Transcriptional Profiling of Human Dendritic Cell Populations and Models - Unique Profiles of In Vitro Dendritic Cells and Implications on Functionality and Applicability

Abstract

BackgroundDendritic cells (DCs) comprise heterogeneous populations of cells, which act as central orchestrators of the immune response. Applicability of primary DCs is restricted due to their scarcity and therefore DC models are commonly employed in DC-based immunotherapy strategies and in vitro tests assessing DC function. However, the interrelationship between the individual in vitro DC models and their relative resemblance to specific primary DC populations remain elusive.ObjectiveTo describe and assess functionality and applicability of the available in vitro DC models by using a genome-wide transcriptional approach.MethodsTranscriptional profiling was performed with four commonly used in vitro DC models (MUTZ-3-DCs, monocyte-derived DCs, CD34-derived DCs and Langerhans cells (LCs)) and nine primary DC populations (dermal DCs, LCs, blood and tonsillar CD123+, CD1c+ and CD141+ DCs, and blood CD16+ DCs).ResultsPrincipal Component Analysis showed that transcriptional profiles of each in vitro DC model most closely resembled CD1c+ and CD141+ tonsillar myeloid DCs (mDCs) among primary DC populations. Thus, additional differentiation factors may be required to generate model DCs that more closely resemble other primary DC populations. Also, no model DC stood out in terms of primary DC resemblance. Nevertheless, hierarchical clustering showed clusters of differentially expressed genes among individual DC models as well as primary DC populations. Furthermore, model DCs were shown to differentially express immunologically relevant transcripts and transcriptional signatures identified for each model DC included several immune-associated transcripts.ConclusionThe unique transcriptional profiles of in vitro DC models suggest distinct functionality in immune applications. The presented results will aid in the selection of an appropriate DC model for in vitro assays and assist development of DC-based immunotherapy.

MUTZ-3 Langerhans Cell maturation and CXCL12 independent migration in reconstructed human gingiva

“…We now show that in oral mucosa, in contrast to skin, maturing of GE-LC, in addition to becoming CD1a -, became mRNA CD83 + . This is in line with our previous findings and those of others who reported HLA-DR + DC subsets in human tonsils that lack CD1a expression (Summers et al, 2001;Lundberg et al, 2013), indicating that migration to the draining lymph nodes cannot be ruled out. Most likely the loss of CD1a upon migration is caused by differences in the LP in GE vs. dermis in the SE, consistent with our previous observations that the phenotype of migrating DC is highly plastic and dependent on the cytokine balance in the tissue microenvironment (de Gruijl et al, 2006;Ouwehand et al, 2011a).…”

Aryl hydrocarbon receptor contributes to the MEK/ERK-dependent maintenance of the immature state of human dendritic cells

“…44 Thus, although the transcriptional profile of in vitro-generated MDDCs is more related to that of CD1c 1 blood DCs than to other primary DC populations, 46 the gene expression signature of MDDCs is also considerably skewed by IL-4 and, interestingly, it reveals a significantly higher expression of the prototypical AhR-target gene CYP1B1 than other in vitrogenerated DC populations. 46 Therefore, it is conceivable that differentiation-driving cytokines might determine the level of expression or activation of AhR in CD1c 1 DCs and MDDCs, thus explaining the cell-context dependency of the ERK-AhR link that we have observed.…”

Human XCR1+ Dendritic Cells Derived In Vitro from CD34+ Progenitors Closely Resemble Blood Dendritic Cells, Including Their Adjuvant Responsiveness, Contrary to Monocyte-Derived Dendritic Cells

“…Only MoDC (27) and Langerhans cells (LC) (28) have been generated reproducibly in vitro by many teams. Gene expression profiling and/or ontogenic studies showed that MoDC, Langerhans cell, and DC lines strikingly differ from LT-DC subsets and are more similar to monocytes or macrophages (4,(29)(30)(31)(32). Cultures of CD34 + progenitors with FLT3-ligand (FLT3-L) or thrombopoietin (TPO) have been reported to yield pDC, CD141 + CLEC9A + DC, or CD1c + DC (11,33,34).…”