I need to identify dendritic cells in a lymph nodes. If both lymphocytes and DC express CD11c is it possible to discriminate it from lymphocytes by physycs parameter?
CD11c is definitely expressed by lymphocytes. It is a common misconception that it is a DC-specific marker. Check out the data browser at www.immgen.org for specifics (the gene name for CD11c is itgax) - but it's now well described that CD11c is expressed by NK cells, activated T cells, a subset of regulatory CD8+ T cells, gdT cells, and certain macrophage populations. And probably others! Hope that helps.
It depends on the kind of DCs you want. If you need DCs and pDCs, and if you use CD11c+ selection, you might be excluding your CD11c- pDCs from your samples. Activated lymphocytes may express CD11c, in special conditions, like disease, because CD11c is not a mere cell marker, but it is an adhesion molecule (among other functions), and cells may increase and decrease their expression depending on their needs. Otherwise, you may use DC-SIGN as a marker of imature DCs, which lymphocytes are not supposed to express. Everything will also depend on the methodology you want to use. Do you need to identify your cells in lymph nodes by immunohistochemistry, flow cytometry, magnetic beads separation (MACS/Miltenyi) or something else?
CD11c is definitely expressed by lymphocytes. It is a common misconception that it is a DC-specific marker. Check out the data browser at www.immgen.org for specifics (the gene name for CD11c is itgax) - but it's now well described that CD11c is expressed by NK cells, activated T cells, a subset of regulatory CD8+ T cells, gdT cells, and certain macrophage populations. And probably others! Hope that helps.
Thank you for your answer. I'm agree with Anne and Joao!
I'm tracking a fluorescent anitigen following nasal immunization, so i need to identify antigen bearing dendritic cells by flow cyctometry. I'm using CD11c and MHC II to identify it, gating on cells with high FSC and SSC excluding lymphocytes population. Do you think that it is a good strategy?
If you are looking for DC then I would suggest that you go for double staining of CD11c and DEC 205. DEC 205 is a good DC marker.
Gennaro and Aritri, I think you are both right. You can use the strategy of CD11c/MHC staining to identify the cells, gating on FSCxSSC parameters. Further on your analysis, you can add DEC205, to detect immature DCs, and other molecules, like CD80/CD83/CD86/DC-SIGN, and anything else you would like to measure. If you use human cells, you can also stain them with CD14, to be sure you're not getting a high monocytes/macrophages contamination in your samples.
Just read an interesting paper on the subject. Nat Rev Immunol. ; 12(3): 191–200 Maeker et al "Standardizing immunophenotyping for the Human Immunology
Project". Great schematic in figure 2.
I downloaded and have not gotten around to reading Nat Protoc. 2010 ; 5(2): 357–370 Fung et al "Multiplexed immunophenotyping of human antigen-presenting
cells in whole blood by polychromatic flow cytometry". I am admittedly fond of the idea of moving towards standardization.
BTW, CD11c is expressed in hairy cell leukemia malignant cells.
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