RT Journal Article
T1 Study of Oligonucleotides Access and Distribution in Human Peripheral Blood Mononuclear Cells.
A1 Fernandez-Delgado, Manuel
A1 Sendra, Luis
A1 Herrero, Maria Jose
A1 Olivera-Pasquini, Gladys G
A1 Batista-Duharte, Alexander
A1 Aliño, Salvador F
K1 CD8+ treg
K1 FOXP3
K1 PBMC
K1 Treg
K1 Cell uptake
K1 Flow cytometry
K1 Fluorescent labeling
K1 Oligonucleotides
K1 Quenching
AB Therapeutic oligonucleotides have achieved great clinical interest since their approval as drug agents by regulatory agencies but their access and distribution in blood cells are not completely known. We evaluated by flow cytometry the ability of short fluorescent scramble oligonucleotides (ON*) to access human peripheral blood mononuclear cells (PBMC) after incubating with ON* during 1 h and 7 days of culture follow-up 'in vitro'. Blood samples were treated with chemically modified oligonucleotides (phosphorothioate backbone and 2' O-Me ends) to resist nuclease digestion under culture conditions. The ON* internalization was determined after discarding the membrane-associated fluorescence by trypan blue quenching. Whereas the oligonucleotide accessed neutrophils and monocytes rapidly, achieving their maximum in 1 h and 24 h, respectively, lymphocytes required 7 days to achieve the maximum (80% of cells) transfection. The ON*ability to access lymphocyte types (T, B, and NK) and T cell subtypes (CD4+, CD8+, and CD4-CD8-) were similar, with T cells being more accessible. Regulatory CD4+ and CD8+ T cells were classified in low and high Foxp3 expressers, whose expression proved not to alter the ON* internalization during the first hour, achieving 53% of CD4+Foxp3+ and 40% of CD8+Foxp3+ cells. Our results contribute to understanding and improving the management of therapeutic ONs.
PB MDPI
YR 2022
FD 2022-05-19
LK http://hdl.handle.net/10668/21158
UL http://hdl.handle.net/10668/21158
LA en
NO Fernández-Delgado M, Sendra L, Herrero MJ, Olivera-Pasquini GG, Batista-Duharte A, Aliño SF. Study of Oligonucleotides Access and Distribution in Human Peripheral Blood Mononuclear Cells. Int J Mol Sci. 2022 May 23;23(10):5839
DS RISalud
RD Apr 19, 2025