Single-color Multitarget Flow Cytometry Using Monoclonal Antibodies Labeled with Different Intensities of the Same Fluorochrome
2012; 32(3): 171-176
Korean J Lab Med 2011; 31(3): 148-153
Published online July 1, 2011 https://doi.org/10.3343/kjlm.2011.31.3.148
Copyright © Korean Journal of Laboratory Medicine.
Mijeong Im, M.D.1, Hyojin Chae, M.D.1, Taehoon Kim, M.D.3, Hun-Hee Park, M.D.4, Jihyang Lim, M.D.2, Eun-Jee Oh, M.D.2, Yonggoo Kim, M.D.2, Yeon-Joon Park, M.D.2, and Kyungja Han, M.D.2
Department of Laboratory Medicine1, Graduate School and Departments of Laboratory Medicine2 and Internal Medicine3, The Catholic University of Korea, College of Medicine, Seoul; Department of Clinical Laboratory4, Ansan College, Ansan, Korea
Correspondence to: Kyungja Han, M.D.
Department of Laboratory Medicine, College of Medicine, The Catholic University of Korea, Seoul St. Mary’s Hospital, 505 Banpo-dong, Seocho-gu, Seoul 137-701, Korea
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This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
Background: Since the recent introduction of radioimmunotherapy (RIT) using antibodies against cluster of differentiation (CD) 45 for the treatment of lymphoma, the clinical significance of the CD45 antigen has been increasing steadily. Here, we analyzed CD45 expression on lymphocyte subsets using flow cytometry in order to predict the susceptibility of normal lymphocytes to RIT.
Methods: Peripheral blood specimens were collected from 14 healthy individuals aged 25-54 yr. The mean fluorescence intensity (MFI) of the cell surface antigens was measured using a FACSCanto II system (Becton Dickinson Bioscience, USA). MFI values were converted into antibody binding capacity values using a Quantum Simply Cellular microbead kit (Bangs Laboratories, Inc., USA).
Results: Among the lymphocyte subsets, the expression of CD45 was the highest (725,368±42,763) on natural killer T (NKT) cells, 674,030±48,187 on cytotoxic/suppressor T cells, 588,750±48,090 on natural killer (NK) cells, 580,211±29,168 on helper T (Th) cells, and 499,436±21,737 on B cells. The Th cells and NK cells expressed a similar level of CD45 (P=0.502). Forward scatter was the highest in NKT cells (P<0.05), whereas side scatter differed significantly between each of the lymphocyte subsets (P<0.05). CD3 expression was highest in the Th and NKT cells.
Conclusions: NKT cells express the highest levels of CD45 antigen. Therefore, this lymphocyte subset would be most profoundly affected by RIT or pretargeted RIT. The monitoring of this lymphocyte subset during and after RIT should prove helpful.
Keywords: Antibody binding capacity, CD45, Lymphocyte subset