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POSITIVE SELECTION OF Tac‐ (CD25) POSITIVE CELLS FOLLOWING T‐CELL ACTIVATION: USE OF IMMUNOMAGNETIC SEPARATION AND IMPLICATIONS FOR T‐CELL CLONING
Author(s) -
Lundin K. E. A.,
Ovigstad E.,
Sollid L. M.,
Gjertsen H. A.,
Gaudernack G.,
Thorsby E.
Publication year - 1989
Publication title -
international journal of immunogenetics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.41
H-Index - 47
eISSN - 1744-313X
pISSN - 1744-3121
DOI - 10.1111/j.1744-313x.1989.tb00461.x
Subject(s) - immunomagnetic separation , cloning (programming) , il 2 receptor , population , antigen , microbiology and biotechnology , biology , antibody , clone (java method) , chemistry , t cell , immunology , biochemistry , immune system , gene , medicine , computer science , programming language , environmental health
SUMMARY 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 monclonal 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.