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Abstract
Anti-tumour effector cells were generated through 4 days culture of normal C57BL/6 splenocytes in a medium containing concanavalin A supernatant and then fractionated with Dolichos biflorus lectin (DBA) into DBA+ (agglutinable with DBA) and DBA- (non-agglutinable with DBA) cells. The DBA- cells, infused intravenously into mice together with B16 melanoma cells, or adoptively transferred into mice 3 days after the injection of B16 cells, caused a marked decrease in the number of lung nodules, while the DBA+ cells exerted no effect. On the other hand, the DBA+ cells exhibited higher cytolytic activity in vitro than the DBA- cells in short-term 51Cr-release assays. Then, we analysed the mechanism of the strong anti-tumour activity of DBA- cells in vivo. We found that DBA- cells showed higher response to recombinant interleukin-2 (rIL-2) than DBA+ cells and proliferated very well with a small amount of IL-2. In addition, DBA- cells adhered more strongly to lung endothelial cells than DBA+ cells in response to rIL-1 or rTNF. Furthermore, DBA- cells produced larger amounts of macrophage activating factor (MAF) including IFN-gamma when cultured with B16 melanoma. Taken together, our results show that DBA- cells are effective in reducing experimental pulmonary metastases not only by the direct lytic activity but also by the indirect killing activity through the activated macrophage.
Highlights
Recent studies have shown that murine splenocytes or human peripheral blood lymphocytes, on incubation in vitro in the presence of IL-2, acquire the ability to lyse a variety of fresh syngeneic murine and autologous human tumour cells in short-term 5'Cr-release assays
Since we previously demonstrated that alloreactive CTL (Yamazaki et al, 1983) and Con A sup-cultured splenocytes from X5563 tumour-bearing C3H/HeN mice (Okada et al, 1986) could be enriched in the cell fraction having affinity for DBA (DBA+ cells), when examined in vitro against X5563 cells in a 4 h 51Cr release assay, we investigated whether or not Con A sup-cultured splenocytes from normal mice were enriched in the DBA +fraction in the present study
Splenocytes were cultured for 4 days in the presence of 10% Con A sup (v/v) and separated into DBA+ and DBA- fractions by means of rosette formation using DBA coupled sheep red blood cells (SRBC)
Summary
Recent studies have shown that murine splenocytes or human peripheral blood lymphocytes, on incubation in vitro in the presence of IL-2, acquire the ability to lyse a variety of fresh syngeneic murine and autologous human tumour cells in short-term 5'Cr-release assays. These lymphokine-activated killer (LAK) cells are distinct from natural killer (NK) cells or classical cytotoxic T-cells, and are lytic toward allogenic tumour target cells from murine and human sources as well (Grimm et al, 1982, 1983; Grimm & Rosenberg, 1983; Rosenstein et al, 1984)
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