A direct method for the calculation of alloreactive CD4+ T cell precursor frequency1
H Noorchashm, YK Lieu, SY Rostami, HK Song… - …, 1999 - journals.lww.com
H Noorchashm, YK Lieu, SY Rostami, HK Song, SAS Greeley, S Bazel, CF Barker, A Naji
Transplantation, 1999•journals.lww.comBackground. Direct measurement of the precursor frequency of alloreactive CD4+ T cells
has been impossible due to the lack of a specific means of determining the absolute number
of daughter cells generated with each division in a repertoire of stimulated T cells. Methods.
Responder lymphocytes were fluorescently labeled and adoptively transferred into
irradiated allogeneic stimulator mice or incubated in vitro with irradiated stimulator
splenocytes. After a 65-to 70-hr stimulation period, responder cells were analyzed by flow …
has been impossible due to the lack of a specific means of determining the absolute number
of daughter cells generated with each division in a repertoire of stimulated T cells. Methods.
Responder lymphocytes were fluorescently labeled and adoptively transferred into
irradiated allogeneic stimulator mice or incubated in vitro with irradiated stimulator
splenocytes. After a 65-to 70-hr stimulation period, responder cells were analyzed by flow …
Abstract
Background.
Direct measurement of the precursor frequency of alloreactive CD4+ T cells has been impossible due to the lack of a specific means of determining the absolute number of daughter cells generated with each division in a repertoire of stimulated T cells.
Methods.
Responder lymphocytes were fluorescently labeled and adoptively transferred into irradiated allogeneic stimulator mice or incubated in vitro with irradiated stimulator splenocytes. After a 65-to 70-hr stimulation period, responder cells were analyzed by flow cytometry.
Results.
The precursor frequency of dividing CD4+ T cells was determined both in vivo and in vitro. The observed number of alloreactive daughter cells generated with each round of division was used to calculate the frequency of alloantigen-specific CD4+ T cells.
Conclusions.
A novel method for the direct calculation of the frequency of alloreactive CD4+ T cells is described. This technique allows the determination of changes in the frequency of alloreactive T cells that might underlie tolerance to alloantigens.
Lippincott Williams & Wilkins