Human basophils express CD22 without expression of CD19

• Published in: Cytometry (New York, N.Y.) N.Y.), 1999-11, Vol.37 (3), p.178-183
• Main Authors: Han, Kyungja, Kim, Yonggoo, Lee, Jehoon, Lim, Jihyang, Lee, Kyo Young, Kang, Chang Suk, Kim, Won Il, Kim, Byung Kee, Shim, Sang In, Kim, Sun Moo
• Format: Article
• Language: English
• Subjects: Antigens, CD - metabolism; Antigens, CD19 - metabolism; Antigens, Differentiation, B-Lymphocyte - metabolism; B-Lymphocytes - cytology; basophil; Basophils - cytology; Basophils - metabolism; CD19; CD22; Cell Adhesion Molecules; Cell Separation; CML; flow cytometry; Flow Cytometry - methods; Fluorescent Antibody Technique, Indirect; Humans; Immunophenotyping - methods; Lectins; Leukemia, Myelogenous, Chronic, BCR-ABL Positive - blood; Leukemia, Myelogenous, Chronic, BCR-ABL Positive - metabolism; Leukocyte Count - methods; Sialic Acid Binding Ig-like Lectin 2
• ISSN: 0196-4763; 1097-0320
• DOI: 10.1002/(SICI)1097-0320(19991101)37:3<178::AID-CYTO3>3.0.CO;2-Z

Background: Even modern automatic cell counters cannot count basophils precisely. Therefore, we need a rapid, accurate, precise, and easy method for counting basophils. Methods: Using flow cytometry, basophils (CD22+/CD19‐) and B cells (CD22+/CD19+) were counted. Within a large lymphocyte light scatter gate, % basophils (G%baso) and % B cells (G%B) were determined from the total count. Another method of analysis was to make two regions (R1 for basophils and R2 for B cells) and to determine in those the % basophils (R1%baso) and % B cells (R2%B) without gating. The flow cytometric basophil counts of the blood of 21 normal controls and 43 chronic myelogenous leukemia (CML) patients were compared with manual basophil count (Ma%baso) and basophil count by Coulter electronic cell counter (Hialeah, FL) (Auto%baso). CD22+/CD19‐ cells were sorted by a FACSCalibur (Becton Dickinson, San Jose, CA). Results: The G%baso of all samples was 4.66 ± 5.35%, and R1%baso was 4.23 ± 4.88%, and they were well‐correlated (r = 0.996, P < 0.001). The G%B of all samples was 1.55 ± 1.68%, and R2%B was 1.59 ± 1.67%, and they were also well‐correlated (r = 0.993, P < 0.001). Their correlation was better in normal controls than in CML. G%baso was well‐correlated to Ma%baso (r = 0.827) and Auto%baso (r = 0.806), and R1%baso was well‐correlated to Ma%baso (r = 0.831) but showed poor correlation to Auto%baso (r = 0.734). Auto%baso revealed the poorest correlation to Ma%baso (r = 0.692). The sorted CD22+/CD19‐ cells were all basophils (99.48 ± 0.30%), and they revealed CD13, CD33, and dim CD45 expression, whereas CD3, CD14, CD16, and HLA‐DR were not detected on them. Conclusions: We discovered a specific marker combination to identify basophils (CD22+/CD19‐), and we suggest that flow cytometric analysis using these markers is an easy, reliable, and accurate method of basophil counting. Cytometry 37:178–183, 1999. © 1999 Wiley‐Liss, Inc.