Abstract
The modification of CD4+ T cells with exogenous nucleic acids or proteins is a critical step in several research and therapeutic applications, such as HIV studies and cancer immunotherapies. However, efficient cell transfections are not always easily achieved when working with these primary hard-to-transfect cells. While the modification of T cells is typically performed by viral transduction or electroporation, their use is associated with safety issues or cytotoxicity. Vapor nanobubble (VNB) photoporation with sensitizing gold nanoparticles (AuNPs) has recently emerged as a new technology for safe and flexible cell transfections. In this work, we evaluated the potential of VNB photoporation as a novel technique for the intracellular delivery of macromolecules in primary human CD4+ T cells using fluorescent dextrans as model molecules. Our results show that VNB photoporation enables efficient delivery of fluorescent dextrans of 10 kDa in Jurkat (>60% FD10+ cells) as well as in primary human CD4+ T cells (±40% FD10+ cells), with limited cell toxicity (>70% cell viability). We also demonstrated that the technique allows the delivery of dextrans that are up to 500 kDa in Jurkat cells, suggesting its applicability for the delivery of biological macromolecules with a wide range of molecular weights. Altogether, VNB photoporation represents a promising technique for the universal delivery of macromolecules in view of engineering CD4+ T cells for use in a wide variety of research and therapeutic applications.
Highlights
Adoptive T cell transfer represents a promising, emerging approach for cancer immunotherapy and HIV therapy [1]
We demonstrated that the technique allows the delivery of dextrans that are up to 500 kDa in Jurkat cells, suggesting its applicability for the delivery of biological macromolecules with a wide range of molecular weights
vapor nanobubbles (VNB) photoporation represents a promising technique for the universal delivery of macromolecules in view of engineering CD4+ T cells for use in a wide variety of research and therapeutic applications
Summary
Adoptive T cell transfer represents a promising, emerging approach for cancer immunotherapy and HIV therapy [1]. High electric fields can result in a drasticporation decrease in viability; recently, microfluidics for transient mechanical membrane bycell cell squeezing were electroporation has been shown to substantially alter the T cell phenotype and reduce its shown to therapeutic enable theefficacy transfection of different cell types with limited cytotoxicity [27,28]. (c) Upon nanosecond pulsedislaser irradiation, arise from theassociated cell-bound cause physical cell membrane poration This allows the extracellular macromolecules to diffuse into the thermal energy is consumed, the VNBs will collapse and the associated shock waves cause physical the cell cytoplasm. VNBs the can be used for efficient pore formation in cell pressure membranes,waves allowing the can cause is consumed, which bubble violently collapses, producing that intracellular delivery of biological macromolecules [31,32,33]. These results demonstrate the potential of VNB photoporation for the efficient intracellular delivery of macromolecules in primary human CD4+ T cells, thereby providing a very promising basis for future research on the transfection of functional macromolecules in view of CD4+ T-cell engineering
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