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Analysis of tumor infiltrating lymphocytes (TIL) by immunohistochemistry (IHC)

AP Annacarmen Petrizzo
AM Angela Mauriello
AL Antonio Luciano
DR Domenica Rea
AB Antonio Barbieri
CA Claudio Arra
PM Piera Maiolino
MT Marialina Tornesello
VG Vincenzo Gigantino
GB Gerardo Botti
GC Gennaro Ciliberto
FB Franco M. Buonaguro
MT Maria Tagliamonte
LB Luigi Buonaguro
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Explanted tumors were analyzed by IHC. Tumors were formalin-fixed paraffin embedded (FFPE). Sections were stained using standard immunohistochemistry (IHC) techniques. Areas of necrosis or hemorrhage, identified on corresponding H&E-stained sections, were excluded from TIL scoring and two blinded individuals scored each section. 50-µm consecutive sections (3 per mouse) were stained for CD3 (clone SP7, Abcam), GZMB (clone 1, catalogue no. 50134-M08H, Sino Biologinal) and FoxP3 (polyclonal, catalogue no.NB100-39002, Novus Biologicals) and detected by a secondary HRP-conjugated antibody (Poly-HRP-anti-rabbit IgG, ImmunoLogic). Reaction was visualized by incubation with the peroxidase substrate (Vector Nova Red, Vector Laboratories). Each sample was analyzed by counting cells in 10 independent areas.

Copyright and License information:  ©2018 Petrizzo et al.
This is an open-access article distributed under the terms of the Creative Commons Attribution License 3.0 (CC BY 3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

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