Characterization of cells from invaded lymph nodes in patients with solid tumors. Lymphokine requirement for tumor-specific lymphoproliferative response.

Abstract

The specific immune response against the malignant cells was investigated in patients with urinary bladder or larynx cancer. Lymphocytes from lymph nodes that drain the tumor site were tested for their proliferative and cytotoxic capacities against autologous malignant cells isolated from the primary tumor. In no occasion was a proliferative or a cytotoxic response observed. However, when the lymph node cell suspensions were depleted of cells expressing both OKM1 and Leu-7 markers by rosetting with the appropriate mAbs, a proliferative response could be observed. The lymphocytes responded to autologous tumor cells only if IL-2 was added to the cultures. IL-2 alone induced some cell proliferation, which was not, however, comparable to that observed in response to both IL-2 and tumor cells. A panel of allogeneic tumor cells consistently failed to stimulate OKM1-, Leu-7- cells in vitro. Response to autologous tumor cells was not caused by HLA-encoded molecules, as occurs in the autologous mixed lymphocyte reaction, since OKM1-, Leu-7- cells failed to be stimulated by autologous non-T cells. A proliferative response was observed only with cells from lymph nodes that had been classified as invaded by malignant cells according to histopathologic criteria. Cells from noninvaded lymph nodes consistently failed to respond. Cells stimulated with autologous tumor cells could be expanded in short-term lines by continuous addition of IL-2 and malignant cells. One of these lines, which comprised mainly T8+ cells, was stimulated to proliferate only by autologous tumor cells, and its proliferative response was inhibitable by anti-class I and not by anti-class II mAbs. This line showed lytic capacities against autologous malignant targets, while it was inefficient against all of the other allogeneic cells tested. In another set of experiments, the mechanisms whereby exogenous IL-2 had to be added to the cultures to sustain a proliferative response against neoplastic cells were investigated. When cocultured with autologous malignant cells, OKM1-, Leu-7- lymphocytes expressed IL-2 receptors, as could be assessed by anti-Tac fluorescent staining. Under these culture conditions, these cells did not produce IL-2, and no proliferation was observed. Addition of purified IL-1 to the cultures induced IL-2 production and cell proliferation. It is concluded that metastatic lymph nodes contain a T cell population that can be detected in a proliferative assay when both suppressor cells are removed and the appropriate molecular signals are supplied.

Lymphokine-activated killer cell function of peripheral blood mononuclear cells, spleen cells and regional lymph node cells in gastric cancer patients

“…The coexistence of suppressor cells in LNC has been reported by several authors [15,35]. Cozzolino et at.…”

“…Cozzolino et at. [35] showed that lymph nodes of patients with solid tumours contained a higher proportion of lymphocytes with CDS^, CDlIb*, CD57^ surface phenotype that had suppressor properties. In the present study, however, CD8'CDIIb' [36,37] cells were few in all cell populations, and remained at low levels after culture with IL-2.…”

Interleukin‐1 and interleukin‐2 control granulocyte‐ and granulocyte‐macrophage colony‐stimulating factor gene expression and cell proliferation in cultured acute myeloblastic leukemia

“…The patients were classified according to the FAB scheme (Bennett et al, 1976) as follows: 4 patients with MI AML, 6 with M2, 6 with M4, and 5 with M5. In all experiments, leukemic cells were depleted of T cells by rosetting with neuraminidase-treated sheep red blood cells and depleted of monocytes by plastic adherence at 37" C. The percentage of cells expressing the p55 subunit of IL-2 receptor was assessed by flow cytometry of cells stained with anti-CD25 MAbs (anti-Tac) (Cozzolino et al, 1987).…”

Suppression of the generation of lymphokine-activated killer (LAK) cells by serum-free supernatants of in vitro maintained tumour cell lines

“…A number of reports have demonstrated that supernatants derived from tumour cell lines can inhibit lymphocyte proliferation and cytokine production (Werkmeister et al, 1980;Meischner et al, 1986;Ebert et al, 1987;Pommier et al, 1987;Farram et al, 1982;Hersey et al, 1983). However, the effects of these supernatants on the generation of cytotoxic responses has rarely been examined (Cozzolino et al, 1987 33258. In experiments in which the tumour cells were added directly to the LAK generation cultures, they were obtained from the confluent monolayers after a brief trypsinisation followed by six washes in complete medium, before re-suspension at a cell density appropriate to a final lymphocyte:tumour cell ratio of 50:1, as previously described (Guillou et al, 1989 Cytotoxicity assay This was a standard 4 h 5"Cr release assay as previously described (Monson et al, 1987).…”

“…A number of reports have demonstrated that supernatants derived from tumour cell lines can inhibit lymphocyte proliferation and cytokine production (Werkmeister et al, 1980; Meischner et al, 1986;Ebert et al, 1987;Pommier et al, 1987;Farram et al, 1982;Hersey et al, 1983). However, the effects of these supernatants on the generation of cytotoxic responses has rarely been examined (Cozzolino et al, 1987 …”