Autologous Bone Marrow Transplantation - Blog Science Connections

426 Purged Marrow Engraftment in Neuroblastoma
marrow and autologous transplantation in the treatment of 30 patients with
disseminated neuroblastoma, giving special attention to factors that influence
marrow engraftment. These studies were carried out at five institutions—the
University of Florida College of Medicine in Gainesville; Brooke Army Medical
Center, Fort Sam Houston, TX; Cook Children's Medical Center, Fort Worth, TX;
Montreal Children's Hospital; and the Cleveland Clinic, Cleveland, OH—on a
Pediatric Oncology Group pilot protocol. Purging was performed at the
University of Florida.
PATIENTS AND METHODS
Thirty patients 1-14 years old (median age, 4 years) with disseminated
(stage D) neuroblastoma were enrolled in the study. All had been initially treated
with combinations of cyclophosphamide, Adriamycin (doxorubicin), cisplatin,
and teniposide or etoposide. Cyclophosphamide, vincristine, and etoposide
were most often used for reinduction following relapse. Equal numbers
underwent transplantation in initial remission (group 1) and in subsequent
remission (group 2). All patients received high-dose melphalan (60 mg/m 2 i.v.
daily for 3 days), total body irradiation (200 cGy twice daily for 3 days, with or
without irradiation for local lesions of 150 cGy twice daily for 5 days), followed by
autologous bone marrow infusions.
Bone marrow was harvested when restaging indicated that patients were in
clinical remission (grossly normal aspirate and biopsy specimens of >75%
cellularity). Bone marrow (10-15 ml/kg) was aspirated (with the patient under
general anesthesia) from the posterior iliac crests bilaterally using syringes
primed with 1056 by volume of phenol red-free medium 199 containing
preservative-free heparin at a concentration of 20 U/ml. The bone marrow was
filtered through a 400-jum steel mesh screen followed by filtering through a
100-jum steel mesh screen and transferred to 600-ml blood bags. A minimum
of 1 x 10 8
nucleated cells/kg body weight was collected. All bone marrow was
immunomagnetically purged within 12 hours of collection and before cryopreservation
in liquid nitrogen. Bone marrow from referring centers (for 16
patients) was sealed in blood transfer packs, wrapped loosely in insulating
material, and placed on wet ice packs for transportation by air to the University
of Florida for purging. Purged bone marrow was cryopreserved and returned in
liquid nitrogen.
Six monoclonal antibodies (MAbs)—UJ13A, 223.8,181.4, HI 1, Thy 1, and
127.11—were used. They have been demonstrated to bind selectively to
neuroblastoma cells in bone marrow (1). All were purified on immobilized
Protein A or by fast-performance liquid chromatography and titrated by indirect
immunofluorescence on tissue culture neuroblastoma cell lines.
Paramagnetic microspheres, which are monodisperse polystyrene beads
of 4.5-/um diameter, contain about 2056 magnetite by weight. Before use they
were dispersed in water, sonicated briefly, and sterilized by incubation in 70%