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Four-color Fluorescence Immunohistochemistry of T-cell Subpopulations in Archival Formalin-fixed, Paraffin-embedded Human Oropharyngeal Squamous Cell Carcinoma Samples
- Source :
- Punt, S, Baatenburg de Jong, R J & Jordanova, E S 2017, ' Four-color Fluorescence Immunohistochemistry of T-cell Subpopulations in Archival Formalin-fixed, Paraffin-embedded Human Oropharyngeal Squamous Cell Carcinoma Samples ', Journal of Visualized Experiments, no. 125 . https://doi.org/10.3791/55589, Journal of Visualized Experiments. MYJoVE Corporation, ISSUE=125;ISSN=1940-087X;TITLE=Journal of Visualized Experiments
- Publication Year :
- 2017
- Publisher :
- MyJove Corporation, 2017.
-
Abstract
- The four-color fluorescence immunohistochemistry (IHC) technique is a method to quantify cell populations of interest while taking into account their relative distribution and their localization in the tissue. This technique has been extensively applied to study the immune infiltrate in various tumor types. The tumor microenvironment is infiltrated by immune cells that are attracted to the tumor site. Different immune cell populations have been found to play different roles in the tumor microenvironment and to have a different impact on the outcome of disease. This manuscript describes the use of multiparameter fluorescence IHC on oropharyngeal squamous cell carcinoma (OPSCC) as an example. This technique can be extended to other tissue samples and cell types of interest. In the presented study, we analyzed the intraepithelial and stromal compartment of a large OPSCC cohort (n = 162). We focused on total T lymphocytes (CD3(+)), immunosuppressive regulatory T cells (Tregs; i.e., FoxP3(+)), and T helper 17 (Th17) cells (i.e., IL-17(+)CD3(+)) using a nuclear counterstain to distinguish tumor epithelium from stroma. A high number of T cells was found to be correlated with improved disease-free survival in patients with a low number of intratumoral IL-17(+) non-T cells. This suggests that IL-17(+) non-T cells may be correlated with a poor immune response in OPSCC, which is in agreement with the correlation described between IL-17 and poor survival in cancer patients. Currently, novel multiparameter fluorescence IHC techniques are being developed using up to 7 different fluorochromes and will enable the more precise characterization and localization of immune cells in the tumor microenvironment.
- Subjects :
- Male
Cancer Research
Pathology
medicine.medical_specialty
Cell type
Stromal cell
CD3 Complex
T-Lymphocytes
General Chemical Engineering
CD3
T cell
Video Recording
Kaplan-Meier Estimate
Biology
T-Lymphocytes, Regulatory
Disease-Free Survival
General Biochemistry, Genetics and Molecular Biology
Immune system
medicine
Humans
Tumor microenvironment
Paraffin Embedding
General Immunology and Microbiology
General Neuroscience
Interleukin-17
FOXP3
Forkhead Transcription Factors
Middle Aged
Immunohistochemistry
Oropharyngeal Neoplasms
medicine.anatomical_structure
Microscopy, Fluorescence
Carcinoma, Squamous Cell
biology.protein
Th17 Cells
Female
Subjects
Details
- ISSN :
- 1940087X
- Database :
- OpenAIRE
- Journal :
- Journal of Visualized Experiments
- Accession number :
- edsair.doi.dedup.....a24ea2984cf6f52ef28412330796a5f6