Substance P increases in vitro lymphokine-activated-killer (LAK) cell cytotoxicity against fresh colorectal cancer cells

Abstract

Gut-associated lymphoid cells are modulated by several gut hormones. We postulated that lymphokine-associated-killer (LAK) cell cytotoxicity of lymphocytes isolated from the gut mucosa may be increased by substance P (SP). Intestinal lamina propria mononuclear cells (LPMC) and colonic cancer cells were isolated from operative specimens by successive mechanical and enzymatic dissociation methods. Effector LAK cells were induced by culturing LPMC with recombinant interleukin-2 at a concentration of 250 U/ml. Substance P (10 −5, M) was added to the culture medium. Targets consisted of fresh colon cancer cells, HT-29 (cultured human colon cancer cell line), and control cell lines. After 4 days of incubation, cytotoxicity was measured using a 4-h 51Cr release assay. LAK cells alone showed moderate cytotoxicity against HT-29 and none against fresh colon cancer cells. LAK cells generated in the presence of substance P showed moderate cytotoxicity against HT-29 and strong cytotoxicity against fresh colorectal cancer cells. The percentage of cytotoxicity ± SEM at various effector to target ratios was [(∗) denotes P < 0.05 compared with above]: Effector Target N 5:1 10:1 25:1 50:1 LAK HT-29 7 2.8±0.5 11.6±1.0 12.3±2.3 17.8±4.2 LAK +SP HT-29 11 1.2±0.5 4.1±0.9 9.3±1.0 16.9±3.1 LAK +SP Fresh Ca 11 5.6 ± 1.8 12.6 ± 3.3 38.3 ± 6.3∗ 59.4 ± 9.5∗ LAK Fresh Ca 11 0.0 ± 0.0∗ 0.1 ± 0.1∗ 0.1±0.1∗ 0.5 ± 0.5∗ We conclude that substance P significantly increases 1 LAK cell cytotoxicity against fresh colon cancer cells, but not against cultured cells.