Reduced expression of CD27 by collagenase treatment: implications for interpreting b cell data in tissues

PloS One
Chanjuan ShenXiaolei Wang

Abstract

Surface markers have been used to identify distinct cell subpopulations and to delineate various stages of maturation or activation of lymphocytes. In particular CD27 is used for delineation of naïve and memory B cell populations, and is readily detected by flow cytometry. We here used flow cytometry to examine the expression of CD27 on lymphocytes isolated from various tissues of rhesus macaques, and found its expression was consistently low to absent on intestinal cell suspensions. However, immunohistochemistry revealed abundant CD27+ cells in intestinal tissue sections. Further investigation showed the marked loss of CD27 expression on processed intestinal cells was due to collagenase digestion of intestinal tissues, yet CD27 expression was recoverable within hours of cell isolation. By combining confocal microscopy, we confirmed that only a fraction of B cells express CD27, in contrast to expression on all T cells from tissues examined including the gut. Taken together, our results suggest that CD27 may be a memory marker for B cells, but not for T cells, since essentially all CD3 T cells expressed CD27. In summary, it is important to consider the influence of isolation procedures on cell surface expression of phenotypic ma...Continue Reading

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Citations

Sep 9, 2017·AIDS Research and Human Retroviruses·Martin TrapecarShomyseh Sanjabi
Sep 27, 2018·American Journal of Veterinary Research·Wei Duan, Mandi J Lopez
Jan 29, 2020·American Journal of Reproductive Immunology : AJRI·Kevin A StonerSharon L Achilles

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Methods Mentioned

BETA
flow cytometry
density gradient centrifugation
FCS
confocal microscopy

Software Mentioned

GraphPad
GraphPad Prism
Adobe Photoshop

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