Studies on Single-Cell Adhesion Probability between Lymphocytes and Endothelial Cells with Micropipette Technique

An in vitro model with micropipette technique was used to investigate single-cell adhesion probability between lymphocytes and endothelial cells. The basal adhesion probability between lymphocytes and endothelial cells was low and was significantly increased when either lymphocytes were activated by phytohemagglutinin (PHA) or endothelial cells were stimulated by tumor necrosis factor. The adhesion probability of lymphocytes to human umbilical vein endothelial cells was similar to that of lymphocytes to human brain microvascular endothelial cells (HB-MVEC). However, lymphocyte adhesion probability was higher in HB-MVEC than in mouse brain microvascular endothelial cells (MB-MVEC) under both resting and activated conditions. Furthermore, lymphocytes preincubated with monoclonal antibodies to lymphocyte function-associated antigen-1 (LFA-1) or HB-MVEC preincubated with monoclonal antibodies to intercellular adhesion molecule 1 (ICAM-1) significantly down-regulated the adhesion probability between lymphocytes and endothelial cells, indicating that the adhesion probability is related to the expression of LFA-1 on lymphocytes and to the expression of ICAM-1 on endothelial cells. Lymphocytes isolated from patients with cerebral stroke exhibited increased adhesion probability to HB-MVEC as compared with lymphocytes from healthy donors. Preincubation of lymphocytes with tetramethylpyrazine (TMP), an extract from a Chinese traditional herb, effectively inhibited the adhesion probability to HB-MVEC, suggesting that TMP has a potential therapeutic value. These results indicate that the micropipette technique is a useful model for investigating single-cell adhesion probability between lymphocytes and endothelial cells in vitro.