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Isolation of Human Lymphatic Endothelial Cells by Multi-parameter Fluorescence-activated Cell Sorting
Published on: May 1, 2015
Distinct recirculating and non-recirculating B-lymphocyte pools in the peripheral blood are defined by coordinated
A J Young1, W L Marston, M Dessing
1Basel Institute for Immunology, Basel, Switzerland. young@bii.ch
Insights
Peripheral blood lymphocytes (PBLs) show distinct migration patterns. Over a third of PBLs are excluded from lymphatic recirculation, highlighting limitations in using blood samples to assess the immune system.
Area of Science:
- Immunology
- Cell Biology
- Hematology
Background:
- Lymphocyte recirculation between blood, tissues, and lymph is crucial for immune responses.
- Understanding lymphocyte migration is key to interpreting immune system status.
Purpose of the Study:
- To compare the functional and phenotypic properties of lymphocytes from blood versus lymph.
- To identify distinct lymphocyte populations within peripheral blood and their migratory behavior.
Main Methods:
- Isolation and analysis of lymphocytes from peripheral blood and lymph.
- Functional assays to assess migration competence.
- Phenotypic analysis using cell surface markers like CD21 and L-selectin.
Main Results:
- Lymph-borne lymphocytes readily entered the lymphatic recirculation pathway.
- Over one-third of peripheral blood lymphocytes (PBLs) were excluded from lymphatic recirculation.
- Non-recirculating PBLs, primarily B cells, lacked CD21 and L-selectin, unlike recirculating B cells.
Conclusions:
- Peripheral blood contains distinct lymphocyte pools with differing recirculation capabilities.
- Blood sampling is insufficient for directly assessing the recirculating immune system.
- Interpreting clinical and experimental data on blood lymphocytes requires considering these migration differences.
Abstract:
The continual recirculation of lymphocytes between the blood, tissues, and lymph is essential for the coordination and dissemination of immune responses. We have compared the functional and phenotypic properties of lymphocytes isolated from blood and lymph, the two major migratory populations. Lymph-borne lymphocytes migrated readily into the lymphatic recirculation pathway, but greater than one third of all peripheral blood lymphocytes (PBLs) were excluded from the lymphatic circuit and showed an enhanced migration to the spleen. Phenotypic analysis showed that most non-recirculating PBLs were B cells. The migration competence of B cells correlated with the surface expression of CD21 and L-selectin; recirculating B cells expressed both of these molecules, whereas non-recirculating B cells lacked both antigens. These results establish that blood contains distinct pools of lymphocytes that differ in their recirculation competence. Clearly, blood sampling is not an efficient method to directly measure the status of the recirculating immune system, and implies important constraints and restrictions in the interpretation of experimental or clinical data that include phenotypic and quantitative analyses of blood lymphocytes.
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