Identification and immunocytochemical analysis of DCNP1, a dendritic cell-associated nuclear protein.
Dendritic cells (DCs) are potent antigen-presenting cells (APCs). Among so-called professional APCs, only DCs can activate naive T cells to initiate immune response. To better understand molecular mechanisms underlying unique functions of DCs, we searched for genes specifically expressed in human DCs, using PCR-based cDNA subtraction in conjunction with differential screening. cDNAs generated from CD34(+) stem cell-derived CD1a(+) DC were subtracted with cDNA from monocytes and used for generation of a cDNA library. The cDNA library was differentially screened to select genes expressed in DCs more abundantly than in monocytes. We identified a gene encoding a protein composed of 244 amino acids, which we designated as DCNP1 (dendritic cell nuclear protein 1). In Northern blot analysis, DCNP1 mRNA was highly expressed in mature DCs and at a lower level in immature DCs. In contrast, monocytes and B cells do not express the gene. In multiple human tissue Northern blot analysis, expression of DCNP1 was detected in brain and skeletal muscle. To examine subcellular localization of DCNP1, we performed immunofluorescence analysis using an anti-DCNP1 polyclonal antibody and found the molecule to be localized mainly in the perinucleus. In an immunohistochemical analysis, we compared the expression of DCNP1 with CD68, a marker for DCs and macrophages, in spleen, lymph node, liver, and brain. While DCNP1-positive cells showed a similar tissue distribution to CD68-positive cells, the number of DCNP1-positive cells was much smaller than that of CD68-positive cells. Our findings are consistent with the proposal that DCNP1 is specifically expressed in DCs.
Masuda
Masako
M
Division of Immunogenetics, Kumamoto University Graduate School of Medical Sciences, Kumamoto, 860-0811, Japan.
Senju
Satoru
S
Fujii Si
Shin-ichiro
S
Terasaki
Yasuhiro
Y
Takeya
Motohiro
M
Hashimoto Si
Shin-ichi
S
Matsushima
Kouji
K
Yumoto
Eiji
E
Nishimura
Yasuharu
Y
D000595:Amino Acid Sequence; D015703:Antigens, CD / Q000032:analysis; D018952:Antigens, CD34 / Q000032:analysis; D015214:Antigens, Differentiation, Myelomonocytic / Q000032:analysis; D001483:Base Sequence; D002478:Cells, Cultured; D003001:Cloning, Molecular; D003713:Dendritic Cells / Q000737:chemistry / Q000276:immunology*; D015723:Gene Library; D006801:Humans; D007150:Immunohistochemistry; D016130:Immunophenotyping; D008856:Microscopy, Fluorescence; D008969:Molecular Sequence Data; D009000:Monocytes / Q000737:chemistry; D009687:Nuclear Proteins / Q000032:analysis / Q000235:genetics* / Q000276:immunology; D012333:RNA, Messenger / Q000096:biosynthesis; D014018:Tissue Distribution; D000097382:CD68 Molecule
D015703:Antigens, CD; D018952:Antigens, CD34; D015214:Antigens, Differentiation, Myelomonocytic; D009687:Nuclear Proteins; D012333:RNA, Messenger; C067980:CD68 antigen, human; D000097382:CD68 Molecule; C446761:DCANP1 protein, human