Human peripheral blood monuclear cells transfected with messenger RNA stimulate antigen-specific cytotoxic T-lymphocytes in vitro
Open accessAbstract
The efficiency of test vaccines needs to be evaluated by quantification of the triggered cellular immune response. Usually, for these assays, autologous target cells expressing the vaccine antigen are required. In the context of messenger RNA (mRNA)-based vaccinations, the target cells used for the read-out are mRNA-transfected monocyte-derived dendritic cells (Mo-DCs). Their production typically requires samples of 100 ml blood from the patients, and limits the number of assays that can be performed. We show here that fresh peripheral blood mononuclear cells (PBMCs) can be transfected with mRNA by electroporation. Such cells are as efficient as mRNA-transfected Mo-DCs for their ability to activate memory T cells in vitro. Thus, mRNA-transfected PBMCs are a convenient replacement of mRNA-transfected Mo-DCs for the in vitro monitoring of natural or vaccine-induced immune responses.
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Vectors used currently for preclinical and clinical studies resort either to UTRs of β-globin or to autochthonous UTRs of the antigen they deliver, which are not further characterized in terms of their impact on RNA stability.
“Vectors used currently for preclinical and clinical studies resort either to UTRs of -globin 5,14,23 or to autochthonous UTRs of the antigen they want to deliver, 12 which are not further characterized in terms of their impact on RNA stability.”
From Modification of antigen-encoding RNA increases stability, translational efficacy, and T-cell stimulatory capacity of dendritic cells (2006) · cites this paper as [5]
The Messenger RNA section (page unavailable) says the constructs used in this study contain UTRs from globin mRNAs, supporting this paper as an example of globin-UTR use. It does not specify β-globin or characterize the UTRs’ impact on RNA stability. (LLM, conf=0.82)
Experimental — accuracy not yet measuredAntigen-encoding RNA constructs used currently for preclinical studies have poly(A) tails measuring 30 to 70 nucleotides, whereas those administered in clinical trials measure 64 nucleotides.
“Antigen-encoding RNA constructs used currently for preclinical studies have poly(A) tails measuring 30 to 70 nucleotides in length, 4,5 whereas those administered in clinical trials measure 64 nucleotides in length.8,12”
From Modification of antigen-encoding RNA increases stability, translational efficacy, and T-cell stimulatory capacity of dendritic cells (2006) · cites this paper as [5]
In vitro-transcribed RNA used in such studies typically consists of a 5' cap structure followed by the sequence encoding the antigen of interest and a poly(A) tail 30 to 70 nucleotides in length.
“In vitro-transcribed RNA generated from plasmid templates and used in such studies typically consists of a 5Ј cap structure followed by the sequence encoding the antigen of interest and a poly(A) tail 30 to 70 nucleotides in length.4,5”
From Modification of antigen-encoding RNA increases stability, translational efficacy, and T-cell stimulatory capacity of dendritic cells (2006) · cites this paper as [5]
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Holtkamp, S., Kreiter, S., Selmi, A., Simon, P., Koslowski, M., Huber, C., Türeci, O., & Sahin, U. (2006) · Blood · doi:10.1182/blood-2006-04-015024 · Cited by 625 · see citing works
Weide, B., Pascolo, S., Scheel, B., Derhovanessian, E., Pflugfelder, A., Eigentler, T. K., Pawelec, G., Hoerr, I., Rammensee, H.-G., & Garbe, C. (2009) · Journal of Immunotherapy · doi:10.1097/cji.0b013e3181a00068 · Cited by 378 · see citing works
Self-citationResults of the First Phase I/II Clinical Vaccination Trial With Direct Injection of mRNA
Weide, B., Carralot, J.-P., Reese, A., Scheel, B., Eigentler, T. K., Hoerr, I., Rammensee, H.-G., Garbe, C., & Pascolo, S. (2008) · Journal of Immunotherapy · doi:10.1097/cji.0b013e31815ce501 · Cited by 261 · see citing works
Self-citationLong-term storage of DNA-free RNA for use in vaccine studies
Jones, K. L., Drane, D., & Gowans, E. J. (2007) · BioTechniques · doi:10.2144/000112593 · Cited by 115 · see citing works
Establishing Preferred Product Characterization for the Evaluation of RNA Vaccine Antigens
Poveda, C., Biter, A. B., Bottazzi, M. E., & Strych, U. (2019) · Vaccines · doi:10.3390/vaccines7040131 · Cited by 54 · see citing works
- Authors
- Teufel, R., Carralot, J.-P., Scheel, B., Probst, J., Walter, S., Jung, G., Hoerr, I., Rammensee, H.-G., & Pascolo, S.
- Affiliations
- CureVac (Germany) (DE)
- University of Tübingen (DE)
- Corresponding author
- Pascolo, S.
- Venue
- Cellular and Molecular Life Sciences
- Published
- 2005-07-07
- Volume
- 62
- Issue
- 15
- Pages
- 1755-1762
- ISSN
- 1420-682X (ISSN-L)
- 1420-9071
- PMC
- PMC11139132
- OpenAlex
- W2037813830
- Cited by
- 22
- References
- 0
- Citations by year
- 2024: 1 · 2023: 3 · 2021: 1 · 2019: 1 · 2015: 1 · 2012: 3
- FWCI
- 0.95
- Type
- journal article
- Language
- en
- Open access
- green
- www.ncbi.nlm.nih.gov
- Licence
- www.springer.com/tdm (tdm)
- Locations
- Cellular and Molecular Life Sciences (publishedVersion)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- Topics
- Life Sciences › Immunology and Microbiology › Immunology › Immunotherapy and Immune Responses
- Life Sciences › Immunology and Microbiology › Immunology › T-cell and B-cell Immunology
- Life Sciences › Immunology and Microbiology › Immunology › Immune responses and vaccinations
- Keywords
- peripheral blood mononuclear cells, electroporation, mRNA transfection, antigen-specific cytotoxic T lymphocytes, monocyte-derived dendritic cells, vaccine-induced immune response
- MeSH terms
- Antigens, Cells, Cultured, Humans, Immunologic Memory, Leukocytes, Mononuclear, Proteins, RNA, Messenger, T-Lymphocytes, Cytotoxic, Transfection, Histocompatibility Antigens Class I, Cryopreservation, Antigen Presentation, Electroporation, Coculture Techniques
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