Positive selection of Tac- (CD25) positive cells following T-cell activation. Use of immunomagnetic separation and implications for T-cell cloning.
We have investigated if positive selection for cells expressing activation antigens, which appear on the cell surface during T-lymphocyte activation, could be used for cloning purposes. For this purpose, we used paramagnetic, monodisperse Dynabeads coated with anti-Tac monoclonal antibody, which recognizes CD25 (interleukin-2 receptor light chain). After the first 6-12 h of a primary response, depletion of Tac+ cells could largely abrogate the specific response. This indicated that the specifically responding cells were found among the Tac+ population. T-cell cloning was thus performed on Tac+ blasts positively selected after 18 h of a primary response, at day 6 of a primary response or during secondary stimulation, and gave a high percentage of specific clones. This method is thus a good alternative to established techniques.
Lundin
K E
KE
Institute of Transplantation Immunology, National Hospital, Oslo, Norway.
Ovigstad
E
E
Sollid
L M
LM
Gjertsen
H A
HA
Gaudernack
G
G
Thorsby
E
E
D002469:Cell Separation / Q000379:methods; D002999:Clone Cells / Q000276:immunology; D006801:Humans; D007158:Immunologic Techniques; D008213:Lymphocyte Activation; D008212:Lymphocyte Depletion; D008280:Magnetics; D015375:Receptors, Interleukin-2; D013601:T-Lymphocytes / Q000276:immunology*
D015375:Receptors, Interleukin-2