title: Double immunoenzymatic APAAP staining for the detection of leukemia-associated immunophenotypes.
abstract: Detection of unusual or aberrant cell immunophenotype with flow cytometry is the basis for the immunologic recognition of minimal residual disease (MRD) in patients with acute leukemia (AL). In this study, we have shown that the double immunocytochemical alkaline phosphatase antialkaline phosphatase (APAAP) staining technique also makes possible the detection of leukemic cells with unusual (leukemic) combinations of antigens (ULCA) both at diagnosis and during follow-up of patients with ULCA+ AL. The applicability of double APAAP was analyzed on bone marrow (BM) samples obtained from 12 patients (8 with AML, 3 with ALL, and 1 with undifferentiated acute leukemia [AUL]) randomly chosen from a larger group of 22 ULCA+ patients treated at our center in a 3-year period (22% observed ULCA+ AL frequency). The percentages of ULCA+ BM cells before chemotherapy were in the range of 5%-60%, which dropped to 0%-7% in 10 patients who achieved remission (range 0%-7%, p < 0.01). However, these cells could also be found 60 days after the initiation of therapy, ranging from 0%-2% of all nucleated cells. In 2 of 10 patients who achieved remission, 2% ULCA+ BM cells were found on days 35 and 60 after initiation of chemotherapy, and this finding was followed by relapse on days 110 and 270. However, the other 8 patients remained in remission despite positive finding of ULCA+ BM cells ranging from 0.2%-2% on at least one occasion. In 2 patients with AML FAB-M3 and cytomorphologic remission, the finding of ULCA+ cells by double APAAP correlated with the molecular finding of PML/RARalpha junction. These results indicate that double APAAP staining can identify leukemic cells in samples with a cytomorphologic pattern consistent with remission, but its applicability in detection of MRD awaits additional studies on a larger number of patients with ULCA+ AL.
authors.affiliation: Department of Cytology and Immunology of the Clinical Institute of Laboratory Diagnosis, Zagreb University Clinical Hospital Center, Croatia.
authors.forename: M
authors.initials: M
authors.lastname: Sucić
authors.forename: D
authors.initials: D
authors.lastname: Boban
authors.forename: M
authors.initials: M
authors.lastname: Marković-Glamocak
authors.forename: B
authors.initials: B
authors.lastname: Uzarević
authors.forename: R
authors.initials: R
authors.lastname: Zadro
authors.forename: M
authors.initials: M
authors.lastname: Petrovecki
authors.forename: S
authors.initials: S
authors.lastname: Ries
authors.forename: K
authors.initials: K
authors.lastname: Gjadrov
authors.forename: M
authors.initials: M
authors.lastname: Golemović
authors.forename: A
authors.initials: A
authors.lastname: Stavljenić-Rukavina
authors.forename: B
authors.initials: B
authors.lastname: Labar
authors.forename: D
authors.initials: D
authors.lastname: Batinić
mesh_terms: D000208:Acute Disease; D000293:Adolescent; D000328:Adult; D000469:Alkaline Phosphatase / Q000276:immunology*; D015703:Antigens, CD / Q000032:analysis; D000951:Antigens, Neoplasm / Q000032:analysis; D000971:Antineoplastic Combined Chemotherapy Protocols / Q000627:therapeutic use; D001853:Bone Marrow / Q000473:pathology; D002648:Child; D005260:Female; D006801:Humans; D007124:Immunoenzyme Techniques; D016130:Immunophenotyping / Q000379:methods*; D007938:Leukemia / Q000145:classification / Q000188:drug therapy / Q000276:immunology / Q000473:pathology*; D008297:Male; D008875:Middle Aged; D018365:Neoplasm, Residual; D014411:Neoplastic Stem Cells / Q000276:immunology*; D012074:Remission Induction
chemical_list: D015703:Antigens, CD; D000951:Antigens, Neoplasm; D000469:Alkaline Phosphatase