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,
H. PHILLIP KOEFFLER
,
JUNE E. BISHOP and
ANTHONY W. NORMAN
Division of Biomedical Sciences and the Department of Biochemistry, University of California Riverside, California 92521
The Division of Hematology, the Department of Medicine, University of California Los Angeles, California 90024
Address all correspondence and requests for reprints to: Prof. Anthony W. Norman, Department of Biochemistry, University of California, Riverside, California 92521.
Cultured normal human pulmonary alveolar macrophages and peripheral blood monocyte-derived macro-phages were studied for their capacity to metabolize [3H]25-hydroxyvitamin D3 (25OHD3). Incubation of macrophages with bacterial lipopolysaccharide (LPS) resulted in the conversion of [3H]25OHD3 to a more polar vitamin D3 metabolite (up to 15 pmol/106 cells). Untreated macrophages did not synthesize this metabolite. Several findings suggested that the metabolite was the biologically active form of vitamin D3, namel y 1,25-dihydroxyvitamin D3 [1,25-(OH)2D3]. (1) The metabolite comigrated with chemically synthesized 1,25-(OH)2D3 on four different high performance liquid chromatographic systems. (2) The metabolite had the same affinity for the chick intestinal 1,25-(OH)2D3 receptor as authentic 1,25-(OH)2D3. (3) The biological activity of the macrophage metabolite in vivo (stimulation of intestinal calcium absorption and bone calcium mobilization in rachitic chicks) was identical to the activity of chemically synthesized 1,25-(OH)2D3. The LPS-stimulated synthesis of the 1,25-(OH)2D3-like compound by macrophages was dose dependent in a linear fashion; a half-maximal response was typically found with 100–200 ng LPS/106 cells. Polymyxin B abolished the effects of LPS on 25OHD3 metabolism in macrophages. Our data suggest that LPS-stimulated macrophages can modulate, on a local level, the function of l,25-(OH)2D3-responsive cells by releasing the l,25-(OH)2D3-like metabolite.
* This work was supported by USPHS Grants CA-26038, CA-33936, and AM-14750.
Supported by the Deutsche Forschungs gemeinschaft (Re 613/1-1).
Supported by a Career Development Award, the Murray Geisler Memorial Foundation, and the Louis Fagin Leukemia Research Foundation and is a member of the Jonsson Cancer Center.
Received March 10, 1986.
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