Quantitative Immunohistochemical Analysis of Immune Markers

After IHC staining, all TMA slides were scanned and evaluated through Pannoramic Viewer (3DHISTECH, Budapest, HUNGARY). CD73 expression was evaluated by IHC score. As previously described, the IHC score was generated by multiplying the percentage of immunoreactive cells by their corresponding staining intensity [18 (link)]. The mean optic density (MOD) was used to quantify the staining density of HHLA2 assisted by the Densitoquant and HistoQunant module from 3DHISTECH [31 (link)]. CD8 + TILs, Foxp3 + Tregs, CD68 + , and CD163 + TAMs were calculated manually. For each patient, five independent microscopic fields (400 ×) of the immune cell-infiltrating area were selected and counted manually by 2 investigators blinded to the patient information. The discrepancy between investigators was resolved together. The cutoff values were determined by R for optimal survival separation.

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Sun B.Y., Yang Z.F., Wang Z.T., Liu G., Zhou C., Zhou J., Fan J., Gan W., Yi Y, & Qiu S.J. (2023). Integrative analyses identify CD73 as a prognostic biomarker and immunotherapeutic target in intrahepatic cholangiocarcinoma. World Journal of Surgical Oncology, 21, 90.

Publication 2023

Pannoramic Viewer Densitoquant

Cd163 Cd73 Cell Microscopic Optic Patient Tams Tils

Corresponding Organization : Fudan University

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Variable analysis

independent variables

CD73 expression (IHC score)
HHLA2 staining density (MOD)

dependent variables

CD8+ TILs
Foxp3+ Tregs
CD68+
CD163+ TAMs

control variables

Microscopic fields (400x) of immune cell-infiltrating area

controls

Positive control: Not specified
Negative control: Not specified

Annotations

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