. Author manuscript; available in PMC: 2023 Dec 8.

Published in final edited form as: Eur J Immunol. 2022 Oct 27;53(11):e2249816. doi: 10.1002/eji.202249816

Table 13.

Proposed guidelines to ensure of the identity of the DC types generated in vitro, as compared to cells of the monocyte/macrophage lineages

Cell type	Phenotypic identity^a)	Positive transcriptomic signature^b)	Negative transcriptomic signature^c)	Key functions that can be tested in vitro	Additional characteristics	References
cDC1	HLA-DR⁺ CD11c⁺ CD33⁺ CD141⁺ CLEC9A⁺ XCR1⁺ CADM1⁺ BTLA^hi	*C1orf54, CADM1, CLEC9A, CLNK, CPNE3, GCSAM, NAAA, WDFY4, RAB32, XCR1, BATF3, BTLA, HLA-DOB, ID2, IDO1, IRF8*	NN	• IFN-III production upon TLR3 stimulation. • IL-12p70 production upon TLR8 stimulation. • High efficacy for cross-presentation of (dead-) cell-associated Ag.	• BATF3- and IRF8-dependent differentiation.	[78, 89, 95, 96, 106–110]
cDC2	HLA-DR⁺ CD11c⁺ CD33⁺ CD1c⁺ CLEC10A⁺ FCER1A⁺ CD5lo_to_hi BTLAlo_to_hi CD14– CD163⁻ CX3CR1⁺ CD88⁻	*CD1C, CD1E, CLEC10A, FCER1A, BTLA, CD5, FLT3, HLA-DOB, IRF4*	*C3AR1, C5AR1, CD14, CD163, S100A8, S100A9, FCN1, VCAN*	• IL-12p70 production upon triggering of CD40 and TLR8, IL-23 production. • Functional polarization of CD4⁺ T cells toward Th17 or Th2?	• IRF8-dependent differentiation.	[96, 101, 105, 107–111]
DC3	HLA-DR⁺ CD11c⁺ CD33⁺ CD1c⁺ CD163⁺ CD14lo_to_hi CD5– BTLA⁻ CD88⁻	*CD14, CD163, CD1C, CLEC10A, FCER1A, S100A8, S100A9, CDKN1A, CLEC4E, F13A1, FCN1, LMNA, VCAN*	*C1QA, C1QB, C1QC, C3AR1, C5AR1, MERTK*	• TGFβ and IL-1β production. • T_RM differentiation.	• Differentiation does not require as high IRF8 expression as for cDC2.	[101, 111, 112]
ASDC	HLA-DR⁺ CD11c^low/+ CD33⁺ CD123^low/+ CLEC4C⁺ SIGLEC6⁺ AXL⁺ CX3CR1⁺ CD2⁺ CD5⁺	*AXL, CD123, SIGLEC1, SIGLEC6, TCF4, BCL11A, KLF12, LYZ*	*GZMB, LILRA4, NLRP7, PACSIN1*	• Lack of IFN-I/III production upon TLR7 or TLR9 stimulation • High antigen presentation capacity, similar to that of cDC	• Dendritic morphology • TCF4-dependent differentiation. • High susceptibility to HIV-1 infection	[107–109, 113]
pDC	HLA-DR⁺ CD11c⁻ CD33⁻ CD123⁺ CLEC4C⁺ CD2⁻ CD5⁻	**CD123, CLEC4C, GZMB, IRF7, LILRA4, NLRP7, PACSIN1, TCF4, TLR7, TLR9*, BCL11A, EPHB1, PTPRS, RUNX2, SPIB*	*ZBTB46*	• IFN-I/III production upon TLR7 or TLR9 stimulation. • Lower steady state antigen presentation capacity than cDC, but that is strongly boosted upon adequate stimulation.	• Plasmacytoid morphology. • TCF4-dependent differentiation.	[95, 105, 107–109, 114, 115]
MoDC	HLA-DR⁺ CD11c^hi CD11b^hi CD172a^hi CD163⁻ CD1a⁺ CD206low/+ CD209low/+	*CD1A, CD1C, CD209, FCER1A, FCER2, FCGR2A, FCGR2B, FCGR2C, FCGR3A, IRF4, ITGAM, LILRB2, MRC1, S100A8, S100A9, SIRPA, STAB1, TLR2, TLR4, VCAN, CD14, CD226, CD68, CLEC10A, CSF1R, EBI3, LAMP3, MMP12*	*C1QA, C1QB, C1QC, CD163, FLT3, MERTK*	• Production of high levels of TNF, CXCL1, CCL7 and IL-10 in response to TLR4 triggering. • IL-23 production upon CD40 triggering. • Functional polarization of CD4⁺ T cells toward Th17 or Tfh (or Th2?).	• Dendritic morphology • IRF4-dependent differentiation.	[30, 85, 89, 105, 116, 117]
LC	HLA-DR⁺ CD11c^hi CD11b⁺ CD1a⁺ CD207⁺ E-Cad⁺	*CD207, EPCAM, ABCC4, AXIN2, BMPR1A, MICAL2, MYO6, TJP1*	NN	• High efficacy for CD8⁺ T cell priming.	• Birbeck granules.	[87, 106]
CD14⁺ DDC	HLA-DR⁺ CD11c⁺ CD11b^hi CD1a⁻ CD14^hi CD207⁻ E-Cad⁻	*CD14, CEBPB, MAFB, MSR1, TLR4*		• Functional polarization of CD4⁺ T cells toward Tfh.	• Dendritic morphology.	[87, 106]
cMo	CD88⁺ CD11b^hi CD172a^hi CD14⁺ CD163⁻ CD206⁻	*CD14, CEBPB, FCN1, LYZ, S100A8, S100A9, VCAN*	*C1QA, C1QB, C1QC, CD163, FLT3, MERTK, ZBTB46*	• Direct B cell stimulation. • Low phagocytosis compared to macrophages • Low antigen presentation ability compared to cDC	• Non-dendritic and non foamy morphology	[105, 110]
MoMac	CD88⁺ CD11b^hi CD172a^hi CD16⁺ CD163⁺ CD206⁺ CD1a⁻	*APOE, C1QA, C1QB, C1QC, C3AR1, C5AR1, CD163, FCGR3A, FOLR2, MERTK*	NN	• High phagocytosis. • Production of high levels of TNF in response to TLR4 triggering.	• Foam cell morphology. • MAFB-dependent differentiation.	[30, 85, 105, 110]

^a)

The minimal set of phenotypic markers recommended to identify the cell types is highlighted in bold. – −

^b)

The minimal set of positive marker genes recommended to identify the cell types is highlighted in bold.

^c)

The minimal set of negative marker gene recommended to discriminate the target cell type from the other types of mononuclear phagocytes that harbor the closest gene expression profiles is highlighted in bold. NN, not necessary, the use of negative marker genes is not necessary.