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Antigenic differences between subsets of peripheral blood lymphocytes differing in their right angle light scatter in
R Rabinowitz1, E Granot, R Deckelbaum
1Paul Ehrlich Center for the Study of Normal and Leukemic White Blood Cells, Hebrew University-Hadassah Medical School, Jerusalem, Israel.
Insights
Flow cytometry side scatter (SSC) effectively distinguishes lymphocyte subsets in children and adults. High SSC identifies activated T cells and NK cells, while low SSC indicates naive T cells, aiding immune status assessment.
Area of Science:
- Immunology
- Cell Biology
Background:
- Peripheral blood lymphocytes (PBL) comprise diverse subpopulations with distinct functions.
- Differentiating these subpopulations is crucial for understanding immune responses and states of activation.
Purpose of the Study:
- To investigate the utility of light scatter properties in flow cytometry for discriminating lymphocyte subsets.
- To analyze differences in cell surface marker expression between high and low side scatter (SSC) lymphocyte populations in children and adults.
Main Methods:
- Flow cytometry was used to analyze peripheral blood lymphocytes (PBL) from children and adults.
- Lymphocytes were gated based on high or low light side scatter (SSC).
- Expression of cell surface markers including CD4, CD8, CD57, CD29, and CD45RA was quantified.
Main Results:
- CD4+ (helper T) cells were more frequent in low SSC PBL, while CD57+ (NK) cells were enriched in high SSC PBL.
- Activated/memory T cells (CD4+CD29+ and CD8+CD29+) were significantly more abundant in high SSC populations.
- Naive T cells (CD4+CD45RA+) were predominantly found in low SSC lymphocytes in adults.
Conclusions:
- Light side scatter (SSC) is a valuable parameter for improving the discrimination of lymphocyte subpopulations by flow cytometry.
- SSC analysis aids in assessing the activation state of T cells and identifying specific lymphocyte subsets.
Abstract:
The expression of various cell surface markers on peripheral blood lymphocytes (PBL) of young children and of adults was determined by flow cytometry among cells with either high or low light side scatter (SSC). In adults and in children, CD4+ lymphocytes (helper T cells) were more abundant among PBL with low SSC than among PBL with high SSC. The proportion of CD57+ (Leu-7+) cells (NK cells and a subset of CD8+ T cells) was significantly elevated among high SSC PBL, while that of CD8+ PBL (cytotoxic suppressor T cells) was only slightly elevated. CD4+ and CD8+ lymphocytes which coexpressed the CD29 marker, characteristic for activated or memory T cells, were significantly more abundant among lymphocytes with high SSC. In adults, the proportion of CD4+ CD45RA+ lymphocytes, naive T cells, was significantly higher among low SSC cells. The present study indicates that determination of the SSC of lymphocyte subsets by flow cytometry can improve the discrimination among lymphocyte subpopulations and contribute to assessment of their state of activation.