Pre-clinical characterization of GMP grade CCL21-gene modified dendritic cells for application in a phase I trial in Non-Small Cell Lung Cancer

被引：39

作者：

Baratelli, Felicita ^{[1
]}

Takedatsu, Hiroko ^{[1
]}

Hazra, Saswati ^{[1
]}

Peebles, Katherine ^{[1
]}

Luo, Jie ^{[1
]}

Kurimoto, Pam S. ^{[1
]}

Zeng, Gang ^{[4
]}

Batra, Raj K. ^{[1
,3
]}

Sharma, Sherven ^{[1
,3
]}

Dubinett, Steven M. ^{[1
,2
,3
]}

Lee, Jay M. ^{[1
,5
]}

机构：

[1] Univ Calif Los Angeles, Lung Canc Res Program, Jonsson Comprehens Canc Ctr, Div Pulm & Crit Care Med,Dept Med, Los Angeles, CA 90095 USA

[2] Univ Calif Los Angeles, Geffen Sch Med, Dept Pathol & Lab Med, Los Angeles, CA 90095 USA

[3] Vet Affairs Greater Los Angeles Healthcare Syst, Mol Med Lab, Los Angeles, CA 90073 USA

[4] Univ Calif Los Angeles, Geffen Sch Med, Dept Urol, Los Angeles, CA 90095 USA

[5] Univ Calif Los Angeles, David Geffen Sch Med, Dept Surg, Div Cardiothorac Surg, Los Angeles, CA 90095 USA

来源：

JOURNAL OF TRANSLATIONAL MEDICINE | 2008年 / 6卷 / 1期

基金：

美国国家卫生研究院;

关键词：

D O I：

10.1186/1479-5876-6-38

中图分类号：

R-3 [医学研究方法]; R3 [基础医学];

学科分类号：

1001 [基础医学];

摘要：

Background: Our previous studies have demonstrated that transduction of human dendritic cells (DC) with adenovirus encoding secondary lymphoid chemokine, CCL21, led to secretion of biologically active CCL21 without altering DC phenotype or viability. In addition, intratumoral injections of CCL21-transduced DC into established murine lung tumors resulted in complete regression and protective anti-tumor immunity. These results have provided the rationale to generate a clinical grade adenoviral vector encoding CCL-21 for ex vivo transduction of human DC in order to assess intratumoral administration in late stage human lung cancer. Methods: In the current study, human monocyte-derived DC were differentiated by exposure to GM-CSF and IL-4 from cryopreserved mononuclear cells obtained from healthy volunteers. Transduction with clinical grade adenoviral vector encoding CCL21 (1167 viral particles per cell) resulted in secretion of CCL21 protein. Results: CCL21 protein production from transduced DC was detected in supernatants (24-72 hours, 3.5-6.7 ng/4-5 x 10(6) cells). DC transduced with the clinical grade adenoviral vector were > 88% viable (n = 16), conserved their phenotype and maintained integral biological activities including dextran uptake, production of immunostimulatory cytokines/chemokines and antigen presentation. Furthermore, supernatant from CCL21-DC induced the chemotaxis of T2 cells in vitro. Conclusion: Viable and biologically active clinical grade CCL21 gene-modified DC can be generated from cryopreserved PBMC.

引用

页数：17

共 47 条

[1]

Immature dendritic cells phagocytose apoptotic cells via αvβ5 and CD36, and cross-present antigens to cytotoxic T lymphocytes [J].