| Budget Amount *help |
¥1,600,000 (Direct Cost: ¥1,600,000)
Fiscal Year 1996: ¥600,000 (Direct Cost: ¥600,000)
Fiscal Year 1995: ¥1,000,000 (Direct Cost: ¥1,000,000)
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| Research Abstract |
L-selectin is a lymphocyte homing receptor and two high endothelial venule (HEV)-associate ligands for L-selectin, known as GlyCAM-1 and CD34, have been identified. Using colloidal gold conjugated-recombinant L-selectin (LSIG), antibodies against CD34 and GlyCAM-1, and an anti-peripheral node addressin mAb (MECA79), we performed morphological mapping of L-selectin ligands in peripheral lymph nodes. For MECA-79 and LSIG,intense labeling was detected on the luminal surface and cytoplasm of the high endothelial cells, and the fibrous layr of the perivascular connective tissue (PVC) of the HEV.The L-selectin ligand in the HEV-PVC,through which extravasated lymphocytes pass into the parenchyma of the lymph node, was found to be neither CD34 nor GlyCAM-1 and another one.
In order to measure the changes in secreted plasma GlyCAM-1 levels, we performed sandwich ELISA assay after inflammatory stimulus. BALB/c mice were injected with complete Freund adjuvant in the hind footpads. GlyCAM-1 was increased at 3 h and reached peak levels at 12 h and gradually decreased thereafter. Levels of EGTA-sensitive L-selectin bound to the plasma GlyCAM-1 changed in similar time course and reached peak at 12 h after and began to decrease. These findings show that GlyCAM-1 release is enhanced by inflammatory stimulation and also suggest that released plasma GlyCAM-1 may trap, at least in part, soluble L-selectin shed from stimulatecd leukocytes to neutralize with each other.
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