TY - JOUR
T1 - ELISA for quantitation of l-selectin shed from leukocytes in vivo
AU - Spertini, Olivier
AU - Schleiffenbaum, Boris
AU - White-Owen, Cathy
AU - Ruiz, Phillip
AU - Tedder, Thomas F.
N1 - Funding Information:
This work was supported by grants from the National Institutes of Health, CA-34183, AI-26872, and CA-54464. O.S. and B.S. were supported by grants from the Swiss National Foundation for Scientific Research. TFT is a Scholar of the Leukemia Society of America.
PY - 1992/11/25
Y1 - 1992/11/25
N2 - l-selectin is a cell surface receptor on granulocytes, lymphocytes and monocytes that is responsible for the initial attachment of leukocytes to endothelium. The extracellular domain of l-selectin is proteolytically shed from leukocytes following cellular activation in vitro. The shed form of l-selectin (sl-selectin) is functionally active and at high concentrations can inhibit leukocyte attachment to endothelium. Therefore, an ELISA was developed to quantitate the levels of sl-selectin in biological fluids, biopsy specimens and during recombinant protein production. This simple, quantitative sandwich ELISA uses two monoclonal antibodies directed against the extracellular domain of sl-selectin. The assay has a detection range of 5-1300 ng/ml, is precise and sensitive. The ability of this assay to detect sl-selectin in serum, plasma, and culture supernatant fluid was demonstrated and it was used to quantitate circulating sl-selectin in normal and patient sera. Patients with sepsis and HIV infection showed markedly elevated sl-selectin levels in serum. Thus, the ELISA should prove useful both for laboratory purposes as well as in the diagnostic evaluation of patients with inflammatory diseases.
AB - l-selectin is a cell surface receptor on granulocytes, lymphocytes and monocytes that is responsible for the initial attachment of leukocytes to endothelium. The extracellular domain of l-selectin is proteolytically shed from leukocytes following cellular activation in vitro. The shed form of l-selectin (sl-selectin) is functionally active and at high concentrations can inhibit leukocyte attachment to endothelium. Therefore, an ELISA was developed to quantitate the levels of sl-selectin in biological fluids, biopsy specimens and during recombinant protein production. This simple, quantitative sandwich ELISA uses two monoclonal antibodies directed against the extracellular domain of sl-selectin. The assay has a detection range of 5-1300 ng/ml, is precise and sensitive. The ability of this assay to detect sl-selectin in serum, plasma, and culture supernatant fluid was demonstrated and it was used to quantitate circulating sl-selectin in normal and patient sera. Patients with sepsis and HIV infection showed markedly elevated sl-selectin levels in serum. Thus, the ELISA should prove useful both for laboratory purposes as well as in the diagnostic evaluation of patients with inflammatory diseases.
KW - ELISA
KW - l-Selectin
KW - Leukocyte
KW - Receptor
KW - Shedding
UR - http://www.scopus.com/inward/record.url?scp=0026498829&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0026498829&partnerID=8YFLogxK
U2 - 10.1016/0022-1759(92)90017-N
DO - 10.1016/0022-1759(92)90017-N
M3 - Article
C2 - 1385536
AN - SCOPUS:0026498829
VL - 156
SP - 115
EP - 123
JO - Journal of Immunological Methods
JF - Journal of Immunological Methods
SN - 0022-1759
IS - 1
ER -