Figure 1
Characterisation of lymphocytes. In 15 colon cancer patients, tumour-draining, sentinel lymph nodes were identified by peritumoural injection of a blue tracer. Specimens from the tumour (CC), sentinel node (SN) and nonsentinel node (LN) and stained with haematoxylin–eosin (left panels) (40 ×). Data from patient no 1 with a Duke's B tumour (A) and patient no 12 with Duke's C (B). Arrows indicate the presence of metastatic colon cancer cells in a sentinel node (B, left panel, SN). Single cell suspensions from the specimens were stained with antibodies against CD4 and the activation marker CD69 and analysed using flow cytometry (middle panels), the percentage of double positive activated CD4 T-helper cells are indicated in the upper right corner. Cell suspensions in triplicates were incubated with a 10- and 100-fold dilutions of autologous tumour homogenate in a day 5–7 proliferation assay. Cells were pulsed 18 h before harvesting with 1 μCi 3H-Thymidine. Proliferation data from day 5 (A) and day 6 (B), respectively, are shown. Error bars indicate s.e.m. (right panels).
