PGE2 Is Crucial for the Generation of FAST Whole- Tumor-Antigens Loaded Dendritic Cells Suitable for Immunotherapy in Glioblastoma
Dendritic cells (DC) are the most potent antigen-presenting cells, strongly inducers of T cell-mediated immune responses and, as such, broadly used as vaccine adjuvant in experimental clinical settings. DC are widely generated from human monocytes following in vitro protocols which require 5–7 days...
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| Published in | Pharmaceutics Vol. 12; no. 3; p. 215 |
|---|---|
| Main Authors | , , , , , , , , |
| Format | Journal Article |
| Language | English |
| Published |
Basel
MDPI AG
02.03.2020
MDPI |
| Subjects | |
| Online Access | Get full text |
| ISSN | 1999-4923 1999-4923 |
| DOI | 10.3390/pharmaceutics12030215 |
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| Abstract | Dendritic cells (DC) are the most potent antigen-presenting cells, strongly inducers of T cell-mediated immune responses and, as such, broadly used as vaccine adjuvant in experimental clinical settings. DC are widely generated from human monocytes following in vitro protocols which require 5–7 days of differentiation with GM-CSF and IL-4 followed by 2–3 days of activation/maturation. In attempts to shorten the vaccine’s production, Fast-DC protocols have been developed. Here we reported a Fast-DC method in compliance with good manufacturing practices for the production of autologous mature dendritic cells loaded with antigens derived from whole tumor lysate, suitable for the immunotherapy in glioblastoma patients. The feasibility of generating Fast-DC pulsed with whole tumor lysate was assessed using a series of small-scale cultures performed in parallel with clinical grade large scale standard method preparations. Our results demonstrate that this Fast protocol is effective only in the presence of PGE2 in the maturation cocktail to guarantee that Fast-DC cells exhibit a mature phenotype and fulfill all requirements for in vivo use in immunotherapy approaches. Fast-DC generated following this protocol were equally potent to standard DC in inducing Ag-specific T cell proliferation in vitro. Generation of Fast-DC not only reduces labor, cost, and time required for in vitro clinical grade DC development, but can also minimizes inter-preparations variability and the risk of contamination. |
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| AbstractList | Dendritic cells (DC) are the most potent antigen-presenting cells, strongly inducers of T cell-mediated immune responses and, as such, broadly used as vaccine adjuvant in experimental clinical settings. DC are widely generated from human monocytes following in vitro protocols which require 5–7 days of differentiation with GM-CSF and IL-4 followed by 2–3 days of activation/maturation. In attempts to shorten the vaccine’s production, Fast-DC protocols have been developed. Here we reported a Fast-DC method in compliance with good manufacturing practices for the production of autologous mature dendritic cells loaded with antigens derived from whole tumor lysate, suitable for the immunotherapy in glioblastoma patients. The feasibility of generating Fast-DC pulsed with whole tumor lysate was assessed using a series of small-scale cultures performed in parallel with clinical grade large scale standard method preparations. Our results demonstrate that this Fast protocol is effective only in the presence of PGE
2
in the maturation cocktail to guarantee that Fast-DC cells exhibit a mature phenotype and fulfill all requirements for in vivo use in immunotherapy approaches. Fast-DC generated following this protocol were equally potent to standard DC in inducing Ag-specific T cell proliferation in vitro. Generation of Fast-DC not only reduces labor, cost, and time required for in vitro clinical grade DC development, but can also minimizes inter-preparations variability and the risk of contamination. Dendritic cells (DC) are the most potent antigen-presenting cells, strongly inducers of T cell-mediated immune responses and, as such, broadly used as vaccine adjuvant in experimental clinical settings. DC are widely generated from human monocytes following in vitro protocols which require 5-7 days of differentiation with GM-CSF and IL-4 followed by 2-3 days of activation/maturation. In attempts to shorten the vaccine's production, Fast-DC protocols have been developed. Here we reported a Fast-DC method in compliance with good manufacturing practices for the production of autologous mature dendritic cells loaded with antigens derived from whole tumor lysate, suitable for the immunotherapy in glioblastoma patients. The feasibility of generating Fast-DC pulsed with whole tumor lysate was assessed using a series of small-scale cultures performed in parallel with clinical grade large scale standard method preparations. Our results demonstrate that this Fast protocol is effective only in the presence of PGE2 in the maturation cocktail to guarantee that Fast-DC cells exhibit a mature phenotype and fulfill all requirements for in vivo use in immunotherapy approaches. Fast-DC generated following this protocol were equally potent to standard DC in inducing Ag-specific T cell proliferation in vitro. Generation of Fast-DC not only reduces labor, cost, and time required for in vitro clinical grade DC development, but can also minimizes inter-preparations variability and the risk of contamination.Dendritic cells (DC) are the most potent antigen-presenting cells, strongly inducers of T cell-mediated immune responses and, as such, broadly used as vaccine adjuvant in experimental clinical settings. DC are widely generated from human monocytes following in vitro protocols which require 5-7 days of differentiation with GM-CSF and IL-4 followed by 2-3 days of activation/maturation. In attempts to shorten the vaccine's production, Fast-DC protocols have been developed. Here we reported a Fast-DC method in compliance with good manufacturing practices for the production of autologous mature dendritic cells loaded with antigens derived from whole tumor lysate, suitable for the immunotherapy in glioblastoma patients. The feasibility of generating Fast-DC pulsed with whole tumor lysate was assessed using a series of small-scale cultures performed in parallel with clinical grade large scale standard method preparations. Our results demonstrate that this Fast protocol is effective only in the presence of PGE2 in the maturation cocktail to guarantee that Fast-DC cells exhibit a mature phenotype and fulfill all requirements for in vivo use in immunotherapy approaches. Fast-DC generated following this protocol were equally potent to standard DC in inducing Ag-specific T cell proliferation in vitro. Generation of Fast-DC not only reduces labor, cost, and time required for in vitro clinical grade DC development, but can also minimizes inter-preparations variability and the risk of contamination. Dendritic cells (DC) are the most potent antigen-presenting cells, strongly inducers of T cell-mediated immune responses and, as such, broadly used as vaccine adjuvant in experimental clinical settings. DC are widely generated from human monocytes following in vitro protocols which require 5–7 days of differentiation with GM-CSF and IL-4 followed by 2–3 days of activation/maturation. In attempts to shorten the vaccine’s production, Fast-DC protocols have been developed. Here we reported a Fast-DC method in compliance with good manufacturing practices for the production of autologous mature dendritic cells loaded with antigens derived from whole tumor lysate, suitable for the immunotherapy in glioblastoma patients. The feasibility of generating Fast-DC pulsed with whole tumor lysate was assessed using a series of small-scale cultures performed in parallel with clinical grade large scale standard method preparations. Our results demonstrate that this Fast protocol is effective only in the presence of PGE2 in the maturation cocktail to guarantee that Fast-DC cells exhibit a mature phenotype and fulfill all requirements for in vivo use in immunotherapy approaches. Fast-DC generated following this protocol were equally potent to standard DC in inducing Ag-specific T cell proliferation in vitro. Generation of Fast-DC not only reduces labor, cost, and time required for in vitro clinical grade DC development, but can also minimizes inter-preparations variability and the risk of contamination. Dendritic cells (DC) are the most potent antigen-presenting cells, strongly inducers of T cell-mediated immune responses and, as such, broadly used as vaccine adjuvant in experimental clinical settings. DC are widely generated from human monocytes following in vitro protocols which require 5−7 days of differentiation with GM-CSF and IL-4 followed by 2−3 days of activation/maturation. In attempts to shorten the vaccine's production, Fast-DC protocols have been developed. Here we reported a Fast-DC method in compliance with good manufacturing practices for the production of autologous mature dendritic cells loaded with antigens derived from whole tumor lysate, suitable for the immunotherapy in glioblastoma patients. The feasibility of generating Fast-DC pulsed with whole tumor lysate was assessed using a series of small-scale cultures performed in parallel with clinical grade large scale standard method preparations. Our results demonstrate that this Fast protocol is effective only in the presence of PGE2 in the maturation cocktail to guarantee that Fast-DC cells exhibit a mature phenotype and fulfill all requirements for in vivo use in immunotherapy approaches. Fast-DC generated following this protocol were equally potent to standard DC in inducing Ag-specific T cell proliferation in vitro. Generation of Fast-DC not only reduces labor, cost, and time required for in vitro clinical grade DC development, but can also minimizes inter-preparations variability and the risk of contamination. |
| Author | Parati, Eugenio Agostino Frigerio, Simona Pellegatta, Serena Gatti, Laura Nava, Sara Finocchiaro, Gaetano Lisini, Daniela Pogliani, Simona Bersano, Anna |
| AuthorAffiliation | 1 Cell Therapy Production Unit—UPTC and Cerebrovascular Unit, Fondazione IRCCS Istituto Neurologico Carlo Besta, 20133 Milan, Italy; daniela.lisini@istituto-besta.it (D.L.); simona.frigerio@istituto-besta.it (S.F.); simona.pogliani@istituto-besta.it (S.P.); laura.gatti@istituto-besta.it (L.G.); anna.bersano@istituto-besta.it (A.B.); eugenio.parati@istituto-besta.it (E.A.P.) 2 Unit of Molecular Neuro-Oncology, Fondazione IRCCS Istituto Neurologico Carlo Besta, 20133 Milan, Italy; serena.pellegatta@istituto-besta.it (S.P.); gaetano.finocchiaro@istituto-besta.it (G.F.) 3 Laboratory of Brain Tumor Immunotherapy, Fondazione IRCCS Istituto Neurologico Carlo Besta, 20133 Milan, Italy |
| AuthorAffiliation_xml | – name: 2 Unit of Molecular Neuro-Oncology, Fondazione IRCCS Istituto Neurologico Carlo Besta, 20133 Milan, Italy; serena.pellegatta@istituto-besta.it (S.P.); gaetano.finocchiaro@istituto-besta.it (G.F.) – name: 3 Laboratory of Brain Tumor Immunotherapy, Fondazione IRCCS Istituto Neurologico Carlo Besta, 20133 Milan, Italy – name: 1 Cell Therapy Production Unit—UPTC and Cerebrovascular Unit, Fondazione IRCCS Istituto Neurologico Carlo Besta, 20133 Milan, Italy; daniela.lisini@istituto-besta.it (D.L.); simona.frigerio@istituto-besta.it (S.F.); simona.pogliani@istituto-besta.it (S.P.); laura.gatti@istituto-besta.it (L.G.); anna.bersano@istituto-besta.it (A.B.); eugenio.parati@istituto-besta.it (E.A.P.) |
| Author_xml | – sequence: 1 givenname: Sara surname: Nava fullname: Nava, Sara – sequence: 2 givenname: Daniela orcidid: 0000-0002-7436-8401 surname: Lisini fullname: Lisini, Daniela – sequence: 3 givenname: Simona orcidid: 0000-0001-7078-075X surname: Frigerio fullname: Frigerio, Simona – sequence: 4 givenname: Simona surname: Pogliani fullname: Pogliani, Simona – sequence: 5 givenname: Serena orcidid: 0000-0001-7804-9486 surname: Pellegatta fullname: Pellegatta, Serena – sequence: 6 givenname: Laura orcidid: 0000-0001-6751-5031 surname: Gatti fullname: Gatti, Laura – sequence: 7 givenname: Gaetano surname: Finocchiaro fullname: Finocchiaro, Gaetano – sequence: 8 givenname: Anna surname: Bersano fullname: Bersano, Anna – sequence: 9 givenname: Eugenio Agostino surname: Parati fullname: Parati, Eugenio Agostino |
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| CitedBy_id | crossref_primary_10_1016_j_canlet_2021_04_018 crossref_primary_10_1039_D3BM00702B crossref_primary_10_1007_s12094_024_03830_9 crossref_primary_10_3390_ijms222212339 |
| Cites_doi | 10.1186/s12967-015-0528-7 10.1007/s12026-017-8931-1 10.1080/2162402X.2017.1412901 10.1182/blood.V97.11.3466 10.1007/s00262-008-0489-2 10.4049/jimmunol.1701024 10.1007/s00011-006-0068-y 10.1007/s10616-011-9375-4 10.1016/j.jcyt.2018.06.006 10.1371/journal.pone.0052301 10.1084/jem.186.9.1603 10.3389/fimmu.2018.03176 10.1182/blood-2009-12-258038 10.1073/pnas.75.10.5132 10.1016/j.coi.2017.01.002 10.1111/j.1365-2567.2003.01764.x 10.3171/2019.5.JNS181762 10.1097/CCO.0000000000000135 10.1007/s00262-011-1023-5 10.1016/S0140-6736(05)60748-9 10.4049/jimmunol.165.6.3492 10.1016/S0369-8114(03)00096-8 10.4049/jimmunol.170.8.4069 10.4049/jimmunol.150.12.5321 10.1038/cr.2016.157 10.1016/j.it.2016.09.006 10.1016/j.immuni.2012.12.001 10.1182/blood-2001-12-0360 10.1016/j.molmed.2004.04.001 10.1084/jem.179.4.1109 10.1016/j.cellimm.2009.09.003 10.1182/blood.V99.8.2897 |
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| SubjectTerms | Antigens Cell culture Cytokines Dendritic cells fast protocol Flow cytometry gbm Immune system Immunotherapy Lymphocytes pge2 |
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| Title | PGE2 Is Crucial for the Generation of FAST Whole- Tumor-Antigens Loaded Dendritic Cells Suitable for Immunotherapy in Glioblastoma |
| URI | https://www.proquest.com/docview/2373332914 https://www.proquest.com/docview/2371853058 https://pubmed.ncbi.nlm.nih.gov/PMC7150800 https://doaj.org/article/673bf26dde334825b20098d77ad28e94 |
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