TY - JOUR
T1 - Multiplexed immunofluorescence analysis and quantification of intratumoral PD-1+ Tim-3+ CD8+ T cells
AU - Granier, Clémence
AU - Vinatier, Emeline
AU - Colin, Elia
AU - Mandavit, Marion
AU - Dariane, Charles
AU - Verkarre, Virginie
AU - Biard, Lucie
AU - El Zein, Rami
AU - Lesaffre, Corinne
AU - Galy-Fauroux, Isabelle
AU - Roussel, Hélène
AU - De Guillebon, Eléonore
AU - Blanc, Charlotte
AU - Saldmann, Antonin
AU - Badoual, Cécile
AU - Gey, Alain
AU - Tartour, Éric
N1 - Publisher Copyright:
© 2018 Journal of Visualized Experiments.
PY - 2018/2/8
Y1 - 2018/2/8
N2 - Immune cells are important components of the tumor microenvironment and influence tumor growth and evolution at all stages of carcinogenesis. Notably, it is now well established that the immune infiltrate in human tumors can correlate with prognosis and response to therapy. The analysis of the immune infiltrate in the tumor microenvironment has become a major challenge for the classification of patients and the response to treatment. The co-expression of inhibitory receptors such as Program Cell Death Protein 1 (PD1; also known as CD279), Cytotoxic T Lymphocyte Associated Protein 4 (CTLA-4), T-Cell Immunoglobulin and Mucin Containing Protein-3 (Tim-3; also known as CD366), and Lymphocyte Activation Gene 3 (Lag-3; also known as CD223), is a hallmark of T cell exhaustion. We developed a multiparametric in situ immunofluorescence staining to identify and quantify at the cellular level the co-expression of these inhibitory receptors. On a retrospective series of frozen tissue of renal cell carcinomas (RCC), using a fluorescence multispectral imaging technology coupled with an image analysis software, it was found that co-expression of PD-1 and Tim-3 on tumor infiltrating CD8+ T cells is correlated with a poor prognosis in RCC. To our knowledge, this represents the first study demonstrating that this automated multiplex in situ technology may have some clinical relevance.
AB - Immune cells are important components of the tumor microenvironment and influence tumor growth and evolution at all stages of carcinogenesis. Notably, it is now well established that the immune infiltrate in human tumors can correlate with prognosis and response to therapy. The analysis of the immune infiltrate in the tumor microenvironment has become a major challenge for the classification of patients and the response to treatment. The co-expression of inhibitory receptors such as Program Cell Death Protein 1 (PD1; also known as CD279), Cytotoxic T Lymphocyte Associated Protein 4 (CTLA-4), T-Cell Immunoglobulin and Mucin Containing Protein-3 (Tim-3; also known as CD366), and Lymphocyte Activation Gene 3 (Lag-3; also known as CD223), is a hallmark of T cell exhaustion. We developed a multiparametric in situ immunofluorescence staining to identify and quantify at the cellular level the co-expression of these inhibitory receptors. On a retrospective series of frozen tissue of renal cell carcinomas (RCC), using a fluorescence multispectral imaging technology coupled with an image analysis software, it was found that co-expression of PD-1 and Tim-3 on tumor infiltrating CD8+ T cells is correlated with a poor prognosis in RCC. To our knowledge, this represents the first study demonstrating that this automated multiplex in situ technology may have some clinical relevance.
KW - Checkpoint inhibitors
KW - Digital imaging
KW - Immunofluorescence technique
KW - Immunology
KW - In situ single cell analysis
KW - Issue 132
KW - Multiplex in situ
KW - Multispectral imaging
KW - PD-1
KW - Renal cell carcinoma
KW - Tim-3
KW - Tumor microenvironment
UR - http://www.scopus.com/inward/record.url?scp=85042036124&partnerID=8YFLogxK
U2 - 10.3791/56606
DO - 10.3791/56606
M3 - Article
C2 - 29553498
AN - SCOPUS:85042036124
SN - 1940-087X
VL - 2018
JO - Journal of Visualized Experiments
JF - Journal of Visualized Experiments
IS - 132
M1 - e56606
ER -