G. Mathé

Bone Marrow Graft in Man after Conditioning by Antilymphocytic Serum

Allogeneic bone marrow grafts carried out after previous administration of antilymphocytic serum alone were attempted in 16 patients. Of these, six had acute myeloblastic leukaemia, four acute lymphoblastic leukaemia, and one a blast cell crisis in polycythaemia vera. Ten of these patients were in an overt phase of the disease and resistant to chemotherapy, while nine had complete agranulocytosis. In five of these patients erythrocyte and leucocyte antigenic markers demonstrated the establishment of the graft. One patient had thalassaemia major, and four others had aplasia of the bone marrow, in one case due to chloramphenicol poisoning and in another to virus hepatitis. The grafts were successful in the last two patients and transformed their clinical condition. No signs of early acute secondary disease were noted in any of the patients, either when the donor had been given antilymphocytic serum or when he was untreated. The grafts had no adoptive immunotherapeutic effect on the acute leukaemia. These observations have clearly shown that antilymphocytic serum has an immunosuppressive effect in man when it is used alone.

Adjuvant treatment of node-positive breast cancer with cyclophosphamide, doxorubicin, fluorouracil, and vincristine versus cyclophosphamide, methotrexate, and fluorouracil: final report after a 16-year median follow-up duration.

PURPOSE To determine the long-term impact on disease-free survival (DFS) and overall survival (OS) of adjuvant anthracycline-based chemotherapy, when prospectively compared by random allocation with standard cyclophosphamide, methotrexate, and fluorouracil (CMF) in node-positive (N+) breast cancer patients. PATIENTS AND METHODS Two hundred forty-nine patients with N+ breast cancer, recruited from eight French cancer centers, were randomized to receive 12 monthly cycles of adjuvant chemotherapy, either CMF (n = 112) or doxorubicin, vincristine, cyclophosphamide, and fluorouracil (AVCF) (n = 136). All had a negative metastatic work-up before inclusion, which was stratified by accrual center, tumor stage (International Union Against Cancer [UICC]), and menopausal status. RESULTS No severe adverse effect related to grade 4 (World Health Organization [WHO]) toxicity was observed. There was no difference in second primary tumor incidence between the two arms. The treatment given was 88% of planned for AVCF and 75% for CMF in both premenopausal and menopausal patients. With a median follow-up time of 16 years (range, 13 to 17), the OS and DFS rates are significantly longer in the AVCF arm (56% v 41% [P = .01] for OS, and 53% v 36% [P = .006] for DFS). These differences are significant, irrespective of tumor stage (T1 to T2 v T3 to T4), and remain positive in patients with or without postoperative locoregional radiotherapy (55% of cohort). When analyzed according to menopausal status, the differences remain significant only for premenopausal patients. CONCLUSION This set of mature controlled data confirms the added value of anthracycline-based combination adjuvant therapy for N+ breast cancer patients when compared with CMF, with both regimens given for 1 year.

Two epipodophyllotoxin derivatives, VM 26 and VP 16213, in the treatment of leukemias, hematosarcomas, and lymphomas

The preliminary results obtained with VM 26 and VP 16213 (two semisynthetic derivatives from epipodophyllotoxin) on leukemias and hematosarcomas are described. These compounds are phase M dependent, myelostatic agents whose toxicity can be controlled by intermittent administration. VM 26 proved capable of inducing regression in Hodgkins disease, reticulosarcomas, and lymphosarcomas. Its most remarkable effect was the total regression of neoplastic pleural effusions by systemic administration. VP 16213 seemed to be effective on the same hematosarcomas, but less often. Its most striking action was its oncostatic effect in acute monocytoid leukemia and in the monocytoid component of acute myelomonocytoid leukemia.

In vivo immunostimulation by tuftsin

SummaryTuftsin, a physiological basic tetrapeptide known for its capacity to stimulate the phagocytic activity of macrophages and polymorphonuclear leucocytes, was examined for its immunostimulating properties when injected systemically into mice.Tuftsin was able to potentiate the antibody response to a thymus-dependent antigen (TNP-KLH) when injected 3, 7 or 10 days before the antigen. The response to the same hapten coupled to a thymus-independent carrier (LPS) was stimulated on day 1 and 3 but slightly depressed on day 7.The peptide rendered macrophages highly cytostatic for tumor cells but the activation process required 7 days to develop. In contrast, enhancement of antibody-dependent cell-mediated cytotoxic (ADCC) activity of spleen cells was observed throughout the period of observation (14 days).Tuftsin did not induce nonspecific suppressor cells as the response of normal spleen cells to mitogens in vitro was not depressed when cultivated in the presence of tuftsin-treated spleen cells.These observations suggest that tuftsin acts as an immunostimulant which may be useful in cancer immunotherapy.

NEW ANTIGENS IN HUMAN LEUKÆMIC CELLS, AND ANTIBODY IN THE SERUM OF LEUKÆMIC PATIENTS

Abstract 51 patients with leukaemia or lymphoma, were subjected, before treatment, to a search for serum antibodies against their own leukaemic cells. Four tests were used: cytotoxicity, immunofluorescence on living cells, complement fixation, and immune adherence. The results of these tests were compared with those of electron microscopy for the presence of virus-like particles. In 12 patients, at least one of these immunological tests was positive, whilst virus-like particles were found in only 5. In 4 other patients, particles were found when immunological tests were negative. There was no strict correlation between the results of the various immunological tests. The most sensitive test was immune adherence. These results suggest that there are new antigens on the human leukaemic cells and that possible active antibodies against these antigens are present in the serum of these patients. These findings should certainly encourage further work to be carried out in this field of investigation, which has tended, until now, to be considered as rather negative.

Effect of BCG on hematopoietic stem cells: Experimental and clinical study

SummaryIn (DBA/2×C57Bl/6) F1 mice the i.v. injection of 1 mg of living BCG does not increase the total number of CFU/s per femur, but a marked increase in the percentage of CFU/s in S phase is noted as early as the 8th hr. BCG injected i.v. also increases the absolute number of colony-forming units in agar per femur. The effect of BCG appears quite different from the known effect of bacterial endotoxin, and in particular it does not induce a significant increase in the level of CSF. The administration of BCG 24 hrs after treatment with a single dose of 200 mg/kg of cyclophosphamide significantly reduces the time of hematologic restoration, but the same dose of BCG given after a lethal dose of total body irradiation does not increase survival time in mice. These different effects of BCG seem to be related to the role of BCG in stimulating the multiplication maturation pool of the bone marrow without producing any increase in the reserve pool.

Studies on the mechanisms of action of BCG

SummaryBacillus Calmette-Guérin was given intravenously (i.v.) or subcutaneously (s.c.), 14 days before antigenic or mitogenic stimulation.When given i.v., BCG increased plaque-forming cell production against thymus-dependent antigens (sheep red blood cells and a hapten coupled to hemocyanin) and against a thymus-independent antigen (the same hapten coupled to polymerized flagellin). BCG given s.c. was generally ineffective in potentiating humoral responses.In contrast, a marked depression of spleen cell reactivity in vitro to a T-cell mitogen, concanavalin A (Con A) but an increased response to a B-cell mitogen, dextran sulfate, were observed after BCG i.v., and to a lesser extent after BCG s.c. This inhibitory effect was reversed after removal of cells adherent to nylon or plastic. Normal spleen cell reactivity to Con A was strongly depressed when the cells were cultivated in presence of unfractionated or plastic-adherent BCG-treated spleen cells, suggesting that BCG induces suppressor cells which inhibit T-cell responses in vitro. Similar impairment of T-cell functions in vivo was not observed since BCG given i.v. significantly increased the intensity of delayed-type hypersensitivity to picryl chloride.BCG also induced macrophage activation, measured by macrophage tumoricidal activity in vitro, more strongly and more rapidly when given i.v. than s.c.Modifications of the distribution of lymphocytes from the circulation to the organs were observed. An early increase of lymphocyte migration to the spleen and a long-lasting depression of cell migration to mesenteric lymph nodes and bone marrow were seen after BCG given i.v. When given s.c., BCG induced a progressive increase of cell migration to the draining lymph nodes concomitant with a depression of migration to mesenteric nodes.These results demonstrate the complexity of the mode of action of BCG, but many of its effects on immune responses may result from a primary action on macrophages.

Hydroxyurea, Leucopheresis, and Splenectomy in Chronic Myeloid Leukaemia at the Problastic Phase

Forty-three patients with chronic myeloid leukaemia have been treated with hydroxyurea in order to be subjected to leucopheresis for white cell transfusions. Hydroxyurea decreases leucocytosis when it is administered and the blood granulocyte number increases soon after the drug is stopped. The survival of the patients is not different from the survival of the patients treated with conventional chemotherapy (busulphan, mitobronitol) and it is superior to the survival of patients treated with external radiotherapy or with 32P. Half of the patients were subjected to splenectomy during first remission for a phase II trial. They were not randomized, but the distribution according to age was similar in the two groups. A slight difference appears in favour of splenectomy so far as survival is concerned, but there were three post-operative deaths out of 18 patients. We conclude that a phase II trial on the value of splenectomy is indicated ethically, but that the patients should be operated on and nursed in a microbiologically controlled environment.

Phase-II trial with vindesine for regression induction in patients with leukemias and hematosarcomas.

SummaryVindesine (VDS) has been submitted to a phase-II trial, the results of which were assessed in terms of regression induction. VDS was given weekly IV in doses of 2 mg/m2 on two consecutive days to 59 patients, 55 of whom were evaluable. A high proportion of complete (36%) and over 50% partial regressions were obtained in acute lymphoid leukemias (ALL) (overall response 63%) whatever the perceptible phase, in blastic crisis of chronic myeloid leukemia (55%), and some responses were recorded in lymphosarcoma (40%). No effect has so far been seen in acute myeloid leukemia or in Hodgkins disease. Malignant neoplasms of the immunoblastic type seem to be particularly sensitive to VDS. Continuous 48 h IV infusion can induce a remission where an IV push administration of the same dose has failed. One remarkable characteristic of VDS is the apparent absence of cross-resistance with VCR: in acute leukemic forms, 55% of patients who failed to obtain remission induction after three weekly injections of VCR (used in combination chemotherapy) achieved a complete or partial remission with VDS. The toxicity was mainly neurologic (paralytic ileus, constipation, paresthesias, loss of reflexes) and hematologic (leukopenia and thrombopenia), and was not more significant than with the other agents: four patients died of infection or hemorrhage.

Inhibition of graft-versus-host reaction by preincubation of the graft with a thymic extract (lymphocyte chalone).

Incubation of parental lymphoid cells with the lymphocyte proliferation inhibitor extracted from the thymus (chalone) prevents them from inducing a normal graft-versus-host reaction as measured by the lymph node enlargement, splenomegaly, or mortality in the recipients. This treatment does not impair their capacity to form splenic colonies in irradiated syngeneic recipients.