After After
Before Before

10130 - CD74, Melanoma, TME

INTRODUCTION
CD74, also called invariant chain, plays a role in MHC class II antigen presentation for immune response. Its expression in tumor cells has a prognostic value as well as immune response signaling in many tumors. Thus, it has a potential role in the regulation of an anti-tumor immune response, and has also been shown to be expressed in inflammatory disorders and other inflammation-associated cancers.

The "10130 - CD74, Melanoma, TME" APP detects nuclei and classifies them as either negative, 1+, 2+ or 3+ based on the CD74 staining expression present in each nucleus' vicinity.

KEYWORDS
CD74, melanoma, skin, HLA class II, MHC class II, cancer, oncology, IHC, tumor micro environment

METHODS
To identify the nuclei, the APP performs a two-stage polynomial blob filtering on a blue-enhanced feature image and delimits them using local linear filtering. Each pixel with DAB staining is classified as low, mid and high based on the intensity and grouped together locally. Each nucleus is then classified based on its surroundings in the order of 3+, 2+, 1+ and negative to emphasize the strongest staining present in each nucleus’ vicinity.

QUANTITATIVE OUTPUT VARIABLES
The output variables obtained from this protocol include:

  • Number and percentage of negative, 1+, 2+, and 3+ cells in both stromal and tumor tissue
  • H-score = (% of 3+)x3 + (% of 2+)x2 + (% of 1+) for both stromal and tumor tissue

AUXILIARY APPS (included) 
APP: 01 Detect TumorStroma
The auxiliary APP: “01 Detect TumorStroma” is used for automatic tumor and stromal tissue detection. The analysis APP will then provide results for tumor and stromal tissue separately.

WORKFLOW
Step 1: Load and run the APP “01 Detect TumorStroma” for tumor and stromal tissue identification. Manually correct the result if needed.
Step 2: Load and run the APP “02 CD74 Analysis” for the quantification of cells.

STAINING PROTOCOL
There is no staining protocol available.

ADDITIONAL INFORMATION
The APP utilizes the Visiopharm Engine™ and Viewer software modules, where Engine™ offers an execution platform to expand processing capability and speed of image analysis. The Viewer allows a fast review together with several types of image adjustment properties ex. outlining of regions, annotations and direct measures of distance, curve length, radius, etc.

By adding the Author™ module the APP can be customized to fit other purposes. Author™ offers a comprehensive and dedicated set of tools for creating new fit-for-purpose analysis APPs, and no programming experience is required.

REFERENCES
There are no references available.

RUO
Figure 1
Figure 1
Nuclei surrounded by CD74 negative, low and intermediate staining.
Figure 2
Figure 2
Nuclei classified as either negative (blue), 1+ (yellow) or 2+ (orange) based on the CD74 staining present in each nucleus’ vicinity.
Figure 3
Figure 3
Nuclei surrounded by intermediate and high CD74 staining staining.
Figure 4
Figure 4
Nuclei classified as either 2+ (orange) or 3+ (red) based on the CD74 staining present in each nucleus’ vicinity.
Figure 5
Figure 5
Sample with stromal and tumor tissue.
Figure 6
Figure 6
Automated identification of stromal tissue (shown with a red region of interest (ROI)) and tumor tissue (shown with a blue ROI). The APP outputs, e.g. H-score, is given for the stromal region and tumor region separately.