Achieving dendritic cell subset-specific targeting in vivo by site-directed conjugation of targeting antibodies to nanocarriers

The major challenge of nanocarrier-based anti-cancer vaccination approaches is the targeted delivery of antigens and immunostimulatory agents to cells of interest, such as specific subtypes of dendritic cells (DCs), in order to induce robust antigen-specific anti-tumor responses. An undirected cell and body distribution of nanocarriers can lead to unwanted delivery to other immune cell types like macrophages reducing the vaccine efficacy. An often-used approach to overcome this issue is the surface functionalization of nanocarriers with targeting moieties, such as antibodies, mediating cell type-specific interaction. Numerous studies could successfully prove the targeting efficiency of antibody-conjugated carrier systems in vitro, however, most of them failed when targeting DCs in vivo that is partly due to cells of the reticuloendothelial system unspecifically clearing nanocarriers from the blood stream via Fc receptor ligation. Therefore, this study shows a surface functionalization strategy to site-specifically attach antibodies in an orientated direction onto the nanocarrier surface. Different DC-targeting antibodies, such as anti-CD11c, anti-CLEC9A, anti-DEC205 and anti-XCR1, were conjugated to the nanocarrier surface at their Fc domains. Anti-mouse CD11c antibody-conjugated nanocarriers specifically accumulated in the targeted organ (spleen) over time. Additionally, antibodies against CD11c and CLEC9A proved to specifically direct nanocarriers to the targeted DC subtype, conventional DCs type 1. In conclusion, site-directed antibody conjugation to nanocarriers is essential in order to avoid unspecific uptake by non-target cells while achieving antibody-specific targeting of DC subsets. This novel conjugation technique paves the way for the development of antibody-functionalized nanocarriers for DC-based vaccination approaches in the field of cancer immunotherapy.

Download Full-text

Achieving dendritic cell subset-specific targeting in vivo by site-directed conjugation of targeting antibodies to nanocarriers

Nano Today ◽

10.1016/j.nantod.2022.101375 ◽

2022 ◽

Vol 43 ◽

pp. 101375

Author(s):

Johanna Simon ◽

Michael Fichter ◽

Gabor Kuhn ◽

Maximilian Brückner ◽

Cinja Kappel ◽

...

Keyword(s):

Dendritic Cell ◽

Cell Subset ◽

Dendritic Cell Subset ◽

Specific Targeting

Download Full-text

The CD8+ Dendritic Cell Subset Selectively Endocytoses Dying Cells in Culture and In Vivo

Journal of Experimental Medicine ◽

10.1084/jem.20020161 ◽

2002 ◽

Vol 195 (10) ◽

pp. 1289-1302 ◽

Cited By ~ 451

Author(s):

Tomonori Iyoda ◽

Susumu Shimoyama ◽

Kang Liu ◽

Yoshiki Omatsu ◽

Yuji Akiyama ◽

...

Keyword(s):

T Cells ◽

Cd8 T Cells ◽

Ex Vivo ◽

Osmotic Shock ◽

Cell Subset ◽

Fluorescent Labeling ◽

Dendritic Cell Subset ◽

Dying Cells

Dendritic cells (DCs) are able in tissue culture to phagocytose and present antigens derived from infected, malignant, and allogeneic cells. Here we show directly that DCs in situ take up these types of cells after fluorescent labeling with carboxyfluorescein succinimidyl ester (CFSE) and injection into mice. The injected cells include syngeneic splenocytes and tumor cell lines, induced to undergo apoptosis ex vivo by exposure to osmotic shock, and allogeneic B cells killed by NK cells in situ. The CFSE-labeled cells in each case are actively endocytosed by DCs in vivo, but only the CD8+ subset. After uptake, all of the phagocytic CD8+ DCs can form major histocompatibility complex class II–peptide complexes, as detected with a monoclonal antibody specific for these complexes. The CD8+ DCs also selectively present cell-associated antigens to both CD4+ and CD8+ T cells. Similar events take place with cultured DCs; CD8+ DCs again selectively take up and present dying cells. In contrast, both CD8+ and CD8− DCs phagocytose latex particles in culture, and both DC subsets present soluble ovalbumin captured in vivo. Therefore CD8+ DCs are specialized to capture dying cells, and this helps to explain their selective ability to cross present cellular antigens to both CD4+ and CD8+ T cells.

Download Full-text

Migration ofAntigen-Specific T Cells Away from CXCR4-Binding Human ImmunodeficiencyVirus Type 1gp120

Journal of Virology ◽

10.1128/jvi.78.10.5184-5193.2004 ◽

2004 ◽

Vol 78 (10) ◽

pp. 5184-5193 ◽

Cited By ~ 18

Author(s):

Diana M. Brainard ◽

William G. Tharp ◽

Elva Granado ◽

Nicholas Miller ◽

Alicja K. Trocha ◽

...

Keyword(s):

T Cells ◽

T Cell ◽

Immune Cell ◽

Active Movement ◽

Target Cells ◽

Cell Mediated Immunity ◽

Cell Trafficking ◽

Hiv 1

ABSTRACT Cell-mediated immunity depends in part on appropriate migration and localization of cytotoxic T lymphocytes (CTL), a process regulated by chemokines and adhesion molecules. Many viruses, including human immunodeficiency virus type 1 (HIV-1), encode chemotactically active proteins, suggesting that dysregulation of immune cell trafficking may be a strategy for immune evasion. HIV-1 gp120, a retroviral envelope protein, has been shown to act as a T-cell chemoattractant via binding to the chemokine receptor and HIV-1 coreceptor CXCR4. We have previously shown that T cells move away from the chemokine stromal cell-derived factor 1 (SDF-1) in a concentration-dependent and CXCR4 receptor-mediated manner. Here, we demonstrate that CXCR4-binding HIV-1 X4 gp120 causes the movement of T cells, including HIV-specific CTL, away from high concentrations of the viral protein. This migratory response is CD4 independent and inhibited by anti-CXCR4 antibodies and pertussis toxin. Additionally, the expression of X4 gp120 by target cells reduces CTL efficacy in an in vitro system designed to account for the effect of cell migration on the ability of CTL to kill their target cells. Recombinant X4 gp120 also significantly reduced antigen-specific T-cell infiltration at a site of antigen challenge in vivo. The repellant activity of HIV-1 gp120 on immune cells in vitro and in vivo was shown to be dependent on the V2 and V3 loops of HIV-1 gp120. These data suggest that the active movement of T cells away from CXCR4-binding HIV-1 gp120, which we previously termed fugetaxis, may provide a novel mechanism by which HIV-1 evades challenge by immune effector cells in vivo.

Download Full-text

The Delivery of Biologically Active Agents into the Nuclei of Target Cells for the Purposes of Translational Medicine

Acta Naturae ◽

10.32607/actanaturae.11049 ◽

2020 ◽

Vol 12 (4) ◽

pp. 47-56

Author(s):

A. S. Sobolev

Keyword(s):

Targeted Delivery ◽

Translational Medicine ◽

Intracellular Transport ◽

Biologically Active ◽

Target Cells ◽

Subcellular Compartment ◽

Core Technology ◽

Dividing Cells

Development of vehicles for the subcellular targeted delivery of biologically active agents is very promising for the purposes of translational medicine. This review summarizes the results obtained by researchers from the Laboratory of Molecular Genetics of Intracellular Transport, Institute of Gene Biology RAS, which allowed them to design the core technology: modular nanotransporters. This approach ensures high efficacy and cell specificity for different anti-cancer agents, as they are delivered into the most vulnerable subcellular compartment within the cells of interest and makes it possible for antibody mimetics to penetrate into a compartment of interest within the target cells (diving antibodies). Furthermore, polyplexes, complexes of polycationic block copolymers of DNA, have been developed and characterized. These complexes are efficient both in vitro and in vivo and demonstrate predominant transfection of actively dividing cells.

Download Full-text

Engineered extracellular vesicle decoy receptor-mediated modulation of the IL6 trans-signalling pathway in muscle

10.1101/2020.06.09.142216 ◽

2020 ◽

Author(s):

Mariana Conceição ◽

Laura Forcina ◽

Oscar P. B. Wiklander ◽

Dhanu Gupta ◽

Joel Z. Nordin ◽

...

Keyword(s):

Targeted Delivery ◽

Therapeutic Potential ◽

Extracellular Vesicle ◽

Signalling Pathway ◽

Peptide Sequence ◽

Target Cells ◽

Dimerization Domain ◽

Decoy Receptor ◽

Stat3 Phosphorylation

AbstractThe cytokine interleukin 6 (IL6) is a key mediator of inflammation that contributes to skeletal muscle pathophysiology. IL6 activates target cells by two different mechanisms, the classical and transsignalling pathways. While classical signalling is associated with the anti-inflammatory activities of the cytokine, the IL6 trans-signalling pathway mediates chronic inflammation and is therefore a target for therapeutic intervention. Extracellular vesicles (EVs) are natural, lipid-bound nanoparticles, with potential as targeted delivery vehicles for therapeutic macromolecules. Here, we engineered EVs to express IL6 signal transducer (IL6ST) decoy receptors to selectively inhibit the IL6 trans-signalling pathway. The potency of the IL6ST decoy receptor EVs was optimized by inclusion of a GCN4 dimerization domain and a peptide sequence derived from syntenin-1 which targets the decoy receptor to EVs. The resulting engineered EVs were able to efficiently inhibit activation of the IL6 transsignalling pathway in reporter cells, while having no effect on the IL6 classical signalling. IL6ST decoy receptor EVs, were also capable of blocking the IL6 trans-signalling pathway in C2C12 myoblasts and myotubes, thereby inhibiting the phosphorylation of STAT3 and partially reversing the anti-differentiation effects observed when treating cells with IL6/IL6R complexes. Treatment of a Duchenne muscular dystrophy mouse model with IL6ST decoy receptor EVs resulted in a reduction in STAT3 phosphorylation in the quadriceps and gastrocnemius muscles of these mice, thereby demonstrating in vivo activity of the decoy receptor EVs as a potential therapy. Taken together, this study reveals the IL6 trans-signalling pathway as a promising therapeutic target in DMD, and demonstrates the therapeutic potential of IL6ST decoy receptor EVs.

Download Full-text

Cell-specific and targeted delivery of RNA moieties

10.1101/2020.01.03.893818 ◽

2020 ◽

Author(s):

Aditi Bhargava ◽

Peter Ohara ◽

Luc Jasmin

Keyword(s):

Nucleic Acids ◽

Targeted Delivery ◽

Immune Cell ◽

Neuronal Cell ◽

Cell Types ◽

Specific Cell ◽

Chemically Modified ◽

Covalently Linked

AbstractDelivery of therapeutic moieties to specific cell types, such as neurons remains a challenge. Genes present in neurons are also expressed in non-neuronal cell types such as glia where they mediate non-targeted related functions. Thus, non-specific targeting of these proteins/channels has numerous unwanted side effects, as is the case with current small molecules or drug therapies. Current methodologies that use nanoparticles, lipid-mediated uptake, or mannitol in conjunction with lipids to deliver double-stranded RNA (dsRNA) have yielded mixed and unreliable results. We used a neuroanatomical tracer (B subunit of Cholera Toxin (CTB)) that binds to the ganglioside receptors (GM1) expressed on cells, including primary sensory neurons to deliver encapsulated dsRNA. This approach greatly improved delivery of dsRNA to the desired cells by enhancing uptake, reducing vehicle-mediated toxicity and protecting nucleotides from degradation by endonucleases. The delivery complex is internalized, and once inside the cell, the dsRNA naturally dissociates itself from the carrier complex and is very effective in knocking down cognate targets, both in vivo and in vitro. Past methods have used CTB-fusion proteins or chemically modified oligos or DNA moieties that have been covalently conjugated to CTB. Furthermore, CTB conjugated to an antigen, protein, or chemically modified nucleic acid is a potent activator of immune cell (T and B cells, macrophages) response, whereas CTB admixed with antigens or unmodified nucleic acids does not evoke this immune response. Importantly, in our method, the nucleic acids are not covalently linked to the carrier molecules. Thus, our method holds strong potential for targeted delivery of therapeutic moieties for cell types expressing GM1 receptors, including neuronal cell types.

Download Full-text

Efficient Hepatic Delivery of Drugs: Novel Strategies and Their Significance

BioMed Research International ◽

10.1155/2013/382184 ◽

2013 ◽

Vol 2013 ◽

pp. 1-20 ◽

Cited By ~ 34

Author(s):

Nidhi Mishra ◽

Narayan Prasad Yadav ◽

Vineet Kumar Rai ◽

Priyam Sinha ◽

Kuldeep Singh Yadav ◽

...

Keyword(s):

Targeted Delivery ◽

Surface Membrane ◽

Whole Body ◽

Prolonged Treatment ◽

Target Cells ◽

Nonparenchymal Cells ◽

Drug Molecules ◽

Body Distribution ◽

Herbal Medications ◽

Site Of Action

Liver is a vital organ responsible for plethora of functions including detoxification, protein synthesis, and the production of biochemicals necessary for the sustenance of life. Therefore, patients with chronic liver diseases such as viral hepatitis, liver cirrhosis, and hepatocellular carcinoma need immediate attention to sustain life and as a result are often exposed to the prolonged treatment with drugs/herbal medications. Lack of site-specific delivery of these medications to the hepatocytes/nonparenchymal cells and adverse effects associated with their off-target interactions limit their continuous use. This calls for the development and fabrication of targeted delivery systems which can deliver the drug payload at the desired site of action for defined period of time. The primary aim of drug targeting is to manipulate the whole body distribution of drugs, that is, to prevent distribution to non-target cells and concomitantly increase the drug concentration at the targeted site. Carrier molecules are designed for their selective cellular uptake, taking advantage of specific receptors or binding sites present on the surface membrane of the target cell. In this review, various aspects of liver targeting of drug molecules and herbal medications have been discussed which elucidate the importance of delivering the drugs/herbal medications at their desired site of action.

Download Full-text

In VivoIntradermal Delivery of Bacteria by Using Microneedle Arrays

Infection and Immunity ◽

10.1128/iai.00406-18 ◽

2018 ◽

Vol 86 (9) ◽

Cited By ~ 4

Author(s):

Courtney E. Chandler ◽

Erin M. Harberts ◽

Tim Laemmermann ◽

Qin Zeng ◽

Belita N. Opene ◽

...

Keyword(s):

Targeted Delivery ◽

Immune Cell ◽

Serum Levels ◽

Dependent Manner ◽

Microbial Diseases ◽

Content Type ◽

Biologically Relevant ◽

Francisella Novicida ◽

Microneedle Arrays

ABSTRACTInfectious diseases propagated by arthropod vectors, such as tularemia, are commonly initiated via dermal infection of the skin. However, due to the technical difficulties in achieving accurate and reproducible dermal deposition, intradermal models are less commonly used. To overcome these limitations, we used microneedle arrays (MNAs), which are micron-scale polymeric structures, to temporarily disrupt the barrier function of the skin and deliver a bacterial inoculum directly to the dermis of an animal. MNAs increase reliability by eliminating leakage of the inoculum or blood from the injection site, thereby providing a biologically relevant model for arthropod-initiated disease. Here, we validate the use of MNAs as a means to induce intradermal infection using a murine model of tularemia initiated byFrancisella novicida. We demonstrate targeted delivery of the MNA bolus to the dermal layer of the skin, which subsequently led to innate immune cell infiltration. Additionally,F. novicida-coated MNAs were used to achieve lethality in a dose-dependent manner in C57BL/6 mice. The immune profile of infected mice mirrored that of establishedF. novicidainfection models, consisting of markedly increased serum levels of interleukin-6 and keratinocyte chemoattractant, splenic T-cell depletion, and an increase in splenic granulocytes, together confirming that MNAs can be used to reproducibly induce tularemia-like pathogenesis in mice. When MNAs were used to immunize mice using an attenuatedF. novicidamutant (F. novicidaΔlpxD1), all immunized mice survived a lethal subcutaneous challenge. Thus, MNAs can be used to effectively deliver viable bacteriain vivoand provide a novel avenue to study intradermally induced microbial diseases in animal models.

Download Full-text

Regional induction of adhesion molecules and chemokine receptors explains disparate homing of human B cells to systemic and mucosal effector sites: dispersion from tonsils

Blood ◽

10.1182/blood-2004-12-4630 ◽

2005 ◽

Vol 106 (2) ◽

pp. 593-600 ◽

Cited By ~ 79

Author(s):

Finn-Eirik Johansen ◽

Espen S. Baekkevold ◽

Hege S. Carlsen ◽

Inger Nina Farstad ◽

Dulce Soler ◽

...

Keyword(s):

Chemokine Receptors ◽

Dna Analysis ◽

Immune Cell ◽

Female Genital Tract ◽

Cell Subset ◽

Immunoglobulin M ◽

Respiratory Pathogens ◽

Upper Airways ◽

Direct Tracking

Abstract Ethical constraints restrict direct tracking of immune-cell migration throughout the human body in vivo. We, therefore, used deletion of the immunoglobulin M (IgM) heavy-chain constant-gene (Cμ) segment as a marker to provide a dispersal signature of an effector B-cell subset (IgD+IgM-CD38+) induced selectively in human tonsils. By DNA analysis, the Cμ deletion identified dissemination of such blasts and their plasma-cell progeny to peripheral blood, lymph nodes, and bone marrow, as well as to mucosae and glands of the upper airways. Also the endocervix was often positive, while the small intestine was mainly negative, as could be expected from the identified homing-molecule profile of the marker cells, with relatively low levels of integrin α4β7 and CC chemokine receptor 9 (CCR9). Of further importance for vaccine design, the circulating cells expressed abundantly CD62L (L-selectin) and CCR7, which provided a mechanism for integration of respiratory and systemic immunity. Most mucosal vaccines are at present administered perorally, and our results suggested that the nasal route is no alternative for vaccination against rotavirus or other small-intestinal infections in humans. However, immunization of nasopharynx-associated lymphoid tissue clearly appears preferable to target respiratory pathogens and may to some extent also protect against infections of the female genital tract. (Blood. 2005;106:593-600)

Download Full-text