In vitro generation of tumour-specific lymphocyte reactivity to colonic carcinoma cells

Abstract

Purified tumour cells and normal mucosa cells from fresh human colorectal cancer resection specimens, and T-cell-enriched autologous peripheral blood lymphocytes, were mixed in short-term (6 day) mixed lymphocyte-tumour cell (MLTC) microcultures. Lymphocyte stimulation was measured by 3H-thymidine uptake, and a stimulation index (SI = [lymphocytes vs tumour cells (cpm)-tumour cells (cpm)]/[lymphocytes (cpm)] greater than 3 was regarded as significant. Significant lymphocyte reactivity was found in 10/15 patients with colon carcinoma. However, 1 patient with autologous tumour reactivity, also showed significant stimulation against autologous normal mucosa cells, suggesting tumour-associated reactivity. Maximum stimulation occurred most frequently at a lymphocyte:tumour cell ratio of 2:1 and with nylon wool-passaged lymphocytes.

The Sir David Cuthbertson Medal Lecture 1992

“…Before starting feeding and after 24 h of feeding blood samples were obtained for 'fasted' and 'fed' serum. The tumours were obtained at the time of surgery and by a mixture of mechanical and enzymic digestion a suspension of cancer cells was obtained (Kuppner et al 1987). These cells were incubated in a medium enriched with serum from the same patient taken in either the fed or fasted state and the rate of cellular protein synthesis was determined by the incorporation of [3H]phenylalanine into the cancer cells.…”

Tumour-associated proliferative responses in vitro of regional lymph nodes draining solid cancers in man

“…Lymphocytes were isolated from regional (non-involved) tumour-draining lymph nodes using a mechanical disaggregation technique [6]. The cell suspensions were enriched for T cells by depleting adherent monocytes/macrophages and B cells as described elsewhere [12]. The non-adherent T-enriched T-LNL were resuspended at 1 x 10 6 cells/ml in tissue-culture medium consisting of RMPI 1640 (Gibco, Paisley, UK) supplemented with penicillin and streptomycin (Glaxo, Greenford, UK; 100 U/tal and 100 txg/ml, respectively) and also with 20% heat-inactivated (56°C, 30 min) human serum (Flow, Herts, UK).…”

“…We have shown recently that lymphocytes, isolated from blood, are able to show in vitro-a tumour-specific reactivity when co-cultured with autologous colorectal tumour cells [12].…”

“…There are a substantial amount of published data concerning the generation in vitro of lymphocytes specifically stimulated against tumour-associated antigens. The majority of the latteL however, investigated peripheral blood lymphocytes (PBL) only, co-cultured with autologous tumour cells [7,12,17,18]. In some of these studies [17], as well as in others [9,24], tumour-specific cytotoxic *Part of this project was supported by a grant from Cancer Research Campaign Offprint requests to: T. Mainou-Fowler cells were generated.…”

“…We have used, as detailed previously [12], autologous non-malignant cells, derived from tissue from which the tumour has originated, to delineate tumour specificity. There are a substantial amount of published data concerning the generation in vitro of lymphocytes specifically stimulated against tumour-associated antigens.…”