Breaking Entry-and Species Barriers: LentiBOOST® Plus Polybrene Enhances Transduction Efficacy of Dendritic Cells and Monocytes by Adenovirus 5

Due to their ability to trigger strong immune responses, adenoviruses (HAdVs) in general and the serotype5 (HAdV-5) in particular are amongst the most popular viral vectors in research and clinical application. However, efficient transduction using HAdV-5 is predominantly achieved in coxsackie and adenovirus receptor (CAR)-positive cells. In the present study, we used the transduction enhancer LentiBOOST® comprising the polycationic Polybrene to overcome these limitations. Using LentiBOOST®/Polybrene, we yielded transduction rates higher than 50% in murine bone marrow-derived dendritic cells (BMDCs), while maintaining their cytokine expression profile and their capability to induce T-cell proliferation. In human dendritic cells (DCs), we increased the transduction rate from 22% in immature (i)DCs or 43% in mature (m)DCs to more than 80%, without inducing cytotoxicity. While expression of specific maturation markers was slightly upregulated using LentiBOOST®/Polybrene on iDCs, no effect on mDC phenotype or function was observed. Moreover, we achieved efficient HAdV5 transduction also in human monocytes and were able to subsequently differentiate them into proper iDCs and functional mDCs. In summary, we introduce LentiBOOST® comprising Polybrene as a highly potent adenoviral transduction agent for new in-vitro applications in a set of different immune cells in both mice and humans.

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Breaking Entry- and Species Barriers: LentiBOOST® Plus Polybrene Enables High Transduction Efficacy of Dendritic Cells and Monocytes by Adenovirus 5

10.21203/rs.3.rs-701427/v1 ◽

2021 ◽

Author(s):

Astrid Strack ◽

Andrea Deinzer ◽

Christian Thirion ◽

Silke Schrödel ◽

Jan Dörrie ◽

...

Keyword(s):

Dendritic Cells ◽

Viral Vectors ◽

T Cell Proliferation ◽

Murine Bone Marrow ◽

Cytokine Expression Profile ◽

Adenoviral Transduction ◽

Human Dendritic Cells ◽

Adenovirus Receptor ◽

Adenovirus 5

Abstract Due to their ability to trigger strong immune responses, adenoviruses (HAdVs) in general and the serotype5 (HAdV5) in particular are amongst the most popular viral vectors in research and clinical application. However, efficient transduction using HAdV5 is predominantly achieved in coxsackie and adenovirus receptor (CAR)-positive human cells. In the present study, we used the transduction enhancer LentiBOOST® comprising the polycationic Polybrene to overcome these limitations. Using LentiBOOST®/ Polybrene, we yielded transduction rates higher than 50% in murine bone marrow derived dendritic cells (BMDCs), while maintaining their cytokine expression profile and their capability to induce T-cell proliferation. In human dendritic cells (DCs), we increased the transduction rate from 22% in immature (i)DCs or 43% in mature (m)DCs to more than 80%, without inducing cytotoxicity. While expression of specific maturation markers was slightly upregulated using LentiBOOST®/ Polybrene on iDCs, no effect on mDC phenotype or function was observed. Moreover, we achieved efficient HAdV5 transduction also in human monocytes and were able to subsequently differentiate them into proper iDCs and functional mDCs. In summary, we introduce LentiBOOST® comprising Polybrene as a highly potent adenoviral transduction agent for new in vitro applications in a set of different immune cells in both mice and humans.

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Faculty Opinions recommendation of Ascaris suum infection downregulates inflammatory pathways in the pig intestine in vivo and in human dendritic cells in vitro.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.732114816.793551813 ◽

2018 ◽

Author(s):

Thomas Nutman

Keyword(s):

Dendritic Cells ◽

Ascaris Suum ◽

Inflammatory Pathways ◽

Human Dendritic Cells

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Effects of Staphylococcus aureus Bacteriophage K on Expression of Cytokines and Activation Markers by Human Dendritic Cells In Vitro

Viruses ◽

10.3390/v10110617 ◽

2018 ◽

Vol 10 (11) ◽

pp. 617 ◽

Cited By ~ 5

Author(s):

Helen Freyberger ◽

Yunxiu He ◽

Amanda Roth ◽

Mikeljon Nikolich ◽

Andrey Filippov

Keyword(s):

Staphylococcus Aureus ◽

Dendritic Cells ◽

Inflammatory Response ◽

Adaptive Immune Response ◽

Human Monocyte ◽

Activation Markers ◽

Mhc I ◽

Adaptive Immune ◽

Human Dendritic Cells

A potential concern with bacteriophage (phage) therapeutics is a host-versus-phage response in which the immune system may neutralize or destroy phage particles and thus impair therapeutic efficacy, or a strong inflammatory response to repeated phage exposure might endanger the patient. Current literature is discrepant with regard to the nature and magnitude of innate and adaptive immune response to phages. The purpose of this work was to study the potential effects of Staphylococcus aureus phage K on the activation of human monocyte-derived dendritic cells. Since phage K acquired from ATCC was isolated around 90 years ago, we first tested its activity against a panel of 36 diverse S. aureus clinical isolates from military patients and found that it was lytic against 30/36 (83%) of strains. Human monocyte-derived dendritic cells were used to test for an in vitro phage-specific inflammatory response. Repeated experiments demonstrated that phage K had little impact on the expression of pro- and anti-inflammatory cytokines, or on MHC-I/II and CD80/CD86 protein expression. Given that dendritic cells are potent antigen-presenting cells and messengers between the innate and the adaptive immune systems, our results suggest that phage K does not independently affect cellular immunity or has a very limited impact on it.

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Regulatory perspective on in vitro potency assays for human dendritic cells used in anti-tumor immunotherapy

Cytotherapy ◽

10.1016/j.jcyt.2018.07.006 ◽

2018 ◽

Vol 20 (11) ◽

pp. 1289-1308 ◽

Cited By ~ 2

Author(s):

CHARLOTTE DE Wolf ◽

MARJA VAN DE BOVENKAMP ◽

MARCEL HOEFNAGEL

Keyword(s):

Dendritic Cells ◽

Tumor Immunotherapy ◽

Regulatory Perspective ◽

Human Dendritic Cells

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Murine bone marrow-derived dendritic cells as a potential in vitro model for predictive identification of chemical sensitizers

Toxicology Letters ◽

10.1016/j.toxlet.2007.09.012 ◽

2007 ◽

Vol 175 (1-3) ◽

pp. 89-101 ◽

Cited By ~ 13

Author(s):

E PEPIN ◽

M GOUTET ◽

M BAN

Keyword(s):

Bone Marrow ◽

Dendritic Cells ◽

In Vitro Model ◽

Murine Bone Marrow ◽

Vitro Model

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The novel adjuvant CoVaccineHT™ increases the immunogenicity of cell-culture derived influenza A/H5N1 vaccine and induces the maturation of murine and human dendritic cells in vitro

Vaccine ◽

10.1016/j.vaccine.2009.09.015 ◽

2009 ◽

Vol 27 (49) ◽

pp. 6833-6839 ◽

Cited By ~ 17

Author(s):

R. Bodewes ◽

M.M. Geelhoed-Mieras ◽

J.G.M. Heldens ◽

J. Glover ◽

B.N. Lambrecht ◽

...

Keyword(s):

Cell Culture ◽

Dendritic Cells ◽

Influenza A ◽

The Novel ◽

Human Dendritic Cells ◽

H5n1 Vaccine

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Water Extract of Cordyceps militaris Enhances Maturation of Murine Bone Marrow-Derived Dendritic Cells in Vitro

Biological and Pharmaceutical Bulletin ◽

10.1248/bpb.29.354 ◽

2006 ◽

Vol 29 (2) ◽

pp. 354-360 ◽

Cited By ~ 29

Author(s):

Gi-Young Kim ◽

Woo-Shin Ko ◽

Jae-Yoon Lee ◽

Jeong-Ok Lee ◽

Chung-Ho Ryu ◽

...

Keyword(s):

Bone Marrow ◽

Dendritic Cells ◽

Water Extract ◽

Cordyceps Militaris ◽

Murine Bone Marrow

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HIV inhibits CD4+ T-cell proliferation by inducing indoleamine 2,3-dioxygenase in plasmacytoid dendritic cells

Blood ◽

10.1182/blood-2006-07-034785 ◽

2006 ◽

Vol 109 (8) ◽

pp. 3351-3359 ◽

Cited By ~ 201

Author(s):

Adriano Boasso ◽

Jean-Philippe Herbeuval ◽

Andrew W. Hardy ◽

Stephanie A. Anderson ◽

Matthew J. Dolan ◽

...

Keyword(s):

Cell Proliferation ◽

Dendritic Cells ◽

T Cell ◽

Functional Impairment ◽

Plasmacytoid Dendritic Cells ◽

Cd4 T Cell ◽

T Cell Proliferation ◽

Type I ◽

Indoleamine 2,3 Dioxygenase

AbstractInfection with the human immunodeficiency virus type-1 (HIV) results in acute and progressive numeric loss of CD4+ T-helper cells and functional impairment of T-cell responses. The mechanistic basis of the functional impairment of the surviving cells is not clear. Indoleamine 2,3-dioxygenase (IDO) is an immunosuppressive enzyme that inhibits T-cell proliferation by catabolizing the essential amino acid tryptophan (Trp) into the kynurenine (kyn) pathway. Here, we show that IDO mRNA expression is elevated in peripheral blood mononuclear cells (PBMCs) from HIV+ patients compared with uninfected healthy controls (HCs), and that in vitro inhibition of IDO with the competitive blocker 1-methyl tryptophan (1-mT) results in increased CD4+ T-cell proliferative response in PBMCs from HIV-infected patients. We developed an in vitro model in which exposure of PBMCs from HCs to either infectious or noninfectious, R5- or X4-tropic HIV induced IDO in plasmacytoid dendritic cells (pDCs). HIV-induced IDO was not inhibited by blocking antibodies against interferon type I or type II, which, however, induced IDO in pDCs when added to PBMC cultures. Blockade of gp120/CD4 interactions with anti-CD4 Ab inhibited HIV-mediated IDO induction. Thus, induction of IDO in pDCs by HIV may contribute to the T-cell functional impairment observed in HIV/AIDS by a non–interferon-dependent mechanism.

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