Pre-clinical characterization of GMP grade CCL21-gene modified dendritic cells for application in a phase I trial in Non-Small Cell Lung Cancer
<p>Abstract</p> <p>Background</p> <p>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 viabi...
| Main Authors: | , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
BMC
2008-07-01
|
| Series: | Journal of Translational Medicine |
| Online Access: | http://www.translational-medicine.com/content/6/1/38 |
| _version_ | 1818515083446190080 |
|---|---|
| author | Sharma Sherven Batra Raj K Zeng Gang Kurimoto Pam S Luo Jie Peebles Katherine Hazra Saswati Takedatsu Hiroko Baratelli Felicita Dubinett Steven M Lee Jay M |
| author_facet | Sharma Sherven Batra Raj K Zeng Gang Kurimoto Pam S Luo Jie Peebles Katherine Hazra Saswati Takedatsu Hiroko Baratelli Felicita Dubinett Steven M Lee Jay M |
| author_sort | Sharma Sherven |
| collection | DOAJ |
| description | <p>Abstract</p> <p>Background</p> <p>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 <it>ex vivo </it>transduction of human DC in order to assess intratumoral administration in late stage human lung cancer.</p> <p>Methods</p> <p>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.</p> <p>Results</p> <p>CCL21 protein production from transduced DC was detected in supernatants (24–72 hours, 3.5–6.7 ng/4–5 × 10<sup>6 </sup>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 <it>in vitro</it>.</p> <p>Conclusion</p> <p>Viable and biologically active clinical grade CCL21 gene-modified DC can be generated from cryopreserved PBMC.</p> |
| first_indexed | 2024-12-11T00:24:23Z |
| format | Article |
| id | doaj.art-b0f8895749a64c3bafd5a8941de1912c |
| institution | Directory Open Access Journal |
| issn | 1479-5876 |
| language | English |
| last_indexed | 2024-12-11T00:24:23Z |
| publishDate | 2008-07-01 |
| publisher | BMC |
| record_format | Article |
| series | Journal of Translational Medicine |
| spelling | doaj.art-b0f8895749a64c3bafd5a8941de1912c2022-12-22T01:27:37ZengBMCJournal of Translational Medicine1479-58762008-07-01613810.1186/1479-5876-6-38Pre-clinical characterization of GMP grade CCL21-gene modified dendritic cells for application in a phase I trial in Non-Small Cell Lung CancerSharma ShervenBatra Raj KZeng GangKurimoto Pam SLuo JiePeebles KatherineHazra SaswatiTakedatsu HirokoBaratelli FelicitaDubinett Steven MLee Jay M<p>Abstract</p> <p>Background</p> <p>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 <it>ex vivo </it>transduction of human DC in order to assess intratumoral administration in late stage human lung cancer.</p> <p>Methods</p> <p>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.</p> <p>Results</p> <p>CCL21 protein production from transduced DC was detected in supernatants (24–72 hours, 3.5–6.7 ng/4–5 × 10<sup>6 </sup>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 <it>in vitro</it>.</p> <p>Conclusion</p> <p>Viable and biologically active clinical grade CCL21 gene-modified DC can be generated from cryopreserved PBMC.</p>http://www.translational-medicine.com/content/6/1/38 |
| spellingShingle | Sharma Sherven Batra Raj K Zeng Gang Kurimoto Pam S Luo Jie Peebles Katherine Hazra Saswati Takedatsu Hiroko Baratelli Felicita Dubinett Steven M Lee Jay M Pre-clinical characterization of GMP grade CCL21-gene modified dendritic cells for application in a phase I trial in Non-Small Cell Lung Cancer Journal of Translational Medicine |
| title | Pre-clinical characterization of GMP grade CCL21-gene modified dendritic cells for application in a phase I trial in Non-Small Cell Lung Cancer |
| title_full | Pre-clinical characterization of GMP grade CCL21-gene modified dendritic cells for application in a phase I trial in Non-Small Cell Lung Cancer |
| title_fullStr | Pre-clinical characterization of GMP grade CCL21-gene modified dendritic cells for application in a phase I trial in Non-Small Cell Lung Cancer |
| title_full_unstemmed | Pre-clinical characterization of GMP grade CCL21-gene modified dendritic cells for application in a phase I trial in Non-Small Cell Lung Cancer |
| title_short | Pre-clinical characterization of GMP grade CCL21-gene modified dendritic cells for application in a phase I trial in Non-Small Cell Lung Cancer |
| title_sort | pre clinical characterization of gmp grade ccl21 gene modified dendritic cells for application in a phase i trial in non small cell lung cancer |
| url | http://www.translational-medicine.com/content/6/1/38 |
| work_keys_str_mv | AT sharmasherven preclinicalcharacterizationofgmpgradeccl21genemodifieddendriticcellsforapplicationinaphaseitrialinnonsmallcelllungcancer AT batrarajk preclinicalcharacterizationofgmpgradeccl21genemodifieddendriticcellsforapplicationinaphaseitrialinnonsmallcelllungcancer AT zenggang preclinicalcharacterizationofgmpgradeccl21genemodifieddendriticcellsforapplicationinaphaseitrialinnonsmallcelllungcancer AT kurimotopams preclinicalcharacterizationofgmpgradeccl21genemodifieddendriticcellsforapplicationinaphaseitrialinnonsmallcelllungcancer AT luojie preclinicalcharacterizationofgmpgradeccl21genemodifieddendriticcellsforapplicationinaphaseitrialinnonsmallcelllungcancer AT peebleskatherine preclinicalcharacterizationofgmpgradeccl21genemodifieddendriticcellsforapplicationinaphaseitrialinnonsmallcelllungcancer AT hazrasaswati preclinicalcharacterizationofgmpgradeccl21genemodifieddendriticcellsforapplicationinaphaseitrialinnonsmallcelllungcancer AT takedatsuhiroko preclinicalcharacterizationofgmpgradeccl21genemodifieddendriticcellsforapplicationinaphaseitrialinnonsmallcelllungcancer AT baratellifelicita preclinicalcharacterizationofgmpgradeccl21genemodifieddendriticcellsforapplicationinaphaseitrialinnonsmallcelllungcancer AT dubinettstevenm preclinicalcharacterizationofgmpgradeccl21genemodifieddendriticcellsforapplicationinaphaseitrialinnonsmallcelllungcancer AT leejaym preclinicalcharacterizationofgmpgradeccl21genemodifieddendriticcellsforapplicationinaphaseitrialinnonsmallcelllungcancer |