Influence of nanoparticle-mediated transfection on proliferation of primary immune cells in vitro and in vivo

Abstract

IntroductionOne of the main obstacles in the widespread application of gene therapeutic approaches is the necessity for efficient and safe transfection methods. For the introduction of small oligonucleotide gene therapeutics into a target cell, nanoparticle-based methods have been shown to be highly effective and safe. While immune cells are a most interesting target for gene therapy, transfection might influence basic immune functions such as cytokine expression and proliferation, and thus positively or negatively affect therapeutic intervention. Therefore, we investigated the effects of nanoparticle-mediated transfection such as polyethylenimine (PEI) or magnetic beads on immune cell proliferation.MethodsHuman adherent and non-adherent PBMCs were transfected by various methods (e.g. PEI, Lipofectamine® 2000, magnetofection) and stimulated. Proliferation was measured by lymphocyte transformation test (LTT). Cell cycle stages as well as expression of proliferation relevant genes were analyzed. Additionally, the impact of nanoparticles was investigated in vivo in a murine model of the severe systemic immune disease GvHD (graft versus host disease).ResultsThe proliferation of primary immune cells was influenced by nanoparticle-mediated transfection. In particular in the case of magnetic beads, proliferation inhibition coincided with short-term cell cycle arrest and reduced expression of genes relevant for immune cell proliferation. Notably, proliferation inhibition translated into beneficial effects in a murine GvHD model with animals treated with PEI-nanoparticles showing increased survival (pPEI = 0.002) most likely due to reduced inflammation.ConclusionThis study shows for the first time that nanoparticles utilized for gene therapeutic transfection are able to alter proliferation of immune cells and that this effect depends on the type of nanoparticle. For magnetic beads, this was accompanied by temporary cell cycle arrest. Notably, in GvHD this nonspecific anti-proliferative effect might contribute to reduced inflammation and increased survival.

<p>Virucidal Action Against Avian Influenza H5N1 Virus and Immunomodulatory Effects of Nanoformulations Consisting of Mesoporous Silica Nanoparticles Loaded with Natural Prodrugs</p>

“…92 As nanoformulations containing a concentration equivalent to 25 µg/mL of prodrug inhibited immune T lymphocyte proliferation, they have a beneficial effect as anti-inflammatory or immunomodulation factors, leading to the activation of different molecular signaling pathways in the development of diseases, such as autoimmune diseases, bacterial and viral infection, and cancers. This effect may be due to the drug carrier nanostructure of MSNs mediating the inhibition of lymphocytes 93 and associated with the prodrug activities, such as antioxidant activity. 94 …”

Charge-Altering Releasable Transporters Enable Specific Phenotypic Manipulation of Resting Primary Natural Killer Cells

“…This diverse repertoire is stable over time 59 , but can be rapidly and dramatically modulated by a number of common in vitro interventions, including low-dose (2 ng/mL) IL-15 40 . We hypothesized that established transfection techniques may alter NK cell proteomic phenotype 60 .…”

Emerging Roles of 1D Vertical Nanostructures in Orchestrating Immune Cell Functions

“…Of note, NW‐treated T cells maintained high viability and proliferative capacity, with no discernible increase in key activation markers (Figure 6j); [ 36 ] this overcame significant limitations of conventional viral and nonviral methods that may induce unexpected immunogenicity and altered cellular function. [ 121 ]…”