Quantitation of surface CD14 on human monocytes and neutrophils

P. Antal-Szalmás, Jos A G Van Strijp, Annemarie J L Weersink, Jan Verhoef, Kok P M Van Kessel

Research output: Contribution to journalArticle

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Abstract

The absolute number of membrane expressed CD14, the most important endotoxin receptor, on human monocytes and neutrophils shows remarkable variation in the literature. To quantify these numbers two fluorescence methods using fluorescein isothiocyanate (FITC)-labeled monoclonal antibodies (mAb) were applied. A commercially available set of standard beads was used in flow cytometry to quantitate CD14 with eight different mAbs. Independent from their isotype the various mAbs showed minor differences and indicated that peripheral blood monocytes expressed 99,500-134,600 (115,400 ± 10,600) and neutrophils 1,900-4,400 (3,300 ± 800) CD14 receptors. There was no significant difference in CD14 expression on leukocytes in unprocessed freshly obtained whole blood and after a Ficoll isolation procedure. However, a short temperature shift resulted in a 1.3- to 1.6-fold up-regulation of CD14. The results obtained with the reference beads were verified with fluorescence Scatchard analysis and spectrofluorometry using mAb 26ic-FITC and showed 109,500 CD14 per monocyte and 6,700 CD14 per neutrophil. For comparison the number of CD14 on the monocytic THP-1 cells and Fcγ-receptors on human leukocytes were determined using the reference beads and flow cytometry and gave results comparable to published data. Our data indicate that resting human monocytes express roughly 110,000 CD14 molecules on their surface using a simple fluorometric assay. Correct determination of the number of CD14 and other cell surface receptors is of importance in the monitoring of septic patients.

Original languageEnglish
Pages (from-to)721-728
Number of pages8
JournalJournal of Leukocyte Biology
Volume61
Issue number6
Publication statusPublished - Jun 1997

Fingerprint

Monocytes
Neutrophils
Fluorescein
Flow Cytometry
Leukocytes
Fluorescence
Monoclonal Antibodies
Ficoll
Fc Receptors
Fluorescence Spectrometry
Cell Surface Receptors
Physiologic Monitoring
Up-Regulation
Temperature
Membranes
isothiocyanic acid

Keywords

  • Anti-CD14
  • Flow cytometer
  • Scatchard analysis
  • Spectrofluorometer
  • Standard beads

ASJC Scopus subject areas

  • Cell Biology

Cite this

Antal-Szalmás, P., Van Strijp, J. A. G., Weersink, A. J. L., Verhoef, J., & Van Kessel, K. P. M. (1997). Quantitation of surface CD14 on human monocytes and neutrophils. Journal of Leukocyte Biology, 61(6), 721-728.

Quantitation of surface CD14 on human monocytes and neutrophils. / Antal-Szalmás, P.; Van Strijp, Jos A G; Weersink, Annemarie J L; Verhoef, Jan; Van Kessel, Kok P M.

In: Journal of Leukocyte Biology, Vol. 61, No. 6, 06.1997, p. 721-728.

Research output: Contribution to journalArticle

Antal-Szalmás, P, Van Strijp, JAG, Weersink, AJL, Verhoef, J & Van Kessel, KPM 1997, 'Quantitation of surface CD14 on human monocytes and neutrophils', Journal of Leukocyte Biology, vol. 61, no. 6, pp. 721-728.
Antal-Szalmás P, Van Strijp JAG, Weersink AJL, Verhoef J, Van Kessel KPM. Quantitation of surface CD14 on human monocytes and neutrophils. Journal of Leukocyte Biology. 1997 Jun;61(6):721-728.
Antal-Szalmás, P. ; Van Strijp, Jos A G ; Weersink, Annemarie J L ; Verhoef, Jan ; Van Kessel, Kok P M. / Quantitation of surface CD14 on human monocytes and neutrophils. In: Journal of Leukocyte Biology. 1997 ; Vol. 61, No. 6. pp. 721-728.
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