Cloning and characterisation of the chicken orthologue of dendritic cell-lysosomal associated membrane protein (DC-LAMP).
A cDNA encoding the chicken orthologue of dendritic cell-lysosomal associated membrane protein (DC-LAMP)/CD208 was cloned by RT-PCR from RNA isolated from mature chicken bone marrow-derived dendritic cells (chBM-DCs). The cloned chicken DC-LAMP (chDC-LAMP) cDNA consists of 1281 nucleotides encoding an open reading frame of 426 amino acids (aa). Comparison of the deduced aa sequence of DC-LAMP with orthologous proteins from human and mouse revealed 27 and 24% identity, respectively. The predicted chDC-LAMP protein shares the characteristic features of LAMP family members. ChDC-LAMP mRNA, unlike its mammalian orthologues, was expressed in a wide range of tissues, at highest levels in the lung. Lymphoid tissues including thymus, spleen, bursa, ceacal tonsil and Meckel's diverticulum had high chDC-LAMP mRNA expression levels. ChDC-LAMP mRNA was expressed in all splenocyte subsets with the highest expression in Bu-1(+) B cells and KUL01(+) cells, which would include macrophages and DC. ChDC-LAMP mRNA was highly expressed in chBM-DC, whereas expression levels in chicken monocyte-derived macrophages (chMo-Mac) and the HD11 macrophage cell line were significantly lower. Following CD40L stimulation, chDC-LAMP mRNA expression levels were up-regulated in mature chBM-DC, chMo-Mac and HD11 cells whereas lipopolysaccharide (LPS) only up-regulated chDC-LAMP mRNA expression levels in chBM-DC. ChDC-LAMP is not solely expressed on chicken DC but can be used as a marker to differentiate between immature and mature DC.
Institute for Animal Health, Compton, Berkshire, UK. zhiguang.wu@bbsrc.ac.uk
Zhiguang
Z
Wu
Tuanjun
T
Hu
Colin
C
Butter
Pete
P
Kaiser
D000595:Amino Acid Sequence; D000818:Animals; D002645:Chickens / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D003001:Cloning, Molecular; D017124:Conserved Sequence; D003713:Dendritic Cells / Q000276:immunology / Q000378:metabolism; D006801:Humans; D051907:Lysosomal Membrane Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D008264:Macrophages / Q000276:immunology / Q000378:metabolism; D008969:Molecular Sequence Data; D010802:Phylogeny; D016415:Sequence Alignment
D051907:Lysosomal Membrane Proteins