The tumor microenvironment (TME) is a complex network of cells, signaling molecules and extracellular matrix components. The molecular interactions taking place in the TME can be detected using the Naveni® PPI assays whereas immunofluorescence can be simultaneously applied to detect other proteins (or markers) of interest. Below is a description of co-stains that have been validated together with various Naveni® interaction assays.
Mapping the tumor microenvironment with combined in situ proximity ligation and immunofluorescence assays
The tumor microenvironment (TME) is a complex network of cells, signaling molecules and extracellular matrix components. The molecular interactions taking place in the TME can be detected using the Naveni® PPI assays whereas immunofluorescence can be simultaneously applied to detect other proteins (or markers) of interest. Below is a description of co-stains that have been validated together with various Naveni® interaction assays.
Ki67
Antigen Kiel 67 is an established prognostic and predictive indicator for the assessment of cancer biopsies. It is strongly associated with tumor cell proliferation and growth and its expression is correlated with metastasis and the aggressiveness of the tumor.
Detection of CD8/MHC-I interaction (magenta) in hepatocellular carcinoma with co-staining of Ki67 (cyan). Nuclei in blue.
Ki67
Antigen Kiel 67 is an established prognostic and predictive indicator for the assessment of cancer biopsies. It is strongly associated with tumor cell proliferation and growth and its expression is correlated with metastasis and the aggressiveness of the tumor.
Detection of CD8/MHC-I interaction (magenta) in hepatocellular carcinoma with co-staining of Ki67 (cyan). Nuclei in blue.
CD3
Cluster of differentiation 3 is a T-cell co-receptor which is present on the surface of all T cells. It provides the first signal that initiates T-cell activation and determines the specificity of the immune response. CD3 is present at all stages of T-cell development and is therefore a specific general T-cell marker.
CD8a
Cluster of Differentiation 8a is a cell surface glycoprotein found on most cytotoxic T-lymphocytes that mediates efficient cell-cell interactions within the immune system. CD8a plays a role in T-cell development and activation of mature T-cells, and hence the CD8 molecule is a marker for cytotoxic T-cell population.
Detection of CD8/MHC-I interaction (yellow) in tonsil with co-staining of CD3 (magenta). Nuclei in blue.
CD8a
Cluster of Differentiation 8a is a cell surface glycoprotein found on most cytotoxic T-lymphocytes that mediates efficient cell-cell interactions within the immune system. CD8a plays a role in T-cell development and activation of mature T-cells, and hence the CD8 molecule is a marker for cytotoxic T-cell population.
CD3
Cluster of differentiation 3 is a T-cell co-receptor which is present on the surface of all T cells. It provides the first signal that initiates T-cell activation and determines the specificity of the immune response. CD3 is present at all stages of T-cell development and is therefore a specific general T-cell marker.
Detection of CD8/MHC-I interaction (yellow) in tonsil with co-staining of CD3 (magenta). Nuclei in blue.
CD20
B-lymphocyte antigen CD20 is expressed on the surface of all B-cells beginning at the pro-B phase and progressively increasing in concentration until maturity. CD20 remains present on the cells of most B-cell neoplasms and is absent on otherwise similar appearing T-cell neoplasms, therefore it is very useful in differentiating conditions such as B-cell lymphomas and leukemias from T-cell lymphomas.
Detection of PD1/PD-L1 interaction (cyan) in Hodgkin lymphoma with co-stainings of CD3 (yellow) and CD20 (magenta).
CD20
B-lymphocyte antigen CD20 is expressed on the surface of all B-cells beginning at the pro-B phase and progressively increasing in concentration until maturity. CD20 remains present on the cells of most B-cell neoplasms and is absent on otherwise similar appearing T-cell neoplasms, therefore it is very useful in differentiating conditions such as B-cell lymphomas and leukemias from T-cell lymphomas.
Detection of PD1/PD-L1 interaction (cyan) in Hodgkin lymphoma with co-stainings of CD3 (yellow) and CD20 (magenta).
Pan-cytokeratin antibodies
Pan-cytokeratin antibodies target a group of proteins of the cytoskeletal intermediate filaments that enable cells to withstand mechanical stress. In addition, cytokeratin expression patterns are increasingly utilized to differentiate various types of epithelial malignancies, hence studying cytokeratin expression by immunohistochemistry techniques is a tool widely used for tumor diagnosis and characterization.
Detection of PD1/PD-L1 interaction (cyan) in non-small cell lung cancer with co-stainings of CD8 (yellow) and pan-cytokeratin (magenta).
Pan-cytokeratin antibodies
Pan-cytokeratin antibodies target a group of proteins of the cytoskeletal intermediate filaments that enable cells to withstand mechanical stress. In addition, cytokeratin expression patterns are increasingly utilized to differentiate various types of epithelial malignancies, hence studying cytokeratin expression by immunohistochemistry techniques is a tool widely used for tumor diagnosis and characterization.
Detection of PD1/PD-L1 interaction (cyan) in non-small cell lung cancer with co-stainings of CD8 (yellow) and pan-cytokeratin (magenta).
In situ proximity ligation assay and immunofluorescence co-staining. Illustration of the TME, showing a protein interaction detected by the Naveni® in situ proximity ligation technology and individual protein markers detected by immunofluorescence.
In situ proximity ligation assay and immunofluorescence co-staining. Illustration of the TME, showing a protein interaction detected by the Naveni® in situ proximity ligation technology and individual protein markers detected by immunofluorescence.
How to detect molecular interactions
Use our products NaveniFlex Cell or NaveniFlex Tissue.
How to detect molecular interactions
Use our products NaveniFlex Cell or NaveniFlex Tissue.
NaveniFlex Cell
The assay is designed to be used with a mouse and rabbit primary antibody pair. Detection with TEX615 fluorophore is included.
NaveniFlex Tissue
The assay is designed to be used with a mouse and rabbit primary antibody pair. Detection with Atto647N fluorophore is included.
NaveniFlex Cell
The assay is designed to be used with a mouse and rabbit primary antibody pair. Detection with TEX615 fluorophore is included.
NaveniFlex Tissue
The assay is designed to be used with a mouse and rabbit primary antibody pair. Detection with Atto647N fluorophore is included.
References
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- Li LT, Jiang G, Chen Q, Zheng JN. Ki67 is a promising molecular target in the diagnosis of cancer (review). Mol Med Rep. 2015 Mar;11(3):1566–72.
- Menon AP, Moreno B, Meraviglia-Crivelli D, Nonatelli F, Villanueva H, Barainka M, Zheleva A, van Santen HM, Pastor F. Modulating T Cell responses by targeting CD3. Cancers (Basel). 2023 Feb;15(4):1189.
- Pavlasova G, Mraz M. The regulation and function of CD20: an “enigma” of B-cell biology and targeted therapy. Haematologica. 2020 Jun;105(6):1494–506.
- Cooper K, Leong AS-Y. Manual of diagnostic antibodies for immunohistology. 2nd ed. London: Greenwich Medical Media; 2003. ISBN 978-1-84110-100-2.
- Herrmann H, Bär H, Kreplak L, Strelkov SV, Aebi U. Intermediate filaments: from cell architecture to nanomechanics. Nat Rev Mol Cell Biol. 2007 Jul;8(7):562–73.
- Dabbs DJ. Diagnostic Immunohistochemistry: Theranostic and Genomic Applications. 3rd ed. New York: Saunders; 2010.