A novel system for simultaneous in vivo tracking and biological assessment of leukemia cells and ex vivo generated leukemia-reactive cytotoxic T cells.
To determine the mechanisms by which adoptive immunotherapy could reduce lethality to acute myelogenous leukemia (AML), a novel technique was developed to track both leukemic blasts and adoptively transferred cytotoxic T cells (CTLs) independently and simultaneously in mice. To follow the fate of ex vivo generated anti-AML-reactive CTLs, splenocytes obtained from enhanced green fluorescent protein transgenic mice were cocultured with AML lysate-pulsed dendritic cells, which subsequently were expanded by exposure to anti-CD3/CD28 monoclonal antibody-coated magnetic microspheres. To track AML cells, stable transfectants of C1498 expressing DsRed2, a red fluorescent protein, were generated. Three factors related to CTLs correlated with disease-free survival: (a). CTL L-selectin expression. L-Selectin high fractions resulted in 70% disease-free survival, whereas L-selectin low-expressing CTLs resulted in only 30% disease-free survival. (b). Duration of ex vivo expansion (9 versus 16 days). Short-term expanded CTLs could be found at high frequency in lymphoid organs for longer than 4 weeks after transfer, whereas long-term expanded CTLs were cleared from the system after 2 weeks. Duration of expansion correlated inversely with L-selectin expression. (c). CTL dose. A higher dose (40 versus 5 x 10(6)) resulted in superior disease-free survival. This survival advantage was achieved with short-term expanded CTLs only. The site of treatment failure was mainly the central nervous system where no CTLs could be identified at AML sites.
Sauer
Martin G
MG
University of Minnesota Cancer Center and Department of Pediatrics, Minneapolis, Minnesota 55455, USA.
Ericson
Marna E
ME
Weigel
Brenda J
BJ
Herron
Michael J
MJ
Panoskaltsis-Mortari
Angela
A
Kren
Betsy T
BT
Levine
Bruce L
BL
Serody
Jon S
JS
June
Carl H
CH
Taylor
Patricia A
PA
Blazar
Bruce R
BR
D000818:Animals; D002465:Cell Movement / Q000276:immunology; D006801:Humans; D016219:Immunotherapy, Adoptive / Q000379:methods*; D019041:L-Selectin / Q000096:biosynthesis / Q000276:immunology; D007951:Leukemia, Myeloid / Q000276:immunology* / Q000628:therapy*; D008221:Lymphoid Tissue / Q000276:immunology; D008546:Melanoma, Experimental / Q000276:immunology; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology* / Q000378:metabolism
D019041:L-Selectin