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Published on: December 2, 2010
Detection of soluble CR3 (CD11b/CD18) by time-resolved immunofluorometry
Gitte Krogh Nielsen1, Thomas Vorup-Jensen
1Biophysical Immunology Laboratory, Department of Biomedicine, The Lundbeck Foundation Nanomedicine Center for Individual Management of Tissue Damage and Regeneration, Aarhus University, Aarhus, Denmark.
Insights
Researchers developed new assays to measure soluble complement receptor 3 (CR3) shedding. This advancement aids in understanding CR3
Area of Science:
- Immunology
- Cell Biology
- Biochemistry
Background:
- Complement receptor 3 (CR3), composed of CD11b and CD18 alpha and beta chains, is crucial for immune responses like cell migration, adhesion, and phagocytosis.
- Evidence suggests ectodomain shedding of CR3 contributes to its biological functions, with soluble forms detected in body fluids.
Purpose of the Study:
- To develop quantitative assays for monitoring the shedding of soluble CR3 components, specifically CD11b and CD18.
- To enable the detection of soluble CD11b and CD18 in human and murine plasma or serum.
Main Methods:
- Development of time-resolved immunofluorometric assays (TRIFMA) for sensitive detection of soluble CD11b and CD18.
- Application of TRIFMA to analyze plasma or serum samples from human and murine sources.
Main Results:
- Successful development of TRIFMAs for quantifying soluble CD11b and CD18.
- TRIFMAs offer superior dynamic range and reproducibility compared to traditional ELISA methods.
Conclusions:
- The developed TRIFMAs provide a robust tool for studying CR3 shedding.
- These assays can advance the understanding of CR3 and other cell adhesion molecule shedding in inflammatory diseases and related animal models.
Abstract:
In the cell membrane complement receptor 3 (CR3) consists of one alpha chain (CD11b) and one beta chain (CD18). CR3 participates in many immunological processes, especially those involving cell migration, adhesion, and phagocytosis of complement-opsonized microbes. Recent findings of soluble CR3 in body fluids and in culture supernatant from experiments in vitro point to the involvement of ecto domain shedding as a part of the CR3 biology. To monitor such shedding on a quantitative basis, we have developed time-resolved immunofluorometric assays (TRIFMA) to detect soluble CD11b and CD18 in plasma or serum of either human or murine origin. Compared with most enzyme-linked immunosorbent assays methodologies, TRIFMA possesses prominent advantages, including better dynamic range and reproducibility. These assays may contribute to the understanding of the role of shedding of CR3 and other cell adhesion molecules in human disease and animal models involving inflammation.
