Current state of clinical immunotherapy.
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Biomedical subjects
Publications and source records attributed to M F Woodruff.
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Growth of isogeneic transplants of a spontaneous murine adenocarcinoma, which is virtually devoid of tumour-specific transplantation antigens, is inhibited by i.v. injection of C. parvum 3 days after tumour inoculation, or by mixing a small dose of C. parvum with the tumour inoculum. Moreover, the therapeutic effect of cyclophosphamide, followed by i.v. or i.p. injection of C. parvum 5 days later, on established transplants of the same tumour is greater than that of cyclophosphamide alone. These findings are consistent with the hypothesis that in both situations (i.e. before the appearance of a palpable tumour and after reduction of an established tumour transplant with cyclophosphamide) the effect of C. parvum is largely due to activation of macrophages or macrophage precursors. They have the important practical implication that adjuvant therapy with C. parvum may be of value, even with tumours which are devoid of TSTA.
A comparison of the growth of a number of BALB/c tumours in BALB/c, BALB/c.nu/ + (heterozygous nude) and BALB/c.nu (homozygous) nude mice has shown that a majority of BALB/c lymphoid tumours grow at a significantly reduced rate in BALB/c.nu mice. The three non-lymphoid BALB/c tumours (carcinomas and sarcomas) tested and a minority of lymphoid tumours, however, grew as well, or better, in the BALB/c.nu mice. Furthermore a correlation, without exception to date, has emerged from a comparison between the susceptibility of a tumour cell line to in vitro lysis by spleen cells from syngeneic nude mice and its growth in vivo in syngeneic nude mice. Only those lymphoid tumour cell lines lysed in vitro show the reduced growth rate in syngeneic nude as compared to syngeneic normal mice. Tumour cell lines, both lymphoid and non-lymphoid, which are resistant to in vitro lysis do not show the reduced in vivo growth rate in syngeneic nude mice. Both the in vivo and in vitro effect can be partially abolished by irradiating the nude mice prior to use, suggesting that a radiation-sensitive non-T-cell surveillance of lymphoid tumours is operating in nude mice. While the identity of the antigens is still unknown, the specificity of the phenomena suggest that C-type RNA oncogenic viral determined antigens are involved.
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The effect of systemic or local injection of Corynebacterium parvum at the tumor site on the growth of various murine tumors was studied in intact and congenitally athymic BALB/c mice. Systemic injection of C. parvum usually had a marked antitumor effect in both types of mouse. Two lymphomas, which regressed spontaneously in untreated intact mice but not in athymic mice, grew progressively in intact mice given systemic C. parvum, though their growth was inhibited in similarly treated athymic mice. Local injection into the site of the tumor markedly inhibited tumor growth in intact mice but was without effect in athymic mice. C. parvum was believed to exert its antitumor effects by two different mechanisms, only one of which was T-cell dependent. The mechanism not dependent on T-cells was particularly activated by systemic C. parvum injection.
The radiosensitivity of three forms of response to injection of C. parvum in mice has been investigated. The increase in phagocytin index evoked by IP or IV injection of 0.7 mg C. parvum (but not that evoked by 1.4 mg) was reduced but not abolished in mice given 350-500 rad whole-body irradiation 4 days before C. parvum injection. Irradiation (500-1,000 rad) 4 days after C. parvum injection had no such effect. The antitumour cytotoxicity in vitro of PE and spleen cells from C. parvum-treated mice was abolished by irradiation of the cell donor (400-800 rad) 4 days before C. parvum injection, but was not reduced by irradiation (800 rad) of the cell donor 4 days after C. parvum injection or of the effector cells in vitro. The antitumour response to systemic (IP) injection of C. parvum was reduced by 350-500 rad whole-body irradiation, irrespective of whether this was given 4 days before or 4 days after C. parvum injection. The response to intratumour injection of C. parvum was even more radiosensitive. It has been suggested in previous papers from this laboratory that the antitumour effect of C. parvum is related to its capacity to stimulate macrophage activity, although in addition T lymphocytes are necessary for local injection to be effective and non-T lymphocytes may be concerned in the response to both local and systemic injection. The present results in no way conflict with this view. They suggest in addition that the effect of C. parvum on the macrophage system is, to a considerable extent, due to stimulation of macrophage precursors to differentiate into actively phagocytic and cytotoxic mature cells.
The indications for transplantation, complications, management and results in 127 consecutive patients who had received renal transplants between October 1960 and December 1974 are presented. In 2 cases the donor was an identical twin, in 22 a living relative other than a twin, in 2 an unrelated individual in whom nephrectomy was performed for therapeutic reasons and in 101 a cadaver. At the end of the period under review there were 37 patients alive with grafts which had functioned for 6 months or more; of these, 28 had survived for more than 2 years, 13 for more than 5 years, 6 for more than 8 years and 2 for more than 12 years. Thirty-four patients were employed or doing routine housework or receiving full-time education. One patient fathered a child 4 years after transplantation; another gave birth to a healthy infant 3 years after transplantation.
S.c. injection of tumour cells or small pieces of tumour irradiated to a dose of 22,000 rad evoked resistance to live challenge with the same tumour (a CBA strain fibrosarcoma induced with methylcholanthrene) 14 days later. This resistance was, however, over-ridden if the challenging inoculum was sufficiently large, and did not develop if the cells were irradiated to 100,000 rad. The resistance evoked by injection of 10(6) irradiated tumour cells was impaired by i.p. injection of 1-4 mg C. parvum 5 days before, and virtually abolished by a similar injection 11 days after, the irradiated cells. The effect of s.c. injection of a mixture of 10(6) irradiated cells and C. parvum 14 days before live challenge depended on the dose of C. parvum. With 0-7 mg the development of resistance was largely but not completely abrogated; 0-35 mg resulted in a lesser degree of abrogation, and 0-09 mg or 0-02 mg had little or no effect.
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.
Detailed serological studies have been undertaken in a small group of cancer patients receiving nonspecific immunotherapy with Corynebacterium parvum (C. parvum). These patients included 4 cases of recurrent malignant melanoma, 2 of stomach cancer and 2 of recurrent breast cancer. They all received an initial i.v. infusion of 20 mg of a formol killed suspension of C. parvum followed by 2 mg (i.m.) at weekly intervals for 10-11 weeks. This protocol consistently resulted in an increase in the circulating IgG levels of all patients but had a variable effect on their IgA, IgM and IgE levels. Increases in the concentration of all 4 IgG subclasses contributed to the overall increase in IgG levels and these changes ranked IgG2 greater than IgG1 greater than IgG3 = IgG4. It also had an inconsistent effect upon the levels of alpha-macroglobulin in pregnancy but the levels of normal serum alpha2-macroglobulin were virtually unchanged. Pre-existing antibodies to C. parvum were noted in all the patients. Titres rose appreciably following C. parvum administration and remained at high, though fluctuating levels, throughout the 100-day period of observation. Absorption studies suggested that the development of antibodies to C. parvum accounted in part for the increased IgG levels noted following this form of therapy. The significance of these changes in relation to the possible anti-tumour effect of C. parvum is discussed.
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Various strains of anaerobic coryneforms and the closely related Propionibacteria have been compared in vivo with respect to their anti-tumour activity. Their effectiveness has been correlated with their serological relationship and to some extent with their ability to stimulate the lymphoreticular system. Organisms belonging to Corynebacterium acnes groups I and II and C. avidum group IV were active anti-tumour agents, although of varying effectiveness. These strains are serologically closely related and all produce a soluble cross-reacting antigen. The single C. granulosum group III strain which we tested, an unclassified coryneform, and the classical Propionibacteria did not cross-react with the main group and had little or no anti-tumour activity. At the high dose (0.7 mg) we used, all strains, whether they inhibited tumour development or not, enhanced clearance of colloidal carbon and stimulated production of an inflammatory peritoneal exudate; at lower dosage the results were too variable to permit valid comparison. At the higher dose anti-tumour activity of a strain appeared to correlate best with ability to produce splenomegally and decrease red cell volume in the blood.
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Cells from the spleen, lymph node, peripheral blood and peritoneal exudate of mice treated with C. parvum were tested for their ability to inhibit tumour growth in vitro. The peritoneal exudate cells from C. parvum treated mice were extremely effective in inhibiting tumour growth whereas the spleen and peripheral blood cells were only moderately so. In contrast, the lymph node cells caused only a modest inhibition of tumour growth at a very high effector to target cell ratio. Spleen cells from normal mice also exerted a moderate anti-tumour effect.
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The inhibitory effect of an i.v. or i.p. injection of C. parvum on intrastrain transplants of a mammary carcinoma in A/HeJ mice has been confirmed, and it has been shown further that C. parvum inhibits the growth of transplants of sarcomata induced with methylcholanthrene both in this strain (members of which lack the fifth component of complement) and in CBA mice (which are not complement deficient). In experiments with the mammary carcinoma, 2 injections of C. parvum on days + 3 and + 9 were more effective than a single injection on day + 3; injections on days + 3 and + 6, or + 3 and + 12, appeared to be marginally less effective than on days + 3 and + 9, but the difference was not statistically significant.Development of the CBA sarcoma was inhibited to about the same extent if, instead of treating the mouse with C. parvum, the tumour cells were pre-incubated with anti-tumour globulin (ATG) in the absence of complement prior to inoculation, and the effect of combining these procedures was much greater than that of either alone. Pre-incubation with ATG had a similar but less marked effect on the development of the mammary carcinoma but had no effect on the A/HeJ sarcoma. Injection (i.v.) of ATG did not inhibit the growth of any of the tumours in these experiments and possible reasons for this are discussed.