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KMID : 0578319950050060569
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Molecules and Cells
1995 Volume.5 No. 6 p.569 ~ p.578
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Identification of Peripheral blood Dendritic Cell-specific Class Ⅱ-related Genes by Differetial cDNA Hybridization
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Bae, Yong-Soo
Kim, Kyoung-Joo/Langhoff, Erik
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Abstract
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We report here two peripheral blood dendritic cell (PBDC)-specific class II-related antigen genes identified by differential screening of the PBDC-cDNA library. Dendritic cells (DC) were isolated and purified from the peripheral blood leukocytes (PBL) by employing negative selection procedure and monoclonal antibody sorting with magnetic beads. The DC-cDNA library was constructed using a λgt11 system. The cDNA library was screened by two steps of differential hybridization with [^(32)P]-labeled DC-cDNA probe, and T-and monocyte-cDNA probes. Isolated clones which were potentially specific to DC were sequenced and then searched for their sequence homology in Genbank database. Novel or interesting genes were confirmed for their specificity to DC by checking the relative amounts of the mRNA expressed in T-cell, B-cells, monocytes and DC by polyperase chain reaction (PCR) and Nothern blot hybridization. Through these experiments, two MHC classs II-related genes, DMαa and DMβ, were found to be expressed only in DC among the PBL. Biological functions of these genes remain to be solved by producing monoclonal antibodies against these gene products.
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KEYWORD
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