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KMID : 0578319960060060684
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Molecules and Cells
1996 Volume.6 No. 6 p.684 ~ p.691
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Blood Dendritic Cells Express Detectable Amounts of Class Ⅱ-related DM protein on the Cell Surgaces
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Kim, Kyoung-Ju
Jung, Hyun-Ho/Bae, Yong-Soo
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Abstract
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We have shown that the HLA-DM molecules are exclusively expressed only in blood dendritic cells among the primary periphral blood leukocytes by differential cDNA hybridization (Bae et al., 1995). This experiment was initiated to identify the expression pattern of DM in primary dendritic cells. Extracellular domains of DMα and β were cloned into the pRSET vector and expressed in E. coli JM109(DE3) and BL2l(DE3). Recombinant DM protein purified from a Ni^(+)-NTA agarose column was used to generate polyclonal and monoclonal antibodies in Balb/c mice. Eyen though having some detectable cross-reactivity, polyclonal antibody showed a remarkable binding capacity not only to recombinant but also to wild type DM molecules. Particularly noteworthy is that intact dendritic cells were deeply stained by the DM polyclonal antibody, which strongly suggests that unlike other reports of endosomal location of DM, DM is located, at least in considerable amounts, on the surface of dendritic cells. We raised 4 monoclonal antibodies against recombinant DMα and 1 for recombinant DMβ. However, none of the 5 monoclonal antibodies generated against recombinant DM showed any reactivity to wild type DM in Raji or dendritic cells. It is discussed in the text.
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