Animal cells and systems

The Korean Society for Integrative Biology (한국통합생물학회)
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Expression Analysis of the Ligand to Ly-6E.1 Mouse Hematopoietic Stem Cell Antigen

  • Hwang, Dae-Youn (Department of Biology and College of Natural Sciences, Chungnam National University) ;
  • Min, Dul-Lei (Department of Biochemistry and College of Natural Sciences, Chungnam National University) ;
  • Sonn, Chung-Hee (Department of Biochemistry and College of Natural Sciences, Chungnam National University) ;
  • Chang, Mi-Ra (Department of Biochemistry and College of Natural Sciences, Chungnam National University) ;
  • Lee, Mi-Hyun (Department of Biochemistry and College of Natural Sciences, Chungnam National University) ;
  • Paik, Sang-Gi (Department of Biology and College of Natural Sciences, Chungnam National University) ;
  • Kim, Young-Sang (Department of Biochemistry and College of Natural Sciences, Chungnam National University)
  • Published : 1997.03.01
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Abstract

Ly-6E.1 antigen was proposed as a regulatory molecule of T lymphocyte activation, a hematopoietic stem cell marker, a memory cell marker, and an adhesion molecule. Though there were several reports suggesting the presence of Ly-6 ligand, the characterization of the ligand was not yet performed, As an attempt to screen the expression of Ly-6E.1 ligand, we prepared a probe for detecting Ly-6E.1 ligand by producing a fusion protein between Ly-6E.1 and $hlgC_{r1}$, A mammalian cell expression vector with Ly-6E.$1/hlgC_{r1}$ chimeric cDNA was transfected in SP2/0-Ag14 myeloma cells, and stable transfectants were selected. The fusion protein was produced as a dimer and maintained the epitopes for monoclonal antibodies specific for Ly-6E.1 and for anti-human lgG antibody. The purified fusion protein through Gammabind G column was used for FACS analyses for the expression of Ly-6E.1 ligand. The fusion protein interacted with several cell lines originating from B cells, T cells, or monocytes. The fusion Protein also strongly stained bone marrow, lymph node, and spleen cells, but thymic cells weakly, if any. The staining was more obvious in C57BL/6 $(Ly-6^b)$ than Balb/c $(Ly-6^a)$ mice. These results suggest that the interaction of Ly-6E.1 with Ly-6E.1 ligand may function both in the stem cell environment and in the activation of mature lymphocytes. The fusion protein may be a valuable tool in characterization of biochemical properties of the Ly-6E.1 ligand and, further, in isolating its cDNA.

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References

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