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LPS induced translocation of NF-B occurs only in a subpopulation of CD14-positive mononuclear cells

Laboratory for Experimental Internal Medicine, Academic Medical Center, University of Amsterdam, Amsterdam, The Netherlands, T.tenhove{at}amc.uva.nl

Margriet J.B.M. Vervoordeldonk

Laboratory for Experimental Internal Medicine, Academic Medical Center, University of Amsterdam, Amsterdam, The Netherlands

Pascale E.P. Dekkers

Laboratory for Experimental Internal Medicine, Academic Medical Center, University of Amsterdam, Amsterdam, The Netherlands

Pieter H. Reitsma

Laboratory for Experimental Internal Medicine, Academic Medical Center, University of Amsterdam, Amsterdam, The Netherlands

Sander J.H. van Deventer

Laboratory for Experimental Internal Medicine, Academic Medical Center, University of Amsterdam, Amsterdam, The Netherlands

CD14-receptor occupancy is a potent activator of the ubiquitous transcription factor NF-B. In resting cells, NF-B exists as a heterodimer in the cytoplasm bound to its inhibitor IB. Upon stimulation with lipopolysaccharide (LPS), NF-B translocates to the nucleus. By means of immunohistochemistry, nuclear translocation of the p65 subunit was detected in LPS-stimulated mononuclear cells at the single-cell level. Double-staining experiments showed nuclear translocation of p65 in only a fraction (< 50%) of the LPS-stimulated CD14-positive cells. In vivo, similar results were obtained. In blood from healthy volunteers who were infused with LPS, only a fraction of the CD14-positive cells showed translocation of the p65 subunit. In conclusion, we identified the translocation of NF-B after LPS stimulation at the single cell level in vitro and in vivo. Surprisingly, only a subpopulation of CD14-positive cells showed translocation. These data indicate that the mere expression of CD14 alone is insufficient for LPS-responsiveness.

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