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Calcein: a novel marker for lymphocytes which enter lymph nodes
1Division of Cell Biology, John Curtin School of Medical Research, Australian National University, Canberra.
Cytometry
|January 1, 1992
Summary
Calcein dye uptake by lymphocytes identifies a subpopulation that preferentially homes to lymph nodes. This "dull" fluorescence phenotype correlates with lymph node homing receptor expression, suggesting membrane properties are key for HEV transmigration.
Area of Science:
- Immunology
- Cell Biology
- Biophysics
Background:
- Lymphocyte homing to lymphoid organs is crucial for immune surveillance.
- High endothelial venules (HEV) are specialized blood vessels facilitating lymphocyte entry into lymph nodes.
- Previous studies identified cell surface antigens as markers for lymphocyte-HEV binding.
Purpose of the Study:
- To investigate calcein dye uptake as a novel marker for lymphocyte homing.
- To characterize lymphocyte subpopulations based on calcein fluorescence intensity.
- To correlate calcein fluorescence with in vivo migration patterns and homing receptor expression.
Main Methods:
- Lymphocyte labeling with calcein and other fluorescein-based dyes.
- In vivo lymphocyte migration studies in mice.
- Analysis of lymphocyte fluorescence intensity and homing receptor (MEL-14) expression.
Main Results:
- Calcein differentially labels lymphocytes into "dull" and "bright" populations.
- "Dull" lymphocytes preferentially home to lymph nodes, including the popliteal lymph node (PLN).
- "Bright" lymphocytes are excluded from HEV-dependent entry but enter the spleen.
- A high percentage (76.5%) of "dull" lymphocytes express the PLN homing receptor MEL-14.
Conclusions:
- Calcein uptake serves as a marker for the lymph node-homing lymphocyte subpopulation.
- Differential calcein fluorescence reflects distinct lymphocyte migration behaviors.
- Calcein's utility suggests it may indicate membrane properties essential for HEV passage.
