An efficient and high yield method for isolation of mouse dendritic cell subsets

Pooja Arora, Steven A. Porcelli

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

Dendritic cells (DCs) are professional antigen-presenting cells primarily responsible for acquiring, processing and presenting antigens on antigen presenting molecules to initiate T-cell-mediated immunity. Dendritic cells can be separated into several phenotypically and functionally heterogeneous subsets. Three important subsets of splenic dendritic cells are plasmacytoid, CD8αPos and CD8αNeg cells. The plasmacytoid DCs are natural producers of type I interferon and are important for anti-viral T cell immunity. The CD8αNeg DC subset is specialized for MHC class II antigen presentation and is centrally involved in priming CD4 T cells. The CD8αPos DCs are primarily responsible for cross-presentation of exogenous antigens and CD8 T cell priming. The CD8αPos DCs have been demonstrated to be most efficient at the presentation of glycolipid antigens by CD1d molecules to a specialized T cell population known as invariant natural killer T (iNKT) cells. Administration of Flt-3 ligand increases the frequency of migration of dendritic cell progenitors from bone marrow, ultimately resulting in expansion of dendritic cells in peripheral lymphoid organs in murine models. We have adapted this model to purify large numbers of functional dendritic cells for use in cell transfer experiments to compare in vivo proficiency of different DC subsets.

Original languageEnglish (US)
Article numbere53824
JournalJournal of Visualized Experiments
Volume2016
Issue number110
DOIs
StatePublished - Apr 1 2016

Fingerprint

Dendritic Cells
T-cells
Antigens
T-Lymphocytes
Antigen Presentation
CD1d Antigen
Cross-Priming
CD8 Antigens
Interferons
Interferon Type I
Natural Killer T-Cells
Molecules
Glycolipids
Histocompatibility Antigens Class II
Antigen-Presenting Cells
Cellular Immunity
Immunity
Bone
Bone Marrow
Ligands

Keywords

  • 33D1
  • Antigen presenting cell
  • CD1d
  • CD8α
  • DEC205
  • Dendritic cells
  • Immunology
  • Infection
  • Issue 110
  • Plasmacytoid DC

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Chemical Engineering(all)
  • Immunology and Microbiology(all)
  • Neuroscience(all)

Cite this

An efficient and high yield method for isolation of mouse dendritic cell subsets. / Arora, Pooja; Porcelli, Steven A.

In: Journal of Visualized Experiments, Vol. 2016, No. 110, e53824, 01.04.2016.

Research output: Contribution to journalArticle

@article{9d44fd3fccb349f1b15749f93fa05488,
title = "An efficient and high yield method for isolation of mouse dendritic cell subsets",
abstract = "Dendritic cells (DCs) are professional antigen-presenting cells primarily responsible for acquiring, processing and presenting antigens on antigen presenting molecules to initiate T-cell-mediated immunity. Dendritic cells can be separated into several phenotypically and functionally heterogeneous subsets. Three important subsets of splenic dendritic cells are plasmacytoid, CD8αPos and CD8αNeg cells. The plasmacytoid DCs are natural producers of type I interferon and are important for anti-viral T cell immunity. The CD8αNeg DC subset is specialized for MHC class II antigen presentation and is centrally involved in priming CD4 T cells. The CD8αPos DCs are primarily responsible for cross-presentation of exogenous antigens and CD8 T cell priming. The CD8αPos DCs have been demonstrated to be most efficient at the presentation of glycolipid antigens by CD1d molecules to a specialized T cell population known as invariant natural killer T (iNKT) cells. Administration of Flt-3 ligand increases the frequency of migration of dendritic cell progenitors from bone marrow, ultimately resulting in expansion of dendritic cells in peripheral lymphoid organs in murine models. We have adapted this model to purify large numbers of functional dendritic cells for use in cell transfer experiments to compare in vivo proficiency of different DC subsets.",
keywords = "33D1, Antigen presenting cell, CD1d, CD8α, DEC205, Dendritic cells, Immunology, Infection, Issue 110, Plasmacytoid DC",
author = "Pooja Arora and Porcelli, {Steven A.}",
year = "2016",
month = "4",
day = "1",
doi = "10.3791/53824",
language = "English (US)",
volume = "2016",
journal = "Journal of Visualized Experiments",
issn = "1940-087X",
publisher = "MYJoVE Corporation",
number = "110",

}

TY - JOUR

T1 - An efficient and high yield method for isolation of mouse dendritic cell subsets

AU - Arora, Pooja

AU - Porcelli, Steven A.

PY - 2016/4/1

Y1 - 2016/4/1

N2 - Dendritic cells (DCs) are professional antigen-presenting cells primarily responsible for acquiring, processing and presenting antigens on antigen presenting molecules to initiate T-cell-mediated immunity. Dendritic cells can be separated into several phenotypically and functionally heterogeneous subsets. Three important subsets of splenic dendritic cells are plasmacytoid, CD8αPos and CD8αNeg cells. The plasmacytoid DCs are natural producers of type I interferon and are important for anti-viral T cell immunity. The CD8αNeg DC subset is specialized for MHC class II antigen presentation and is centrally involved in priming CD4 T cells. The CD8αPos DCs are primarily responsible for cross-presentation of exogenous antigens and CD8 T cell priming. The CD8αPos DCs have been demonstrated to be most efficient at the presentation of glycolipid antigens by CD1d molecules to a specialized T cell population known as invariant natural killer T (iNKT) cells. Administration of Flt-3 ligand increases the frequency of migration of dendritic cell progenitors from bone marrow, ultimately resulting in expansion of dendritic cells in peripheral lymphoid organs in murine models. We have adapted this model to purify large numbers of functional dendritic cells for use in cell transfer experiments to compare in vivo proficiency of different DC subsets.

AB - Dendritic cells (DCs) are professional antigen-presenting cells primarily responsible for acquiring, processing and presenting antigens on antigen presenting molecules to initiate T-cell-mediated immunity. Dendritic cells can be separated into several phenotypically and functionally heterogeneous subsets. Three important subsets of splenic dendritic cells are plasmacytoid, CD8αPos and CD8αNeg cells. The plasmacytoid DCs are natural producers of type I interferon and are important for anti-viral T cell immunity. The CD8αNeg DC subset is specialized for MHC class II antigen presentation and is centrally involved in priming CD4 T cells. The CD8αPos DCs are primarily responsible for cross-presentation of exogenous antigens and CD8 T cell priming. The CD8αPos DCs have been demonstrated to be most efficient at the presentation of glycolipid antigens by CD1d molecules to a specialized T cell population known as invariant natural killer T (iNKT) cells. Administration of Flt-3 ligand increases the frequency of migration of dendritic cell progenitors from bone marrow, ultimately resulting in expansion of dendritic cells in peripheral lymphoid organs in murine models. We have adapted this model to purify large numbers of functional dendritic cells for use in cell transfer experiments to compare in vivo proficiency of different DC subsets.

KW - 33D1

KW - Antigen presenting cell

KW - CD1d

KW - CD8α

KW - DEC205

KW - Dendritic cells

KW - Immunology

KW - Infection

KW - Issue 110

KW - Plasmacytoid DC

UR - http://www.scopus.com/inward/record.url?scp=84964856839&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84964856839&partnerID=8YFLogxK

U2 - 10.3791/53824

DO - 10.3791/53824

M3 - Article

C2 - 27166856

AN - SCOPUS:84964856839

VL - 2016

JO - Journal of Visualized Experiments

JF - Journal of Visualized Experiments

SN - 1940-087X

IS - 110

M1 - e53824

ER -