A simplified and rapid limiting dilution assay of T lymphocytes.
We report here the development of an alternative limiting dilution assay (LDA) of T lymphocytes (T cells). Blood mononuclear cells were first stimulated for 60 hr with PHA and then cultured in microwells in the presence of recombinant interleukin-2 without feeder cells. After 4 days of culture, wells were scored for proliferation. Clonal expansion of T cells followed the single-hit model of the Poisson distribution. The progenitor frequency (f) in the mononuclear cells and E-rosette-positive cells from normal donors were 0.082 +/- 0.025 (n = 12) and 0.236 +/- 0.029 (n = 5), respectively, but this was markedly decreased in patients who underwent marrow-ablative chemotherapy and autografts with blood hematopoietic stem cells. This LDA system should be of value in routine use for the evaluation of T cell proliferative activities.
Takaue
Y
Y
Department of Pediatrics, University of Tokushima, Japan.
Watanabe
T
T
Kawano
Y
Y
Saitoh
S
S
Hirao
A
A
Matsunaga
K
K
Abe
T
T
Kuroda
Y
Y
Ninomiya
T
T
Himeno
K
K
D016026:Bone Marrow Transplantation; D006412:Hematopoietic Stem Cells; D006801:Humans; D007158:Immunologic Techniques; D007958:Leukocyte Count; D008213:Lymphocyte Activation; D013601:T-Lymphocytes / Q000276:immunology*