A novel system for simultaneous in vivo tracking and biological assessment of leukemia cells and ex vivo generated leukemia-reactive cytotoxic T cells

• Published in: Cancer research (Chicago, Ill.) Ill.), 2004-06, Vol.64 (11), p.3914-3921
• Main Authors: SAUER, Martin G, ERICSON, Marna E, BLAZAR, Bruce R, WEIGEL, Brenda J, HERRON, Michael J, PANOSKALTSIS-MORTARI, Angela, KREN, Betsy T, LEVINE, Bruce L, SERODY, Jon S, JUNE, Carl H, TAYLOR, Patricia A
• Format: Article
• Language: English
• Subjects: Animals; Antineoplastic agents; Biological and medical sciences; Cell Movement - immunology; Humans; Immunotherapy, Adoptive - methods; L-Selectin - biosynthesis; L-Selectin - immunology; Leukemia, Myeloid - immunology; Leukemia, Myeloid - therapy; Lymphoid Tissue - immunology; Medical sciences; Melanoma, Experimental - immunology; Mice; Mice, Inbred C57BL; Mice, Transgenic; Pharmacology. Drug treatments; T-Lymphocytes, Cytotoxic - immunology; T-Lymphocytes, Cytotoxic - metabolism; Tumors
• ISSN: 0008-5472; 1538-7445
• DOI: 10.1158/0008-5472.CAN-03-3991

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.