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Human BDCA2+CD123+CD56+ dendritic cells (DCs) related to blastic plasmacytoid dendritic cell neoplasm represent a unique myeloid DC subset.

Yu H, Zhang P, Yin X, Yin Z, Shi Q, Cui Y, Liu G, Wang S, Piccaluga PP, Jiang T, Zhang L - Protein Cell (2015)

Bottom Line: In addition, CD56(+) DCs cluster together with mDCs but not pDCs by genome-wide transcriptional profiling.These data suggest that the CD56(+) DCs represent a novel mDC subset mixed with some pDC features.However, we demonstrated that BPDCN is closer to CD56(+) DCs than pDCs by global gene-expression profiling.

View Article: PubMed Central - PubMed

Affiliation: Key Laboratory of Immunity and Infection, Institute of Biophysics, Chinese Academy of Sciences, Beijing, 100101, China.

ABSTRACT
Dendritic cells (DCs) comprise two functionally distinct subsets: plasmacytoid DCs (pDCs) and myeloid DCs (mDCs). pDCs are specialized in rapid and massive secretion of type I interferon (IFN-I) in response to nucleic acids through Toll like receptor (TLR)-7 or TLR-9. In this report, we characterized a CD56(+) DC population that express typical pDC markers including CD123 and BDCA2 but produce much less IFN-I comparing with pDCs. In addition, CD56(+) DCs cluster together with mDCs but not pDCs by genome-wide transcriptional profiling. Accordingly, CD56(+) DCs functionally resemble mDCs by producing IL-12 upon TLR4 stimulation and priming naïve T cells without prior activation. These data suggest that the CD56(+) DCs represent a novel mDC subset mixed with some pDC features. A CD4(+)CD56(+) hematological malignancy was classified as blastic plasmacytoid dendritic cell neoplasm (BPDCN) due to its expression of characteristic molecules of pDCs. However, we demonstrated that BPDCN is closer to CD56(+) DCs than pDCs by global gene-expression profiling. Thus, we propose that the CD4(+)CD56(+) neoplasm may be a tumor counterpart of CD56(+) mDCs but not pDCs.

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CD56+DCs adopt certain characteristics attributed to pDCs. (A) The expression of BDCA4 and ILT7 in different DC subpopulations from PBMC were analyzed by FACS. Data shown are one representative of 3 donors. (B) CD123+CD56− (pDC), CD123+CD56+ (CD56+) and CD123−CD11C+ (mDC) were purified from PBMC, E2-2, Spi-B and TLR9 expression in those three DC populations were analyzed by RT-PCR. Error bars represent standard derivation from duplicated PCR samples of same donor. Data shown are one representative of 3 donors. (C) Purified DCs were stimulated with CpG B for 24 h, and the expression of TNFα were analyzed by ELISA. Data shown are summarized from 3 independent experiments. (D) Lineage depleted PBMCs were stimulated with CpG B for 6 h, then stimulated cells were collected and stained with Lin, HLA-DR, CD123, CD11C and CD56 followed by intracellular staining of TNFα. The Lin− HLA-DR+ cells were divided into CD123+CD56− (pDC), CD123+CD56+ (CD56+) and CD123− CD11C+ (mDC) subsets, TNFα expression in each DC subset were analyzed. (E) Summarized data of (D) from 5 independent donors
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Fig5: CD56+DCs adopt certain characteristics attributed to pDCs. (A) The expression of BDCA4 and ILT7 in different DC subpopulations from PBMC were analyzed by FACS. Data shown are one representative of 3 donors. (B) CD123+CD56− (pDC), CD123+CD56+ (CD56+) and CD123−CD11C+ (mDC) were purified from PBMC, E2-2, Spi-B and TLR9 expression in those three DC populations were analyzed by RT-PCR. Error bars represent standard derivation from duplicated PCR samples of same donor. Data shown are one representative of 3 donors. (C) Purified DCs were stimulated with CpG B for 24 h, and the expression of TNFα were analyzed by ELISA. Data shown are summarized from 3 independent experiments. (D) Lineage depleted PBMCs were stimulated with CpG B for 6 h, then stimulated cells were collected and stained with Lin, HLA-DR, CD123, CD11C and CD56 followed by intracellular staining of TNFα. The Lin− HLA-DR+ cells were divided into CD123+CD56− (pDC), CD123+CD56+ (CD56+) and CD123− CD11C+ (mDC) subsets, TNFα expression in each DC subset were analyzed. (E) Summarized data of (D) from 5 independent donors

Mentions: CD56+ DCs expressed BDCA2 and CD123, which were considered as hallmarks of pDCs (Fig. 1B). In order to further characterize the relationship between CD56+ DCs and pDCs, we examined more pDC-specific genes at both RNA and protein levels in other DC subsets. And we found that BDCA4 and ILT7, two additional pDC specific surface markers, were expressed on CD56+ DCs but not mDCs (Fig. 5A). Both E2-2 and SpiB, the two important transcription factors for pDC lineage development, were expressed in CD56+ DCs at slightly lower levels than pDCs in the RNA-seq data (Table S4) and confirmed by RT-PCR (Fig. 5B, left two panels). It is worth to pointing out that the expression levels of E2-2 and SpiB in CD56+ DCs were much higher than in mDCs (Fig. 5B, left two panels). In addition, pDC highly expressed genes IRF7, BCL11A and CD2AP (Marafioti et al., 2008) were also expressed in CD56+ DCs at a slightly lower level compared to pDCs (Table S4). However, other pDC specific genes, such as Granzyme B, TCL1A (Herling et al., 2003), PACSIN1 (Esashi et al., 2012) and BAD-LAMP (Defays et al., 2011), were expressed at least 10 times higher in pDCs than in CD56+ DCs in RNA-seq data (Table S4).Figure 5


Human BDCA2+CD123+CD56+ dendritic cells (DCs) related to blastic plasmacytoid dendritic cell neoplasm represent a unique myeloid DC subset.

Yu H, Zhang P, Yin X, Yin Z, Shi Q, Cui Y, Liu G, Wang S, Piccaluga PP, Jiang T, Zhang L - Protein Cell (2015)

CD56+DCs adopt certain characteristics attributed to pDCs. (A) The expression of BDCA4 and ILT7 in different DC subpopulations from PBMC were analyzed by FACS. Data shown are one representative of 3 donors. (B) CD123+CD56− (pDC), CD123+CD56+ (CD56+) and CD123−CD11C+ (mDC) were purified from PBMC, E2-2, Spi-B and TLR9 expression in those three DC populations were analyzed by RT-PCR. Error bars represent standard derivation from duplicated PCR samples of same donor. Data shown are one representative of 3 donors. (C) Purified DCs were stimulated with CpG B for 24 h, and the expression of TNFα were analyzed by ELISA. Data shown are summarized from 3 independent experiments. (D) Lineage depleted PBMCs were stimulated with CpG B for 6 h, then stimulated cells were collected and stained with Lin, HLA-DR, CD123, CD11C and CD56 followed by intracellular staining of TNFα. The Lin− HLA-DR+ cells were divided into CD123+CD56− (pDC), CD123+CD56+ (CD56+) and CD123− CD11C+ (mDC) subsets, TNFα expression in each DC subset were analyzed. (E) Summarized data of (D) from 5 independent donors
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Related In: Results  -  Collection

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Fig5: CD56+DCs adopt certain characteristics attributed to pDCs. (A) The expression of BDCA4 and ILT7 in different DC subpopulations from PBMC were analyzed by FACS. Data shown are one representative of 3 donors. (B) CD123+CD56− (pDC), CD123+CD56+ (CD56+) and CD123−CD11C+ (mDC) were purified from PBMC, E2-2, Spi-B and TLR9 expression in those three DC populations were analyzed by RT-PCR. Error bars represent standard derivation from duplicated PCR samples of same donor. Data shown are one representative of 3 donors. (C) Purified DCs were stimulated with CpG B for 24 h, and the expression of TNFα were analyzed by ELISA. Data shown are summarized from 3 independent experiments. (D) Lineage depleted PBMCs were stimulated with CpG B for 6 h, then stimulated cells were collected and stained with Lin, HLA-DR, CD123, CD11C and CD56 followed by intracellular staining of TNFα. The Lin− HLA-DR+ cells were divided into CD123+CD56− (pDC), CD123+CD56+ (CD56+) and CD123− CD11C+ (mDC) subsets, TNFα expression in each DC subset were analyzed. (E) Summarized data of (D) from 5 independent donors
Mentions: CD56+ DCs expressed BDCA2 and CD123, which were considered as hallmarks of pDCs (Fig. 1B). In order to further characterize the relationship between CD56+ DCs and pDCs, we examined more pDC-specific genes at both RNA and protein levels in other DC subsets. And we found that BDCA4 and ILT7, two additional pDC specific surface markers, were expressed on CD56+ DCs but not mDCs (Fig. 5A). Both E2-2 and SpiB, the two important transcription factors for pDC lineage development, were expressed in CD56+ DCs at slightly lower levels than pDCs in the RNA-seq data (Table S4) and confirmed by RT-PCR (Fig. 5B, left two panels). It is worth to pointing out that the expression levels of E2-2 and SpiB in CD56+ DCs were much higher than in mDCs (Fig. 5B, left two panels). In addition, pDC highly expressed genes IRF7, BCL11A and CD2AP (Marafioti et al., 2008) were also expressed in CD56+ DCs at a slightly lower level compared to pDCs (Table S4). However, other pDC specific genes, such as Granzyme B, TCL1A (Herling et al., 2003), PACSIN1 (Esashi et al., 2012) and BAD-LAMP (Defays et al., 2011), were expressed at least 10 times higher in pDCs than in CD56+ DCs in RNA-seq data (Table S4).Figure 5

Bottom Line: In addition, CD56(+) DCs cluster together with mDCs but not pDCs by genome-wide transcriptional profiling.These data suggest that the CD56(+) DCs represent a novel mDC subset mixed with some pDC features.However, we demonstrated that BPDCN is closer to CD56(+) DCs than pDCs by global gene-expression profiling.

View Article: PubMed Central - PubMed

Affiliation: Key Laboratory of Immunity and Infection, Institute of Biophysics, Chinese Academy of Sciences, Beijing, 100101, China.

ABSTRACT
Dendritic cells (DCs) comprise two functionally distinct subsets: plasmacytoid DCs (pDCs) and myeloid DCs (mDCs). pDCs are specialized in rapid and massive secretion of type I interferon (IFN-I) in response to nucleic acids through Toll like receptor (TLR)-7 or TLR-9. In this report, we characterized a CD56(+) DC population that express typical pDC markers including CD123 and BDCA2 but produce much less IFN-I comparing with pDCs. In addition, CD56(+) DCs cluster together with mDCs but not pDCs by genome-wide transcriptional profiling. Accordingly, CD56(+) DCs functionally resemble mDCs by producing IL-12 upon TLR4 stimulation and priming naïve T cells without prior activation. These data suggest that the CD56(+) DCs represent a novel mDC subset mixed with some pDC features. A CD4(+)CD56(+) hematological malignancy was classified as blastic plasmacytoid dendritic cell neoplasm (BPDCN) due to its expression of characteristic molecules of pDCs. However, we demonstrated that BPDCN is closer to CD56(+) DCs than pDCs by global gene-expression profiling. Thus, we propose that the CD4(+)CD56(+) neoplasm may be a tumor counterpart of CD56(+) mDCs but not pDCs.

Show MeSH
Related in: MedlinePlus