Presentation is loading. Please wait.

Presentation is loading. Please wait.

Dissolving Microneedle Delivery of Nanoparticle-Encapsulated Antigen Elicits Efficient Cross-Priming and Th1 Immune Responses by Murine Langerhans Cells 

Published byArleen Matthews Modified over 6 years ago

Similar presentations

Presentation on theme: "Dissolving Microneedle Delivery of Nanoparticle-Encapsulated Antigen Elicits Efficient Cross-Priming and Th1 Immune Responses by Murine Langerhans Cells "— Presentation transcript:

1 Dissolving Microneedle Delivery of Nanoparticle-Encapsulated Antigen Elicits Efficient Cross-Priming and Th1 Immune Responses by Murine Langerhans Cells  Marija Zaric, Oksana Lyubomska, Candice Poux, Mary L. Hanna, Maeliosa T. McCrudden, Bernard Malissen, Rebecca J. Ingram, Ultan F. Power, Christopher J. Scott, Ryan F. Donnelly, Adrien Kissenpfennig  Journal of Investigative Dermatology  Volume 135, Issue 2, Pages (February 2015) DOI: /jid Copyright © 2015 The Society for Investigative Dermatology, Inc Terms and Conditions

2 Figure 1 All skin-derived dendritic cell (DC) subsets present antigen to CD4+ T cells, but Langerhans cells (LCs) are the major population capable of cross-priming CD8+ T cells ex vivo.Mice (n=20) were immunized through microneedles (MNs) with ovalbumin (OVA)-encapsulated nanoparticles, and following 24 hours or 72 hours defined subsets of DCs from auricular lymph nodes were purified by FACS. 5 × 104 OVA-specific CD8+ or CD4+ T cells were cocultured in triplicates with 5 × 103 DCs of each subset for 60 hours. (a) Representative histograms of carboxyfluorescein diacetate succinimidyl ester (CFSE) dilutions of OT-I cell proliferation. (b) Bar graphs representing the percentage of proliferating OT-I T cells. (c) Representative histograms of CFSE dilutions of OT-II cell proliferation. (d) Bar graphs representing the percentage of proliferating OT-II T cells. Data have been pooled from two separate experiments. Journal of Investigative Dermatology  , DOI: ( /jid ) Copyright © 2015 The Society for Investigative Dermatology, Inc Terms and Conditions

3 Figure 2 LCs are the major skin dendritic cell (DC) subset capable of antigen cross-presentation in vivo.LanghDTR mice that were either untreated (diphtheria toxin (DT-)) or treated with DT once (DT+) or twice (DT++), received in vivo 106 carboxyfluorescein diacetate succinimidyl ester (CFSE)-labeled, CD45.1 OT-I cells. Mice were immunized with blank nanoparticles (b-NPs) or ovalbumin-encapsulated nanoparticles (OVA-NPs) the following day and skin-draining LNs were harvested 7 days later. (a) Percentages and total numbers of CD45.1+ OT-I cells recovered within each experimental group are shown. (b) CFSE and expression of IFN-γ among CD8+CD45.1+ cells (left panel) and total numbers of IFN-γ-producing OT-I cells within each group are shown (right panel). (c) Percentages and total numbers of endogenous OVA257–264/H2Kb+CD8+ T cells detected in the auricular LNs among different experimental groups are shown. All flow cytometric dot plots represent an individual mouse from each experimental group obtained in one out of three separate experiments. Journal of Investigative Dermatology  , DOI: ( /jid ) Copyright © 2015 The Society for Investigative Dermatology, Inc Terms and Conditions

4 Figure 3 Langerhans cells promote T-helper (Th)1 and Th17 CD4+ T-cell differentiation.Diphtheria toxin (DT)-untreated or -treated LanghDTR mice received intravenous 106 CD45.1 OT-II cells. Mice were immunized with blank nanoparticles (b-NPs) or ovalbumin-encapsulated nanoparticles (OVA-NPs) the following day and skin-draining lymph nodes (LNs) were harvested 7 days later. (a) Percentages and total numbers of CD45.1+ CD4+T cells recovered within each experimental group are shown. (c) Total LN cells were restimulated with phorbol 12 myristate 13 acetate-Ionomycin prior to intracellular staining. The expression of IFN-γ and IL-4 among CD4+ CD45.1+ gated cells is shown. (c) The same as in b, but the expression of IL-17 and Foxp3 among different experimental groups is shown. (d) Total numbers (mean±SEM, n=6/group) of IL-4, IFN-γ, IL-17, or Foxp3 expressing OT-II cells within each group. Journal of Investigative Dermatology  , DOI: ( /jid ) Copyright © 2015 The Society for Investigative Dermatology, Inc Terms and Conditions

5 Figure 4 Langerhans cells are necessary for efficient anti-tumor responses.LanghDTR mice were diphtheria toxin (DT) treated or untreated and immunized with blank nanoparticles (b-NPs) or ovalbumin-encapsulated nanoparticles (OVA-NPs) microneedles (MNs). Three weeks later, mice were inoculated subcutaneously with B16.OVA melanoma cells. (a) Tumor volume measurements (±SEM, n=9/group) over 18 days among experimental groups. (b) As in a, but percentages of survival among different groups are shown. (c) Percentages of OVA-specific, among tumor infiltrating CD8+ T cells, between experimental groups are shown. (d) LanghDTR mice, DT treated or untreated, were injected subcutaneously with 105 B16.OVA cells, and 4 and 9 days later immunized with b-NPs or OVA-NPs. Tumor volumes (±SEM) among different treatment groups over 15 days post inoculation are shown. (e) As in d, but percentages of survival among treatment groups (n=9/group) are shown. Journal of Investigative Dermatology  , DOI: ( /jid ) Copyright © 2015 The Society for Investigative Dermatology, Inc Terms and Conditions

6 Figure 5 Langerhans cells are required for effective anti-viral immunity (a).LanghDTR mice, either diphtheria toxin (DT) untreated or treated, were immunized with blank nanoparticles (b-NPs) or ovalbumin-encapsulated nanoparticles (OVA-NPs) microneedles (MNs). Mice were challenged intranasally with SeVOVATdT 3 weeks later. Survival among experimental groups following SeVOVATdT challenge (n=8–10/group) is shown. (b) Representative dot-plots indicating percentages of SeVOVATdT-positive cells in total lung cell suspensions between groups 6 days post infection. (c) Bars indicating percentage of SeVOVATdT-positive cells in total lung cell suspensions between groups (n=10/group). (d) Viral titers in lung homogenates between groups at 6 days post infection. (e) Percentages of OVA-specific CD8+T cells infiltrating the lungs between immunization groups. (f) IFN-γ and/or Granzyme B production following phorbol 12 myristate 13 acetate/Ionomycin restimulation of OVA-specific CD8+T cells infiltrating the lungs, as analyzed 6 days post SeVOVATdT challenge. Journal of Investigative Dermatology  , DOI: ( /jid ) Copyright © 2015 The Society for Investigative Dermatology, Inc Terms and Conditions

7 Figure 6 CD207+CD103+ dermal DCs (dDCs) preferentially cross-present soluble antigen delivered following microneedle (MN) immunization.Mice (n=20/group) were immunized through MNs with ovalbumin-encapsulated nanoparticles (OVA-NPs) or soluble OVA, and following 72 hours defined subsets of dendritic cells (DCs) from auricular lymph nodes of both immunization groups were purified by FACS. 3–5 × 104 OVA-specific CD8+ T cells were cocultured in triplicates with 3–5 × 103 DCs of each subset for 60 hours. (a) Representative histograms of carboxyfluorescein diacetate succinimidyl ester (CFSE) dilutions of OT-I cell proliferation among both immunization groups are shown. (b) Bar graphs representing the percentage of proliferating OT-I T cells. Data have been pooled from two separate experiments and are represented as mean±SEM. LC, Langerhans cell. Journal of Investigative Dermatology  , DOI: ( /jid ) Copyright © 2015 The Society for Investigative Dermatology, Inc Terms and Conditions

Download ppt "Dissolving Microneedle Delivery of Nanoparticle-Encapsulated Antigen Elicits Efficient Cross-Priming and Th1 Immune Responses by Murine Langerhans Cells "

Similar presentations

Impaired gp100-Specific CD8+ T-Cell Responses in the Presence of Myeloid-Derived Suppressor Cells in a Spontaneous Mouse Melanoma Model  David G. Mairhofer,

Impaired gp100-Specific CD8+ T-Cell Responses in the Presence of Myeloid-Derived Suppressor Cells in a Spontaneous Mouse Melanoma Model  David G. Mairhofer,
Impaired gp100-Specific CD8+ T-Cell Responses in the Presence of Myeloid-Derived Suppressor Cells in a Spontaneous Mouse Melanoma Model  David G. Mairhofer,

Impaired gp100-Specific CD8+ T-Cell Responses in the Presence of Myeloid-Derived Suppressor Cells in a Spontaneous Mouse Melanoma Model  David G. Mairhofer,
Shamim A. Mollah, Joseph S. Dobrin, Rachel E

Shamim A. Mollah, Joseph S. Dobrin, Rachel E
Volume 22, Issue 4, Pages e4 (October 2017)

Volume 22, Issue 4, Pages e4 (October 2017)
Critical Role for Skin-Derived Migratory DCs and Langerhans Cells in TFH and GC Responses after Intradermal Immunization  Clément Levin, Olivia Bonduelle,

Critical Role for Skin-Derived Migratory DCs and Langerhans Cells in TFH and GC Responses after Intradermal Immunization  Clément Levin, Olivia Bonduelle,
Dermal CD14+ Dendritic Cell and Macrophage Infection by Dengue Virus Is Stimulated by Interleukin-4  Evelyne Schaeffer, Vincent Flacher, Vasiliki Papageorgiou,

Dermal CD14+ Dendritic Cell and Macrophage Infection by Dengue Virus Is Stimulated by Interleukin-4  Evelyne Schaeffer, Vincent Flacher, Vasiliki Papageorgiou,
Identification of CD3+CD4−CD8− T Cells as Potential Regulatory Cells in an Experimental Murine Model of Graft-Versus-Host Skin Disease (GVHD)  Fumi Miyagawa,

Identification of CD3+CD4−CD8− T Cells as Potential Regulatory Cells in an Experimental Murine Model of Graft-Versus-Host Skin Disease (GVHD)  Fumi Miyagawa,
Cutaneous RANK–RANKL Signaling Upregulates CD8-Mediated Antiviral Immunity during Herpes simplex Virus Infection by Preventing Virus-Induced Langerhans.

Cutaneous RANK–RANKL Signaling Upregulates CD8-Mediated Antiviral Immunity during Herpes simplex Virus Infection by Preventing Virus-Induced Langerhans.
Priti Agarwal, Mehdi Rashighi, Kingsley I. Essien, Jillian M

Priti Agarwal, Mehdi Rashighi, Kingsley I. Essien, Jillian M
Dermal Vγ4+ γδ T Cells Possess a Migratory Potency to the Draining Lymph Nodes and Modulate CD8+ T-Cell Activity through TNF-α Production  Satoshi Nakamizo,

Dermal Vγ4+ γδ T Cells Possess a Migratory Potency to the Draining Lymph Nodes and Modulate CD8+ T-Cell Activity through TNF-α Production  Satoshi Nakamizo,
by Daniel L. Barber, Katrin D. Mayer-Barber, Lis R. V

by Daniel L. Barber, Katrin D. Mayer-Barber, Lis R. V
EpCAM Expressed by Murine Epidermal Langerhans Cells Modulates Immunization to an Epicutaneously Applied Protein Antigen  Takeshi Ouchi, Gaku Nakato,

EpCAM Expressed by Murine Epidermal Langerhans Cells Modulates Immunization to an Epicutaneously Applied Protein Antigen  Takeshi Ouchi, Gaku Nakato,
Upregulation of Inflammatory Cytokines and Oncogenic Signal Pathways Preceding Tumor Formation in a Murine Model of T-Cell Lymphoma in Skin  Xuesong Wu,

Upregulation of Inflammatory Cytokines and Oncogenic Signal Pathways Preceding Tumor Formation in a Murine Model of T-Cell Lymphoma in Skin  Xuesong Wu,
Volume 31, Issue 5, Pages (November 2009)

Volume 31, Issue 5, Pages (November 2009)
Volume 34, Issue 1, Pages (January 2011)

Volume 34, Issue 1, Pages (January 2011)
HLA-A*0201+ Plasmacytoid Dendritic Cells Provide a Cell-Based Immunotherapy for Melanoma Patients  Caroline Aspord, Marie-Therese Leccia, Dimitri Salameire,

HLA-A*0201+ Plasmacytoid Dendritic Cells Provide a Cell-Based Immunotherapy for Melanoma Patients  Caroline Aspord, Marie-Therese Leccia, Dimitri Salameire,
Nanoparticle-Based Targeting of Vaccine Compounds to Skin Antigen-Presenting Cells By Hair Follicles and their Transport in Mice  Brice Mahe, Annika Vogt,

Nanoparticle-Based Targeting of Vaccine Compounds to Skin Antigen-Presenting Cells By Hair Follicles and their Transport in Mice  Brice Mahe, Annika Vogt,
Volume 14, Issue 4, Pages (February 2016)

Volume 14, Issue 4, Pages (February 2016)
Lung dendritic cells induce TH17 cells that produce TH2 cytokines, express GATA-3, and promote airway inflammation  Marianne Raymond, PhD, Vu Quang Van,

Lung dendritic cells induce TH17 cells that produce TH2 cytokines, express GATA-3, and promote airway inflammation  Marianne Raymond, PhD, Vu Quang Van,
Self-Antigen Presentation by Keratinocytes in the Inflamed Adult Skin Modulates T-Cell Auto-Reactivity  Michael Meister, Amel Tounsi, Evelyn Gaffal, Tobias.

Self-Antigen Presentation by Keratinocytes in the Inflamed Adult Skin Modulates T-Cell Auto-Reactivity  Michael Meister, Amel Tounsi, Evelyn Gaffal, Tobias.

Similar presentations

Ads by Google