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KMID : 0578319930030030319
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Molecules and Cells
1993 Volume.3 No. 3 p.319 ~ p.325
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Increased Cytosolic Free Calcium Concentration Following HCMV Infection of Human Embryo Kung Cells
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Kang, Jyung-Hee
yoo, Cheong-Hee/Lee, Chan-Hee
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Abstract
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Calcium response and the role of Call in HCMV infection were investigated using Fura-2/ANI as a fluorescent Cal+ indicator. Infection of human fibroblasts with human cytomegalo,,irus (HCMV) resulted in a gradual increase in cytosolic free Cal+ concentration ([Ca2-]i), and this increase was dependent on the amount of infectious virus. The effects of low Ca´-´ medium, Cat+-specific chelator EGTA, a specific Cal+ channel blocker nifedipine, and an intracellular Cal+ chelator BAPTA/AM on HCMV-induced increase in [Ca2+]i were examined. Reduced extracellular Ca" concentration by treating HCMV-infected cells with either low Ca2- medium or EGTA resulted in a diminution of the HCMV-induced [Ca2+]i increase at 6 h after virus infection (p.i.). Treatment with nifedipine also reduced [Ca2+]i in HCMV-infected fibroblasts. Reduction of [Ca"]i by BAPTA/AM suggests the contribution of intracellular Ca´-´ as well as extracellular Ca" to the HCMV-induced [Ca2*]i increase. Although both extracetlular and intracellular Call source appear to be important in HCMV-induced increase in [Ca2-]i, their relative order of participation during HCMV infection might be different. Data obtaind from differential addition or removal of nifedipine or BAPTA/AM in HCMV-infected cells implicate that extracellular Call is responsible for the [Ca2+]i increase in the first 3 h (0 to 3 h p.i.) period, while intracellular Ca" in the subsequent 3 It period (3 to 6 h p.i.) of HCMV infection. Treatment of cells transfected with plasmid containing HCMV major immediate early (MIE) promoter fused with a reporter gene with Ca2+ ionophore stimulated the expression of HCMV MIE gene.
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