Genetic engineering of cytotytic T lymphocytes for adoptive T-cell therapy of neuroblastoma

被引:43
作者
Gonzalez, S [1 ]
Naranjo, A [1 ]
Serrano, LM [1 ]
Chang, WC [1 ]
Wright, CL [1 ]
Jensen, MC [1 ]
机构
[1] City Hope Natl Med Ctr, Div Mol Med, Beckman Res Inst, Dept Pediat Hematol Oncol, Duarte, CA 91010 USA
关键词
adoptive therapy; cytolytic T cells; genetic engineering; neuroblastoma;
D O I
10.1002/jgm.489
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
Background Disease relapse is the leading cause of mortality for children diagnosed with disseminated neuroblastoma. The adoptive transfer of tumor-specific T cells is an attractive approach to target minimal residual disease following conventional therapies. We describe here the genetic engineering of human cytotoxic T lymphocytes (CTL) to express a chimeric immunoreceptor for re-directed HLA-independent recognition of neuroblastoma. Methods The CE711 chimeric immunoreceptor was constructed by PCR splice overlap extension and is composed of a single-chain antibody extracellular domain (scFv) derived from the L1-CAM-specific murine CE7 hybridoma fused to human IgG1 hinge-Fc, the transmembrane portion of human CD4, and the cytoplasmic tail of huCD3-zeta chain (scFvFc:zeta). Primary human T cells were genetically modified by naked DNA electrotransfer of plasmid expression vector CE7R-pMG then analyzed by Western blotting, flow cytometry for CE711 expression and cell surface trafficking, 4-h chromium release assay for re-directed neuroblastoma lysis, and ELISA for tumor-specific activation of cytokine production. Results CE711 is expressed as an intact chimeric protein that trafficks to the cell surface as a type I transmembrane protein. Primary human CE7R-expressing CD8(+) CTL clones specifically recognize human neuroblastoma tumor cells and are activated for tumor cell lysis and T(c)1 cytokine production. Conclusions These data demonstrate the utility of CE711 for re-directing the effector function of CTL to neuroblastoma and have provided the rationale to initiate a FDA-authorized (BB-IND#9149) pilot clinical trial to establish the feasibility and safety of adoptive transfer of autologous CE7R(+)CD8(+) CTL clones to children with recurrent/refractory neuroblastoma. Copyright (C) 2004 John Wiley Sons, Ltd.
引用
收藏
页码:704 / 711
页数:8
相关论文
共 35 条
[1]   Eradication of human hepatic and pulmonary melanoma metastases in SCID mice by antibody-interleukin 2 fusion proteins [J].
Becker, JC ;
Pancook, JD ;
Gillies, SD ;
Mendelsohn, J ;
Reisfeld, RA .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1996, 93 (07) :2702-2707
[2]  
Bolhuis RLH, 1998, ADV EXP MED BIOL, V451, P547
[3]   IL-2 adenovector-transduced autologous tumor cells reduce antitumor immune responses in patients with neuroblastoma [J].
Bowman, L ;
Grossmann, M ;
Rill, D ;
Brown, M ;
Zhong, WY ;
Alexander, B ;
Leimig, T ;
Coustan-Smith, E ;
Campana, D ;
Jenkins, J ;
Woods, D ;
Kitchingman, G ;
Vanin, E ;
Brenner, M .
BLOOD, 1998, 92 (06) :1941-1949
[4]   Prospective evaluation of the International Neuroblastoma Staging System (INSS) and the International Neuroblastoma Response Criteria (INRC) in a multicentre setting [J].
Castel, V ;
García-Miguel, P ;
Cañete, A ;
Melero, C ;
Navajas, A ;
Ruíz-Jiménez, JI ;
Navarro, S ;
Badal, MD .
EUROPEAN JOURNAL OF CANCER, 1999, 35 (04) :606-611
[5]   Anti-GD2 antibody treatment of minimal residual stage 4 neuroblastoma diagnosed at more than 1 year of age [J].
Cheung, NKV ;
Kushner, BH ;
Cheung, IY ;
Kramer, K ;
Canete, A ;
Gerald, W ;
Bonilla, MA ;
Finn, R ;
Yeh, SJ ;
Larson, SM .
JOURNAL OF CLINICAL ONCOLOGY, 1998, 16 (09) :3053-3060
[6]  
COLDMAN AJ, 1980, CANCER, V46, P1896, DOI 10.1002/1097-0142(19801015)46:8<1896::AID-CNCR2820460833>3.0.CO
[7]  
2-Y
[8]  
Darcy PK, 1998, EUR J IMMUNOL, V28, P1663, DOI 10.1002/(SICI)1521-4141(199805)28:05<1663::AID-IMMU1663>3.0.CO
[9]  
2-L
[10]  
DORR U, 1993, EUR J NUCL MED, V20, P431