Panning T cells on vascular endothelial cell monolayers: a rapid method for enriching naive T cells.
A key functional/phenotypic difference between naive and memory T cells is the ability of memory and activated T cells to home to sites of inflammation by adhering to vascular endothelial cells. To determine if this trait could be used to separate naive T cells from memory T cells, CD4+ T cells were incubated with monolayers of IFN-gamma-primed vascular endothelial cells after which the phenotypic and functional characteristics of the nonadherent population were assayed. The nonadherent population 1) contained a five-fold decrease in the frequency of cells displaying the CD44(high)/CD45RB(low) "memory" phenotype and 2) responded well to allostimulation but displayed a reduced ability to respond to immobilized anti-CD3 antibody and, when isolated from ovalbumin-immunized mice, displayed a reduced recall response to ovalbumin in vitro. These studies demonstrate that rwo brief incubations of T cells with monolayers of IFN-gamma-primed endothelial cells can significantly enrich for naive T cells as determined by both phenotypic and functional analyses.
Hoellman
J R
JR
Department of Microbiology, Quillen College of Medicine at East Tennessee State University, Johnson City, USA.
Suttles
J
J
Stout
R D
RD
D000818:Animals; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology*; D002448:Cell Adhesion; D002469:Cell Separation / Q000379:methods*; D016207:Cytokines; D004730:Endothelium, Vascular; D007156:Immunologic Memory; D066298:In Vitro Techniques; D007371:Interferon-gamma / Q000494:pharmacology; D008213:Lymphocyte Activation; D051379:Mice; D008810:Mice, Inbred C57BL; D013601:T-Lymphocytes / Q000502:physiology*
D016207:Cytokines; D007371:Interferon-gamma