MIF, Melanoma, TME

Macrophage migration Inhibitory Factor (MIF) is a dominant cytokine that acts through CD74 to promote cell proliferation, migration and survival pathways in both immune and epithelial cell types. Its binding to CD74 initiates survival pathways and cell proliferation. It is known as a pleiotropic proinflammatory protein for which the effects include the promotion of cytokine expression and inhibition of apoptosis.

The “10131 – MIF, Melanoma, TME” APP detects nuclei and classifies them as either negative, 1+, 2+ or 3+ based on the MIF staining expression present in each nucleus’ vicinity.

Figure 1

Nuclei surrounded by MIF intermediate and high staining.

Figure 2

Nuclei classified as 2+ (orange) or 3+ (red) based on the MIF staining present in each nucleus’ vicinity.

Figure 3

Nuclei surrounded by MIF negative and low staining.

Figure 4

Nuclei classified as negative (blue) or 1+ (yellow) based on the MIF staining present in each nucleus’ vicinity.

Figure 5

Stromal tissue surrounded by tumor tissue.

Figure 6

Automated identification of stromal tissue (shown with a red region of interest (ROI)) and tumor tissue. The APP outputs, e.g. H-score, is given for the stromal region and tumor region separately.

Details

Auxiliary APPs

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.

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

Workflow

Step 1: Load and run the APP “01 Detect TumorStroma” for tumor and stromal tissue identification. Manually correct result if needed.

Step 2: Load and run the APP “02 MIF Analysis” for the quantification of cells.

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.

Staining Protocol

There is no staining protocol available.

Keywords

MIF, macrophage inhibitory factor, melanoma, skin, CD74, cancer, oncology, IHC, tumor micro environment

References

LITERATURE
There are currently no references.