Classification Of Naive, Effector And Memory T Cells

April 17, 2018

Surgery
George Washington University, Washington DC
Post-doctoral Fellow

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Company:

Invitrogen Antibodies

Product Name:

CD62L (L-Selectin) Monoclonal Antibody (DREG-56 (DREG56)), FITC

Catalog Number:

11-0629-42

Naive, memory and effector T cells can be differentiated according to a homing receptor, selectins and specific naive /memory/effector markers such as CD45RA/CD45RO. Naive CD4/CD8 T cells are CD45RA+, CD62L+, CCR7+ and CD45RO– whereas central memory T cells are CD45RA- CD45RA–CD45RO+CCR7+CD62L+, effector memory T cells are CD45RA–CD45RO+CCR7–CD62L–, and effector cells are CD45RA+CD45RO–CCR7–CD62L–. Thus central memory (Tcm) and Effector memory (Tem) can be differentiated on the basis of CD62L and CCR7 expression as both are CD45RA– (negative). Here we show that on the basis of CD45RA negative or positive, we can differentiate various T cell subtype using CD62L and CCR7.

Experimental Design and Results Summary

Applications

Flow Cytometry

Sample

Human PBMC

Primary Incubation

Human PBMC's were stained with CD62L FITC; CD45RA (percpcy5.5), CCR7 (APC); CD8 (PEcy7); CD4 ( APCcy7) and CD3 (e450). all antibodies were diluted to a final concentration of 1:100 (each) and mixed in a cocktail of FACS buffer.PBMC's were stained with the antibody cocktail mix for 30 at room temperature. After staining cells were washed with FACS staining buffer and finally re-suspended in desired volume of FACS buffer (300-500ul per sample). Cell aquisition was done in a 3 laser, 8 color flow cytometer and data analysed by flowjo software.

Blocking Agent

N/A

Secondary Incubation

N/A

Tertiary Incubation

N/A

Detection

Flurochrome conjugated

Results Summary

Distinct staining was obtained for all antibodies on the basis of which T cell memory subtypes were differentiated. Briefly, T cells were gated on CD45RA positive and negative and further subgated on CD62L and CCR7 to identify, naive, Tcm, Tem and T effector populations.

DOI or PMID #

N/A

Additional Notes

N/A

Related Categories

Image Gallery

Summary

The Good

Good surface staining achieved by the recommended antibodies

The Bad

nothing to mention

The Bottom Line

Gating on CD45RA negative cells, and then differentiating on the basis of CD62L and CCR7 expression results in proper analysis of T memory subtypes.

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