Comparative Quantitative Analysis of Cluster of Differentiation 45 Antigen Expression on Lymphocyte Subsets

Abstract

BackgroundSince 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.MethodsPeripheral 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).ResultsAmong 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.ConclusionsNKT 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.

Phagocytosis of a Model Human Immunodeficiency Virus Target by Human Breast Milk Leukocytes Is Predominantly Granulocyte-Driven When Elicited by Specific Antibody

“…Granulocytes were defined relative to the monocytes as SSC high /CD45 intermediate and found to comprise 17.9-47.8% of CD45 + cells. 26,53 Although these sample characteristics varied immensely (as previously re-ported 21,26 ) and did not correlate with the number of days postpartum at which the samples were obtained, the earliest sample analyzed, acquired 7 days after birth-previously defined as late colostral/early transitional milk-was the most cell dense and most replete with leukocytes. 26…”

“…A side scatter (SSC) versus CD45 plot was used to differentiate the major leukocyte classes (granulocytes, monocytes, and lymphocytes) as extensively described. 26,53 Percent of CD45 + cells and percent of each CD45 + subset were averaged from four wells and the data reported (Table 1) as meanstandard error. ADCP activity (bead-positive cells) was measured in the FITC channel.…”

“…1). 26,53 Monocytes were found to comprise 9.1-65.9% of CD45 + cells. Granulocytes were defined relative to the monocytes as SSC high /CD45 intermediate and found to comprise 17.9-47.8% of CD45 + cells.…”

FACS separation of non-compromised forensically relevant biological mixtures

“…Previous research has, however, shown high level expression of CD45 on leukocytes within a relatively narrow range [29], indicating there is little interpersonal variation of CD45 antigen density in healthy adults. Similar work for the expression of CD227 on healthy adult epithelial cells has not been undertaken, although RNA analysis has shown strong expression in cultured oral epithelial cells [30].…”

Stimulation of Chondrogenic Differentiation of Mesenchymal Stem Cells

“…Finally, MSCs co-cultured with CD45-positive cells, show an enhanced expression of specific genes, the markers of chondrogenic phenotype (41). CD45, also known as protein tyrosine phosphatase receptor type C, a type I transmembrane protein, is expressed in differentiated hematopoietic cells (42). In bone marrow, MSCs and hematopoietic stem cells co-exist and can be discerned with the presence of CD45.…”