A direct method for the calculation of alloreactive CD4+ T cell precursor frequency

Hooman Noorchashm, Yen K. Lieu, Susan Y. Rostami, Howard K. Song, Siri A.S. Greeley, Samaneh Bazel, Clyde F. Barker, Ali Naji

Research output: Contribution to journalArticlepeer-review

27 Scopus citations

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 vive 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.

Original languageEnglish (US)
Pages (from-to)1281-1284
Number of pages4
JournalTransplantation
Volume67
Issue number9
DOIs
StatePublished - May 15 1999
Externally publishedYes

ASJC Scopus subject areas

  • Transplantation

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