PMID- 37610279
OWN - NLM
STAT- MEDLINE
DCOM- 20230824
LR  - 20230824
IS  - 2691-1299 (Electronic)
IS  - 2691-1299 (Linking)
VI  - 3
IP  - 8
DP  - 2023 Aug
TI  - Human Dendritic Cell Enrichment and Their Activation of T Cells.
PG  - e873
LID - 10.1002/cpz1.873 [doi]
AB  - Dendritic cells (DCs) enable the immune system to mount and modulate precisely 
      targeted responses to various threats across the organism by bridging innate and 
      adaptive immunity. Historically, DCs have been classified as conventional (cDC) 
      and plasmacytoid (pDC). More recently, cDCs were acknowledged as a heterogenous 
      population composed of several subsets. Examining the functional diversity of 
      cDCs in healthy homeostasis and pathology requires a robust experimental 
      pipeline, beginning with an efficient enrichment protocol in preparation for cell 
      sorting. Unfortunately, several commercial DC enrichment kits were developed 
      before the discovery of the more recently described DC populations. Here, we 
      detail two approaches to enrich human blood DCs or certain DC subsets and an in 
      vitro protocol to examine DC stimulation of naive T cells. The methods employed 
      here overcome many hurdles encountered while enriching human DC subsets. Basic 
      Protocol 1 describes a method that will enrich pDC, Axl Siglec6-DC (AS-DC), cDC1, 
      DC2, DC3, monocytes, and human HLA(+) cells by crosslinking unwanted cells to 
      erythrocytes. Basic Protocol 2 describes the enrichment of pDC, AS-DC, cDC1, and 
      DC2 but not DC3 via a highly efficient negative magnetic selection that is 
      valuable in circumstances where DC3 is not required. Finally, Basic Protocol 3 
      describes a conventional protocol to perform a Mixed leucocyte Reaction (MLR) 
      following the isolation of these DC subsets. These methods detail the advantages 
      and pitfalls when isolating a heterogeneous population of cells. (c) 2023 The 
      Authors. Current Protocols published by Wiley Periodicals LLC. Basic Protocol 1: 
      Human peripheral mononuclear phagocyte enrichment Basic Protocol 2: DC enrichment 
      of pDC, cDC1, AS-DC, and DC2 but not DC3 Basic Protocol 3: Basic mixed lymphocyte 
      reaction protocol with sorted human DC subsets.
CI  - (c) 2023 The Authors. Current Protocols published by Wiley Periodicals LLC.
FAU - Lubin, Ruth
AU  - Lubin R
AD  - Institute of Biomedical and Oral Research, The Hebrew University, Jerusalem, 
      Israel.
FAU - Gvili, Rotem
AU  - Gvili R
AD  - Institute of Biomedical and Oral Research, The Hebrew University, Jerusalem, 
      Israel.
FAU - Hazan, Idit
AU  - Hazan I
AD  - Institute of Biomedical and Oral Research, The Hebrew University, Jerusalem, 
      Israel.
FAU - Yona, Simon
AU  - Yona S
AD  - Institute of Biomedical and Oral Research, The Hebrew University, Jerusalem, 
      Israel.
LA  - eng
PT  - Journal Article
PL  - United States
TA  - Curr Protoc
JT  - Current protocols
JID - 101773894
SB  - IM
MH  - Humans
MH  - *T-Lymphocytes
MH  - *Adaptive Immunity
MH  - Cell Movement
MH  - Cell Separation
MH  - Dendritic Cells
EDAT- 2023/08/23 12:42
MHDA- 2023/08/24 06:42
CRDT- 2023/08/23 09:25
PHST- 2023/08/24 06:42 [medline]
PHST- 2023/08/23 12:42 [pubmed]
PHST- 2023/08/23 09:25 [entrez]
AID - 10.1002/cpz1.873 [doi]
PST - ppublish
SO  - Curr Protoc. 2023 Aug;3(8):e873. doi: 10.1002/cpz1.873.