Expression of IgH Promoter/Enhancer Ly-1 Transgene in Hematopoietic Chimeric Mice Generated by Embryonic Stem Cell Line

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Abstract
We have produced hematopoietic chimeric mice from an embryonic stem (ES) cell line earring Ly-1 cDNA under the control of IgH promoter and enhancer. Various amounts of serum IgM (5–86% of total IgM) in chimeric mice were of ES origin and 30–60% of IgM-positive B cells from the chimeric mice analyzed were reconstituted from ES cells. Using these chimeric mice, the expression of the Ly-1 transgene on lymphoid tissues was examined by polymerase chain reaction assay with primers specific for the transgene, and by cell sorter analysis. Transcription of the Ly-1 transgene was detected in spleen cells, thymocytes and lymph node cells; however, the expression of the Ly-1 molecule was observed only on lipopolysaccharide (LPS)-stimulated splenic IgM-positive B cells but not on resting splenic B cells. There was no significantly increased expression of Ly-1 on splenic T cells and thymocytes. Thus, our findings demonstrate that conventional splenic B cells could express the Ly-1 transgene on their surface in vivo after LPS stimulation. Also discussed is the ES-derived chimeric hematopoietic system.

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