 corresponding LNMs was investigated by IHC. TMAs were prepared following published protocols (Freier et al. 2003 (link); Moratin et al. 2019 (link)).
Slides were stained anti-IDO1 monoclonal antibody (clone D5J4E, Cell Signaling Technology, Cambridge, United Kingdom) at a dilution of 1:400 following the manufacturer’s instructions and were scanned with an Axio Scan.Z1 (Zeiss, Oberkochen, Germany) for further investigation. The TMAs were scored using ZEN (blue edition) program (Zeiss) by three independent observers.
TMAs were assessed by determining the density of IDO1+ immune cells and distinguishing the distribution of IDO1+ immune cells between peritumoural and intratumoural regions.
We used a dataset previously published by Moratin et al. to evaluate the correlation between IDO1 and PD-L1/2 (Moratin et al. 2019 (link)). In situ hybridization for human papillomavirus (HPV)-DNA was performed following published protocols (Kühn et al. 2021 (link); Linxweiler et al. 2015 (link)). Immunohistochemical staining targeting p16 was performed following the manufacturer’s instructions (clone EPR1473, Abcam, Cambridge, United Kingdom).
Slides were stained anti-IDO1 monoclonal antibody (clone D5J4E, Cell Signaling Technology, Cambridge, United Kingdom) at a dilution of 1:400 following the manufacturer’s instructions and were scanned with an Axio Scan.Z1 (Zeiss, Oberkochen, Germany) for further investigation. The TMAs were scored using ZEN (blue edition) program (Zeiss) by three independent observers.
TMAs were assessed by determining the density of IDO1+ immune cells and distinguishing the distribution of IDO1+ immune cells between peritumoural and intratumoural regions.
We used a dataset previously published by Moratin et al. to evaluate the correlation between IDO1 and PD-L1/2 (Moratin et al. 2019 (link)). In situ hybridization for human papillomavirus (HPV)-DNA was performed following published protocols (Kühn et al. 2021 (link); Linxweiler et al. 2015 (link)). Immunohistochemical staining targeting p16 was performed following the manufacturer’s instructions (clone EPR1473, Abcam, Cambridge, United Kingdom).
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