Usefulness of immunocytochemistry for phenotypical analysis of acute leukemia; improved fixation procedure and comparative study with flow cytometry.
We investigated the phenotypes of blast cells of 53 patients with acute leukemia by a modified streptavidin-biotin alkaline phosphatase (SAB-AP) labeling technique, using a panel of monoclonal antibodies [MoAb; anti-CD11b, CD13, CD14, CD33, CD34, CD41, CD3, CD7, CD10, CD19, anti-HLA-DR, and anti-myeloperoxidase (MPO)]. The selection of an optimal fixative solution for each antigen from five options of various combinations of formalin, acetone, methanol, and/or ethanol, successfully conserved cell morphology and improved specific reaction compared with the conventional methods which used a single fixative for multiple antigens. We compared the SAB-AP results with those obtained by flow cytometry (FCM) for surface markers in each case. High concordance rates for both positive and negative results were observed for each marker. However, positive reaction for some markers (anti-CD13, CD14, CD33, and CD34) were often noted only in the cytoplasm by the SAB-AP method, indicating that combination of these two methods is essential for the precise immunophenotyping of poorly differentiated leukemia cells.
Nagai
K
K
Department of Hematology, Nagasaki University School of Medicine, Japan.
Sohda
H
H
Kuriyama
K
K
Kamihira
S
S
Tomonaga
M
M
D000208:Acute Disease; D000469:Alkaline Phosphatase / Q000737:chemistry / Q000276:immunology; D000911:Antibodies, Monoclonal; D014408:Biomarkers, Tumor / Q000032:analysis; D005434:Flow Cytometry; D006801:Humans; D007150:Immunohistochemistry / Q000379:methods*; D016130:Immunophenotyping; D007938:Leukemia / Q000175:diagnosis*
D000911:Antibodies, Monoclonal; D014408:Biomarkers, Tumor; D000469:Alkaline Phosphatase