Phototherapy in the presence of haematoporphyrin derivative has been shown to have a preferential effect on malignant tumours when compared to normal tissue. This communication presents a comparison of the sensitivity to photochemotherapy in vitro of different cell lines. Asynchronous populations of cells were exposed to light in the presence of haematoporphyrin derivative, and found to be inactivated with a comparable efficiency. The lines were of human, Chinese-hamster or mouse origin and had different abilities to form tumours after heterotransplantation into nude mice or transplantation into syngeneic, immunosuppressed mice. Synchronized cells from 4 of the lines showed a similar variation in sensitivity to light throughout the cell cycle. Cells near the middle of interphase showed the highest sensitivity, whilst cells in early G1 were found to be least sensitive towards treatment with haematoporphyrin derivative and light.

The effect of hypoxic misonidazole (MISO) pretreatment on the subsequent heat sensitivity of EMT6 multicellular tumour spheroids has been investigated. Spheroids were grown in static culture and treated when either 200-300 micrometers or 650-800 micrometers in diameter. Pretreatment was carried out in glass spinner vessels containing 100 ml medium with or without 5mM MISO under conditions of hypoxia. Three-hour hypoxic pretreatment with 5mM MISO significantly enhanced the subsequent response of 200 micrometers spheroids to heat at 43 degrees C. Oxic incubation at 37 degrees C between pretreatment and heating caused progressively loss of heat sensitization with time, recovery being almost complete after 6 h. Recovery was inhibited by incubation at 0 degree C during the interval. Preheating 200 micrometers spheroids at 43 degrees C for 2 h increased their subsequent sensitivity to hypoxic MISO at 37 degrees C. These results are discussed in relation to known mechanisms of MISO metabolism.

Cytoplasts and karyoplasts were obtained by ultracentrifugation of Hepatoma D23 cells on a Ficoll gradient containing cytochalasin B. Their nuclear and protein content and their metabolic activity were determined. Three i.p. injections of 2.3 x 10(7) cytoplasts were unable to protect syngeneic WAB/Not rats against an s.c. challenge of 10(4) D23 cells, whereas a similar amount of karyoplasts, or 3 injections of 10(6) irradiated D23 cells, were fully protective. Ability of cytoplasts to act as primary or secondary immunogen were also studied, and compared to that of 0.01% glutaraldehyde-treated cells, 43 degrees C heat-treated cells and 3M KCl-soluble extracts, these preparations also being of weak immunogenicity. Only heat-treated cells behaved as a primary immunogen, whereas none of the preparations provided a secondary stimulation. Moreover, when these preparations were fed in vitro to peritoneal-exudate cells before their injection into rats, cytoplasts and glutaraldehyde-treated cells showed no immunogenicity, whereas heat-treated cells induced full protection against tumour challenge. Therefore, in this tumour model, the in vivo persistence of immunogen and the presence of a nucleus are likely to be crucial in inducing transplantation resistance to tumour.

We observed the growth of 2 sublines of leukaemia L1210 in histocompatible DBA2 mice given 10(3) cells i.p. and studied the protective effect of Corynebacterium parvum (CP). The growth of subline L1210-M was unaffected by pretreatment with CP or admixture with 10(5) peritoneal cells (PC) from CP-treated mice. In contrast, the growth of subline L1210-C was inhibited; CP pretreatment increased the proportion of long-term survivors (70% vs 20%) and admixture with CP-PC prolonged the survival time (59 days vs 49 days; P less than 0.05). In vitro experiments indicated that Sublines M and C were equally sensitive to cytostasis by CP-PC, as measured in a terminal labelling assay (greater than 90% inhibition of proliferation). However, subline C was much more sensitive to cytolysis (18h 125IUDR-release assay) by CP-PC; percentage specific release from L1210-C was at least 90%, whilst from L1210-M it was generally less than 25%. The differential susceptibility of the 2 sublines to cytolytic PC was maintained through 75 passages in culture. The effector cells were considered to be macrophages, because they were adherent, phagocytic, and sensitive to silica. Cytolysis was unrelated to endotoxin contamination, because it was not inhibited by polymyxin B, and was inhibited by pre-incubating PC in culture medium for 24 or 48 h before adding target cells. Thus the relevance of nonspecific macrophage-mediated cytotoxicity in vitro to tumour resistance in vivo may depend on the strength of the cytotoxic reaction.

The concentration of cellular oestrogen receptor (RE) was measured in both the soluble and nuclear-pellet fractions of biopsies from 1,000 breast cancers. Data suggest that functional steroid RE is always in equilibrium between the soluble and nuclear fractions. However, biopsies from only one-third of patients contained detectable amounts of high-affinity RE in both fractions. Thirty patients out of 42 (71%) whose biopsies contained RE in both fractions, showed objective remission after receiving some form of hormonal manipulation as sole treatment. Response rates in the other categories ranged from 9% for those whose biopsies contained no detectable RE to 24% for those who displayed soluble RE alone. The presence of RE in both fractions of primary disease, whereas RE-negativity was maintained during progression from primary to secondary disease. Other aspects of RE status in relation to stage of disease are analysed.

Immune-suppressed mice carrying xenografts of several different types of human germ-cell tumours were injected with a radiolabelled monoclonal antibody (LICR LON/HT13) raised against membrane components of a human germ-cell tumour (HX39). Subsequent assessment of radioactivity in excised organs and tumours showed a selective accretion of antibody in the tumour. Quantitative autoradiography supported the results of radiolocalization observed in vivo in different tumours, and also showed that the antibody localized to viable tumour cells and in close association with their cell membrane. The vascular architecture of tumours was found to be an important factor governing antibody distribution. No localization occurred with radiolabelled normal mouse IgG. Images Fig. 1 Fig. 2 Fig. 3 Fig. 4