A quantitative immunomorphological analysis of actively responding B-(germinal centre) and T-lymphocyte (paracortical) areas in 187 patients with malignant melanoma recurrent in the regional lymph nodes has been made. The results show that tumour involved nodes have significantly increased germinal centre and decreased paracortical response in comparison with uninvolved lymph nodes. Patients treated with surgery alone whose uninvolved nodes had hyperplastic (defined as exceeding the median) germinal centre areas, had significantly shorter survival than those without germinal centre hyperplasia. No association was found between lymph node immunomorphology and survival in BCG treated patients.

Several biologic characteristics of human carcinoembryonic antigen (CEA)-producing colon adenocarcinoma cells grown in vitro were analyzed. Doubling times of exponentially growing cells ranged from 34 to 38 hours for initial cell concentrations of 105 through 7×105/petri dish (4,290-30,030 cells/cm2). The generation time, as analyzed by pulse-labeled mitosis (PLM) techniques and calculated according to the model of Jansson, was 29.3 hours with a coefficient of variation of 22%. Cultures exposed to continuous incubation with tritiated thymidine had a growth fraction of 90%. Compartment distribution calculated in reference to cell cycle transit times measured by PLM techniques was similar to that defined directly by pulse cytophotometry (PCP). PCP analysis of cells in stationary phase of growth revealed a significant fraction of cells with S-phase DNA content, even though the simultaneously defined labeling index was only 1%. Adequate synchronization in S- and G2-phase was achieved by treatment of the cells with 7 mm thymidine for 24 hours. Centrifugal elutriation and mitotic selection techniques were clearly inferior in terms of reproducibility and cell yield. Colony formation was most efficient for cells plated in fresh medium and incubated for 20 days. Cells inoculated into nude mice produced tumor masses that presented morphologic markers similar to those defined for in vitro cells and were capable of synthesizing CEA.

ABSTRACTThe growth kinetics of an established human lymphoma cell line were analyzed by a variety of techniques utilizing various cell inocula (5 x 104 ‐ 5 x 105 cells) dispensed into 60 mm diameter dishes. Techniques included pulse‐labeled mitosis (PLM), continuous labeling with 3H‐TdR, time‐lapse photography (TLP), cell counts by electronic particle counter, and DNA histography obtained by pulse cytophotometry (PCP). There were no significant differences among values determined for any kinetic parameters as a function of cell concentration. the average doubling time of exponentially growing cells, regardless of cell inoculum, was 44.1 hr. the generation time determined by PLM was 31.1 hr with a SD of 4.7 hr. Transit times for each stage were: TG1= 10.6 hr, Ts= 9.9 hr, TG2= 9.9 hr, and Tm= 0.7 hr. Repeated experiments using continuous labeling with 3H‐TdR demonstrated a TG2 of 6.3 hr. the longer value determined by PLM is possibly due to the technical manipulations of this procedure which may delay pulse‐labeled cells from resuming cell cycle transit. Hence, values for cell cycle stages were recalculated to give TG1= 14.1 hr, Ts= 9.9 hr, TG2 = 6.3 hr, and Tm= 0.7 hr. These results were used to compute the size of each cell cycle stage compartment pool and corresponded very closely to values defined directly by PCP. TLP analysis considered only cells that produced colonies of at least thirty‐two cells. Generation times ranged from 8 to 89 hr and showed a positive skewness. the average value measured for 330 divisions was 34.5 hr with a SD of 13.2 hr. Thus, the variance predicted by curve fitting of the PLM data did not correlate with that defined by time‐lapse photography nor did it encompass the range in generation times observed directly by TLP. There was a positive correlation between sister‐sister cell generation times (+0.66) but no relation was noted for mother‐daughter values.

Histological slides of primary tumors and regional lymph nodes from 134 unselected patients operated on for colorectal carcinoma of Dukes' Class B were assessed semiquantitatively for the presence of perivascular lymphocyte cuffing in the muscular layers and pericolic/subserosal fat immediately subjacent to the tumors and for paracortical hyperplasia in the regional lymph nodes. These two immunomorphological features related significantly to each other (p less than 0.05), and their combined presence related signifcantly to favorable disease-free interval (p = 0.02) and to survival (p = 0.04), making possible the identification of a subgroup of approximately one-third of Dukes' B class patients with an estimated better than 85% chance for 5-year recurrence-free survival.

The poor postsurgical prognosis in patients with colorectal cancer of the Dukes' C classification has prompted a clinical trial of adjuvant immunotherapy versus chemoimmunotherapy intended to prolong either the disease-free interval or the overall survival or both. One hundred and twenty-one patients have been entered on this study. Fifty-two patients received BCG alone and 69 patients received the combination of 5-FU and BCG. The disease-free interval and the overall survival were compared with similar parameters in a group of historical controls with similar prognostic characteristics who were operated on in our institution prior to the initiation of the current study. There was no difference as yet between BCG alone and the combination of 5-FU + BCG in terms of both the disease-free interval and the survival. Both treatments, however, had significantly better results than the surgical controls. Adjuvant therapy, especially with BCG is advocated for patients with colorectal carcinoma, Dukes' C class, following potentially curative surgery.