Antigen-specific human T-cell colony formation in a liquid culture system.
We have developed a limiting dilution microculture T-cell cloning technique to generate antigen-specific T-cell clones. Cultures contained peripheral blood T-cells, irradiated autologous mononuclear cells (feeders), human AB serum and recombinant IL-2. Colonies were enumerated at 14 days. Under these conditions the cloning efficiency expressed as the frequency of proliferating cells in response to PHA, ranged from 1/1.07 to 1/3.35. In response to tetanus toxoid, the frequency of proliferating cells ranged from 1/415 to 1/5655. Average colony size ranged from 0.7 x 10(5) to 6.2 x 10(5) cells with a mean T-cell recovery of 2.5 x 10(5) cells per microculture. Restimulation of 14-day tetanus toxoid T-cell colonies revealed marked proliferation to PHA and tetanus toxoid but only background responses to PPD. These studies document the development of a rapid, antigen-specific liquid culture T-cell cloning system which is likely to detect T-cell clones that are representative of the original T-cell population. The clones are of sufficient size to be useful in studies of antigen cross-reactivity.
Keystone
E
E
Department of Medicine, Wellesley Hospital, Toronto, Canada.
Poplonski
L
L
Snow
K M
KM
Paige
C
C
Miller
R G
RG
D002478:Cells, Cultured; D002999:Clone Cells; D000939:Epitopes; D006801:Humans; D007376:Interleukin-2 / Q000276:immunology; D008213:Lymphocyte Activation; D013601:T-Lymphocytes / Q000502:physiology*; D013745:Tetanus Toxoid / Q000276:immunology; D014373:Tuberculin / Q000276:immunology
D000939:Epitopes; D007376:Interleukin-2; D013745:Tetanus Toxoid; D014373:Tuberculin