Effect of local injection of Corynebacterium parvum on the growth of a murine fibrosarcoma

Abstract

Summary.-Local injection (i.e. injection at the site of tumour inoculation) of strains of C. Parvum which have a significant anti-tumour effect when given systemically (i.e. intravenously or, in the case of subcutaneous tumour transplant, intraperitoneally) strongly inhibits the growth of isogeneic transplants of a fibrosarcoma in intact CBA mice but has little or no effect on subcutaneous transplants of the same tumour in T-cell deprived mice. The anti-tumour effect of local injection of C. parvum, unlike that of systemic injection in this particular tumour system, thus appears to be T-cell dependent.PRELIMINARY observations of our own with a mouse fibrosarcoma, and experiments with other mouse tumours recently reported by Likhite and Halpern (1974) and Scott (1974), have shown that intratumour injection of C. parvum may strongly inhibit tumour growth and under some conditions cause complete regression.The present experiments were designed to investigate this phenomenon further, using both subcutaneous and intraperitoneal tumour transplants. To exclude purely mechanical effects we have compared the degree of tumour inhibition resulting from local injection of 3 anaerobic coryneforms which differ markedly in their effect on tumour growth when given systemically, and of an aerobic organism, C. diphtheriae.We have used as tumour hosts both intact and T-cell deprived mice, because in our own experience (Wooodruff and Dunbar, 1972;Woodruff, Dunbar and Ghaffar, 1973), the anti-tumour effect of systemic injections of active strains of C. parvum in respect of cholanthrene induced sarcomata is maintained in T-cell deficient mice, whereas Scott (1974) has reported that the growth of a mastocytoma, is inhibited by intratumour injection of C. parvum in intact mice but not in T-cell deprived mice. MATERIALS AND METHODSMice.-The recipient mice were either intact adult CBA females (20-22 g) or T-cell deprived CBA females prepared as described previously (Woodruff et al., 1973).Tumour. The tumour was originally induced in a female CBA mouse with methylcholanthrene. It was stored in liquid nitrogen after 1]5 transplant generations and was transplanted once more before being used in the experiments. The properties of this tumour have been summarized in a recent review (Woodruff, 1975). It was transplanted subcutaneously (right foreleg) or intraperitoneally, or by both these routes, in the form of a cell suspension prepared with pronase in a dosage (unless otherwise stated) of 104 cells.The mice were inspected and weighed every 2 days. If a subcutaneous tumour was present, its width in 2 directions at rightangles was measured with a caliper and the mean was recorded. Where appropriate, the diameters of subcutaneous tumours were summed, as described previously (Woodruff et al., 1973), up to the day when some of the tumours in control mice were 18-20 mm in diameter, and the group mean sums were compared by Student's t-test. In many cases, however, the differences between groups were sufficiently clearcut to make this unnecessary.

Abrogation of the anti-metastatic activity of C. parvum by antilymphocyte serum

“…In such mice the antimetastatic effects of C. parvum were unchanged. These findings may be comparable with the results obtained by Woodruff and Dunbar (1975) and Scott (1974) on solid tumours in T-cell-deficient mice. However, the depletion of T cells in our experiments was less than in their thymectomized, lethally irradiated and reconstituted mice.…”

“…Furthermore, macrophages from either normal, thymectomized or nude mice, if they have been treated with C. parvum, are cytotoxic for tumour cells in vitro. In contrast, the antitumour activity of C. parvum injected directly into the primary tumour does appear to be dependent on T cells, as this effect is abrogated in T-cell-deprived mice (Scott, 1974;Woodruff and Dunbar, 1975).…”

“…The antimetastatic action of C. parvum was investigated using the Lewis lung carcinoma, a tumour which when grown s.c. metastasizes naturally to the lungs (Simpson-Herren et al, 1974). Reported experiments using primary tumour have indicated that T cell involvement in the anti-tumour response to C. parvum depends upon the route of injection of this vaccine (Woodruff et al, 1973;Scott, 1974;Woodruff and Dunbar, 1975). The importance of T cells in the response of the host against metastases after C. parvum injection was investigated by depleting mice of these cells by thymectomy and irradiation, or by treatment with ALS.…”

Effect of C. parvum and active specific immunotherapy on intracerebral transplants of a murine fibrosarcoma

“…THE GROWTH of subcutaneous isogeneic transplants of a variety of mouse tumours has been shown to be inhibited by both systemic (Woodruff and Boak, 1966;Woodruff and Dunbar, 1973) and local (Likhite and Halpern, 1974;Scott, 1974;Woodruff and Dunbar, 1975) administration of killed vaccines of certain anaerobic corynebacteria and propionibacteria. The present investigation was undertaken to determine whether the growth of intracerebral transplants of one such tumour could be similarly inhibited by systemic or local administration of an active strain of Corynebacterium parvum, either alone or in combination with active specific immunotherapy, partial surgical excision or radiotherapy.…”

“…inhibition when BCG is mixed with the tumour inoculum) is not T-cell dependent (Moore, Lawrence andNisbet, 1976: Pimm andBaldwin, 1976) and probably results from local mobilization and activation of macrophages, and the same is true of contact inhibition of s.c. tumours by C. parvum (Woodruff and Whitehead, unpublished it is just conceivable that this is true also of the response to local injection of C. parvum after tumour inoculation, despite the fact that in the case of s.c. transplants their response has been found to be highly T-cell-dependent (Scott, 1974;Woodruff and Dunbar, 1975;Woodruff and Warner, 1977). The dramatic response of intracerebral transplants to s.c. injection of C. parvum mixed with irradiated tumour cells would seem to imply, however, that such transplants are subject to inhibition as the result of a specific immunological reaction.…”