Hyperthermia increases intercellular adhesion molecule-1 expression and lymphocyte adhesion to endothelial cells.

BACKGROUND

We observed that the synergistic combination of immunotherapy and whole-body hyperthermia is active against large well-vascularized tumors but not microscopic tumors, and we therefore hypothesized that hyperthermia may act on lymphocyte-endothelial cell interactions. We undertook these studies to evaluate the effect of hyperthermia on lymphocyte-endothelial cell adhesion.

METHODS

Cultured human umbilical vein endothelial cells (HUVEC) and normal peripheral blood lymphocytes were used. HUVEC were cultured to confluence. Treatment groups included control, hyperthermia alone (41 degrees C for 2 hours), interferon-gamma (IFN-gamma) alone, or hyperthermia + interferon-gamma. 51Cr-labeled peripheral blood lymphocytes were allowed to adhere to treated HUVEC, and nonadhering cells were washed away. Adherent cells were lysed and counted in a gamma-counter, calculating an adhesion index compared to controls. The experiment was then conducted with the addition of anti-intercellular adhesion molecule (ICAM) antibody. Cell surface ICAM expression was determined with double monoclonal antibody staining and fluorescence-activated cell sorter analysis, and soluble ICAM secretion was determined with enzyme-linked immunosorbent assay in each group.

RESULTS

In a representative experiment, interferon-gamma increased adhesion by a factor of 1.81 (p < 0.05) compared with control and hyperthermia by 1.38 (p < 0.05) and combined treatment by a factor of 2.43 (p < 0.05). Anti-ICAM antibody abrogated the increased adhesion caused by hyperthermia but did not abrogate the effect of interferon-gamma. Although only 26% of control cells expressed ICAM-1 on the cell surface, interferon-gamma increased expression to 53% (p < 0.05), hyperthermia increased expression to 38% (p < 0.05), and combined treatment increased expression to 61% (p < 0.05). Soluble ICAM-1 was not increased 12 hours after treatment, but by 24 hours significant (p < 0.05) differences (control 0.262 ng/ml, IFN alone 1.50, hyperthermia alone 1.57, and combined 2.71) were noted.

CONCLUSIONS

These results suggest that hyperthermia has a significant effect on lymphocyte adhesion to endothelial cells, at least in part mediated by ICAM-1. Cell surface ICAM-1 is increased at 12 hours, and soluble ICAM-1 is increased at 24 hours. These data suggest that hyperthermia may function by increasing lymphocyte adhesion, providing another locus of action to improve clinical results with immunotherapy.