Effects of Injectable Solutions on the Quality of Monocyte-Derived Dendritic Cells for Immunotherapy

Therapeutic vaccines based on monocyte-derived dendritic cells have been shown to be promising strategies and may act as complementary treatments for viral infections, cancers, and, more recently, autoimmune diseases. Alpha-type-1-polarized dendritic cells (aDC1s) have been shown to induce type-1 im...

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Main Authors: Laís Teodoro Da Silva, Bruna Tiaki Tiyo, Silvia de Jesus Mota, Marina Mazzilli Ortega, Gabriela Justamante Handel Schmitz, Noemi Nosomi Taniwaki, Gislene Mitsue Namiyama Nishina, Alberto José da Silva Duarte, Telma Miyuki Oshiro
Format: Article
Language:English
Published: Wiley 2024-01-01
Series:Journal of Immunology Research
Online Access:http://dx.doi.org/10.1155/2024/6817965
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author Laís Teodoro Da Silva
Bruna Tiaki Tiyo
Silvia de Jesus Mota
Marina Mazzilli Ortega
Gabriela Justamante Handel Schmitz
Noemi Nosomi Taniwaki
Gislene Mitsue Namiyama Nishina
Alberto José da Silva Duarte
Telma Miyuki Oshiro
author_facet Laís Teodoro Da Silva
Bruna Tiaki Tiyo
Silvia de Jesus Mota
Marina Mazzilli Ortega
Gabriela Justamante Handel Schmitz
Noemi Nosomi Taniwaki
Gislene Mitsue Namiyama Nishina
Alberto José da Silva Duarte
Telma Miyuki Oshiro
author_sort Laís Teodoro Da Silva
collection DOAJ
description Therapeutic vaccines based on monocyte-derived dendritic cells have been shown to be promising strategies and may act as complementary treatments for viral infections, cancers, and, more recently, autoimmune diseases. Alpha-type-1-polarized dendritic cells (aDC1s) have been shown to induce type-1 immunity with a high capacity to produce interleukin-12p70 (IL-12p70). In the clinical use of cell-based therapeutics, injectable solutions can affect the morphology, immunophenotypic profile, and viability of cells before delivery and their survival after injection. In this sense, preparing a cell suspension that maintains the quality of aDC1s is essential to ensure effective immunotherapy. In the present study, monocytes were differentiated into aDC1s in the presence of IL-4 and GM-CSF. On day 5, the cells were matured by the addition of a cytokine cocktail consisting of IFN-α, IFN-γ, IL-1β, TNF-α, and Poly I:C. After 48 hr, mature aDC1s were harvested and suspended in two different solutions: normal saline and Ringer’s lactate. The maintenance of cells in suspension was evaluated after 4, 6, and 8 hr of storage. Cell viability, immunophenotyping, and apoptosis analyses were performed by flow cytometry. Cellular morphology was observed by electron microscopy, and the production of IL-12p70 by aDC1s was evaluated by ELISA. Compared with normal saline, Ringer’s lactate solution was more effective at maintaining DC viability for up to 8 hr of incubation at 4 or 22°C.
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spelling doaj-art-976efc83127a495e9f5ea424ac482b9f2025-08-20T02:09:51ZengWileyJournal of Immunology Research2314-71562024-01-01202410.1155/2024/6817965Effects of Injectable Solutions on the Quality of Monocyte-Derived Dendritic Cells for ImmunotherapyLaís Teodoro Da Silva0Bruna Tiaki Tiyo1Silvia de Jesus Mota2Marina Mazzilli Ortega3Gabriela Justamante Handel Schmitz4Noemi Nosomi Taniwaki5Gislene Mitsue Namiyama Nishina6Alberto José da Silva Duarte7Telma Miyuki Oshiro8Laboratory of Medical Investigation in Dermatology and Immunodeficiencies (LIM-56)Laboratory of Medical Investigation in Dermatology and Immunodeficiencies (LIM-56)Laboratory of Medical Investigation in Dermatology and Immunodeficiencies (LIM-56)Laboratory of Medical Investigation in Dermatology and Immunodeficiencies (LIM-56)Laboratory of Medical Investigation in Dermatology and Immunodeficiencies (LIM-56)Electron Microscopy NucleusElectron Microscopy NucleusLaboratory of Medical Investigation in Dermatology and Immunodeficiencies (LIM-56)Laboratory of Medical Investigation in Dermatology and Immunodeficiencies (LIM-56)Therapeutic vaccines based on monocyte-derived dendritic cells have been shown to be promising strategies and may act as complementary treatments for viral infections, cancers, and, more recently, autoimmune diseases. Alpha-type-1-polarized dendritic cells (aDC1s) have been shown to induce type-1 immunity with a high capacity to produce interleukin-12p70 (IL-12p70). In the clinical use of cell-based therapeutics, injectable solutions can affect the morphology, immunophenotypic profile, and viability of cells before delivery and their survival after injection. In this sense, preparing a cell suspension that maintains the quality of aDC1s is essential to ensure effective immunotherapy. In the present study, monocytes were differentiated into aDC1s in the presence of IL-4 and GM-CSF. On day 5, the cells were matured by the addition of a cytokine cocktail consisting of IFN-α, IFN-γ, IL-1β, TNF-α, and Poly I:C. After 48 hr, mature aDC1s were harvested and suspended in two different solutions: normal saline and Ringer’s lactate. The maintenance of cells in suspension was evaluated after 4, 6, and 8 hr of storage. Cell viability, immunophenotyping, and apoptosis analyses were performed by flow cytometry. Cellular morphology was observed by electron microscopy, and the production of IL-12p70 by aDC1s was evaluated by ELISA. Compared with normal saline, Ringer’s lactate solution was more effective at maintaining DC viability for up to 8 hr of incubation at 4 or 22°C.http://dx.doi.org/10.1155/2024/6817965
spellingShingle Laís Teodoro Da Silva
Bruna Tiaki Tiyo
Silvia de Jesus Mota
Marina Mazzilli Ortega
Gabriela Justamante Handel Schmitz
Noemi Nosomi Taniwaki
Gislene Mitsue Namiyama Nishina
Alberto José da Silva Duarte
Telma Miyuki Oshiro
Effects of Injectable Solutions on the Quality of Monocyte-Derived Dendritic Cells for Immunotherapy
Journal of Immunology Research
title Effects of Injectable Solutions on the Quality of Monocyte-Derived Dendritic Cells for Immunotherapy
title_full Effects of Injectable Solutions on the Quality of Monocyte-Derived Dendritic Cells for Immunotherapy
title_fullStr Effects of Injectable Solutions on the Quality of Monocyte-Derived Dendritic Cells for Immunotherapy
title_full_unstemmed Effects of Injectable Solutions on the Quality of Monocyte-Derived Dendritic Cells for Immunotherapy
title_short Effects of Injectable Solutions on the Quality of Monocyte-Derived Dendritic Cells for Immunotherapy
title_sort effects of injectable solutions on the quality of monocyte derived dendritic cells for immunotherapy
url http://dx.doi.org/10.1155/2024/6817965
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