Molecular understanding of oxygen tension and patient-variability effects on ex vivo expanded T cells

被引:17
作者
Haddad, H [1 ]
Windgassen, D [1 ]
Ramsborg, CG [1 ]
Paredes, CJ [1 ]
Papoutsakis, ET [1 ]
机构
[1] Northwestern Univ, Dept Chem & Biol Engn, Evanston, IL 60208 USA
关键词
oxidative stress; ROS; cell therapy; microarray; quantitative RT-PCR; ontological analysis;
D O I
10.1002/bit.20166
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
Immunotherapy with ex vivo cultured T cells depends on a large supply of biologically active cells. Understanding the effects of culture parameters is essential for improving the proliferation and efficacy of the expanded cells. Low oxygen tension (5% pO(2)) was previously reported to improve T-cell expansion and alter cellular phenotypic characteristics compared to T cells cultured at 20% pO(2). Here we report the use of DNA-array based transcriptional analysis coupled with protein-level analysis to provide molecular insights into pO(2) and patient-variability effects on expanded primary human T cells. Analysis of seven blood samples showed that reduced pO(2) results in higher expression of genes important in lymphocyte biology, immune function, and cell-cycle progression. 20% pO(2) resulted in higher expression of genes involved in stress response, cell death, and cellular repair. Expression of granzyme A (gzmA) was found to be significantly regulated by oxygen tension with cells at 5% pO(2) having greater gzmA expression than at 20% pO(2). Protein-level analysis of gzmA was consistent with transcriptional analysis. Granzyme K (gzmK) was coexpressed with gzmA, whereas Granzyme B (gzmB) expression was found to precede the expression of both gzmA and gzmK in 15-day cultures. Temporal gene expression patterns for seven blood samples demonstrate that most genes are expressed by all patient samples in similar temporal patterns. However, several patient-specific gene clusters were identified, and one cluster was found to correlate well with cell proliferation and may potentially be used to predict patient-specific T-cell expansion. (C) 2004 Wiley Periodicals, Inc.
引用
收藏
页码:437 / 450
页数:14
相关论文
共 63 条
[11]  
CRAVEN RJ, 1995, CANCER RES, V55, P3969
[12]   Phase I trial of anti-CD3-stimulated CD4+ T cells infusional interleukin-2, and cyclophosphamide in patients with advanced cancer [J].
Curti, BD ;
Ochoa, AC ;
Powers, GC ;
Kopp, WC ;
Alvord, WG ;
Janik, JE ;
Gause, BL ;
Dunn, B ;
Kopreski, MS ;
Fenton, R ;
Zea, A ;
Dansky-Ullmann, C ;
Strobl, S ;
Harvey, L ;
Nelson, E ;
Sznol, M ;
Longo, DL .
JOURNAL OF CLINICAL ONCOLOGY, 1998, 16 (08) :2752-2760
[13]   Regulation of proliferation, differentiation and survival by the IL-3/IL-5/GM-CSF receptor family [J].
de Groot, RP ;
Coffer, PJ ;
Koenderman, L .
CELLULAR SIGNALLING, 1998, 10 (09) :619-628
[14]   INTERLEUKIN-13 IS A B-CELL STIMULATING FACTOR [J].
DEFRANCE, T ;
CARAYON, P ;
BILLIAN, G ;
GUILLEMOT, JC ;
MINTY, A ;
CAPUT, D ;
FERRARA, P .
JOURNAL OF EXPERIMENTAL MEDICINE, 1994, 179 (01) :135-143
[15]   Distinct roles for STAT1, STAT3, and STAT5 in differentiation gene induction and apoptosis inhibition by interleukin-9 [J].
Demoulin, JB ;
Van Roost, E ;
Stevens, M ;
Groner, B ;
Renauld, JC .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1999, 274 (36) :25855-25861
[16]   Genomic expression programs and the integration of the CD28 costimulatory signal in T cell activation [J].
Diehn, M ;
Alizadeh, AA ;
Rando, OJ ;
Liu, CL ;
Stankunas, K ;
Botstein, D ;
Crabtree, GR ;
Brown, PO .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2002, 99 (18) :11796-11801
[17]   Cluster analysis and display of genome-wide expression patterns [J].
Eisen, MB ;
Spellman, PT ;
Brown, PO ;
Botstein, D .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1998, 95 (25) :14863-14868
[18]   Cascades of transcriptional induction during human lymphocyte activation [J].
Ellisen, LW ;
Palmer, RE ;
Maki, RG ;
Truong, VB ;
Tamayo, P ;
Oliner, JD ;
Haber, DA .
EUROPEAN JOURNAL OF CELL BIOLOGY, 2001, 80 (05) :321-328
[19]   Peripheral lymphocytes and intracellular cytokines in C282Y homozygous hemochromatosis patients [J].
Fabio, G ;
Zarantonello, M ;
Mocellin, C ;
Bonara, P ;
Corengia, C ;
Fargion, S ;
Fiorelli, G .
JOURNAL OF HEPATOLOGY, 2002, 37 (06) :753-761
[20]  
FUJIWARA Y, 1995, ONCOGENE, V10, P891