title: High-sensitivity immunocytologic analysis of neuroblastoma cells in paired blood and marrow samples.
abstract: High-sensitivity immunocytochemistry was used to evaluate the relative frequency of neuroblastoma cells in bone marrow and peripheral blood in patients with neuroblastoma (NB). A total of 51 concomitant paired blood and marrow samples (102 total) from 35 patients with NB (age 4 months-31 years; stage 29 stage IV, 4 stage III, 2 stage IVS; 14 at diagnosis, 18 in relapse, 12 during treatment, and 7 off-therapy) were analyzed. Cytospins containing up to 10(6) cells each were prepared using the mononuclear cell (MNC) fraction. For immunocytologic staining, a primary mouse monoclonal anti-GD2 antibody (3F8), a secondary antimouse biotinylated antibody, and a streptavidin-alkaline phosphatase complex were used. A minimum of two cytospins containing a mean of 1.4 x 10(6) total MNCs was analyzed in addition to a negative and a positive control. No circulating tumor cells were detected when the concomitant marrow samples were negative or had <10 positive cells per 106 MNC (23 of 51 samples). Of the 18 marrow samples positive at 10-10,000 cells per 106 MNC, 6 had detectable NB cells in the corresponding blood sample, whereas for marrow samples with >10,000 NB cells per 10(6) MNC (1%), the concomitant blood sample was positive for 9 of the 10. When both marrow and blood samples were positive (15 BM-PB pairs), NB cell frequency was significantly lower in blood, with a mean difference of 2.14 logs (median 2.22, range -0.16-4.8, standard error 0.38). In patients with NB, circulating tumor cell frequencies seem to be substantially lower than in concomitant marrow samples, with a mean difference of >2 logs.
authors.affiliation: Department of Pediatrics, University of Florence, Ospedale Pediatrico A. Meyer, Italy.
authors.forename: L B
authors.initials: LB
authors.lastname: Faulkner
authors.forename: V
authors.initials: V
authors.lastname: Tintori
authors.forename: A
authors.initials: A
authors.lastname: Tamburini
authors.forename: A
authors.initials: A
authors.lastname: Paoli
authors.forename: A
authors.initials: A
authors.lastname: Garaventa
authors.forename: E
authors.initials: E
authors.lastname: Viscardi
authors.forename: F
authors.initials: F
authors.lastname: Tucci
authors.forename: A A
authors.initials: AA
authors.lastname: Lippi
authors.forename: B
authors.initials: B
authors.lastname: De Bernardi
authors.forename: G
authors.initials: G
authors.lastname: Bernini
mesh_terms: D000293:Adolescent; D000328:Adult; D000818:Animals; D001853:Bone Marrow / Q000473:pathology*; D001932:Brain Neoplasms / Q000097:blood / Q000175:diagnosis / Q000473:pathology*; D002648:Child; D002675:Child, Preschool; D006801:Humans; D007150:Immunohistochemistry / Q000379:methods*; D007223:Infant; D051379:Mice; D018365:Neoplasm, Residual; D009447:Neuroblastoma / Q000097:blood / Q000175:diagnosis / Q000473:pathology*; D012008:Recurrence; D012680:Sensitivity and Specificity