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The role of cellular maturation in neutrophil heterogeneity
P J Krause1, M B Todd, W W Hancock
1Department of Pediatrics, Hartford Hospital, CT 06115.
Abstract:
Previous studies have shown that many neutrophil (PMN) characteristics are heterogeneous but the origin of PMN heterogeneity is unknown. It is unclear if PMN functional heterogeneity is secondary to maturational differences or due to distinct subpopulations of cells that possess different functional capacities. The PMN 31D8 antigen is a useful probe for evaluation of PMN subpopulations. The majority of PMNs (approximately 85%) exhibit a high intensity fluorescence after 31D8 monoclonal antibody (MoAb) labeling (31D8 enriched or "bright" PMNs) as determined by flow cytometric analysis. These cells are more functional than cells with low intensity fluorescence (31D8 diminished or "dull" PMNs). Various immunologic, clonogenic and functional techniques were used to study the expression of the 31D8 antigen in HL-60 cells and myeloid cells in order to evaluate antigenic and functional heterogeneity during morphologic maturation. The results of this study indicate that the percentage of 31D8 antigen positive (31D8 antigen enriched and diminished) bone marrow cells increases from 20 +/- 11% in myeloblast cells to 68 +/- 10% in promyelocytes, 93 +/- 2% in myelocytes and 99 +/- 1% in bands and PMNs. 31D8 antigen enriched cells first appear at the myelocyte stage (32 +/- 10%) and increase in bands (52 +/- 13%), marrow PMNs (62 +/- 13%) and peripheral blood PMNs (88 +/- 4%). These data indicate that the heterogeneous expression of 31D8 antigen in PMNs is due, at least in part, to maturational differences within the PMN population and raise the possibility that other heterogeneously expressed PMN characteristics are also maturationally derived. They also suggest that 31D8 antigenic expression may be a more precise indicator of myeloid functional maturation than maturation as identified by cellular morphology.
Insights
Neutrophil (PMN) heterogeneity arises partly from maturational differences, as shown by the 31D8 antigen. This antigen
Area of Science:
- Immunology
- Cell Biology
- Hematology
Background:
- Neutrophil (PMN) populations exhibit functional heterogeneity, but its origin remains unclear.
- It is unknown if this heterogeneity stems from maturational differences or distinct cell subpopulations.
- The 31D8 antigen serves as a probe to evaluate PMN subpopulations and heterogeneity.
Purpose of the Study:
- To investigate the origin of PMN heterogeneity using the 31D8 antigen.
- To evaluate antigenic and functional heterogeneity during myeloid cell maturation.
- To correlate 31D8 antigen expression with cellular morphology and functional capacity.
Main Methods:
- Flow cytometry to analyze 31D8 monoclonal antibody (MoAb) labeling intensity.
- Immunologic, clonogenic, and functional assays on HL-60 cells and myeloid cells.
- Tracking 31D8 antigen expression across different stages of myeloid maturation.
Main Results:
- The percentage of 31D8 antigen-positive cells increases significantly during myeloid maturation.
- 31D8 antigen-enriched cells emerge at the myelocyte stage and increase with maturation.
- Approximately 85% of peripheral blood PMNs are 31D8-enriched ('bright') and more functional.
Conclusions:
- Heterogeneous 31D8 antigen expression in PMNs is largely due to maturational differences.
- Other heterogeneously expressed PMN characteristics may also be maturationally derived.
- 31D8 antigen expression may be a more precise indicator of myeloid functional maturation than morphology.