Co-Expression of Positive Activation Marker CD38 and Negative or Exhaustion Marker PD-1 in CD8+ T Cells

George Washington University, Washington DC
Surgery
Post-doctoral Fellow

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Company:

BioLegend

Product Name:

PE Anti-human CD38 Antibody

Catalog Number:

303506

CD38 is considered a marker of immune activation expressed in T, B and NK cells. The role of CD38 in CD8+ T cell function against viral infection has been recently in focus and co-expression of PD-1 has been termed as a condition which favors progression of disease. Thus, CD8+ T cells with increased co-expression of CD38 and PD-1 have been termed progressor CD8 T cells and the ones with lower levels of co-expression as controller T cells. It should be kept in mind that this is a relative phenomenon and there will certainly be some percentage of co-expression in CD8+ T cells for these two markers at a given time point.

Experimental Design and Results Summary

Application

Flow cytometry

Starting Material

PBMC, lymphocytes

Protocol Overview

PBMC's form human patients infected with virus were thawed. Cells were washed twice with PBS and stained for the following antibodies- CD4, CD8, CD38, PD-1. Staining was done by diluting the antibodies 1:50 and preparing a cocktail for all the samples except the isotype, unstained, and single color controls. Cells were incubated with the antibody cocktail for 30 mins at 4 degree C/ over ice. Cells were washed twice with PBS and re-suspended in FACS buffer for flow cytometric analysis.

Tips

Preparation of cocktail of antibodies is important to standardize results. Prepare cocktails for n+2 number of samples.

Results Summary

Expression of CD38 and PD-1 were clearly visible in CD8+ T cells (gated on CD8+ population). The percentage of co-expression of CD38 and PD-1 was indicative of progressor vs. controller CD8 T cells.

Additional Notes

None

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Summary

The Good

Good florescence, easily distinguishable.

The Bad

None

The Bottom Line

Easy, reliable and repeatable method of surface staining of CD8+ T cells to identify distinct population co-expressing CD38 and PD-1

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