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An enzyme linked immunosorbent assay for human carcinoembryonic antigen-related cell adhesion 8, a biological marker of granulocytes in vivo
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Medical Sciences, Clinical Chemistry.
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2004 (English)In: JIM - Journal of Immunological Methods, ISSN 0022-1759, E-ISSN 1872-7905, Vol. 293, no 1-2, 201-214 p.Article in journal (Refereed) Published
Abstract [en]

Carcinoembryonic antigen-related cell adhesion molecule 8 (CEACAM8), also known as CD66b, NCA-95 and CD67, is a highly glycosylated protein expressed only in neutrophils and eosinophils in humans. The precise function of CEACAM8 remains unclear. As a member of the family of carcinoembryonic antigen (CEA), it may play a role in the interaction between granulocytes or between granulocytes and epithelial cells. We describe here an accurate, specific and reproducible enzyme-linked immunosorbent assay (ELISA) using purified native CEACAM8 as standard for the measurement of CEACAM8 with a detection range of 1-64 microg/l. We also present data on the levels of CEACAM8 in the blood of healthy individuals and patients undergoing surgery, as well as in patients with acute infections. The highly elevated levels of CEACAM8 in the blood of these patients, which are significantly correlated with the surface expression of CEACAM8 on neutrophils and the number of circulating neutrophils, suggest that CEACAM8 could serve as a biological marker for granulocyte activities in vivo.

Place, publisher, year, edition, pages
2004. Vol. 293, no 1-2, 201-214 p.
National Category
Medical and Health Sciences
Identifiers
URN: urn:nbn:se:uu:diva-92101DOI: 10.1016/j.jim.2004.08.009PubMedID: 15541289OAI: oai:DiVA.org:uu-92101DiVA: diva2:165059
Available from: 2004-09-20 Created: 2004-09-20 Last updated: 2017-12-14Bibliographically approved
In thesis
1. CarcinoEmbryonic Antigen-related Cell Adhesion Molecule 8 (CEACAM8): Purification, Characterization, Cellular and Clinical Studies
Open this publication in new window or tab >>CarcinoEmbryonic Antigen-related Cell Adhesion Molecule 8 (CEACAM8): Purification, Characterization, Cellular and Clinical Studies
2004 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

A 95-kDa protein was purified from normal human granulocytes. The protein reacted with a monoclonal antibody against CEACAM8. MALDI-Tof and MS/MS analyses revealed the protein to be a CGM6 gene product. Thus, the protein was proved to be identical to CEACAM8.

An ELISA for CEACAM8 was developed with detection range of 1-64μg/L. Data are presented on the levels of CEACAM8 in the blood of healthy individuals and patients undergoing surgery, as well as in patients with acute infection. The highly elevated levels of CEACAM8 in the blood of these patients were significantly correlated with the surface expression of CEACAM8 on neutrophils and the number of circulating neutrophils, which suggests that CEACAM8 could serve as a biological marker for granulocyte activitiy in vivo.

The cellular content of CEACAM8 in neutrophils was estimated to be 82.4 ± 8.9 ng/106 cells. Subcellular localisation and mobilisation studies showed that the majority of CEACAM8 is present in the secondary granules of human neutrophils, with a small amount on the plasma membranes. Upon stimulation, CEACAM8 translocated to the plasma membranes from the secondary granules and was also released extracellularly (5.5 ± 0.7% of the total content of CEACAM8).

In eosinophils, the cellular content of CEACAM8 was estimated to be 73.8 ± 6.0 ng/106 cells. In these cells, CEACAM8 is mainly stored in secretory vesicles. Upon activation, eosinophils released 5.1 ± 1.1% of the total content of CEACAM8.

Administration of granulocyte colony-stimulating factor (G-CSF) to healthy individuals resulted in an increased content of CEACAM8 in neutrophils on day 1, and decreased on day 4. However, the content of CEACAM8 in light membrane fractions was increased on day 4. The translocation of CEACAM8 observed in vivo after G-CSF administration is probably not directly related to this cytokine but to other cytokines such as TNF-a.

Place, publisher, year, edition, pages
Uppsala: Acta Universitatis Upsaliensis, 2004. 64 p.
Series
Comprehensive Summaries of Uppsala Dissertations from the Faculty of Medicine, ISSN 0282-7476 ; 1372
Keyword
Biochemistry, human neutrophil, human eosinophil, granule proteins, CEA, CEACAM8, G-CSF, ELISA, subcellular fractionation, Biokemi
National Category
Biochemistry and Molecular Biology
Identifiers
urn:nbn:se:uu:diva-4534 (URN)91-554-6031-3 (ISBN)
Public defence
2004-10-15, Rosensalen, entrance 95/96, Akademiska sjukhuset, Uppsala, 13:15
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Available from: 2004-09-20 Created: 2004-09-20Bibliographically approved

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