Rapid quantification of naive alloreactive T cells by TNF-alpha production and correlation with allograft rejection in mice.
Allograft transplantation requires chronic immunosuppression, but there is no effective strategy to evaluate the long-term maintenance of immunosuppression other than assessment of graft function. The ability to monitor naive alloreactive T cells would provide an alternative guide for drug therapy at early, preclinical stages of graft rejection and for evaluating tolerance-inducing protocols. To detect and quantify naive alloreactive T cells directly ex vivo, we used the unique ability of naive T cells to rapidly produce TNF-alpha but not IFN-gamma. Naive alloreactive T cells were identified by the production of TNF-alpha after a 5-hour in vitro stimulation with alloantigen and were distinguished from effector/memory alloreactive T cells by the inability to produce IFN-gamma. Moreover, naive alloreactive T cells were not detected in mice tolerized against specific alloantigens. The frequency of TNF-alpha-producing cells was predictive for rejection in an in vivo cytotoxicity assay and correlated with skin allograft rejection. Naive alloreactive T cells were also detected in humans, suggesting clinical relevance. We conclude that rapid production of TNF-alpha can be used to quantify naive alloreactive T cells, that it is abrogated after the induction of tolerance, and that it is a potential tool to predict allograft rejection.
Brehm
Michael A
MA
Department of Pathology, University of Massachusetts Medical School, Worcester, MA 01655, USA. michael.brehm@umassmed.edu
Mangada
Julie
J
Markees
Thomas G
TG
Pearson
Todd
T
Daniels
Keith A
KA
Thornley
Thomas B
TB
Welsh
Raymond M
RM
Rossini
Aldo A
AA
Greiner
Dale L
DL
D000818:Animals; D006084:Graft Rejection / Q000276:immunology*; D006801:Humans; D007371:Interferon-gamma / Q000096:biosynthesis / Q000097:blood; D007962:Leukocytes / Q000276:immunology; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D008808:Mice, Inbred CBA; D008822:Mice, Transgenic; D016038:Skin Transplantation / Q000276:immunology; D013601:T-Lymphocytes / Q000166:cytology / Q000276:immunology*; D023001:Transplantation Tolerance; D014409:Tumor Necrosis Factor-alpha / Q000096:biosynthesis* / Q000097:blood
D014409:Tumor Necrosis Factor-alpha; D007371:Interferon-gamma