The adhesion of Anti-CD3-Activated mouse T cells to syngeneic colon adenocarcinoma cells is differentially regulated by protein tyrosine kinase-, protein kinase C-, and cAMP-dependent pathways in the effector cell

The adhesion of anti-CD3-activated mouse T cells (AK-T cells) to syngeneic colon adenocarcinoma (MCA-38) cells is mediated principally through the integrin VLA-4 (α₄,β₁). We investigated the signalling pathways through which this adhesive interaction might be regulated. The protein tyrosine kinase inhibitors genistein and methyl 2,5-dihydroxycinnamate (MDHC) markedly inhibited the adhesion of AK-T cells to MCA-38 cells. Furthermore, pretreatment of the AK-T cells alone (but not the MCA-38 targets) with MDHC inhibited adhesion to a comparable extent as when MDHC was present during the assay. Calphostin C, an inhibitor of protein kinase C, also inhibited the adhesion of AK-T cells to MCA-38 monolayers. However, the phosphatidylinositol 3-kinase inhibitor wortmannin failed to alter AK-T cell adhesion to MCA-38 tumour cells. Inhibition of protein kinase A with the R(p) diastereomer of adenosine cyclic 3',5'-phosphorothioate had no effect on adhesion, but the adenylyl cyclase activator forskolin and the cell-permeable cAMP analogues 8-Br-cAMP and dibutyryl-cAMP significantly suppressed adhesion. Pretreatment of AK-T cells alone with forskolin also inhibited adhesion. The adhesion of AK-T cells to MCA-38 tumour targets is therefore promoted by protein tyrosine kinases and protein kinase C, but inhibited by cAMP-dependent pathways, and the predominant location of the regulatory pathways is within the effector cell.