PMID- 33316130
OWN - NLM
STAT- MEDLINE
DCOM- 20210830
LR  - 20240410
IS  - 1934-368X (Electronic)
IS  - 1934-3671 (Print)
IS  - 1934-3671 (Linking)
VI  - 131
IP  - 1
DP  - 2020 Dec
TI  - A Comprehensive Experimental Guide to Studying Cross-Presentation in Dendritic 
      Cells In Vitro.
PG  - e115
LID - 10.1002/cpim.115 [doi]
AB  - Cross-presentation was first observed serendipitously in the 1970s. The 
      importance of it was quickly realized and subsequently attracted great attention 
      from immunologists. Since then, our knowledge of the ability of certain antigen 
      presenting cells to internalize, process, and load exogenous antigens onto MHC-I 
      molecules to cross-prime CD8(+) T cells has increased significantly. Dendritic 
      cells (DCs) are exceptional cross-presenters, thus making them a great tool to 
      study cross-presentation but the relative rarity of DCs in circulation and in 
      tissues makes it challenging to isolate sufficient numbers of cells to study this 
      process in vitro. In this paper, we describe in detail two methods to culture DCs 
      from bone-marrow progenitors and a method to expand the numbers of DCs present in 
      vivo as a source of endogenous bona-fide cross-presenting DCs. We also describe 
      methods to assess cross-presentation by DCs using the activation of primary 
      CD8(+) T cells as a readout. (c) 2020 Wiley Periodicals LLC. Basic Protocol 1: 
      Isolation of bone marrow progenitor cells Basic Protocol 2: In vitro 
      differentiation of dendritic cells with GM-CSF Support Protocol 1: Preparation of 
      conditioned medium from GM-CSF producing J558L cells Basic Protocol 3: In vitro 
      differentiation of dendritic cells with Flt3L Support Protocol 2: Preparation of 
      Flt3L containing medium from B16-Flt3L cells Basic Protocol 4: Expansion of cDC1s 
      in vivo for use in ex vivo experiments Basic Protocol 5: Characterizing resting 
      and activated dendritic cells Basic Protocol 6: Dendritic cell stimulation, 
      antigenic cargo, and fixation Support Protocol 3: Preparation of model antigen 
      coated microbeads Support Protocol 4: Preparation of apoptotic cells Support 
      Protocol 5: Preparation of recombinant bacteria Basic Protocol 7: 
      Immunocytochemistry immunofluorescence (ICC/IF) Support Protocol 6: Preparation 
      of Alcian blue-coated coverslips Basic Protocol 8: CD8(+) T cell activation to 
      assess cross-presentation Support Protocol 7: Isolation and labeling of CD8(+) T 
      cells with CFSE.
CI  - (c) 2020 Wiley Periodicals LLC.
FAU - Sadiq, Barzan A
AU  - Sadiq BA
AD  - The Jill Roberts Institute for Research in Inflammatory Bowel Disease, Weill 
      Cornell Medicine, Cornell University, New York, New York.
FAU - Mantel, Ian
AU  - Mantel I
AD  - The Jill Roberts Institute for Research in Inflammatory Bowel Disease, Weill 
      Cornell Medicine, Cornell University, New York, New York.
AD  - Immunology and Microbial Pathogenesis Program, Weill Cornell Graduate School of 
      Medical Sciences, Weill Cornell Medicine, Cornell University, New York, New York.
FAU - Blander, J Magarian
AU  - Blander JM
AD  - The Jill Roberts Institute for Research in Inflammatory Bowel Disease, Weill 
      Cornell Medicine, Cornell University, New York, New York.
AD  - Immunology and Microbial Pathogenesis Program, Weill Cornell Graduate School of 
      Medical Sciences, Weill Cornell Medicine, Cornell University, New York, New York.
AD  - Department of Microbiology and Immunology, Weill Cornell Medicine, Cornell 
      University, New York, New York.
AD  - Sandra and Edward Meyer Cancer Center, Weill Cornell Medicine, Cornell 
      University, New York, New York.
AD  - Joan and Sanford I. Weill Department of Medicine, Weill Cornell Medicine, Cornell 
      University, New York, New York.
LA  - eng
GR  - R01 AI170897/AI/NIAID NIH HHS/United States
GR  - R01 DK111862/DK/NIDDK NIH HHS/United States
GR  - R01 AI073899/AI/NIAID NIH HHS/United States
GR  - R01 AI170832/AI/NIAID NIH HHS/United States
GR  - R21 AI154294/AI/NIAID NIH HHS/United States
GR  - R01 AI127658/AI/NIAID NIH HHS/United States
GR  - R56 AI073899/AI/NIAID NIH HHS/United States
GR  - R01 AI123284/AI/NIAID NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - Curr Protoc Immunol
JT  - Current protocols in immunology
JID - 9101651
MH  - Animals
MH  - Bone Marrow Cells/cytology
MH  - CD8-Positive T-Lymphocytes/*immunology
MH  - Cell Culture Techniques
MH  - Cell Differentiation
MH  - Cell Proliferation
MH  - Cross-Priming/*immunology
MH  - Dendritic Cells/*cytology/metabolism
MH  - Humans
MH  - Lymphocyte Activation
MH  - Mice
MH  - Microspheres
PMC - PMC9060150
MID - NIHMS1646175
OTO - NOTNLM
OT  - CD8+ T cell
OT  - DC
OT  - MHC-I
OT  - cross-presentation
OT  - cross-priming
OT  - dendritic cells
EDAT- 2020/12/15 06:00
MHDA- 2021/08/31 06:00
PMCR- 2022/05/02
CRDT- 2020/12/14 17:16
PHST- 2020/12/14 17:16 [entrez]
PHST- 2020/12/15 06:00 [pubmed]
PHST- 2021/08/31 06:00 [medline]
PHST- 2022/05/02 00:00 [pmc-release]
AID - 10.1002/cpim.115 [doi]
PST - ppublish
SO  - Curr Protoc Immunol. 2020 Dec;131(1):e115. doi: 10.1002/cpim.115.