Every entry summarises what the study set out to test, what it found and why it changes how we design delivery systems. Browse research and reviews or search the collection.
Kelsey L. Swingle, Margaret M. Billingsley, Sourav K. Bose, Brandon White, Rohan Palanki, Apeksha Dave, Savan K. Patel, Ningqiang Gong, Alex G. Hamilton, Mohamad-Gabriel Alameh, Drew Weissman, William H. Peranteau, Michael J. Mitchell
Congenital protein deficiencies often begin irreversible pathology in utero, but postnatal treatments are limited. In utero mRNA delivery could enable protein replacement before disease onset, yet LNP stability in fetal amniotic fluid and how it affects mRNA delivery had not been previously investigated. LNP A12 was most stable in mouse amniotic fluid; A1 was least stable. A12 had significantly higher bound protein content than A1 (p < 0.0021). - Percent change in PDI inversely correlated with in vitro luciferase.
Hanwen Zhang, Xuexiang Han, Mohamad-Gabriel Alameh, Sarah J. Shepherd, Marshall S. Padilla, Lulu Xue, Kamila Butowska, Drew Weissman, Michael J. Mitchell
LNPs can trigger innate immune responses and inflammation, which can suppress mRNA translation and cause adverse effects. There is a need for LNP formulations that reduce LNP-induced inflammation while maintaining or improving mRNA delivery. C9D1 and C10D0 had similar size, PDI, and >90% mRNA encapsulation; Dex substitution did not impair in vitro transfection or increase cytotoxicity. - Higher Dex substitution (C7D3, C5D5, C3D7, C0D10) reduced.
Arun R. K. Kumar, Yufeng Shou, Brian Chan, Krishna L., Andy Tay
This review examines how viral vectors, electroporation, microfluidic systems, nanoparticles and high-aspect-ratio nanostructures can improve the efficiency, viability and scalability of transfecting primary immune cells for applications such as CAR-T, CAR-NK and gene editing. It highlights the trade-offs between delivery efficiency, cell fitness, cargo flexibility, manufacturing complexity and clinical translation.
Ian C. Miller, Ali Zamat, Lee-Kai Sun, Hathaichanok Phuengkham, Adrian M. Harris, Lena Gamboa, Jason Yang, John P. Murad, Saul J. Priceman, Gabriel A. Kwong
CAR T cell therapy for solid malignancies typically results in poor responses. Systemic administration of immunomodulatory biologics (cytokines, BiTEs) can augment T cell activity but off-target toxicity narrows the therapeutic window. A method to spatially and temporally control transgene expression by engineered T cells at tumor sites—without systemic exposure—could improve safety and efficacy. ### Thermal Switch Engineering | Parameter | Result | |---------------|------------| | Best HSE repeat number | 7 repeats (5H-7H best; 2H-4H lower response) | | Best core promoter | YB (synthetic) — ~60-fold induction.
Julie J.-J. Tang, Alina P. Sung, Mariel J. Guglielmo, Laura N.-G., Doug Redelman, James Smith-Gagen, Diane Hudig
Monoclonal antibody anti-tumor therapies rely on NK cell-mediated antibody-dependent cell-mediated cytotoxicity (ADCC), but patient responses vary widely. While tumor antigen escape is one cause of treatment failure, the role of inter-individual variation in NK cell serial killing capacity—the ability of a single NK cell to kill multiple target cells sequentially—has received little attention. A predictive biomarker for serial ADCC capacity. ### Killing Frequency Variability (E:T=1:4, n=24 donors) | Parameter | All Donors | Females | Males | |---------------|----------------|-------------|-----------| | KF range | 1.1 - 2.2 | 1.1 - 2.2 | 1.2 - 2.1 | | Mean.
Dylan J. Drakes, Sarwish Rafiq, Terence J. Purdon, Andrea V. Lopez, Smita S. Chandran, Christopher A. Klebanoff, Renier J. Brentjens
T-cell receptor (TCR)-modified T-cell gene therapy can target a variety of extracellular and intracellular tumor-associated antigens, yet has had little clinical success. A potential explanation for limited antitumor efficacy is a lack of T-cell activation in vivo, particularly when tumor cells downregulate costimulatory molecules. A method to provide a durable proinflammatory signal to TCR-modified T cells could enhance persistence, effector. ### In Vitro T Cell Activation & Cytotoxicity | Parameter | Control | mIL12 pmel-1 | mIL18 pmel-1 | |---------------|-------------|------------------|------------------| | IFNγ secretion | Baseline | Significantly.
Bradley Vis, Rachel E. Hewitt, Tom P. Monie, Camilla Fairbairn, Suzanne D. Turner, Stephen D. Kinrade, Jonathan J. Powell
Ultrasmall silica nanoparticles (USSN, <10 nm diameter) were recently shown to stimulate T lymphocytes directly at relatively low exposure doses, but the underlying mechanisms and associated cell signaling were unknown. Understanding whether USSN directly engage the T cell receptor (TCR) complex or act through other pathways is critical for both safety assessment and therapeutic translation of these inexpensive, rapidly dissolving nanoparticles. ### USSN Activation of T Cells (CD69 Expression, 24 h) | Cell Type | Control (%) | USSN (800 μM) (%) | Fold Increase | |---------------|-----------------|-----------------------|------------------| | CD4⁺ T cells (n=23).
Coralie M. Backlund, Christopher R. Hango, Lisa M. Minter, Gregory N. Tew
Intracellular protein and antibody delivery is limited by inefficient carriers, high toxicity, and a frequent requirement for serum-free conditions, especially in difficult-to-transfect cells such as immune cells, stem cells, and neurons. A simple noncovalent carrier that works in complete media and across diverse cell types is needed. Commercial carriers: Xfect delivered GFP to all five cell types only in serum-free media; neurons were most challenging. PULSin delivered to all except neurons in serum-free media. ProteoJuice achieved only ~20%.
Janusz Khetan, Madhan Srikar, Daniel B. Forger, Sutapa Barua
The general perception is that cellular uptake of materials is proportional to cell volume (mass) or surface area, following cubic or square relationships with cell radius. However, experimental data on nanoparticle uptake in MDA-MB 231 breast cancer cells show a linear relationship with cell radius—a result that is counterintuitive. A quantitative modeling framework is needed to explain this unexpected correlation and to understand how cell. ### MDA-MB 231 Cell Size Distribution | Parameter | Value | |---------------|-----------| | Mean cell radius (FSC, uncorrected) | ~11 μm | | Mean cell radius (microscopy, corrected) | ~8 μm | | Distribution (σc) | 0.20.
Francesca Giulimondi, Luca Digiacomo, Daniela Pozzi, Sara Palchetti, Elisabetta Vulpis, Anna Laura Capriotti, Riccardo Zenezini Chiozzi, Aldo Laganà, Heinz Amenitsch, Laura Masuelli, Giovanna Peruzzi, Morteza Mahmoudi, Isabella Screpanti, Alessandra Zingoni
Liposome clinical translation is limited by opsonization and rapid clearance by immune cells. Although the protein corona is recognized as the liposome’s “biological identity,” it was unclear whether pre-adsorbed corona proteins could help liposomes avoid capture by circulating leukocytes in whole blood and thereby prolong blood residency. Zeta potential: At high HP, zeta potential of DOTAP, DOPC, and DOPG converged to a common negative plateau of ~ −23 mV, independent of pristine surface charge. - Size: DOTAP size increased at low HP, peaked near charge.
Magnesium ions (Mg²⁺) promote bone regeneration, but their effects are strongly concentration-dependent: too little has minimal effect, while excess Mg²⁺ can impair osteoblast activity and bone metabolism. There is no well-established bone tissue engineering scaffold that precisely controls local Mg²⁺ release. Injectable, biodegradable microspheres that provide tunable Mg²⁺ release could fill irregular bone defects and improve bone regeneration. Tunable Mg²⁺ release: Higher MgO fractions gave faster release and higher initial Mg²⁺ concentrations; higher MgCO₃ fractions gave slower, longer-term sustained release. PMg-III released approximately 30–50 ppm Mg²⁺ in.
Drugs with ideal pharmacokinetic profiles require long half-lives but little organ accumulation. Generally, these factors are contradictory: smaller particles lead to faster excretion and shorter half-lives, while larger particles lead to longer circulation but stronger organ accumulation and potential toxicity. The effect of nanoparticle size on biodistribution has been inconsistent in the literature due to the confounding effect of shape. ### Nanoparticle Size Characterization (DLS) | Nanoparticle | Designed Size | Measured DLS (nm) | |------------------|-------------------|----------------------| | 5 nm square | 5 nm | 5.60 ± 1.01 | | 10 nm square | 10.
Chao Wang, Wujin Sun, Yanqi Ye, Hunter N. Bomba, Zhen Gu
Immune cell/organ therapies based on manipulation, infusion, and implantation of autologous or allogeneic cells/organs are costly, time-consuming, and sometimes limited in clinical effectiveness. Biomimetic materials and strategies offer an alternative by enabling artificial immune cells and lymphoid organs that are ready-to-use and scalable for immunotherapy. --- - PLGA aAPCs with anti-CD3, anti-CD28, pMHC, and encapsulated IL-2 showed stable ligand presentation for ~20 days, significant IFN-γ secretion, and 45-fold T cell expansion; 6–10 μm particles were most effective. - Ellipsoidal PLGA aAPCs were more efficient than spherical particles; nanoellipsoidal aAPCs stimulated stronger in vivo immune responses at reduced protein dose and showed enhanced pharmacokinetics. - Filamentous anti-CD3 polymers induced more robust T cell responses than PLGA microparticles due to structural flexibility and multivalency. - Magnetic nano-aAPCs with a
Susanne Przybylski, Michaela Gasch, Anne Marschner, Marcus Ebert, Alexander Ewe, Gisa Helmig, Nadja Hilger, Stephan Fricke, Susanne Rudzok, Achim Aigner, Jana Burkhardt
Nanoparticle-based gene delivery methods are promising for transfecting immune cells, but their impact on basic immune functions such as proliferation—a cornerstone of cellular immunity—has not been systematically investigated. Understanding whether transfection reagents non-specifically influence immune cell proliferation is critical for both therapeutic applications (where effects may be beneficial or detrimental) and for accurately. ### Proliferation Effects (PBMCs, AON/siRNA Transfection) | Reagent | Magnetic Enhancement | Proliferation Change (vs. Control) | Significance |.
Daniela Schmid, Chun Gwon Park, Christina A. Hartl, Nikita Subedi, Adam N. Cartwright, Regina Bou Puerto, Yiran Zheng, James Maiarana, Gordon J. Freeman, Kai W. Wucherpfennig, Darrell J. Irvine, Michael S. Goldberg
Systemic immune stimulation for cancer immunotherapy is associated with autoimmune-type pathologies, as the doses required to break immune tolerance can invoke undesired host-vs.-host effects. Targeting endogenous immune cell subsets in the circulation—which actively migrate into tumors—could concentrate immunomodulatory drugs on tumor-reactive effector cells, improving both efficacy and safety compared to administration of free drugs or direct. ### Nanoparticle Characterization | Parameter | Uncoated NPs | Isotype NPs | CD8a NPs | |---------------|------------------|-----------------|--------------| | Size (nm) | 267 ± 8 | 269 ± 8 | 273 ± 8 | | Antibody.
Petra Furedi, Zsofia Edit Papay, Kristof Kovacs, Borbala Dalmadi Kiss, Krisztina Ludanyi, Istvan Antal, Imre Klebovich
Topical fungal infections, including ocular mycoses, are increasing, and voriconazole (VCZ) is an effective antifungal but has poor water solubility, limiting liquid ophthalmic formulations. Lipid-based nanoparticles (LNPs/SLNs) may improve VCZ solubility, loading, and topical delivery. Optimized formulation (SLN2): Witespol W35 0.25 g + VCZ 50 mg; particle size 182 ± 4.1 nm, PDI 0.269 ± 0.01, entrapment efficiency 78.79 ± 3.4%. - Five homogenization cycles at 600 bar reduced PDI by ~36.5% compared.
Joshua J. Glass, Daniel Yuen, James Rae, Angus P. R. Johnston, Robert G. Parton, Stephen J. Kent, Robert De Rose
Investigations of nanoparticle targeting to immune cells are conventionally restricted to monocultured cell line models that lack the complexity of primary human blood cell populations, including non-specific association with, and competition from, diverse cell types. A more physiologically relevant system using mixed primary human blood cells is needed to evaluate nanoparticle targeting, particularly for therapeutically relevant non-phagocytic. ### Nanoparticle Characterization | Parameter | Untargeted | αCD4 | αCD20 | |---------------|---------------|----------|-----------| | EM diameter (nm) | 42.7 ± 8.3 | 41.3 ± 8.3 | 45.3 ± 6.8 | | DLS hydrodynamic size.
Zhihe Lu, Jingrong Su, Zhengrong Li, Yuzhu Zhan, Decai Ye
Gemcitabine (GEM) monotherapy for pancreatic cancer has poor tumor specificity, short circulation half-life, and drug resistance. Baicalein (BCL) can downregulate Bcl-2/Mcl-1 and sensitize tumor cells, but it has poor solubility, low bioavailability, and rapid metabolism. A targeted co-delivery system was needed to combine GEM and BCL, improve pharmacokinetics, and enhance antitumor efficacy. HA-GEM-BCL NLCs: size 131.4 ± 3.9 nm, PDI 0.12 ± 0.03, zeta potential −25.1 ± 1.9 mV, GEM EE 85.1 ± 2.3%, BCL EE 82.9 ± 2.4%. - NLCs were stable over 1 month; release was faster at pH 5.0 than pH 7.4. - Cellular uptake.
5-Fluorouracil (5-FU) plus cisplatin (CDDP) is a first-line gastric cancer chemotherapy regimen, but it causes substantial toxicity and 5-FU has poor biopharmaceutical properties. A targeted nanoparticle platform could codeliver both drugs, improve synergy, and reduce systemic toxicity. Optimal HA coating concentration was 0.2% (w/v); higher concentrations (0.3–0.4%) decreased encapsulation efficiency and increased particle size. - HA-FU/C-NLC showed sustained release: >80% 5-FU release at 48 h vs 36 h.
Satyajit Mayor, Robert G. Parton, Julie G. Donaldson
Clathrin-mediated endocytosis (CME) has long been the dominant paradigm for cellular uptake, but many endocytic pathways operate without clathrin. These clathrin-independent endocytic (CIE) pathways are increasingly recognized as distinct routes with different cargo selectivity, molecular machinery, kinetics, and intracellular destinations. The review addresses the need to organize and understand this diversity, especially for small-scale CIE processes that operate without clathrin coats and, in some cases, without dynamin. --- - Caveolae are dynamic endocytic carriers: In interphase cells, ~85% of caveolae were internalized within a 10-min observation period, with lifetimes ranging from <2 s (30% of caveolae) to >7 min. Budded structures were cavin-positive, and dynamin dependence was confirmed. - EHD2 is a negative regulator of caveolar endocytosis: Knockdown of EHD2 increases caveolar budding. EHD2 links caveolae to actin and associa
Ying-Chun Lo, Michael A. Edidin, Jonathan D. Powell
Soluble anti-CD3 antibodies are potent T cell mitogens but activate all T cells regardless of TCR specificity, limiting their therapeutic utility for boosting antigen-specific responses. TCRs cluster into microclusters on the surface of antigen-experienced T cells but remain dispersed on naive T cells. This difference in receptor organization could be exploited to selectively activate only previously activated T cells using anti-CD3 antibodies. ### TCR Clustering (Confocal Microscopy) | T Cell State | Microclusters/Cell | Average Cluster Area (μm²) | Mean Intensity |.
David N. Nguyen, Kerry P. Mahon, Ghania Chikh, Phillip Kim, Hattie Chung, Alain P. Vicari, Kevin T. Love, Michael Goldberg, Steve Chen, Arthur M. Krieg, Jianzhu Chen, Robert Langer, Daniel G. Anderson
TLR7/8 agonists are promising vaccine adjuvants and antivirals, but RNA agonists are unstable and small-molecule agonists cause systemic toxicity. A delivery system specifically designed to deliver immunostimulatory RNA (isRNA) to TLR-expressing cells could enable safer and more localized innate immune activation. 14 lipidoid formulations matched or exceeded Lipofectamine 2000 for isRNA-induced type I IFN in PBMCs; 100-core lipidoids were enriched. - Second-generation LRNP-I and LRNP-II were superior to DOTAP and ND(5)-98-1 for.
Chemo-immunotherapy combinations are limited by systemic immunotoxicity from cancer drugs and rapid clearance of low-molecular-weight drugs after intratumoral injection. A local, biodegradable delivery system was needed to sustain drug/GMCSF exposure at the tumor site, reduce systemic toxicity, and promote tumor antigen cross-presentation and CD8+ T cell-mediated anti-tumor immunity. Tumor growth: CH containing cancer drug + GMCSF significantly reduced TC-1 tumor growth compared with CH–cancer drug alone, CH-GMCSF, or CH alone. CTX was the most potent drug in combination with GMCSF. Fig. 1 reported.
James Harris, Dirk Werling, Jayne C. Hope, Geraldine Taylor, Chris J. Howard
Caveolae and caveolin are cholesterol-rich membrane microdomains involved in endocytosis, cholesterol regulation, and signal transduction, but their presence and function in immune cells have been contentious. This review addresses the need to clarify the distribution and roles of caveolae/caveolin in mammalian immune cells, especially given emerging evidence that they mediate pathogen internalization by antigen-presenting cells and participate in immune-cell signaling. --- 1. Distribution is contentious and context-dependent: Caveolae/caveolin are commonly found in myeloid cells (macrophages, mast cells, dendritic cells, neutrophils) but not consistently in lymphoid cells. More recent evidence suggests they may be present in all immune cell types, with expression/distribution dependent on activation and/or maturation state. 2. Species and cell-type differences in caveolin localization: Human CD26+ and CD21+ peripheral blood lymphocytes
