Langerhans-type and monocyte-derived human dendritic cells have different susceptibilities to mRNA electroporation with distinct effects on maturation and activation: implications for immunogenicity in dendritic cell-based immunotReport as inadecuate




Langerhans-type and monocyte-derived human dendritic cells have different susceptibilities to mRNA electroporation with distinct effects on maturation and activation: implications for immunogenicity in dendritic cell-based immunot - Download this document for free, or read online. Document in PDF available to download.

Journal of Translational Medicine

, 11:166

Immunobiology and immunotherapy

Abstract

BackgroundmRNA electroporation of dendritic cells DCs facilitates processing and presentation of multiple peptides derived from whole antigen, tailored to different HLA molecules. Clinical responses to electroporated moDC vaccines, however, have been suboptimal. Human Langerhans-type DCs LCs are the most potent conventional DC subtype for inducing CD8 cytotoxic T lymphocytes CTLs in vitro. We recently demonstrated that Wilms’ tumor 1 WT1 mRNA-electroporated LCs are superior to moDCs as stimulators of tumor antigen-specific CD8 CTLs, even though they are comparable stimulators of allogeneic T cell proliferative responses. A detailed comparative evaluation of the effects of mRNA electroporation on LCs versus moDCs, however, is needed.

MethodsImmature and partially-matured human moDCs and LCs electroporated with mRNA were compared for transfection efficiency, phenotypic changes, viability, retention of transgene expression after cryopreservation, and immunogenicity. Student t test was used for each pairwise comparison. One-way analysis of variance was used for multiple group comparisons.

ResultsTransfection efficiency after electroporation with enhanced green fluorescent protein eGFP mRNA was higher for immature than for partially-matured moDCs. In contrast, transfection efficiency was higher for partially-matured than for immature LCs, with the additional benefit that electroporation itself increased maturation and activation of CD83HLA-DR LCs but not moDCs. Electroporation did not impair final maturation and activation of either DC subtype, after which both mRNA-electroporated LCs and moDCs were functionally similar in stimulating allogeneic T cell proliferation, a standard assay of DC immunogenicity.

ConclusionsThese findings support mRNA electroporation of DCs, and in particular LCs, as an effective non-viral method to stimulate specific, potent CD8 CTL responses. The differences between LCs and moDCs regarding this form of antigen-loading have important implications for DC-based immunotherapies.

KeywordsmRNA electroporation Dendritic cells Cancer Immunotherapy AbbreviationsAPCAntigen-presenting cell

CTLCytotoxic T lymphocyte

DCDendritic cell

eGFPEnhanced green fluorescent protein

FLT-3-ligandFms-related tyrosine kinase-3-ligand

fluMPFlu matrix peptide

G-CSFGranuloctye colony-stimulating factor

GM-CSFGranuloctye macrophage colony-stimulating factor

HPCHematopoietic progenitor cell

IL-1βInterleukin-1-beta

IL-6Interleukin-6

IL12p70Interleukin-12p70

IL15Interleukin-15

IL15R-alphaInterleukin-15-receptor-alpha

LCLangerhans-type DC

MHCMajor histocompatibility complex

MLRMixed leukocyte reaction

moDCMonocyte-derived DC

NHSNormal human serum

NK cellNatural killer cell

pSTAT5Phosphorylated signal transducer and activator of transcription 5

TGF-βTransforming growth factor-beta

TNF-αTumor necrosis factor-alpha

WT1Wilms’ tumor 1

Electronic supplementary materialThe online version of this article doi:10.1186-1479-5876-11-166 contains supplementary material, which is available to authorized users.

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Author: David J Chung - Emanuela Romano - Katherine B Pronschinske - Justin A Shyer - Milena Mennecozzi - Erin T St Angelo - Ja

Source: https://link.springer.com/







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