PMID: 8938520Oct 1, 1996Paper

A new limiting dilution culture system for the detection of T cell subsets in T cell-depleted marrow grafts

Journal of Hematotherapy
Y KawanishiC A Keever-Taylor

Abstract

T cell depletion (TCD) has been achieved using techniques that cause the inactivation, lysis, or physical removal of T cells from the donor marrow. The clinical results of TCD reflect, in part, the degree of TCD achieved and the subsets that are removed. To better evaluate TCD using the monoclonal antibody (mAb) T10B9, we have performed a series of flow cytometry and mAb blocking studies and have developed a new limiting dilution assay (LDA) that allows the detection of T cell subsets that survive treatment. T cell growth was stimulated with PHA, rIL-2, and irradiated feeder PBMC in a total well volume of 20 microliters. Growth was scored by microscopic examination on days 14-16 of incubation. Immunomagnetic beads coated with mAb were added to the growing wells and incubated, then the plates were fixed to a template of samarium cobalt magnets before washing away nonadherent cells. Wells in which > 50 cells bound > or = 2 beads were scored as positive. Flow cytometry indicated that T10B9 recognized all T cells, but complement-mediated lysis spared a significant proportion of the TCR gamma delta + subset. The epitope recognized by T10B9 on TCR gamma delta + cells appears to be differentially expressed compared with TCR alpha beta...Continue Reading

References

Jul 1, 1992·British Journal of Haematology·P C Doherty
Jan 1, 1990·Journal of Bone and Mineral Research : the Official Journal of the American Society for Bone and Mineral Research·S C GarnerS U Toverud
Jan 1, 1988·Annals of the New York Academy of Sciences·J A Tami, K A Krolick
May 1, 1989·European Journal of Immunology·C TransyE L Reinherz
Sep 1, 1987·Transplantation·R Korngold, J Sprent
Jun 1, 1987·Proceedings of the National Academy of Sciences of the United States of America·M S KrangelM B Brenner
Feb 11, 1988·The New England Journal of Medicine·K J GaskinS F Dorney
Jan 1, 1985·Annals of the New York Academy of Sciences·H C Stary

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