CD3+ T Cell Quantification in Paraffin-Embedded MycCaP Tumors

Paraffin-embedded MycCaP tumors were stained for CD3 expression by immunohistochemistry as described (18 (link)). Sections were stained with primary antibodies (CD3: clone SP7, Abcam), developed using the LSAB⁺ System-HRP (Agilent Technologies, Santa Clara, CA) and Metal Enhanced DAB Substrate Kit DAB metal concentration (Thermo Fisher Scientific, Waltham, MA), imaged using an Olympus BX51 fluorescent microscope (Olympus, Lombard, IL) in combination with SPOT RT analysis software (SPOT Imaging Solutions, Sterling Heights, MI), and quantified by the frequency of CD3⁺ cells per 10× field, counting at least five fields per tumor section per animal by a blinded investigator.

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Olson B.M., Gamat M., Seliski J., Sawicki T., Jeffery J., Ellis L., Drake C.G., Weichert J, & McNeel D.G. (2017). Prostate cancer cells express more androgen receptor (AR) following androgen deprivation, improving recognition by AR-specific T cells. Cancer immunology research, 5(12), 1074-1085.

Publication 2017

CD3: clone SP7 LSAB+ System-HRP BX51 fluorescent microscope

Animal Antibodies Cells Clone Immunohistochemistry Metal Microscope Paraffin Tumor

Corresponding Organization : University of Wisconsin Carbone Cancer Center

Other organizations : Dana-Farber Cancer Institute, Harvard University, Columbia University, University of Wisconsin–Madison

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Variable analysis

independent variables

Immunohistochemical staining for CD3 expression

dependent variables

Frequency of CD3+ cells per 10x field

control variables

Paraffin-embedded MycCaP tumors
Primary antibody (CD3: clone SP7, Abcam)
LSAB+ System-HRP (Agilent Technologies, Santa Clara, CA)
Metal Enhanced DAB Substrate Kit DAB metal concentration (Thermo Fisher Scientific, Waltham, MA)
Olympus BX51 fluorescent microscope (Olympus, Lombard, IL)
SPOT RT analysis software (SPOT Imaging Solutions, Sterling Heights, MI)
Blinded investigator

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