Use of cell affinity chromatography for separation of lymphocyte subpopulations.

We describe the use of affinity chromatography for separation of cell populations that do not differ significantly with respect to gross physical properties such as size, density, or charge. Cell affinity chromatography exploits differences in cell surface macromolecules by passage of mixtures of cell populations through a column containing beads to which are attached chemical ligands with specific binding affinity for particular cell surface receptors. In this article we focus on the application of this concept to separation of mature T lymphocytes from peripheral blood. This serves as a model for the separation of these cells from bone marrow in order to prevent graft-vs.-host disease in bone marrow transplantation. However, the concept of cell affinity chromatography should find more general widespread utility in a variety of biotechnological applications. Thus, we introduce a simple theoretical framework which is necessary in order to understand the results that might be expected in any given situation. Finally, we use this theory to provide a tentative explanation for experimental observation of the effects of temperature and flowrate on the degree of separation achieved for our current pplication.

Treatment of advanced neuroblastoma with supralethal chemotherapy, radiation, and allogeneic or autologous marrow reconstitution.

Ten children with recurrent metastatic (stage IV) neuroblastoma received local radiation therapy, supralethal chemotherapy, and total-body irradiation. Rescue with infusions of either allogeneic (four patients) or autologous (six patients) bone marrow followed. The drugs given to the first two patients were individualized combinations based on previous tumor responses. Both patients died with recurrent tumor three and nine months posttransplant. The eight remaining patients were treated more uniformly with local irradiation, VM-26, doxorubicin, melphalan (L-phenylalanine mustard), and 1,000-rad total-body irradiation in three fractions. Two of these patients had cardiac dysfunction and received no doxorubicin. Three children died in the immediate posttransplant period with disseminated fungal infections. A fourth relapsed and died nine months posttransplant. As of December 1, 1983, two children who received allogeneic marrow grafts have survived in complete remission for 54 and 36 months, and two children who received autologous marrow grafts have survived in complete remission for 35 and 22 months. These results suggest that relapsed metastatic neuroblastoma can be controlled by supralethal combinations of chemotherapy and irradiation coupled with bone-marrow rescue.

Absorption of oral aminoglycosides following bone marrow transplantation.

Four patients with severe gastrointestinal reactions receiving oral "nonabsorbable" antibiotics for gut sterilization following bone marrow transplantation absorbed clinically significant amounts of aminoglycoside (gentamicin and/or tobramycin). Serum concentrations of 2.2, 2.6, 5.8, and 12.0 micrograms/ml were measured. Two of these patients had acute graft versus host reactions and two had severe mucositis following cytoreduction with intensive chemotherapy and irradiation. Nephrotoxicity occurred in the latter patients. One patient was studied in detail. Her hospital course and investigative results are presented. Four additional patients with mild gastrointestinal reactions following cytoreduction did not absorb gentamicin when their toxicity was maximal. Serum aminoglycoside determinations are necessary in patients receiving oral aminoglycosides for gut sterilization following bone marrow transplantation if moderate to severe gastrointestinal reactions occur.

Pulmonary function in patients undergoing bone marrow transplantation.

Pulmonary function was studied prospectively in 25 children with leukemia and aplastic anemia undergoing bone marrow transplantation (BMT). Whereas 11 patients have died, only one did so primarily due to interstitial pneumonia. Fourteen patients (56%) survived a median of at least 36 months. Seventeen patients received pulmonary function tests (PFTs). Four patients transplanted for leukemia in relapse following preparation with a very intensive regimen (cyclophosphamide, 200 mg/kg, total body irradiation, 1,000 rad, BCNU, cytosine arabinoside) developed restrictive lung changes. Patients undergoing BMT for aplastic anemia and leukemia in remission prepared with more commonly used and less intensive regimens maintained normal pulmonary function. As new regimens are devised, PFTs should be utilized to characterize the pulmonary toxicity of these regimens as well.

Total body irradiation as preparation for bone marrow transplantation in treatment of acute leukemia and aplastic anemia.

In an attempt to improve survival while minimizing toxicity, many bone marrow transplant centers are now studying the use of cytoreduction regimens with an increased amount of radiation in single-dose or fractionated-exposure schedules for patients with leukemia and aplastic anemia. In order to review the current results, the literature prior to September, 1982 was surveyed and data were tabulated for each transplant center regarding the number of patients receiving transplants, diagnoses, cytoreducation regimen, clinical status, revission duration, relapse rate, causes of death and incidence of interstitial pneumonia. The incidence and severity of cataracts, growth failure, hypothyroidism and second malignant neoplasms were noted, and the data obtained from the literature search were updated and expanded by telephone questionnaire when possible. Marked variation in the technique of transplantation was found among the participating institutions, making it difficult to determine the contribution of the various TBI doses, dose rates and fractionation schedules to the efficacy and toxicity of the combined regimen. In order to define the risk-benefit ratio of the various TBI regimens more clearly, prospective controlled, randomized studies will be required.

Acyclovir treatment of herpes zoster infections. Use in children undergoing bone marrow transplantation.

Three patients in whom herpes zoster infections developed following bone marrow transplantation were treated with acyclovir. The patients experienced pain relief within 24 hours of starting treatment. The progression of their skin lesions halted within 1, 2, and 4 days of therapy, respectively, and healed completely within two weeks of therapy. Pharmacokinetic studies indicated that acyclovir plasma concentration-time profiles approximated biexponential equations. The drug half-lives were 3.91, 3.83, and 3.40 hours, respectively. Acyclovir was not myelotoxic and may be helpful in aborting varicella-zoster virus infections in bone marrow transplant recipients.

Leukemic optic neuropathy.

The clinical course and ophthalmic manifestations of an eight year old child with acute undifferentiated leukemia and unilateral blindness secondary to leukemic optic nerve head infiltration are described. At autopsy the involved nerve head and peripapillary retina demonstrated massive leukemic cell infiltration and hemorrhagic necrosis. This manifestation of leukemia is quite uncommon and prognosis for life in such cases is poor with existing methods of therapy.

Radiographic features of gastrointestinal graft-vs.-host disease.

Five patients underwent bone-marrow transplantation, and graft-vs.-host disease (GVHD) developed. Radiographic features are described. Three patients showed an abrupt transition at the jejuno-ileal junction from normal small bowel to ileal loops which were narrowed, tubular, separated, edematous, and, generally, without mucosal markings. The transit time for the contrast material to reach the anus from the mouth was 20 to 25 minutes. A fourth patient showed mucosal ulcerations of the transverse and descending colon. The fifth patient with acute, ultimately fatal GVHD had an edematous duodenum and colon, in addition to an edematous, tubular jejunum and ileum without mucosal markings. Transit time through the intestines for the contrast material was rapid.

Polyinosinic acid--polycytidylic acid: inhibition of DNA synthesis stimulated by isoproterenol.

Polyinosinic acid * polycytidylic acid (poly I * poly C) inhibits isoproterenol-stimulated DNA synthesis in salivary glands of mice. A single intraperitoneal injection of 250 micrograms of poly I * poly C inhibits the stimulation of DNA synthesis when given 10 minutes before isoproterenol or at any time during the 20-hour lag period between the injection of isoproterenol and the subsequent DNA synthesis. The inhibition caused by poly I * poly C is not due to a generalized toxic action but seems to be exerted through a relatively selective mechanism. Polyuridylic acid and diethylaminoethyl dextran are less effective in inhibiting isoproterenol-stimulated DNA synthesis.