Role of bone marrow transplantation in 90Y antibody therapy of colon cancer xenografts in nude mice.

The efficacy of bone marrow transplantation (BMT) for the prevention of 90Y toxicity and extension of survival in nude mice with i.p. LS174T carcinomatosis was evaluated. 90Y-labeled monoclonal antibody (MAB) directed against carcinoembryonic antigen (90Y-anti-CEA MAB) at a dose of 120 microCi caused no deaths due to treatment toxicity and increased the duration of animal survival. No long term cures were obtained in these mice. At doses of 160 microCi or more 90Y-anti-CEA MAB led to hematological deaths. Nude mice were given i.p. injections of 10(6) LS174T tumor cells on day 0. On day 7 the mice received 90Y-anti-CEA MAB i.p. at doses of 120-225 microCi. Syngeneic bone marrow cells (10(7) cells) were then injected i.v. into the mice at 1, 3, 5, 7, 10, or 14 days following 90Y treatment. In the absence of BMT, toxic deaths for animals given 175 microCi 90Y were 11 of 24 (46%) with a median survival of 17 days and 13 of 20 (65%) for animals given 225 microCi 90Y with a median survival of 14 days. Animals receiving the same two doses of 90Y and given BMT 5 days following the 90Y treatment showed 0 of 24 (0%) and 0 of 54 (0%) toxicity deaths, respectively. The optimal time of BMT in relation to 90Y therapy was dependent upon the dose of 90Y-anti-CEA MAB (225 microCi, 3-5 days; 175 microCi, 5-14 days). The mean survival in tumor bearing animals was extended from 31.7 +/- 1.2 (SE) to 45.3 +/- 2.0 days by treatment with 120 microCi of 90Y-anti-CEA MAB. By increasing the dose of 90Y-anti-CEA MAB to 225 microCi and undertaking BMT 5 days later the mean survival was further extended to 63.2 +/- 3.6 days (P less than 0.005). BMT administered at the optimal times can prevent toxic deaths and facilitates higher, more effective doses of tumor specific 90Y-MAB.

In vitro determination of red cell alloantibody significance using an assay of monocyte-macrophage interaction with sensitized erythrocytes.

One hundred and forty-eight red cell alloantibodies, of specificities generally considered to be of clinical significance, were studied in vitro for their ability to induce phagocytosis of sensitized red cells by allogeneic mononuclear phagocytes. Results indicate that only 53% of the alloantibodies studied mediated significant phagocytosis in vitro. The percentages for each blood group system were as follows: Kell, 73%; Jka, 32%; Jkb, 67%; D, 75%; E, 60%; Fya, 62%; Yta, 25%; Ge, 22%; and Vel, 25%. Significant phagocytosis was independent of the strength of the indirect antiglobulin test. The percentage of anti-Jka and anti-Fya mediating significant phagocytosis was increased when fresh complement was added during the sensitization procedure and/or red cells homozygous for the antigen in question were used. The in vivo clinical significance or lack of significance was documented for nine alloantibodies; five caused haemolysis and four did not. Those causing in vivo haemolysis mediated in vitro phagocytosis by monocyte-macrophages whereas the antibodies that did not result in haemolysis showed no increased in vitro phagocytosis. Autologous monocytes were more reliable than random allogeneic monocytes in that phagocytosis was increased over that obtained using allogeneic monocyte-macrophages with two of four alloantibodies having documented clinical significance. The use of target red cells homozygous for the antigen in question, the addition of fresh complement in the antibody sensitization procedure, and use of autologous and allogeneic monocyte-macrophages appear necessary for optimal results. Since 47% of those alloantibodies generally considered to be clinically significant failed to mediate phagocytosis in vitro, the monocyte-macrophage assay should not be considered a predictive assay of a given alloantibody's in vivo significance or lack of significance until more extensive correlation of these assays with in vivo red blood cell survival is obtained.

Reticuloendothelial cell function in alpha-methyldopa-induced hemolytic anemia.

2 patients having alpha-methyldopa-induced hemolytic anemia were followed sequentially using an in vitro assay of autologous monocyte-macrophage activity to determine if their reticuloendothelial system (RES) function was abnormal and thus could be related to the mechanism of lysis. RES function was evaluated while the patients were actively hemolyzing and during remission, following discontinuance of the drug. The results indicated that RES activity is normal in patients having hemolytic anemia due to alpha-methyldopa administration. Also, following cessation of drug therapy, the patients' IgG-coated red cells interacted significantly for a prolonged period (4-5 months) with autologous or normal allogeneic monocyte-macrophages. This was associated with a concurrent reticulocytosis and indicates a persistent low-level hemolytic phase throughout this period, even though hemoglobin and hematocrit values remained within the normal ranges. Although levels of IgG sensitizing the patients' red cells were essentially constant during the hemolytic phase and when the patients were in complete remission, significant monocyte-macrophage activity was only evident during the hemolytic period. In an attempt to explain this phenomenon, it is postulated that hemolysis in patients receiving alpha-methyldopa is related to the interaction of drug with red cell membrane proteins which results in a variably expressed 'altered' antigen which is recognized by 'autoantibody'. The proper expression of the Fc portion of the immunoglobin molecule to result in specific recognition by receptors on monocyte-macrophages depends upon the extent of the antigen alteration by alpha-methyldopa. If the drug does not result in appropriate antigen alteration, then, although 'autoantibody' may still bind to the red blood cell, its Fc region is not readily recognized by monocyte-macrophages and little or no erythrophagocytosis occurs.