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Assay of Adhesion Under Shear Stress for the Study of T Lymphocyte-Adhesion Molecule Interactions
Published on: June 29, 2016
Expression and function of intercellular adhesion molecule 1 (ICAM-1) on rat thymic macrophages in culture
Insights
Intercellular adhesion molecule 1 (ICAM-1) is expressed on rat thymic macrophages (TMF). ICAM-1 mediates TMF and thymocyte binding, crucial for cell-cell communication in the thymus.
Area of Science:
- Immunology
- Cell Biology
- Histology
Background:
- Thymic macrophages (TMF) play a role in T-cell development.
- Intercellular adhesion molecule 1 (ICAM-1) is involved in immune cell interactions.
Purpose of the Study:
- To investigate ICAM-1 expression on rat TMF.
- To determine the role of ICAM-1 in TMF-thymocyte adhesion.
Main Methods:
- Monoclonal antibody (mAb) 1A 29 and streptavidin-biotin immunoperoxidase technique for ICAM-1 detection.
- Double immunofluorescence staining to identify macrophage subsets.
- Thymocyte binding assays and inhibition studies with anti-ICAM-1 and anti-LFA-1 mAbs.
Main Results:
- Freshly isolated TMF express high levels of ICAM-1 (~80%), decreasing upon cultivation.
- ICAM-1 is present on both cortical and medullary TMF subsets.
- Recombinant IFN-gamma, IL-1, and TNF-alpha up-regulate ICAM-1, while dexamethasone down-regulates it.
- ICAM-1 mediates thymocyte binding to TMF, confirmed by inhibition studies.
Conclusions:
- ICAM-1 is dynamically expressed on rat TMF and modulated by cytokines and dexamethasone.
- ICAM-1 is a key adhesion molecule mediating TMF-thymocyte interactions, facilitating direct cell-cell communication.
Abstract:
The expression of intercellular adhesion molecule 1 (ICAM-1) was studied on freshly isolated rat thymic macrophages (TMF) and after their cultivation in serum-free medium using monoclonal antibody (mAb) 1A 29 and a streptavidin-biotin immunoperoxidase technique. ICAM-1 was expressed on about 80% of freshly isolated TMF. Upon cultivation, the percentage of ICAM-1+ TMF decreased to about 30-40% in 12-day-old culture. Using double immunofluorescence staining it was found that ICAM-1 was expressed both on cortical (R-MC 40+) and CMZ/medullary (R-MC 43+) macrophage subsets. ICAM-1 was up-regulated on TMF in culture by recombinant IFN-gamma, IL1 and TNF-alpha and was down-regulated by dexamethasone. Syngeneic thymocytes bound to cultivated TMF in a rosette form at both 37 degrees C and 4 degrees C. IFN-gamma treatment did not increase the binding formation. The binding between thymocytes and IFN-gamma-stimulated TMF at 37 degrees C was inhibited by pretreatment of TMF with anti-ICAM-1 mAb or pretreatment of thymocytes with anti-LFA-1 mAb, indicating that ICAM-1 on TMF is one of the ligands involved in TMF/thymocyte adhesion and subsequent direct cell-cell communication.
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