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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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Related in: MedlinePlus

CD56+DCs functionally resemble mDCs. (A) The expression of TLR4 was quantified by RT-PCR with purified DCs. Error bars represent standard derivation from duplicated PCR samples of the same donor. Data shown are from one representative of 3 independent donors. (B) Lineage depleted PBMC were stimulated with LPS for 6 h, then the stimulated cells were collected and stained with surface markers followed by intracellular staining of TNFα. CD123+CD56− (pDC), CD123+CD56+ (CD56+) and CD123− CD11C+ (mDC) in the Lin− HLA-DR+ gate were analyzed for TNFα expression. (C) Summarized data of (B) from 5 independent donors. (D) Purified DCs were stimulated with mixture of LPS, CpG and IFNγ for 6 h, the expression of IL-12 p35 and p40 were analyzed by RT-PCR. Data shown are representative of 3 independent experiments. (E) Purified DCs were stimulated mixture of LPS, CpG and IFNγ for 24 h, and the expression of IL-12 p70 were analyzed by ELISA. Data shown are representative of 2 independent experiments. (F) Expression of HLA-DR and CD86 on different blood DC subsets were analyzed by FACS. (G) Purified allogenic naive T cells were cocultured with different numbers of CD123+CD56− (pDC, circle), CD123+CD56+ (CD56+, square) and CD123−CD11C+ (mDC, triangle) DCs . After 5 days, the cocultured cells were pulsed with 1 mCi [3H] thymidine for 18 h before harvest. Data shown are one representative of 4 experiments
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Fig4: CD56+DCs functionally resemble mDCs. (A) The expression of TLR4 was quantified by RT-PCR with purified DCs. Error bars represent standard derivation from duplicated PCR samples of the same donor. Data shown are from one representative of 3 independent donors. (B) Lineage depleted PBMC were stimulated with LPS for 6 h, then the stimulated cells were collected and stained with surface markers followed by intracellular staining of TNFα. CD123+CD56− (pDC), CD123+CD56+ (CD56+) and CD123− CD11C+ (mDC) in the Lin− HLA-DR+ gate were analyzed for TNFα expression. (C) Summarized data of (B) from 5 independent donors. (D) Purified DCs were stimulated with mixture of LPS, CpG and IFNγ for 6 h, the expression of IL-12 p35 and p40 were analyzed by RT-PCR. Data shown are representative of 3 independent experiments. (E) Purified DCs were stimulated mixture of LPS, CpG and IFNγ for 24 h, and the expression of IL-12 p70 were analyzed by ELISA. Data shown are representative of 2 independent experiments. (F) Expression of HLA-DR and CD86 on different blood DC subsets were analyzed by FACS. (G) Purified allogenic naive T cells were cocultured with different numbers of CD123+CD56− (pDC, circle), CD123+CD56+ (CD56+, square) and CD123−CD11C+ (mDC, triangle) DCs . After 5 days, the cocultured cells were pulsed with 1 mCi [3H] thymidine for 18 h before harvest. Data shown are one representative of 4 experiments

Mentions: TLR expression patterns of pDCs and mDCs are drastically different. pDCs preferentially express TLR7 and TLR9 while mDCs express higher levels of TLR2 and TLR4 (Liu 2005; Cerboni et al., 2013). We found that the expression level of TLR4 (Fig. 4A) and the amount of TNFα produced upon LPS stimulation (Fig. 4B and 4C) were similar between CD56+ DCs and mDCs. Moreover, CD56+ DCs could produce IL-12 at a comparable level to mDCs at both RNA (Fig. 4D) and protein level (Fig. 4E). While pDCs did not produce IL-12 (Fig. 4D and 4E) as previously reported (Kadowaki et al., 2001).


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 functionally resemble mDCs. (A) The expression of TLR4 was quantified by RT-PCR with purified DCs. Error bars represent standard derivation from duplicated PCR samples of the same donor. Data shown are from one representative of 3 independent donors. (B) Lineage depleted PBMC were stimulated with LPS for 6 h, then the stimulated cells were collected and stained with surface markers followed by intracellular staining of TNFα. CD123+CD56− (pDC), CD123+CD56+ (CD56+) and CD123− CD11C+ (mDC) in the Lin− HLA-DR+ gate were analyzed for TNFα expression. (C) Summarized data of (B) from 5 independent donors. (D) Purified DCs were stimulated with mixture of LPS, CpG and IFNγ for 6 h, the expression of IL-12 p35 and p40 were analyzed by RT-PCR. Data shown are representative of 3 independent experiments. (E) Purified DCs were stimulated mixture of LPS, CpG and IFNγ for 24 h, and the expression of IL-12 p70 were analyzed by ELISA. Data shown are representative of 2 independent experiments. (F) Expression of HLA-DR and CD86 on different blood DC subsets were analyzed by FACS. (G) Purified allogenic naive T cells were cocultured with different numbers of CD123+CD56− (pDC, circle), CD123+CD56+ (CD56+, square) and CD123−CD11C+ (mDC, triangle) DCs . After 5 days, the cocultured cells were pulsed with 1 mCi [3H] thymidine for 18 h before harvest. Data shown are one representative of 4 experiments
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Related In: Results  -  Collection

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Fig4: CD56+DCs functionally resemble mDCs. (A) The expression of TLR4 was quantified by RT-PCR with purified DCs. Error bars represent standard derivation from duplicated PCR samples of the same donor. Data shown are from one representative of 3 independent donors. (B) Lineage depleted PBMC were stimulated with LPS for 6 h, then the stimulated cells were collected and stained with surface markers followed by intracellular staining of TNFα. CD123+CD56− (pDC), CD123+CD56+ (CD56+) and CD123− CD11C+ (mDC) in the Lin− HLA-DR+ gate were analyzed for TNFα expression. (C) Summarized data of (B) from 5 independent donors. (D) Purified DCs were stimulated with mixture of LPS, CpG and IFNγ for 6 h, the expression of IL-12 p35 and p40 were analyzed by RT-PCR. Data shown are representative of 3 independent experiments. (E) Purified DCs were stimulated mixture of LPS, CpG and IFNγ for 24 h, and the expression of IL-12 p70 were analyzed by ELISA. Data shown are representative of 2 independent experiments. (F) Expression of HLA-DR and CD86 on different blood DC subsets were analyzed by FACS. (G) Purified allogenic naive T cells were cocultured with different numbers of CD123+CD56− (pDC, circle), CD123+CD56+ (CD56+, square) and CD123−CD11C+ (mDC, triangle) DCs . After 5 days, the cocultured cells were pulsed with 1 mCi [3H] thymidine for 18 h before harvest. Data shown are one representative of 4 experiments
Mentions: TLR expression patterns of pDCs and mDCs are drastically different. pDCs preferentially express TLR7 and TLR9 while mDCs express higher levels of TLR2 and TLR4 (Liu 2005; Cerboni et al., 2013). We found that the expression level of TLR4 (Fig. 4A) and the amount of TNFα produced upon LPS stimulation (Fig. 4B and 4C) were similar between CD56+ DCs and mDCs. Moreover, CD56+ DCs could produce IL-12 at a comparable level to mDCs at both RNA (Fig. 4D) and protein level (Fig. 4E). While pDCs did not produce IL-12 (Fig. 4D and 4E) as previously reported (Kadowaki et al., 2001).

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